Objective To investigate the distribution of CD14C ( 260)T single nucleotide polymorphism (SNP) in the Han population in Chongqing and to test the reliability with this random sample in genetic research. Methods Single round Touchdown PCR (TD PCR) and restriction fragment length polymorphism (RFLP) method were used for genotyping in 110 people from the Han population in Chongqing. Results Frequencies of TT, CT, and CC genotypes were 43 6%, 39 1%, and 17 3%, respectively. The sample distribution was accorded with Hardy Weinberg principle. Conclusion CD14C( 260)T SNP was found in the Han population in Chongqing, and the distribution of alleles was significantly different from that in white population. The sample from an H W equilibrium population can be used in the association study of the systematic inflammatory reaction in the Han population in Chongqing.

Objective To establish an animal model of PCOS and to detect the DNA methylation states of LHR,INSR and AR genes. Methods 24-days-old female Sprague-Dawley (SD) rats were randomly divided into two groups. The rats in the experimental group were given subcutaneous implanting of levonorgestrel silica gel staff and injected 1.5 IU hCG twice daily for 9 days from the 4th day. The rats in the control group were injected with normal saline at the same time. Ovarian morphologic changes,sex hormone levels,fasting serum insulin and glucose were detected. The LHR,INSR and AR genes' DNA methylation patterns were checked by methylation specific PCR in modeling group and control group.Results The ovarian weight and volume in modeling group were higher than those in control group. The ovaries in modeling group showed multiple follicular cysts,and the number of theca cells and interstitial cells increased. Less developed follicles and corpus lutea were seem. The serum level of progesterone,testosterone,luteinizing hormone,fasting insulin and glucose were significantly higher in experimental group than those in control group,so as the LH/FSH ratio and HOMA index. No methylation of LHR and AR genes were found in both groups. The methylation frequency of INSR gene (76.7%) was significantly higher in modelinggroup than that in control group (P

Objective To study the clinical value of HRCT in staging of lung interstitial disease(LID) in connective tissue disease(CTD).Methods 222 patients with CTD confirmed clinically underwent HRCT scan.The staging of LID according to the HRCT features of LID were done and the therapeutic effect was compared between each stage.Results In 222 cases ,64 cases were negative on HRCT as stage 0,158 cases had disseminated LID in different degree,including stage Ⅰ in 107,stage Ⅱ in 36 and stage Ⅲ in 15.There were significant statistically between the therapeutic effect and staging of LID.Conclusion The staging of CTD with LID by HRCT is helpful for judging the extent of LID and clinical treatment.

Objective To identify the efficacy small interfering RNA against Pseudomonas aeruginosa expressing MexA-MexB-OprM efflux pumps.Methods Four siRNA ( siRNA1,siRNA2,siRNA3 and siRNA4) against mexB gene were designed and prepared by electricity transference in vitro.MICs of antibiotic combined with efflux pump inhibitors against multiple resistant strain PAO1 and PAO3 were determined by E-test method.The mRNA expression levels of efflux pump gene (mexB) were quantified by real time fluorescent quantitative PCR.Results siRNA expression vectors were constructed success by enzyme cut method.48 after PAO1 and multiple drug resistant PAO3 transfected with siRNA4,the sensibilities to antibiotic were enhanced.48 after PAO1 and multiple drug resistant PAO3 transfected with siRNAl,siRNA2 and siRNA3,the sensibilities to antibiotic didn't change obviously.48 after PAO1 and multiple drug resistant PAO3 ttransfected with siRNA4,the expression level of mexB was decreased obviously (P ＜ 0.05 ).48 after PAO1 and multiple drug resistant PAO3 transfected with siRNA1,siRNA2 and siRNA3,the expression level of mexB didn't change obviously.Conclusion siRNA against Pseudomonas aeruginosa expressing MexA-MexB-OprM efflux pumps enhanced the sensibility to antibiotic and inhibited the expression of mexB gene.Our results demonstrate the using RNAi may be potential targets for novel therapies directed against treatment of antibiotic-resistant infections.

Objective To investigate the effect of quorum sensing autoinducer 3OC12-HSL of Pseudomonas aeruginosa on the apoptosis of Caco-2 cells and its mechanism. Methods Caco-2 cells were incubated with 3OC12-HSL for 4 h and then examined by MTT method for cytoactivities. Flow cytometry was used to analyze apoptosis rate of Caco-2 cells. Expression of apoptosis associated proteins p-p38/MAPK and NF-κB were detected by Western blot. Results After exposure to 3OC12-HSL for 4 h, cytoactivities of Caco-2 cells was reduced(P<0.05) with dose-dependent pattern, and higher dose of 3OC12-HSL leaded to increasing apoptosis rate of Caco-2 cells(P<0.05). 3OC12-HSL raised expression of apoptosis associated proteins p-p38/MAPK and NF-κB detected by Western blot. Conclusion Quorum sensing autoinducer 3OC12-HSL can effect cytoactivities of Caco-2 cells and may induce its apoptosis by enhancing the expression of p-p38/MAPK and NF-κB protein.

Objective To establish an animal model of PCOS and to detect the DNA methylation states of LHR, INSR and AR genes. Methods 24-days-old female Sprague-Dawley (SD) rats were randomly divided into two groups. The rats in the experimental group were given subcutaneous implanting of levonorgestrel silica gel staff and injected 1.5 IU hCG twice daily for 9 days from the 4th day. The rats in the control group were injected with normal saline at the same time. Ovarian morphologic changes, sex hormone levels, fasting serum insulin and glucose were detected. The LHR, INSR and AR genes' DNA methylation patterns were checked by methylation specific PCR in modeling group and control group. Results The ovarian weight and volume in modeling group were higher than those in control group. The ovaries in modeling group showed multiple follicular cysts, and the number of theca cells and interstitial cells increased. Less developed follicles and corpus lutea were seem. The serum level of progesterone, testosterone, luteinizing hormone, fasting insulin and glucose were significantly higher in experimental group than those in control group, so as the LH/FSH ratio and HOMA index. No methylation of LHR and AR genes were found in both groups. The methylation frequency of INSR gene (76.7%) was significantly higher in modeling group than that in control group (P<0. 001). Conclusion The depression of INSR gene's transcriptional induced by DNA methylation is important in the development of insulin resistance in PCOS.

Objective To explore the biological basis of TCM syndromes from a biomolecules network perspective with qi deficiency syndrome as the breakthrough point. Methods A data dictionary of neuro-endocrine-immune (NEI) related genes and qi deficiency syndrome characterization terminology thesaurus were established. Literature about qi deficiency syndrome characterization was retrieved by using Genclip, to excavate the characteristic NEI gene, thereby to explore different bioactive substances of syndromes. Results The analysis of the genetic data, showed qi deficiency related cluster with the relevance of endocrine, signal transduction, hematopoietic cell and immune deficiencies etc. It is confirmed that the intrinsic biological features of TCM syndrome can effectively identify in the NEI level. Conclusion Literature mining method as a new way to discover syndromes biological indicators has certain feasibility, and it is recommended to be further expanded into other studies on syndromes to validate the universality and reliability of this method.

The therapeutic effectiveness of nutritional support in the treatment of severe chronic hepatitis and posthepatitic cirrhosis was evaluated. 143 patients with severe chronic hepatitis and 83 with posthepatitic cirrhosis were evaluated with SGA for assessing the nutritional status before the treatment. Patients with severe chronic hepatitis were divided into three groups: group A subject to enteral nutrition (EN) and parenteral nutrition (PN), group B subject to comprehensive treatment (CT)+PN; group C subject to CT+EN. The patients with posthepatitic cirrhosis were divided into two groups: group D receiving CT and group E receiving CT+PN+EN. The function of liver and kidney and nutritional status were monitored to assess the therapy in 6 weeks. The results showed before treatment, over 90 % patients had moderate to severe malnutrition. After nutritional support, the liver function (ALT, T-bil) and nutritional status (TP, TC) in group A was improved significantly as compared with that in groups B and C (P<0.05). Compared with group D, the values of TP and Alb were increased significantly in group E (P<0.05), but the levels of ALT, AST and T-bil had no obvious change. It was suggested that most patients with severe chronic hepatitis or posthepatitic cirrhosis had malnutrition to varying degrees. The nutritional support treatment could obviously improve the nutritional status of these patients, and was helpful to ameliorate the liver function of the patients with severe chronic hepatitis. Among the methods of nutritional support treatment, PN combined with EN had the best effectiveness.

In order to investigate the role of the MexA-MexB-OprM efflux pump system in the pathogenesis of Pseudomonas aeruginosa (PA)-induced pulmonary infection, pulmonary infection models were established by intratracheal injection of K767 (wild type), nalB (MexA-MexB-OprM up-regulated mutant), and ΔmexB (knockout) strains, separately. All mice were treated with Meropenem (intraper Δ itoneal injection, 100 mg/kg body weight, twice every day), and strain-related pathology, bacteria count, cytokine level, myeloperoxidase (MPO, indicator of neutrophil recruitment) activity, and macrophage inflammatory protein-2 (MIP-2) expression were evaluated at early (3rd day post-infection) and late (7th and 14th day post-infection) stages of infection. E-test showed that ΔmexB was more significantly Δ sensitive to panipenan (ETP), meropenem (MP) and imipenem (IP) than K767 and nalB strains. There was no significant difference in sensitivity to cefepime (TM) among the three stains. In contrast to the K767 and nalB groups, the ΔmexB group showed decreased bacteria burden over time and less exte Δ nsive pathological change. Additionally, MPO activity and levels of inflammatory cytokines (IL-1b, IL-12, and TNF-α) were increased at the early stage (day 3) and decreased at the later stage (day 14). Serum MIP-2 expression level was steadily increased in all three groups from early to late stages, but significantly higher in ΔmexB group than in K767 and nalB groups ( Δ P<0.05). In conclusion, the MexA-MexB-OprM efflux pump system might play an important role in PA-induced chronic pulmonary infection. High expression of the MexA-MexB-OprM efflux pump could increase antibacterial resistance and promote infection.

Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.