Asian Pacific Journal of Tropical Medicine

179-183

Authors: Amanda Sánchez-Recillas, Paul Mantecón-Reyes, Patricia Castillo-España, Rafael Villalobos-Molina, Maximiliano Ibarra-Barajas, Samuel Estrada-Soto

Objective To assess the relaxant effect of several organic extracts obtained from Agastache mexicana (A. mexicana), Cochlospermum vitifolium (C. vitifolium), Cordia morelosana (C. morelosana), Lepechinia caulescens (L. caulescens) and Talauma mexicana (T. mexicana) used in Mexican traditional medicine for the treatment of several diseases. Methods Extracts were obtained by maceration at room temperature using hexane, dichloromethane and methanol for each plant material. The organic extracts were evaluated ex vivo to determine their relaxant activity on the contractions induced by carbachol (cholinergic receptor agonist, 1 μ mol/L) in isolated rat tracheal rings. Results A total of 15 extracts were evaluated (three for each species). All test samples showed significant relaxant effect, in a concentration-dependent manner, on the contractions induced by 1 μ mol/L carbachol, with exception of extracts from C. morelosana. Active extracts were less potent than theophylline [phosphodiesterase inhibitor, EC50: (28.79±0.82) μg/mL] that was used as positive control. Concentration-response curves revealed that the extracts with more significant effects were dichloromethanic extracts of T. mexicana [Emax: (103.03±3.32)% and EC50: (159.39±3.72) μg/mL) and C. vitifolium [Emax: (106.58±2.42)% and EC50: (219.54±7.61) μg/mL]. Finally, hexanic and dichloromethanic extracts from A. mexicana were fully effective but less potent than T. mexicana and C. vitifolium. Conclusions Less polar extracts obtained from A. mexicana, T. mexicana and C. vitifolium exhibited greater relaxant effect on tracheal rat rings, which allows us to suggest them as sources for the isolation of bioactive molecules with potential therapeutic value in the treatment of asthma.

193-196

Authors: Mohammad Javad Nasiri, Faranak Rezaei, Samin Zamani, Davod Darban-Sarokhalil, Abbas Ali Imani Fooladi, Hasan Shojaei, Mohammad Mehdi Feizabadi

Objective To determine the patterns of resistance to first line anti-tuberculosis (TB) drugs among a collection of Mycobacterium tuberculosis (MTB) isolates from 5 provinces of Iran. Methods A total of the 6 426 clinical specimens from patients suspected of active TB were collected from March 2010 to June 2012. All specimens were subjected for microscopy and culture tests in the TB centers of studies provinces. Drug susceptibility testing to the first line anti-TB drugs for culture positive MTB was performed on Löwenstein-Jensen (LJ) medium using proportion method. Results Of 6 426 clinical specimens, 261 were culture positive for mycobacteria, of which 252 were MTB and 9 were MOTT (mycobacteria other than tuberculosis). Of 252 MTB isolates, 211 (83.7%) were pan-susceptible and 41 (16.3%) were resistant to at least one drug. Resistance was most common to streptomycin, 30 isolates (12.0%), followed by isoniazid, 20 isolates (8.0%), rifampin, 15 isolates (6.0%) and ethambutol, 14 isolates (5.5%). Sixteen (6.3%) MTB isolates were MDR. A clear evidence of heterogeneity amongst the 5 provinces in the proportions with resistance to one or more drugs was observed [χ2; = 12.209 (4 degrees of freedom), P values = 0.015 9]. Conclusions The prevalence of drug resistance in this study area underscoring the need for further enforcement of TB control strategies in the Iran. Drug susceptibility testing for all TB cases to provide optimal treatment, establishing advanced diagnostic facilities for rapid detection of MDR-TB and continuous monitoring of drug resistance are recommended for prevention and control of drug-resistant TB.

206-207

Authors: Ebrahim Miri-Moghaddam, Zakaria Bameri, Mehdi Mohamadi

Objective To determine the distribution of Duffy blood group genotypes in Balouch population as a major ethnic group that living in a sub-tropical area in south East of Iran. Methods In this study, the Duffy blood group FY phenotypes were determined using indirect anti-globulin technique and also genotype by PCR-RFLP in 160 vivax malaria patients and 160 control individuals. Results The results showed that the most common Duffy genotype was FYA/FYB (46.6%) followed by FYA/FYA (15.3%), FYA/FYO (14.4%), FYB/FYO (11.9%), FYB/FYB (10%) and FYO/FYO (1.9%). In case individuals, frequency of FYA, FYB and FYO alleles were 0.471, 0.431 and 0.097, respectively compaired to 0.444, 0.353 and 0.203, respectively in control (non-infected) group. Conclusions This data provide evidence that individuals with the FYA/FYB genotype have higher susceptibility to malaria and there are significant associations between Duffy blood group variants and susceptibility or resistance to vivax malaria.

212-215

Authors: Mert Köroğlu, Bekir Erol, Cemil Gürses, Barış Türkbey, Cem Yunus Baş, Ahmet Şükrü Alparslan, Banu Kale Köroğlu, İclal Erdem Toslak, Bülent Çekiç, Okan Akhan

Objective To demonstrate utility and safety of the puncture aspiration injection and reaspiration (PAIR) technique for outpatients. Methods Percutaneous treatment with US guidance was applied to 33 patients for 44 cysts. Patients treated with the PAIR technique, were outpatients. PAIR and catheterization technique were evaluated for efficacy and safety of procedure and complication rates. Results Thirty-five of 44 cysts were treated with the PAIR and 9 of 44 were treated with the catheterization technique. The success rate of the cysts Gharbi type 1 (CE1) and type 2 (CE3a) treated with the PAIR technique was 100%. In the follow up of 9 cysts treated with the catheterization technique, 2 of them (22%) developed cyst infection and 1 (11%) developed a biliary fistula. Conclusions The PAIR technique was found to be an effective and safe approach in order to treat Gharbi type 1 and type 2 cysts percutaneously for outpatients. It has a very low complication rate in comparison with the catheterization technique. So every effort should be made to finish the treatment with PAIR technique.

221-225

Authors: Bao-Nan Liu, Bo-Xiang Han, Feng Liu

Objective To explore the neuroprotective effect of pAkt and HIF-1 α on ischemia rats. Methods The rat model of cerebral ischemia which induced by permanent middle cerebral artery occlusion was established, Silybin were given respectively. The behavior was measured by modified Longa method, brain water content were measured by the dry-wet method. Infarct volume was measured by image analysis method, Akt, HIF-1 α, Bcl-2, Bax, NF-κ B protein expressions were detected by Western blotting. The Akt, HIF-1 α, Bcl-2, Bax, NF-κ B mRNA expression were detected by RT-PCR. Results The control group, low-dose silibinin group and high-dose silibinin group showed paralytic of the left body of rats in various degrees, the brain water content increased and different infarction size. There was no abnormal of the neurobehavioral assessment and no cerebral infarction in the blank group. Compared with the control group, there was no significant improvement of neurological function (t=1.341, P=0.188) or significant changes of the infarct volume (t=1.737, P=0.091) in the low-dose silibinin group, while there was significantly improvement of the neurological function in the high dose silibinin group (t=12.979, P<0.001), and the infarct volume was significantly reduced (t=23.503, P<0.001), the difference had statistically significant. The brain water content of lesion side of the control group increased (t=43.536, P<0.001), while the brain water content of lesion side of the low-dose silibinin group and the high-dose silybin group were significantly reduced (t=25.571, P<0.001; t= 42.426, P<0.001). The differences were statistical significance. The p-Akt 473, p-Akt 308, HIF-1 α, Bax, NF-κ B protein and the Akt, Bax, NF-κ B mRNA expression were increased of the control group, while the Bcl-2 protein and mRNA expression were decreased, the differences were statistically significant (P<0.05), there was no significant change of the Akt protein expression and HIF-1 α mRNA in the control group (P>0.05). In the high dose silybin group, the p-Akt 473, p-Akt 308, HIF-1α, Bcl-2 protein and Akt, Bcl-2 mRNA expression were increased, while the Bax, NF-κ B protein and Bax, NF-κ B mRNA expression were decreased, the differences were statistically significant (P<0.05), there was no significant change of the Akt protein expression and HIF-1 α mRNA in the high dose silybin group (P>0.05). Conclusions pAkt, HIF-1 α have neuroprotective effect on ischemia rats.

231-236

Authors: Jun-Jun Liang, Wei Xue, Li-Zhi Lou, Cheng Liu, Zhao-Fen Wang, Qing-Guo Li, Shao-Hua Huang

Objective To establish rabbit model of restenosis after carotid endarterectomy surgery, and to study tissue inflammatory cytokines (TNF-α, IL-6) involved in restenosis. Methods A total of 32 rabbits were randomly divided into two groups: model group and control group. The right common carotid artery in rabbits was damaged by carotid endar terectomy in model group. The tissues were harvested at different time points respectively, the pathological changes of the vascular wall after operation were observed at different time points. The changes of expression of tissue vascular wall inflammatory cytokines (TNF-α, IL-6) at different time points after the surgery was observed by RT-PCR, and the changes of serum inflammatory cytokines (TNF-α, IL −6) were detected by ELISA. Results The new intima appeared after 7 days of the injury and reached the peak on 28 d which is uneven and significantly thicker than the control group (P<0.01). The tissue inflammatory cytokines (TNF-α, IL-6) were significantly increased after the rabbit common carotid artery injury, which was significant difference compared with normal control group (P<0.05). Conclusions The tissue inflammatory factors significantly increase after the rabbit carotid artery injury, which suggests the mutual concurrent effects of inflammatory cytokines can result in the proliferation of vascular restenosis.

241-243

Authors: Xiao-Yi Yu, Wei Wu, Xiao-He Lu

Objective To observe the influence of the the transforming growth factor β1 (TGF-β1) eye drops on rabbit aqueous humor TGF-β1 concentration, and to analyze the best drug concentration. Methods A total of 30 New Zealand white rabbits were randomly divided into 5 groups with 6 in each. Rabits in control group had PBS eye drops, group A, B, C, D adopted TGF-β1 eye drops at 0.5, 1.0, 2.0, 4.0 mg/L, respectively, 4 times a day. Aqueous humor of right eye was extracted 1 week after administration to detect concentration changes of TGF-β1 by ELISA; rabbits in fpur hroups adopted 2.0 mg/L eye drops to left eyes 4 times a day, 0.2 mL aqueous humor was extracted left eye at the scheduled time point 0, 30 min, 2 h, 4 h, 24 h for testing, the slit lamp was used to observe the cornea, chamber and lens. Results No obvious pathological changes in conjunctiva, cornea, rabbit conjunctival, anterior chamber, and the lens was found. Concentration of TGF-β1 in rabbit aqueous humor in C, D group was significantly higher than the control group (P<0.05). Conclusions TGF-β1 eye drops at 2.0 mg/L, 4.0 mg/L can significantly increase concentration of TGF-β1 in rabbit aqueous humor, withe good ocular surfac permeability.

250-252

Authors: Kushal Naha, Barkur Ananthakrishna Shastry, Kavitha Saravu

A 19-year-old Asian Indian female presented with productive cough since the past one month and low grade fever since the past two weeks. She was diagnosed with pulmonary tuberculosis and treated with antitubercular drugs. Subsequently, delayed cultures of bronchoalveolar lavage fluid grew Burkholderia pseudomallei (B. pseudomallei). On follow up the patient reported significant subjective improvement and ESR progressively returned to normal. In summary, this case report raises two distinct and equally intriguing roles for B. pseudomallei, i.e. respiratory colonization and spontaneously resolving pulmonary infection. The pathogenic potential of B. pseudomallei, the etiologic agent of melioidosis, is well known. Confirmation of either colonization or spontaneous resolution, would potentially spare many patients unnecessary and expensive therapy with broad-spectrum antibiotics, and contribute to more rational usage of antibiotics, especially in co-infection with Mycobacterium tuberculosis and B. pseudomallei-two bacterial diseases with closely similar clinical, radiologic and histopathologic features.

93-96

Authors: Afshar A Aghaei, Y Rassi, I Sharifi, H Vatandoost, HR Mollaie, M.A Oshaghi, MR Abai, S Rafizadeh

Objective To identify the Leishmania species in infected sand flies by Real-time PCR coupled with HRM analysis. Methods Real-time PCR coupled with HRM analysis targeting the first internal transcribed spacer (ITS1) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish Leishmania species in sand flies specimens. Results Three out of 115 females of Phlebotomus sergenti (P. sergenti) (2.6%) were positive to Leishmania tropica (L. tropica). Conclusions This is the first report on P. sergenti as the main and proven vector of anthroponitic cutaneous leishmaniasis in Dehbakri County using Real-time PCR coupled with HRM analysis. This method is rapid, sensitive and specific for diagnosing of parasites in infected Sand flies and ideal for large scale genotyping projects.

105-112

Authors: Kilani-Jaziri Soumaya, Ghedira Zied, Nasr Nouha, Krifa Mounira, Ghedira Kamel, Franca Dijoux Marie Genviève, Ghedira Chekir Leila

Objective To evaluate in vitro antioxidant and apoptotic activities of Cyperus rotundus (C. rotundus). Methods The phytochemical study and the antioxidant activities of both methanol and aqueous extracts from C. rotundus aerial part were determined. In addition, these extracts were also investigated for their cytotoxic and apoptotic activities. The major compound of the methanol extract was isolated. Both methanol and aqueous extracts (300, 150, and 50 μg/mL) were evaluated for their antioxidant activity by the xanthine/xanthine oxidase assay system. However, 16, 8, and 4 mg/mL of each extract were tested to investigate their OH. formation scavenging potential. Aqueous extract (800, 400, and 200 μg/mL) and methanol extract (350, 175, and 88 μg/mL) were tested against lipid peroxidation, induced by 75 μM H2O2. The cytotoxicity (by MTT assay) and cell DNA fragmentation of both extracts were evaluated towards K562 and L1210 cell lines. The major compound was obtained from the butanol fraction of methanol extract and its structure was determined by RMN spectroscopic analysis. Results The methanol and aqueous extracts showed respectively, 88% and 19% inhibition of xanthine oxidase activity. Yet, the same extracts inhibited lipid peroxidation by 61.5% and 42.0%, respectively. Both extracts inhibited OH. formation by 27.1% and 25.3%, respectively. Only methanol extract induced DNA degradation. Orientin was determined as the major compound isolated from the butanol fraction of methanol extract. Conclusions It appears that C. rotundus extracts exhibit a potential use as a natural antioxidant and an apoptosis inducer.

119-123

Authors: K Thima, H Mori, R Praevanit, S Mongkhonmu, J Waikagul, D Watthanakulpanich

Objective To obtain the prevalence with clinical symptoms of Cyclospora cayetanensis (C. cayetanensis), a coccidian protozoan parasite, in Thailand which is the cause of an intestinal infection characterized by sporadic-to-frequent explosive diarrhea. Methods In a field survey conducted by the Faculty of Tropical Medicine, Mahidol University, as part of the existing parasite-control program, a total of 2 540 faecal samples from villagers in Nan Province, Thailand, were collected and examined to determine the prevalence and clinical characteristics of parasitic infections. Results Twelve cases of C. cayetanensis infection were found during faecal examination of schoolchildren aged 5-12 years. None exhibited obvious clinical symptoms, especially evidence of diarrhea; 5 of 12 had loose faeces, one reported frequent symptoms of abdominal discomfort, and another had pale conjunctiva with low hematocrit. The children were generally asymptomatic. Conclusions This finding confirms a public-health issue with potentially serious consequences whereby children can be exposed to an environment contaminated with food-and water-borne transmitted oocysts, and can hence become infected with C. cayetanensis.

130-135

Authors: Ying Zheng, Xue-Gang Li, Qiu-Zhen Wang, Ai-Guo Ma, Ib Christian Bygbjerg, Yong-Ye Sun, Yong Li, Ming-Ci Zheng, Xi Wang

Objective To investigate whether there is an association between diameter of bacille Calmette-Guérin (BCG) scars and effect of purified protein derivative (PPD) reaction and to determine whether vitamin A (VA) combined vitamin D (VD) supplementation influences the immune response to BCG revaccinated in Chinese infants. Methods A cross-section and 3-month community-randomised trial was conducted. A total of 5 629 infants at 3, 6 and 12 months of age in Junan County of China were examined for BCG scar formation. Then, 597 revaccinated infants were randomly assigned to supplementation (n=307) and control (n=290) groups. The supplementation group were daily assigned to 1 500 IU VA and 500 IU VD for 3 months. Then all infants were subjected to skin test with PPD. Results The diameter of BCG scars was positively correlated with diameter of skin indurations of PPD (r=0.17, P<0.05) in the 5 629 infants. The rate of positive response to PPD was higher in the supplementation group than in the control group (96.1% versus 89.7%, P<0.05, prevalence ratio 1.07, 95% CI 1.02-1.12). The prevalence ratio of PPD response for the supplementation group compared with that for the control group was 1.07 (95% CI 1.01-1.13) for the males and 1.08 (95% CI 1.00-1.17) for the females. For the supplementation group, the males got larger tuberculin induration than the females [(0.73±0.21) cm versus (0.67±0.20) cm, P<0.05) after intervention. Conclusions The diameter of BCG scars was effectively correlated with PPD response, which indicates BCG scar formation may be an useful tool to evaluate the effect of tuberculosis prevention. VA combined VD supplementation may play an immuno-regulatory role in BCG revaccination. This may contribute to the prevention of childhood tuberculosis.

141-143

Authors: Fang Chen, Ye-Jiang Zhou

Objective To analyze the hepatic protection of n-acetyl cysteine (NAC) on severe acute pancreatitis (SAP). Methods SD rats were randomly divided into control group, SAP group and NAC group. SAP AHO method was adopted to establish the model, 2 h after modeling, rats in NAC group had intraperitoneal injection of NAC (200 mg/kg). Ten rats from each group were sacrificed in every 6 and 12 h at different time points respectively. Liver damage, liver function and serum amylase, AST, ALT and malondialdehyde (MDA) were determined. Results Serum amylase, AST, ALT and MDA content in SAP, NAC group at each time point were significantly higher in the control group (P<0.05), serum amylase, AST, ALT and MDA content in NAC group rats were lower in the SAP group significantly (P<0.05); Microscopic examination showed that the liver injury in rats and the NAC group significantly reduced in the SAP group. Conclusions NAC provides effective protection against liver damage to SAP, protective from SAP liver injury.

149-152

Authors: Shan Bao, Shu-Ying Yang, Zhuo-Ri Li, Ge-Bo Wen

Objective To screen, identify, and compare the serum biomarkers between anovulatory dysfunctional uterine bleeding (ADUB) and ovulatory dysfunctional uterine bleeding (ODUB) in Lizu females. Methods The subjects included 128 ADUB patients, 63 ODUB patients, and 93 controls. The serum and supernate of the subjects' mense were collected and stored at −80 °C until use. Differential proteins in the sera of three groups were screened using surface-enhanced laser desorption ionization time-of-flight mass spectrometry. The screened proteins were then identified by tricine-SDS-PAGE gel and spectrometry. Protein expression levels in the menses of ADUB, ODUB, and control subjects were determined using ELISA, RT-PCR, and Western blotting. SPSS 14.1 was used for statistical analysis and chart drawing (α = 0.05). Results Three differential protein peaks with peak values of 11.80, 13.59, and 14.68 km/z were screened and identified as serum amyploid protein A (SAA), vascular endothelial growth factor, and vitamin K epoxide reductase, respectively. The SAA was highly expressed in the menses of ADUB and ODUB patients but poorly expressed in the controls. The vascular endothelial growth factor was highly expressed in the menses of ODUB and controls but poorly expressed in ADUB patients. Meanwhile, the vitamin K epoxide reductase was highly expressed in the menses of ADUB and control subjects but poorly expressed in ODUB patients. Conclusions The SAA is the common serum biomarker of ADUB and ODUB. ADUB may be related to angiogenesis impairment, whereas ODUB may be associated with blood coagulation disruption.

160-163

Authors: Xiao-Hua Hu, Chuan-Zhu Lu, Min Li, Cai-Hong Zhang, Hua Zhang

Objective To explore how to choose the center locations to build the medical and health supplies reserve among many island towns. Methods The center locations were selected from 18 towns Hainan Province, it's maximum service range (distance) was required to reach the minimum, or to minimize. Results Three scenarios were considered, the center locations included only one town, two towns, three towns. By the use of graph theory and MATLAB programming, a mathematical model was established to obtain the shortest distance and the shortest path between arbitrary two towns. Conclusions We find out the center sites under certain conditions, and determine the specific service ranges of the center sites.

933-939

Authors: B Senthilkumar, K Anbarasu, D Senbagam, M Rajasekarapandian

Objective To develop attenuated strains of Salmonella enterica serovar Typhi (S. typhi) for the candidate vaccine by osmolar stress. Methods S. typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in Namakkal, Tamil Nadu, India. Both strains were grown in LB (Luria Bertani) medium supplemented with various concentration of NaCl (0.1–0.7M) respectively. The effect of osmolar stress was determined at molecular level by PCR using MGR 06 and MGR 07 primers corresponding to ompR with chromosomal DNA of S. typhi SS3 and SS5 strains. Attenuation by osmolar stress results in deletion mutation of the S. typhi strains was determined by agglutination assays, precipitation method, SDS PAGE analysis and by animal models. Results The 799 bp amplified ompR gene product from wild type S. typhi SS3 and SS5 illustrate the presence of virulent gene. Interestingly, there was only a 282 bp amplified product from S. typhi SS3 and SS5 grown in the presence of 0.5, 0.6 and 0.7 M NaCl. This illustrates the occurrence of deletion mutation in ompR gene at high concentration of NaCl. Furthermore, both the wild-type and mutant S. typhi outer membrane SDS-PAGE profile reveals the differences in the expression of ompF, ompC and ompA proteins. In mice, wild type and mutant strains lethal dose (LD50) were determined. The mice died within 72 h when both the wild type strains were injected intraperitoneally with 3 log CFU.mL−1. When the mice were injected with the mutants in same dosage, no clinical symptoms were observed; whereas the serum antibody titre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response. These results suggest that S. typhi SS3 and SS5 may be used as good candidate strains for the development of live attenuated vaccine against salmonellosis. Conclusions This study demonstrates that the S. typhi strains were attenuated and could be good vaccine candidates in future.

946-951

Authors: Pritom Chowdhury, Siraj Ahmed Khan, Rashmee Topno, Prafulla Dutta, R.N.S. Yadav, Jagadish Mahanta

Objective To assess the best suitable condition for virus inactivation, and to study the immunogenic potential and protective efficacy of a circulating West Nile virus (WNV) strain in Assam. Methods Bulk preparation of circulating WNV: WNIRGC07 (GeneBank ID: HQ246154), was undertaken in a bioreactor using cytodex-1. Virus Inactivation was done in three different conditions; 22 °C, 4 °C and room temperature. The virus preparations were evaluated for antigenicity by ELISA and toxicity by cell proliferation kit. Virus efficacy was done in-vivo on swiss albino mice against standard Indian WNV and Japanese encephalitis virus (JEV) strain. Humoral and cell mediated immune response was evaluated in mice sera by ELISA and neutralization assay. Results Inactivation at 22 °C was found to be more suitable in terms of less toxicity and high antigenicity. The same was selected to study the immune response and efficacy in mice. It induced neutralizing antibody titre of 1: 625 and high IgG response. In vivo experiment showed 100% protective efficacy against WNV and 20.8% cross protective efficacy against JEV. Further assessment of cellular immunity through immunized mice revealed augmentation of high levels of pro-inflammatory cytokines and moderate levels of anti-cytokines indicating a mixed balance of Th1 and Th2 response. Conclusions Findings suggest that formalin inactivated Indian WNV strain has a good immunogenic potential. This is the first study on assessment of immunogenic potential of a lineage 5 strain of WNV. Our study reveals that it would be a promising and effective candidate for vaccine studies which warrants further evaluation.

956-961

Authors: Abbas Khani, Monireh Heydarian

Objective To test fumigant and repellent properties of sesquiterpene-rich essential oil from Teucrium polium subsp. capitatum (L.). Methods The fumigant toxicity test was performed at (27±1)°C, (65±5)% relative humidity, and under darkness condition and 24 h exposure time. The chemical composition of the isolated oils was examined by gas chromatography-mass spectrometry. Results The major compounds were α-cadinol (46.2%), caryophyllene oxide (25.9%), α muurolol epi (8.1%), cadalene (3.7%) and longiverbenone (2.9%). In all cases, considerable differences in mortality of insect to essential oil vapor were observed in different concentrations and exposure times. Callosobruchus maculatus (C. maculates) (LC50=148.9 μL/L air) was more susceptible to the tested plant product than Teucrium castaneum (T. castaneum) (LC50=360.2 μL/L air) based on LC50 values. In the present investigation, the concentration of 3 μL /mL acetone showed 60% and 52% repellency against T. casteneum and C. maculatus adults, respectively. Conclusions The results suggests that sesquiterpene-rich essential oils from the tested plant could be used as a potential control agent for stored-product insects.

968-976

Authors: Thirunavukkarasu Santhoshkumar, Abdul Abdul Rahuman, Chidambaram Jayaseelan, Govindasamy Rajakumar, Sampath Marimuthu, Arivarasan Vishnu Kirthi, Kanayairam Velayutham, John Thomas, Jayachandran Venkatesan, Se-Kwon Kim

Objective To determine the efficacies of antibacterial and antioxidant activities of aqueous leaf extract of Psidium guajava mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs). Methods Synthesized TiO2 NPs were tested by disc diffusion method against against human pathogenic bacteria. The total antioxidant activity and phenolic content (Folin-Ciocalteau method) of synthesized TiO2 NPs and aqueous plant extract were determined. The scavenging radicals were estimated by DPPH method. The synthesized TiO2 NPs were characterized by XRD, FTIR, FESEM and EDX. Results FTIR spectra of synthesized TiO2 NPs exhibited prominent peaks at 3 410 cm−1 (alkynes), 1 578 cm−1, 1 451 cm−1 (alkanes), and 1 123 cm−1(C-O absorption). The morphological characterization of synthesized TiO2 NPs was analysed by FESEM which showed spherical shape and clusters with an average size of 32.58 nm. The maximum zone of inhibition was observed in the synthesized TiO2 NPs (20 μg/mL) against Staphylococcus aureus (25 mm) and Escherichia coli (23 mm). The synthesized TiO2 NPs showed more antibacterial activity than the standard antibiotic disk, tetracycline which drastically reduces the chances for the development of antibiotics resistance of bacterial species. The plant aqueous extract and synthesized TiO2 NPs were found to possess maximum antioxidant activity when compared with ascorbic acid. The content of phenolic compounds (mg/g) in leaf aqueous extract and synthesized TiO2 NPs were found to be 85.4 and 18.3 mgTA/g, respectively. Conclusions Green synthesized TiO2 NPs provides a promising approach can satisfy the requirement of large-scale industrial production bearing the advantage of low-cost, eco-friendly and reproducible.

985-990

Authors: Bao-Qin Liu, Xin Gong, Zhe Jin

Objective To investigate the effects of traditional Chinese medicine, Danzhi decoction, on the expression angiogenesis factors in human endometrial cells during the sequelae of pelvic inflammatory disease (SPID) and explore the role of Danzhi decction in improving the blood stasis microenvironment of SPID. Methods A three-dimensional (3D) co-culture system including human vascular endothelial cells (VECs), endometrial stromal cells and glandular epithelial cells was established in vitro and treated with Danzhi decoction, sterilized water and aspirin respectively. A Milliplex multifunctional liquid chip technique was used to measure the expression levels of vascular endothelial growth factor (VEGF)-A/C/D, fibroblast growth factor −1/2, angiopoietin-2, epidermal growth factor (EGF), HB-EGF, bone morphogenetic protein-9, endoglin, endothelin-1, granulocyte colony stimulating factor, hepatocyte growth factor, interleukin-8, follistatin, placenta growth factor and leptin. The location of angiogenesis factors was monitored by immunofluorescence labeling and confocal laser scanning microscope 3D reconstruction. Results Endometrial stromal cells and glandular epithelial cells were isolated and primary cultured for establishing a 3D co-culture system. The levels of VEGF-A/C/D in Danzhi decoction group and aspirin group were significantly lower than those in mock group (P<0.05), while there was no significant difference between Danzhi decoction group and aspirin group (P>0.05). Furthermore, the alterative location of VEGF-A/C/D was observed in the cytoplasm of endometrial glandular epithelial cells. Conclusions Danzhi decoction may inhibit the expression of VEGF in the blood stasis microenvironment of SPID by targeting the cytoplasm of endometrial glandular epithelial cell.

996-999

Authors: Hong-Mei Liu, Kai-Jie Wang

Objective To investigate the therapeutic effect of the intervention treatment with different doses of Captopril on TNF-αcontents in serum of rheumatoid arthritis (RA) rats, and to provide the theoretical proofs for clinical application of Captopril in treatments of rheumatoid diseases. Methods Fifty Wistar rats were randomly divided into 5 groups, namely, Group A, Group B, Group C, Group D, Group E with 10 rats in each group. Injection of Freund's complete adjuvant was employed to establish adjuvant-induced arthritis model in rats. Group A was model group; after model establishment, rats were treated with 20 mL normal saline as placebo (ip.). Rats in Group B were treated with 8 mg/kg cyclophosphamide (ip.). Rats in Group C, D and E were intraperitoneally injected with 30 mg/kg, 100 mg/kg and 300 mg/kg Captopril respectively. Rats in each group were subjected to continuous treatment for 3 weeks, and then sacrificed. Eyeballs of rats were excised and blood was collected. TNF-αcontent in serum were detected using ELISA; each group rats were compared for the hind legs arthrocele. Right ankle tissues of rats were collected to prepare section, and microscopic observation of pathological changes was performed. Results TNF-αcontent in serum of Group A rats was significantly higher than that of rats in other 4 groups (P<0.05). TNF-αcontent in serum of Group B rats was significantly lower compared with that of rats in Groups C, D and E. The highest TNF-αcontent in serum of rats treated with Captopril was found in Group C, followed by Groups D and E (P<0.05). Right ankle arthrocele of rats in Groups B, C, D and E in early stage showed no statistical difference compared with that of Group A rats (P>0.05). From Day 8, ankle arthrocele of rats in Groups B, C, D and E was obviously relieved compared with that of Group A rats; the anti-inflammatory effects were gradually enhanced with the extension of medication time. Treatments of Groups C, D and E showed significant activities against tardive arthrocele; the degree of ankle arthrocele in rats of these three groups was lower than that of Group A rats (P<0.01). Histological observation showed that large amount of inflammatory cells and plasmocyte infiltration was found in ankle synovial tissues of Group A rats. Relief of hyperaemia and edema of right ankle synovial tissues as well as significant decrease in synoviocyte layer hyperplasia, intra-articular inflammatory cells infiltration and cartilago articularis damage degree etc. were observed in Groups B, C, D and E. Conclusions Intervention treatment with Captopril can effectively reduce the TNF-αcontent in serum of rheumatoid arthritis rats and inhibit the generation of inflammatory factors, so as to achieve the therapeutic effect.

1005-1008

Authors: Tie-Gang Jia, Jian-Qing Zhao, Jian-Hua Liu

Objective To explore the serum levels of IL-32, MMP-9,PCT and CRP in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods A total of 50 patients with AECOPD and 45 cases with acute asthma attack admitted from October 2013 to August 2014 were selected, and the serum levels of IL-32, MMP-9, PCT and CRP were determined and compaed by using Double antibody sandwich Enzyme linked immunosorbent assay, immunofluorescence double antibody sandwich assay and immunoturbidimetry assay. Results Serum levels of IL-32, MMP-9, PCT and CRP were significantly higher in AECOPD group than acute asthma attack group (P<0.05). IL-32 and MMP-9 were negatively correlated with lung function. MMP-9 in AECOPD patients was increased more significantly. Conclusions Serum levels of IL-32 and MMP-9 were negatively correlated with lung function, and the worse the lung function is, the more significant the increase is.

849-853

Authors: Hye-Jin Park

Objective To evaluated the immunomodulatory effect of BRP-4, an acidic polysaccharide from Basella rubra (B. rubra) L on the macrophage activity. Methods Phagocytic activity was determined by the ingestion of Latex Beads-Rabbit IgG-FITC using the fluorescent microscopy and flow cytometry analysis and nitric oxide production was measured using Griess reaction assay. Results An enhanced production of NO was observed at 10 and 100 μg/mL of BRP-4. The phagocytic activity of macrophage was enhanced in BRP-4 treated RAW264.7 cells. BRP-4 combined with concanavalin A (Con A) provided obvious promotion and strengthening of the proliferation of the splenocytes. Conclusions BRP-4, polysaccharide isolated from B. rubra, is suggested to activate macrophage function and stimulate splenocyte proliferation. The strong immunomodulatory activity of BRP-4 confirmed its good potential as an immunotherapeutic adjuvant.

860-866

Authors: Nooran Sherif Elleboudy, Mohammad Mabrouk Aboulwafa, Nadia Abdel-Haleem Hassouna

Objective To study characteristics of phospholipases C (PLCs), their importance for producing microorganisms as well as the potential of their use for industrial purposes. Method PLC from Bacillus cereus (B. cereus) D101 was selected as an example of Gram-positive PLCs and PLC from Pseudomonas aeruginosa (P. aeruginosa) D183 of Gram-negative ones. Enzymes were partially purified by ammonium sulfate precipitation followed by membrane dialysis. Partially purified preparations were used to study effect of different factors on activities as well as in substrate specificity tests which were conducted using a turbidimetric assay method. Results Maximum activity was at pH 7 and 8 and 40 °C for P. aeruginosa PLC, and pH 8-10 and 37 °C for B. cereus PLC. Both PLCs were inhibited by Pi at 5 mM or higher, whereas, PLC from B. cereus only was inhibited by EDTA. Activity of P. aeruginosa PLC was not affected by removing Zn2+ ions from reaction mixture or their replacement with Ca2+, Ba2+, Mg2+ or Mn2+ ions. Vis-à-vis, activity of B. cereus PLC was found to be metal ion dependent. PLCs from both isolates were relatively thermostable and showed maximum affinity toward phosphatidylcholine. Sphingomyelin and phosphatidylethanolamine were not good substrates and phosphatidylinositol, phosphatidylserine, phosphatidylglycerol and cardiolipin could be considered non-substrates. Conclusion Human body physiological conditions could favor activity of P. aeruginosa and B. cereus PLCs. These enzymes may participate in phosphate scavenging and virulence of producing isolates but not in autolysis. PLCs from both isolates are potential candidates for industrial use.

873-878

Authors: Sheng Qiang, Zhen-Fang Du, Min Huang

Objective To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro. Method NDRG2 was harvested by RT-PCR, confirmed by DNA sequencing, and then cloned into the eukaryotic expression vector pIRES2-EGFP, which encodes green fluorescent protein (GFP), to construct pIRES2-EGFP-NDRG2 plasmid. OS-RC-2 cells with NDRG2 negative expression were transfected with pIRES2-EGFP-NDRG2 plasmid. The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes. After colony-forming cell assays, cell proliferation detection and MTT assays, the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells. Cell cycle was determined by flow cytometry. Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle. Results A eukaryotic expression vector pIRES2-EGFP-NDRG2 was successfully constructed. After NDRG2 transfection, the growth of OS-RC-2 cells was inhibited. Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present, and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion. Conclusions NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.

884-888

Authors: Yi Huang, Jiang-bin Chen, Bo Yang, Hui Shen, Jin-Jun Liang, Qiong Luo

Objective To observe the regulatory effects of RhoA/ROCK pathway on the apoptosis of cardiac myocyte induced by anoxia and its mechanism. Methods The model of cardiac myocyte anoxia was established. The beat pulsations and apoptosis rates after 1 h, 3 h, 6 h, 9 h and 12 h of anoxia were recorded and the expressions of RhoA, ROCK1/2, p-PI3K, p-AKT and caspae-3 were detected, too. The apoptosis and the expressions of related proteins were detected after RNAi of RhoA and the inhibition of ROCK by Y-27632. Results The beat pulsations after 1 h, 3 h, 6 h, 9 h and 12 h decreased gradually but the apoptosis rates increased gradually, and the expressions of RhoA, ROCK1/2, p-PI3K, p-AKT and caspase-3 were increasing along with the increasing duration of anoxia. The apoptotic rates after 1 h, 3 h, 6 h, 9 h and 12 h of anoxia were (4.36依0.98)%, (8.36依2.12)%, (15.32依3.62)%, (18.68依4.83)% and (24.56依6.22)%, respectively and decreased more significantly than control group in different time points of anoxia (P<0.05), and the expressions of RhoA, ROCK1/2, p-PI3K, p-AKT and caspase-3 decreased significantly (P<0.05). The apoptosis rate and the expressions of RhoA, ROCK1/2, p-PI3K, p-AKT and caspase-3 decreased significantly (P<0.05) after the inhibition of ROCK by Y-27632 (P<0.05). Conclusions RhoA/ROCK pathway plays a critical role in the regulation of the apoptosis of cardiac myocyte induced by anoxia, which may be accompanied by regulating the activity of PI3K/AKT/Caspase-3 pathway.

895-899

Authors: Zhao-Xun Pan, Hong-Xin Zhang, Ye-Xin Wang, Long-Di Zhai, Wei Du

Objective To observe the effect of recombinant human bone morphogenetic protein 2/poly-lactide-co-glycolic acid (rhBMP-2/PLGA) with core decompression on repair of rabbit femoral head necrosis. Method Bilateral femoral head necrosis models of rabbit were established by steroid injection. A total of 48 rabbits (96 femoral head necrosis) were randomly divided into 4 groups: Group A, control group with12 rabbits, 24 femoral head necrosis; Group B, treated with rhBMP-2/PLGA implantation after core depression, with 12 rabbits, 24 femoral head necrosis; Group C, treated with rhBMP-2 implantation after core depression, with 12 rabbits, 24 femoral head necrosis; Group D treated with core depression group without implantation, with 12 rabbits, 24 femoral head necrosis. All animals were sacrificed after 12 weeks. The ability of repairing bone defect was evaluated by X-ray radiograph. Bone mineral density analysis of the defect regions were used to evaluate the level of ossification. The morphologic change and bone formation was assessed by HE staining. The angiogenesis was evaluated by VEGF immunohistochemistry. Results The osteogenetic ability and quality of femoral head necrosis in group B were better than those of other groups after 12 weeks by X-ray radiograph and morphologic investigation. And the angiogenesis in group B was better than other groups. Group C had similar osteogenetic quality of femoral head necrosis and angiogenesis with group D. Conclusion The treatment of rhBMP-2/PLGA implantation after core depression can promote the repair of rabbit femoral head necrosis. It is a promising and efficient synthetic bone material to treat the femoral head necrosis.

905-908

Authors: Hong-Mei Liu, Yu-Na Guo

Objective To observe the effect of captopril on the tumor necrosis factor-α (TNF-α) level and arterial blood gases in acute lung injury (ALI) induced by HCL in rats, and to analyze its protective mechanism. Methods Fifty Wistar rats were selected and randomly divided into three groups, with 20 rats in Group I and II, respectively and 10 animals in Group III. ALI model was constructed by intratracheal injection of diluted hydrochloric acid (pH=1.25, 1.2 mL/kg). Group I rats received not any treatment after construction of ALI model. Group II rats were treated with captopril (5 mg/kg, i.p.) 5 min after induction of ALI. Group III served as normal control without any treatment. Ninety minutes after construction of ALI model, all the rats were sacrificed. Blood was withdrawn for detection of TNF-α level and arterial blood gases index. And lung tissue slices of the three groups were prepared for observation of pathologic histology changes. Results TNF-α level in serum of Group I and II rats was significantly higher than that in Group III (P<0.05), while TNF-α level in serum of Group II was significantly lower in Group I (P<0.05). PaCO2 level was significantly higher (P<0.05), while PaO2 was significantly lower (P<0.05) in Group I and II rats than those in Group III. PaCO2 was significantly lower (P<0.05) and PaO2 was significantly higher (P<0.05) in Group II than those in Group I. Histological observation showed diffuse congestion and severe edema of lung tissue, obvious thickening and structure damage of alveolar walls and a large amount of neutrophil infiltration in Group I rats. Group II rats showed mild edema of lung tissue; only a small portion of alveolar walls showed thickening and only a few of neutrophil infiltration could be observed. The degree of injury was remarkably slighter than that of Group I rats. Group III rats showed clear lung tissue structure and normal morphology; alveolar walls were uniform and the margin was smooth and few neutrophil could be observed. Conclusions Captopril can significantly reduce serum TNF-α level, elevate PaO2 and reduce PaCO2 in rats with ALI. It has a protective effect on ALI rats.

913-917

Authors: Liu-Hui Wang, Ying Ye, Yi-Qun Zhang, Tao Xiao

Objective To explore the effect of bacilli Galmette-Gurin (BCG)-polysaccharide nuceic acid on atopic dermatitis in mice and its mechanism. Method Forty NC/Nga mice were selected and randomly divided into Group A (model group), Group B (dexamethasone treatment group), Group C (BCG polysaccharide nucleic acid treatment group) and Group D (control group) with 10 mice in each group. Atopic dermatitis model were constructed by applying 2, 4-dinitrochlorobenzene on the skin of the mice. Mice in Group D were treated with acetone solution (100 μL) on the foot pad and abdomen after hair removal at the age of 7 weeks, then on ear skin at the age of 8-13 weeks. For mice in A, B and C groups, 100 μL of acetone solution containing 2, 4-dinitrochlorobenzene was applied to the foot pad and the abdomen at the age of 7 weeks, then on ear skins at the age of 8 to 13 weeks. At the age of 7-13 weeks, mice in Group A and Group D were treated with 100 μL saline (i.p.); mice were given dexamethasone (0.1 mL/kg, i.p.) every other day for 7 weeks in Group B; mice were treated with BCG polysaccharide nucleic acid (0.5 mg/kg, i.p.) every other day for 7 weeks in Group C. The ear thickness was measured every week and the scratching frequency was recorded 1 times for 10 min a week. The mice were sacrificed after the last administration of drugs. IgE, IL-4, IL-10, IL-12 and IFN-γ in the plasma were detected using ELISA, and RT-PCR method was employed to detect the concentrations of IL-4, IL-10, IL-12 and IFN-γ proteins. After HE staining, the lesion degree of inflammation in ear tissue was observed microscopically. Results The ear thickness and scratching frequency of Group A were significantly higher than those in group B, C and D (P<0.05), and there was no significant difference between Group B and C (P>0.05); the concentrations of IgE, IL-4 and IL-10 in the plasma and the expression of IL-4, IL-10 mRNA in the spleen tissues of Group A, B and C were all significantly higher than those of Group D (P<0.05); the concentrations of plasma IL-12 and IFN-γ, and spleen protein expression of IL-12 and IFN-γ in Group C mice were significantly higher than those of Group A (P<0.05). Histological observation showed obvious ear tissue exudation, erythema, swelling, desquamation of skin, and scabbing in Group A. Histopathology of the skin lesion also showed hyperkeratosis, focal-parakeratosis, stratum spinosum hypertrophy, mild sponge-like edema, a large number of lymphocytes along with plasma cell infiltration in dermis, angiectasis and hyperemia in Group A, while degree of ear skin lesion in Group B and D mice was significantly lighter than that of Group A. Conclusions BCG polysaccharide nucleic acid can significantly reduce the serum IgE concentrations, increase the expression of IL-12, IFN-γ protein, correct the imbalance of Thl/Th2 in atopic dermatitis mice, and has obvious inhibitory effect on atopic dermatitis in NC/Nga mice.

922-924

Authors: Özge Altaş, Sabit Sarıkaya, Hakan Saçlı, Onur Yerlikhan, Kaan Kırali

We present a retrospective analysis of three cases of cardiac hydatidosis, who underwent surgery between 2010 and 2012. Two patients had a lesion in the interventricular septum, whereas one patient had the lesion in apicoinferior wall of LV. The diagnosis was made by echocardiography, but magnetic resonance imaging was utilized to assess cyst activity and extend of disease. All patients were placed on cardiopulmonary bypass. No postoperative complication or death occurred. The patients discharged uneventfully and all of them were free from hydatid disease at two years follow-up. We concluded that cardiac hydatid cysts should be removed surgically regardless of their location or extent, even in asymptomatic patients.

761-764

Authors: Yan Zheng, Guo-Rong Wang, Jin-Jing Jia, Su-Ju Luo, Hao Wang, Sheng-Xiang Xiao

Objectives To explore the expressions of c-fos and c-myc in skin lesion of cutaneous squamous cell carcinoma (CSCC). Methods Using retrospective analysis, 73 cases of CSCC were selected from Department of Dermatology, the Second Affiliated Hospital of Xi'an Jiaotong University, which were removed between January 2000 and January 2012. It was considered as experimental group. Meanwhile, 11 cases of normal skin specimens of non tumor patients were selected as control group. The expression level of c-fos and c-myc was compared in the two groups. Results The expressions of c-fos [72.60% (53/73)] and c-myc [83.56% (61/73)] in experimental group were statistically significant (P≤0.05) compared with control group (0%). Expression of c-myc protein was negatively related to differentiation of CSCC. The difference was statistically significant (χ2=7.26, P=0.001<0.05). While expression of c-fos protein was positively related to differentiation of CSCC, which was statistically significant (χ2=7.47, P=0.001 2<0.025). Conclusions The expression level of c-fos and c-myc can be used as an important indicator of CSCC differentiation, and it has closely connection with the differentiated degree, which can guide clinical prognosis.

772-775

Authors: Jian-Bo Che, Zhong-Hua Liu, Hong-Bing Ma, Yong Li, Hui Zhao, Xiao-Hui Li, Wei-Chao Liu, Gong-Ning Shi

Objectives To study the effect of arsenic trioxide (As2O3) combined with ginsenosides Rg3 on inhibiting the NCI-H1299 lung cancer cells and subsistence in nude mice bearing hepatoma. Methods MTT method was used to measure the inhibition effect of As2O3 combined Rg3 on NCI-H1299 cells, and the proliferation inhibiting effect was observed via establishing the transplanted tumor model in vitro. A total of 40 tumor-bearing nude mice were randomly divided into normal saline group, As2O3, Rg3 and As2O3+Rg3 group. Transplantation tumor model of lung cancer in nude mice was constructed, followed by injection of certain concentrations of normal saline, As2O3, ginseng saponin Rg3 and As2O3+Rg3 every day. The survival duration and the tumors size of the mice were recorded and the Kaplan-Meier curve was made; microscopic observation of apoptosis of tumor cells in vivo was done using TUNEL staining. Results After 72 h of injection, inhibition rate of tumor cell in normal saline group, As2O3 group, Rg3 group and As2O3+Rg3 group was (5.66±0.31)%, (65.58±4.75)%, (44.69±3.32)% and (82.67±5.43)%, respectively. Inhibition rate of tumor cell in As2O3 group, Rg3 group and As2O3+Rg3 group was significantly higher than that of normal saline group (P<0.01); inhibition rate of tumor cells of As2O3+Rg3 group was significantly higher than that of the two groups given As2O3 or Rg3 alone (P<0.01). The tumor volume of As2O3 group, Rg3 group and As2O3+Rg3 group shrank to (65.38±3.25)%, (77.68±3.43)% and (42.65±3.55)% of the original, tumor volume of saline group was 1.21 times of the original size (P<0.01); Median survival of saline group, Rg3 group, As2O3 group were significantly shorter than that of As2O3+Rg3 group (P<0.01); co-ordinated intervention ability of As2O3+Rg3 on NCI-H1299 cell was significantly higher than that of As2O3 or Rg3, separately. Conclusions As2O3 combined with Rg3 can significantly inhibit proliferation of NCI-H1299 cells in lung cancer, prolong survival of tumor-bearing nude mice, and promote tumor cell apoptosis, and have significant effect on lung cancer treatment.

780-786

Authors: Chang-Jiang Lei, Chun Yao, Qing-Yun Pan, Hao-Cheng Long, Lei Li, Shu-Ping Zheng, Cheng Zeng, Jian-Bin Huang

Objectives To build GPC3 gene short hairpin interference RNA (shRNA) slow virus vector, observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth, and provide theoretical basis for gene therapy of liver cancer. Methods Hepatocellular carcinoma cell line Huh-7 was transfected by a RNA interference technique. GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR. Targeted GPC3 gene sequences of small interfering RNA (siRNA) PGC-shRNA-GPC3 were restructured. Stable expression cell lines of siRNA were screened and established with the help of liposomes (lipofectamineTM2000) as carrier transfection of human liver cell lines. In order to validate siRNA interference efficiency, GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot. The absorbance value of the cells of blank group, untransfection group and transfection group, the cell cycle and cell apoptosis were calculated, and effects of GPC3 gene on Huh-7 cell proliferation and apoptosis were observed. Results In the liver cancer cell lines Huh-7, GPC3 gene showed high expression. PGC-shRNA-GPC3 recombinant plasmid was constructed successfully via sequencing validation. Stable recombinant plasmid transfected into liver cancer cell lines Huh-7 can obviously inhibit GPC3 mRNA expression level. Conclusions The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.

792-795

Authors: Ying Qin, Ya-Ting Sun, Lin-Xia Xia, Ying-Jie Zhang, Xue-Jun Yang

Objectives To observe the intervention influence and effect of simvastatin on the expression of interleukin 17 (LI17), high mobility group protein 1 (HMGB1) and TLR4 path in Lupus nephritis (LN) rats. Methods A total of 28 BSXSB male mice with LN (16 weeks) were randomly divided into observation group and the comparison group, observation group was given 6 mg•kg−1•d−1 simvastatin in 0.1% PBS lavage for 4 weeks, the comparison group was not given any treatment. Blood urea nitrogen (BUN) level and urine trace albumin (Scr) level of two groups were determined. The expression of IL17, HMGB1 and TLR4 protein was detected using immune histochemical method, and the kidney histological damage was observed. Results BNU, LI17, HMGB1, TLR4 protein and HMGB1 mRNA in observation group was significantly lower than that in control group (P<0.05); There was no statistical difference of Scr level between two groups (P>0.05). Histological observation showed glomerular lesions integral of observation group was obviously lower than that of control group. Conclusions Simvastatin can reduce the expression of IL17, HMGB1 and TLR4 protein in LN mice, thereby can inhibit the autoimmune response as a potential treatment function of LN.

806-810

Authors: Xue-Mei Liu, Yi Feng, Ai-Min Li

Objectives To investigate the nerve protective effect and mechanism of baicalin on newborn rats with hypoxic ischemic brain damage (HIBD). Methods A total of 64 SD newborn rats were randomly divided into control group, model group, nerve growth factor group and baicalin group, with 16 in each group. Left carotid artery ligation method was adopted to establish the HIBD model except for in control group, which was treated with intraperitoneal injection of salin e10 mL/kg for 3 d. After oxygen recovery on hypoxia ischemia rats, intraperitoneal injection of saline 10 mL/kg was adopted in model group for 3 d. Intraperitoneal injection of nerve growth factor injection 50 μg/kg per day was adopted in nerve growth factor group for 3 d; intraperitoneal injection of radix scutellariae 16 mg/kg per day was adopted in baicalin group for 3 d after modeling. Four rats of each group were sacrificed at Day 1, 2, 3, 7 for microscopic observation of pathological morphological changes in brain tissue after HE staining, S-P immunohistochemical method was used for observation of Fas and FasL expression in brain cells. Results Neat structure of cells was observed in control group; edema cells in disordered arrangement was observed in model group, with some cells necrosis and cavity change; tissue injury in nerve growth factor group and baicalin group was significantly lighter than that in model group; Fas and FasL expression in model group, nerve growth factor group and baicalin group were significantly higher than that in control group at different time points (P<0.05); Fas and FasL expression in nerve growth factor group and baicalin group were significantly lower than that in model group at different time points (P<0.05); There was no statistical difference of Fas, FasL expression at each time point between nerve growth factor group and baicalin group (P>0.05). Conclusions Baicalin can reduce expression of Fas and FasL in HIBD rats, inhibit apoptosis of nerve cells, thus achieve the protective effect on HIBD rat nerves.

819-822

Authors: Xiao-Hui Peng, Pei-Yu Liang, Shan-Ji Ou, Xiong-Bing Zu

Objectives To investigate the protective effect of pioglitazone on kidney injury in diabetic rat model and its mechanisms. Methods Forty healthy Sprague Dawley rats were selected and randomly divided into five groups, with 8 rats in each group. Group A served as control group and were administered with sterile citrate buffer (i.p.) as placebo. Groups B, C, D and E rats were injected (i.p.) with streptozotocin to induce type I diabetes. Diabetic rats in Group B were intragastrically administered with sterile saline solution alone. Groups C, D and E rats were intragastrically given pioglitazone hydrochloride suspension at doses of 10, 20, 30 mg/kg per day, respectively. After eight weeks of treatment, all rats were anesthetized and blood was withdrawn from the abdominal aortic for detection of hemoglobin A1c, serum creatinine (SCr) and blood urea nitrogen (BUN) levels. Rats were then sacrificed and the left kidney was excised for calculation of kidney hypertrophy index (KHI), observation of renal pathological changes using light microscope and electron microscope. Mean glomerular cross-sectional areas (MGA), mean glomerular volume (MGV), glomerular basement membrane thickness and foot process fusion ratio were calculated. RT-PCR was employed for detection of podocalyxin (PCX) protein expression. Results Results showed that levels of hemoglobin A1c, BUN, SCr in Groups B, C, D and E rats were significantly higher than those in Group A (P<0.05), while BUN and SCr levels in rats of Groups C, D and E were significantly lower than those in Group B (P<0.05). KHI, MGA and MGV levels were significantly higher in Groups B, C, D and E rats than those in Group A (P<0.05); KHI and MGA levels in Group B rats were significantly higher than those in Groups C, D and E (P<0.05) and MGV in Groups D and E was significantly lower than that in Groups B and C (P<0.05). Histology study showed normal glomerulus structure, morphology, volume, endothelial cells and mesangial cells as well as clear glomerular capillary in Group A rats. Renal mesangial matrix proliferation and expansion of glomerulus cavities in Groups B, C, D and E were observed. However, damage degree in Groups C, D and E were more moderate than that in Group B. Conclusions Pioglitazone can reduce kidney damage in diabetic rats, which may be attributed to its role in increasing glomerular PCX protein expression and inhibiting urinary excretion of PCX, and its effect is dose dependent.

827-831

Authors: Tao Wang, Chang-Liang Zhu, Shuai Wang, Li-Wen Mo, Guo-Dong Yang, Jun Hu, Fan Zhang

Objectives To investigate the role of NLRP3 and NLRP1 inflammasomes signaling pathways in rheumatoid arthritis (RA). Methods A total of 36 patients with RA were selected, peripheral blood mononuclear cell (PBMC) and granulocyte were separated from venous blood. RT-qPCR method was used to detect the expression level and diversity of NLRP3 and NLRP1 in PBMC and granulocyte mRNA in patients with RA, and detect the mRNA expression of downstream factor IL-1α. The correlation between RA and the expression of NLRP3 and NLRP1 was analyzed. Normal 30 cases were set as control group. Results Expression levels of NLRP1, and caspase-1 mRNA in PBMC of RA group were significantly lower than those of control group (P<0.05), while there was no significant difference in expression levels of NLRP3, ASC, IL-1α mRNA between these two groups (P>0.05); NLRP3, caspase-1, and ASC mRNA expression in granulocyte of RA patients were significantly lower than those in control group (P<0.05). There was no correlation between rheumatoid factor and expression levels of NLRP3, ASC, caspase-1 mRNA in RA group (P>0.05); NLRP1, IL-1α mRNA expression level had a negative correlation with anti-rheumatoid factor antibody (P=0.033 2, 0.034 0). Conclusions NLRP3 and NLRP1 inflammasomes signaling pathways are involved in RA inflammatory reaction process as protective factors, and play an important role in RA inflammatory mechanisms.