The association analysis of TLR2 and TLR4 gene with tuberculosis in the Tibetan Chinese population.

BACKGROUND: The present study was undertaken to explore the relationship of Toll-like receptor (TLR) 2, TLR4 genes polymorphisms with Pulmonary tuberculosis (PTB) risk in a sample of Chinese population. METHODS: For this study, we recruited 467 subjects with PTB and 504 healthy subjects from a Tibetan population living in near or in Xi'an, China. Association analyses of single-nucleotide polymorphisms (SNPs) in TLR2 and TLR4 were performed with SPSS Statistics (version 17.0), SNPStats, Haploview (version 4.2), and SHEsis software. RESULTS: The research results that is association analysis of pulmonary tuberculosis show there are two increased-risk SNPs (rs7696323, OR=1.32, 95%CI =1.08-1.62, P= 0.007; rs12377632, OR=1.30, 95%CI =1.09-1.55, P= 0.004) and three decreased-risk SNPs (rs3804099, OR=0.64, 95%CI =0.52-0.79, P= 1.9510-5; rs3804100, OR=0.67, 95%CI =0.54-0.82, P= 0.0001; rs11536889, OR=0.54, 95%CI =0.42-0.69, P= 9.1410-7). CONCLUSIONS: We found that two SNPs are associated with increased PTB risk and three SNPs decreased PTB risk in the Chinese Tibetan population. Our findings demonstrate an association between TLR2 and TLR4 polymorphisms and PTB.

Peginterferon and Chinese herbs exert a combinatorial effect in HBeAg-positive chronic hepatitis B.

INTRODUCTION: Traditional Chinese herbs are widely used for the treatment of chronic hepatitis B (CHB) in China. The aim of this study was to perform a meta-analysis of randomized controlled trials (RCTs) comparing peginterferon therapies with peginterferon plus Chinese herbal therapies in hepatitis B e antigen (HBeAg)-positive CHB patients. METHODOLOGY: The main biomedical databases were searched to identify RCTs that compared the efficiency of peginterferon with peginterferon plus Chinese herbs in CHB patients. RESULTS: The literature search yielded 616 studies, and 8 RCTs (624 patients) matched the selection criteria. Combined therapies of peginterferon plus Chinese herbal therapies were superior to peginterferon therapies alone in achieving the serum HBV DNA clearance rate (64.5% vs. 45.0%), serum HBeAg clearance rate (47.4% vs. 33.5%), and HBeAg seroconversion rates (39.2% vs. 23.1%) at the end of treatment. Combined therapies were more effective than peginterferon alone therapies in the improvement of liver fibrosis related biomarkers, including hyaluronic acid, procollagen type III, type IV collagen, and lamina. Combined therapies also resulted in fewer relapses, fewer adverse events, and more rapid alanine transaminase normalization. CONCLUSIONS: The current evidence suggests that peginterferon plus Chinese herbal therapies were associated with higher virological response than peginterferon alone in HBeAg-positive CHB patients.

Atorvastatin delays the glucose clearance rate in hypercholesterolemic rabbits.

The administration of statin might increase the risk of new-onset diabetes in hypercholesterolemic patients based on the recent clinical evidence. However, the causal relationship must be clarified and confirmed in animal experiments. Therefore, we mimicked hypercholesterolemia by feeding rabbits a high-cholesterol diet (HCD) and performed 16 weeks of atorvastatin administration to investigate the effect of statin on glucose metabolism. The intravenous glucose tolerance test showed that plasma glucose levels in the statin-treated rabbits were consistently higher and that there was a slower rate of glucose clearance from the blood than in HCD rabbits. The incremental area under the curve for glucose in the statin-treated rabbits was also significantly larger than in the HCD rabbits. However, there was no significant difference between the two groups in the intravenous insulin tolerance test. The glucose-lowering ability of exogenous insulin was not impaired by statin treatment in hypercholesterolemic rabbits. The administration of a single dose of statin did not affect glucose metabolism in normal rabbits. The statin also significantly increased the levels of high-density lipoprotein cholesterol, alanine aminotransferase and aspartate transaminase and decreased plasma levels of total cholesterol, triglycerides and low-density lipoprotein cholesterol in the hypercholesterolemic rabbits, whereas it did not affect plasma levels of glucose and insulin. The current results showed that atorvastatin treatment resulted in a significant delay of glucose clearance in hypercholesterolemic rabbits, and this rabbit model could be suitable for studying the effects of statin on glucose metabolism.

The expression of small regulatory RNAs in clinical samples reflects the different life styles of Staphylococcus aureus in colonization vs. infection.

Small RNAs (sRNAs) are involved in the post-transcriptional regulation of metabolic pathways and in responses to stress and virulence. We analyzed the expression levels of five sRNAs of Staphylococcus aureus during human colonization or infection. Total RNA was isolated from nasal carriers, abscesses and cystic fibrosis patients (20 subjects per condition). The expression levels of the sRNAs were measured in the clinical samples and compared with those of the corresponding strains grown in vitro. Five sRNAs were encoded and expressed in all clinical strains in vitro. In vivo, the global expression of the five sRNAs was extremely variable in the abscessed patients, more homogeneous in the cystic fibrosis patients, and highly uniform in the nasal carrier samples. The expression levels of the sRNAs in vivo resembled those obtained at exponential phase or late exponential phase of growth in vitro, for three and one sRNA respectively; while for one sRNA, the expression was always higher in vivo as compared to in vitro growth. The in vitro conditions do not uniformly mimic the in vivo conditions for sRNA expression. Nasal colonization is associated with a unique expression pattern of sRNA that might reflect the commensalism of S. aureus in this niche.

Membrane proteomic analysis comparing squamous cell lung cancer tissue and tumour-adjacent normal tissue.

Lung cancer is the leading cause of cancer-related deaths worldwide. Squamous cell carcinoma is one of the predominant histological subtypes of lung cancer. Detecting lung cancer at an early stage is essential for successful therapy and increasing survival. There are still no satisfactory biomarkers for the early detection of lung cancer. In this study, tumour tissue paired with tumour-adjacent normal bronchial epithelial tissue was obtained from patients with squamous cell lung carcinoma without metastasis. The proteins extracted from the cell membrane were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and were analysed with the Image Master two-dimensional platinum software. Twenty-five significantly different protein spots were selected and identified with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). A total of 19 proteins were successfully identified. Twelve proteins were up-regulated, and seven proteins were down-regulated in the cancerous tissue compared with the tumour-adjacent normal tissue. One up-regulated protein and one down-regulated protein in squamous cell lung carcinoma were verified by Western blot analysis and RT-PCR; the results were consistent with the 2-DE analysis. In conclusion, membrane proteomics identified a number of candidate biomarker proteins that were differentially expressed between squamous cell lung cancer tissue and adjacent normal tissue. These biomarker candidates have the potential to elucidate the underlying pathogenesis of squamous cell lung cancer.

[Bioinformatics analysis on CK8 full length coding sequence and eukaryotic expression in SMMC7721 cells].

AIM: To construct an eukaryotic expression vector containing the coding region of human full length cytokeratin 8 gene and to detect its expression in SMMC7721 cells. METHODS: CK8 cDNA was amplified by RT-PCR and cloned to pMD18-T simple vector. After confirming the sequence, the cDNA was inserted into pEGFP-C1 and the positive clone pEGFP-CK8 was obtained. The recombinant plasmid was transfected into SMMC7721 cells with Lipofectamine(TM);2000 and the expression was detected by fluorescence microscope, real time PCR and Western blot. The physical-chemical properties, signal peptide and functional motifs were predicted by the bioinformatics software. RESULTS: PCR, restriction enzyme digestion and DNA sequencing showed that the recombinant plasmid contained the coding region of full length CK8 gene. Observation under fluorescence microscope and the results of real time PCR and western blot indicated CK8 was over-expressed in SMMC7721 cells. CONCLUSION: The eukaryotic expression vector containing the CK8 gene was successfully constructed and expressed, which provides a basis for the study for biological function of CK8.

Effects of T-2 toxin and selenium on chondrocyte expression of matrix metalloproteinases (MMP-1, MMP-13), α2-macroglobulin (α2M) and TIMPs.

T-2 toxin is regarded as an important etiological factor of Kashin-Beck disease, and supplementation of selenium-salt partly prevents Kashin-Beck disease. The present study investigated the effects of T-2 toxin on the degradation of type II collagen in human chondrocytes in vitro. Human chondrocytes were isolated and cultured on bone matrix gelatin to form an artificial cartilage model in vitro with or without T-2 toxin and selenium. Immunohistochemistry analyses showed that T-2 toxin decreased type II collagen staining and selenium appeared to prevent the decrease in type II collagen induced by T-2 toxin in engineered cartilage. Then, Western blot and RT-PCR analyses showed that an increase in MMP-13 and MMP-1 expressions, and a decrease in the expression of the general endoproteinase inhibitor (α(2)M) were induced by T-2 toxin. Gelatin reverse zymography showed that TIMP-1 and TIMP-2 levels were decreased in a dose-dependent manner after exposure of T-2 toxin. Selenium had a protective role by increasing the level of type II collagen protein through down-regulation of MMP-13 protein and mRNA expression and up-regulation of TIMP-1 and TIMP-2 expressions. These data suggest T-2 toxin induces cartilage matrix degradation by the up-regulation of MMP-13 and TIMP-1, and down-regulation of TIMP-2 and α(2)M expressions.

Interferon plus Chinese herbs are associated with higher sustained virological response than interferon alone in chronic Hepatitis C: a meta-analysis of randomised trials.

BACKGROUND/AIMS: Traditional Chinese herbal therapies are widely used for the treatment of chronic hepatitis C (CHC) in Asia. The aim of this study was to perform a meta-analysis of randomised controlled trials (RCTs) comparing interferon therapies with Chinese herbal therapies and/or interferon plus Chinese herb therapies for the treatment of CHC. METHODS: The Cochrane Central Register of Controlled Trials, Medline, Science Citation Index, EMBASE, China National Knowledge Infrastructure, Wanfang Database and China Biomedical Database were searched to identify RCTs that evaluated the virological response to interferon therapies, Chinese herbal therapies and interferon plus Chinese herb therapies in CHC patients. We statistically combined data using a random-effect meta-analysis according to the intention-to-treat principle. RESULTS: The literature search yielded 770 studies, and 26 RCTs comprising 1905 patients matched the selection criteria. Overall, the sustained virological response (SVR) was significantly higher in patients treated with interferon plus Chinese herbs than in patients treated with interferon alone (49% vs 33%, relative risk, 1.52; 95% confidence interval: 1.23-1.89; p<0.05). Combined therapies of interferon plus Chinese herb therapies were also superior to interferon therapies alone in achieving the end-of-treatment viral response (ETVR), and resulted in fewer relapses, fewer adverse events and more rapid alanine transaminase normalisation. Interferon therapies achieved higher ETVR than Chinese herbal therapies, but they yielded a similar SVR. CONCLUSIONS: The current evidence suggests that combined therapies of interferon plus Chinese herbs yielded a higher SVR, and resulted in fewer relapses and fewer adverse events than interferon therapies.

Human prion protein mutants with deleted and inserted octarepeats undergo different pathways to trigger cell apoptosis.

Octarepeats region sequence is one of the most important characteristics of PrP topology. To explore the mechanism of deleted and inserted octarepeats mutants PrP-caused apoptosis, wild-type PrP (PrP-PG5), and PrP with deleted octarepeats (PrP-PG0) and with four (PrP-PG9) and seven (PrP-PG12) extra octarepeats were transiently induced into SH-SY5Y cell. The results indicated PrP-PG9 and PrP-PG12 mainly retained in fraction of cytoplasm, while PrP-PG5 and PrP-PG0 presented both in cell membrane and cytoplasm. Cells expressing PrP-PG9 and PrP-PG12 were sensitive to endoplasmic reticulum (ER) stimuli, tunicamycin, and brefeldin A. ER-stress-related proteins, Grp94, XBP1, TRAF2, and CHOP, were significantly increased in cells expressing PrP-PG9 and PrP-PG12, while Grp78 increased markedly 12 h and pro-caspase-12 decreased sharply 20 h post-transfection. It indicates that expressions of PrP mutants with inserted octarepeats cause ER stress and lead to cell apoptosis lately. Meanwhile, cellular Cytochrome C increased and Bcl-2 decreased obviously in cells expressing PrP-PG0, indicating triggering a mitochondrial-related apoptosis. These data highlight that PrP mutants in region of octarepeats may undergo different pathways to trigger cell apoptosis, in which PrPs with inserted octarepeats via ER stress and PrP mutant without octarepeats via mitochondrial-related pathway.

Expressions of nucleoside diphosphate kinase (nm23) in tumor tissues are related with metastasis and length of survival of patients with hepatocellular carcinoma.

OBJECTIVE: To evaluate the relationship of expressions of nucleoside diphosphate kinase (nm23) and proliferating cell nuclear antigen (PCNA), as well as apoptosis, with the prognosis of HCC patients by analyzing their pathological and clinical data. METHODS: The expressions of nm23 and PCNA were analyzed by immunohistochemistry and the apoptotic phenomena were detected by TUNEL technique in the liver samples from 43 HCC tissues, 39 para-neoplastic tissues, and 10 normal tissues. The mean apoptosis index (AI) and proliferative index (PI) in individual sample were calculated. RESULTS: As shown by the detection, 32.6% of carcinomas had negative nm23 signal in tumor tissues, whereas all para-neoplastic and normal tissues had positive nm23. The AI in nm23 positive HCC was significantly higher than that in nm23 negative one, with statistical difference (P<0.05). Furthermore, the expressions of nm23, and the values of AI and PI were contrastively analyzed with some main pathological and clinical data of HCC. It revealed that HCC with extrahepatic metastasis showed remarkable correlation with the negative nm23 (P=0.013) and higher PI values of HCC (P=0.015). The disease-free survival in HCC patients with negative nm23 expression was significantly poorer than that in patients with positive nm23 expression. CONCLUSIONS: These data suggest that expressions of nm23 protein in tumor tissues are correlated with occurrences of metastasis and length of survival of the HCC patients, which may be an indicator for their prognosis.

[Correlation between ethambutol-resistance and embB gene mutation in Mycobacterium tuberculosis].

OBJECTIVE: To analyze the characteristics of embB gene mutation in ethambutol-resistant Mycobacterium tuberculosis (M. TB) strains isolated from patients in Xi'an for establishing a rapid method to detect the mutation of embB gene in M. TB. METHODS: The embB gene of 104 Mycobacterium tuberculosis isolates was detected by PCR-Single Strand Conformation Polymorphism (PCR-SSCP) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP). RESULTS: Taking strain H37Rv as a control, the SSCP and RFLP profiles of embB of 35 drug-sensitive strains in 104 Mycobacterium tuberculosis isolates were normal. The display of SSCP profiles in 39 of 69 (56.52%) ethambutol-resistant isolates and the display of RFLP profiles in 19 of 69 (27.54%) ethambutol-resistant isolates were abnormal. CONCLUSION: The EMB-resistance of M. TB strains isolated from patients in Xi'an is related to the mutation of embB gene, PCR-SSCP and PCR-RFLP are sensitive methods for detecting ethambutol-resistance of M. tuberculosis.

[Expression of keratin 8 in carbon tetrachloride-induced liver injury of mice].

OBJECTIVE: To investigate the expression of keratin 8 (K8) in carbon tetrachloride (CCl(4))-induced liver injury of mice. METHODS: Forty ICR mice were divided into four groups. CCl(4) 300 microl/kg body weight in olive oil was injected intraperitoneally for 0, 2, 4, 6 weeks in group A, B, C and D, respectively. Mice were sacrificed 3 d after the last injection and then the vital organs were collected and weighed. RT-PCR and immunohistochemistry methods were used to analyze the expression of keratin 8 in the liver. RESULTS: The ratios of liver and body weight were increased significantly after administration of CCl(4), which were 5.60 %, 6.87%, 7.83 % and 7.76% at 0, 2, 4 and 6 weeks after injection, respectively. The expression of K8 was increased at the 2 w, 4 w and 6 w after CL(4) administration. CONCLUSION: The expression of K8 is positively correlated with the liver injury induced by CCl(4). The accumulation of K8 may be involved in the mechanism of liver injury.

[Cloning and expression of human keratin 8 gene cDNA in E.coli].

AIM: To clone and express human keratin 8 gene cDNA in E.coli. METHODS: Human cytokeratin 8 gene cDNA was amplified by RT-PCR from genomic RNA of human cell line 7721. The amplified cytokeratin 8 gene cDNA was cloned into pMD18-T vector. Then, the CK8 cDNA was amplified by PCR from recombinant plasmid pMD18-CK8, and was subcloned into pET-28a(+) expression vector. The recombinant plasmid pET-28a-CK8 DNA was transformed into E.coli DH5alpha strain. RESULTS: Human cytokeratin 8 gene cDNA was cloned, and the recombinant plasmid pMD18-CK8 was transformed into E.coli. The CK8 cDNA was subcloned into E.coli DH5alpha strain, and successfully expressed in E.coli. CONCLUSION: Human cytokeratin 8 gene cDNA is cloned, and successfully expressed in E.coli, which lay the foundation of further study on the CK8 biological properties and functions.

Separation and identification of differentially expressed nuclear matrix proteins in breast carcinoma forming.

BACKGROUND: Breast carcinoma is one of most prevalent malignant tumors occurring in women. Short of prevention, detection of breast carcinoma at an early, still curable stage would offer the best route to decrease its mortality rates. This highlights the urgent need for suitable biomarkers for early diagnosis and a better understanding of the disease pathogenesis. MATERIAL AND METHODS: NMPs were extracted from normal human breast tissue (Group I), from hyperplastic mammary tissue specimens (Group II), from atypical epithelial hyperplasia specimens (Group III), and from breast carcinoma (Group IV) tissue. Differential proteome profiles were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The different NMPs were analyzed in the National Center for Biotechnology Information (NCBI) database with Mascot software. RESULTS: Well-resolved, reproducible 2-DE profiles of human breast tissues were obtained. Average protein spots were 904 +/- 58, 912 +/- 51, 931 +/- 63, 944 +/- 70 in Group I, Group II, Group III, and Group IV, respectively. Several different proteins were analyzed using mass spectrometry and bioinformation. Of these, 12 were well characterized. Compared to Group I, three proteins were up-regulated in Groups II, III, and IV, including Hsp27, prohibitin, and laminA/C. Upregulation was confirmed using Western blotting and immunohistochemical analysis. The correlation of prohibitin expression with clinicopathological features was also investigated. DISCUSSION: The proteins identified in this study may potentially prove to be useful markers for breast carcinoma diagnosis.

[Meta-analysis on peginterferon plus ribavirin in treatment of hepatitis C virus genotype 1 or 4 infection in HIV patients].

OBJECTIVE: To perform a Meta-analysis on peginterferon with interferon in treatment of HIV patients coinfected with refractory genotype HCV. METHODS: A literature search of Medline was conducted to identify eligible randomized controlled trials. Meta analysis was conducted to evaluate peginterferon and interferon in treatment of coinfected HCV genotype 1 or 4 in HIV patients. RESULT: Six trials of 88 matched the selection criteria. Total 1,131 patients with coinfection of HCV genotype 1 or 4 and HIV were included. Sustain viral response was higher in patients treated with peginterferon plus ribavirin compared with that of interferon plus ribavirin (26 % compared with 8 %) or peginterferon alone (26 % compared with 13 %). Severe adverse effects and withdrawal rates were similar for patients treated with peginterferon and patients treated with interferon. CONCLUSION: Peginterferon plus ribavirin in treatment of patients with coinfection of genotype 1 or 4 HCV and HIV can achieve higher sustain viral response and the likelihoods of serious adverse effects and withdrawal rates are similar to other therapies.

Recent dermatophyte divergence revealed by comparative and phylogenetic analysis of mitochondrial genomes.

BACKGROUND: Dermatophytes are fungi that cause superficial infections of the skin, hair, and nails. They are the most common agents of fungal infections worldwide. Dermatophytic fungi constitute three genera, Trichophyton, Epidermophyton, and Microsporum, and the evolutionary relationships between these genera are epidemiologically important. Mitochondria are considered to be of monophyletic origin and mitochondrial sequences offer many advantages for phylogenetic studies. However, only one complete dermatophyte mitochondrial genome (E. floccosum) has previously been determined. RESULTS: The complete mitochondrial DNA sequences of five dermatophyte species, T. rubrum (26,985 bp), T. mentagrophytes (24,297 bp), T. ajelloi (28,530 bp), M. canis (23,943 bp) and M. nanum (24,105 bp) were determined. These were compared to the E. floccosum sequence. Mitochondrial genomes of all 6 species were found to harbor the same set of genes arranged identical order indicating that these dermatophytes are closely related. Genome size differences were largely due to variable lengths of non-coding intergenic regions and the presence/absence of introns. Phylogenetic analyses based on complete mitochondrial genomes reveals that the divergence of the dermatophyte clade was later than of other groups of pathogenic fungi. CONCLUSION: This is the first systematic comparative genomic study on dermatophytes, a highly conserved and recently-diverged lineage of ascomycota fungi. The data reported here provide a basis for further exploration of interrelationships between dermatophytes and will contribute to the study of mitochondrial evolution in higher fungi.

Anticancer activity of PDSS2, prenyl diphosphate synthase, subunit 2, in gastric cancer tissue and the SGC7901 cell line.

The aim of this study was to assess whether PDSS2 (prenyl diphosphate synthase, subunit 2), a candidate tumor suppressor protein, has a potential anticancer role in human gastric cancer tissue and the SGC7901 gastric cell line. A PDSS2 eukaryotic expression vector was constructed and introduced into SGC7901 cells. The relationship between PDSS2 expression and cell proliferation, cell cycle distribution, and apoptosis in tumor cells was analyzed by RT-PCR, western blotting, the MTT colorimetric assay, flow cytometry, and immunohistochemistry. Increased exogenous PDSS2 expression in vitro is associated with decreased cellular proliferation of the gastric cancer cell line SGC7901. PDSS2 also induced apoptosis in SGC7901 cells by causing cell cycle arrest in the G0/G1 phase. Moreover, a significantly low expression level of PDSS2 protein was found in gastric cancer. Decreased or absent expression of PDSS2 was showed in the gastric tumor biopsy samples analyzed, correlating with cancer differentiation. PDSS2 has potent anticancer activity in gastric cancer tissues and the SGC7901 cell line and is possibly involved in apoptosis in SGC7901 cells.

The correlations between HPV16 infection and expressions of c-erbB-2 and bcl-2 in breast carcinoma.

Breast carcinoma (BC) is a prevalent malignant tumour occurring in women. Many studies have indicated the role of human papilloma virus type 16 (HPV16) in the pathogenesis of BC; however, the correlations of HPV16 infection with the clinicopathologic features of BC and the expressions of c-erbB-2 and bcl-2 have not yet been elucidated. In this study, HPV16 was detected by amplifying the HPV16 E6 gene by the polymerase chain reaction method, and the expressions of c-erbB-2 and bcl-2 in 40 BCs and 20 normal breast tissue samples, obtained from Shaanxi Province, were examined using the streptavidin-peroxidase method with monoclonal antibodies specific to c-erbB-2 and bcl-2. The infection rate of HPV16 E6 and the positive expression rate of c-erbB-2 were significantly higher in the BCs than in the normal tissues (HPV16 E6: 60% vs. 5%; c-erbB-2: 42.5% vs. 5%, P < 0.05). However, the positive expression rate of bcl-2 was significantly lower in the BCs than in the normal tissues (67.5% vs. 95%, P < 0.05). The infection rate of HPV16 did not correlate with any of the pathological features observed (P > 0.05). HPV16 infection correlated with bcl-2 expression (P = 0.015) but not with c-erbB-2 expression (P = 0.747) in the BCs. Interestingly, HPV16 infection correlated with bcl-2 expression in grade I BCs (P = 0.018) but not in grade II-III BCs (P = 0.633). Our data suggest that HPV16 infection is correlated with bcl-2 expression in BCs.

Over-expression of nm23-H1 in HeLa cells provides cells with higher resistance to oxidative stress possibly due to raising intracellular p53 and GPX1.

AIM: To determine whether the antitumor factor nm23 is related with antioxidation. METHODS: Full-length human nm23-H1 was cloned into a mammalianexpressing vector and transiently introduced into HeLa cells. RESULTS: A remarkably low level of reactive oxygen species (ROS) was detected in the cells overexpressing nm23-H1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays found that the cells transfected with a nm23- H1-expressing plasmid had higher viability and stronger resistance to oxidative stress. Immunoprecipitation tests revealed that endogenous nm23-H1 formed a protein complex with p53. Furthermore, the intracellular levels of p53 and p53- regulated gene GPX1 were obviously increased in the cells overexpressing nm23- H1. The downregulation of p53 in the cells overexpressing nm23-H1 resulted in a higher cellular ROS level and lower cell viability. CONCLUSION: The findings suggest that nm23-H1 may act as a cellular protector against oxidative stress, possibly triggering the p53-related antioxidative pathway.

Proteome analysis identifies the role of heat stress in production of progeny HCV in Huh7 cells harboring intact HCV.

OBJECTIVE: The intact hepatitis C virus (HCV) cell culture system has provided a powerful tool for studying the interaction between HCV and host cell. We applied proteomic techniques to globally analyze the protein expression profiles of Huh7 in the absence and presence of HCV, with the aim to elucidate the host cell components response to HCV. METHODS: Proteomic and molecular biology techniques were used for this aim. RESULTS: The expression of many proteins, including regucalcin, centriolin and several keratins, was up-regulated, after HCV transfection. Among the down-regulated expression proteins, the heat-shock protein family was of prime significance. Subsequently, we studied the role of heat stress in the interaction between HCV and host cell, based on proteomics analysis. We found heat stress did not affect the production of HCV in this in vitro cell culture system. CONCLUSIONS: Moreover, this study also provided the global information of proteomic alteration of Huh7 cells in the presence of intact HCV and further improved the understanding of the mechanism of interaction between HCV and host cell. We also draw a conclusion that heat stress had no affect on the production of HCV.