ECSM2, an endothelial specific VE-cadherin binding protein, has a tyrosine phosphorylation site essential to cell migration.

Endothelial cell-specific molecule 2 (ECSM2) is a transmembrane protein located in cell-cell junction of endothelial cells (EC). ECSM2 was determined to play an important role in vascular development, EC migration, apoptosis and proliferation, however, no functional domains were determined in intracellular and extracellular region of ECSM2. In current work, functional domains of ECSM2, the relationship of ECSM2 with other endothelial specific protein such as VE-cadherin and the role of ECSM2 in neovascular diseases were determined. It was shown that the 54th amino acid residue of ECSM2 extracellular domain was a tyrosine phosphorylation site, whose mutation led to the loss of EGF-induced tyrosine phosphorylation and inhibition of cell migration. In primary human umbilical vein endothelial cells, ECSM2 bound with VE-cadherin and VEGF stimulation enhanced their binding. In hepatocellular carcinoma, ECSM2 expression was increased, as compared with para-cancerous tissue. This data firstly revealed the functional sites of ECSM2, the crosstalk between ECSM2 and other endothelial cell specific molecules, the expression of ECSM2 in tissues of angiogenesis diseases, thus providing understanding about ECSM2 in depth.

IRAK-M alters the polarity of macrophages to facilitate the survival of Mycobacterium tuberculosis.

BACKGROUND: Intracellular bacterium, Mycobacterium tuberculosis (M. tb), infects specifically macrophages as host cells. IRAK-M, a member of IRAK family, is a negative regulator in TLR signaling and specifically expresses in monocytes and macrophages. The role of IRAK-M in intracellular growth of M. tb and macrophage polarization was explored, for deeply understanding the pathogenesis of M. tb, the significance of IRAK-M to innate immunity and pathogen-host interaction. METHODS: IRAK-M expression was detected in M. tb infected macrophages and in human lung tissue of pulmonary tuberculosis with immunofluorescence staining, Western blot and immunohistochemistry. IRAK-M knock-down and over-expressing cell strains were constructed and intracellular survival of M. tb was investigated by acid-fast staining and colony forming units. Molecular markers of M1-type (pSTAT1 and iNOS) and M2-type (pSTAT6 and Arg-1) macrophages were detected using Western blot in IRAK-M knockdown U937 cells infected with M. tb H37Rv. U937 cells were stimulated with immunostimulant CpG7909 into M1 status and then infected with M. tb H37Rv. Expression of IRAK-M, IRAK-4 and iNOS was detected with immunofluorescence staining and Western blot, to evaluate the effect of IRAK-M to CpG directed M1-type polarization of macrophages during M. tb infection. Molecules related with macrophage's bactericidal ability such as Hif-1 and phosphorylated ERK1/2 were detected with immunohistochemistry and Western blot. RESULTS: IRAK-M increased in M. tb infected macrophage cells and also in human lung tissue of pulmonary tuberculosis. IRAK-M over-expression resulted in higher bacterial load, while IRAK-M interference resulted in lower bacterial load in M. tb infected cells. During M. tb infection, IRAK-M knockdown induced M1-type, while inhibited M2-type polarization of macrophage. M1-type polarization of U937 cells induced by CpG7909 was inhibited by M. tb infection, which was reversed by IRAK-M knockdown in U937 cells. IRAK-M affected Hif-1 and MAPK signaling cascade during M. tb infection. CONCLUSIONS: Conclusively, IRAK-M might alter the polarity of macrophages, to facilitate intracellular survival of M. tb and affect Th1-type immunity of the host, which is helpful to understanding the pathogenesis of M. tb.

Hypoxia-inducible factor-1alpha regulates autophagy to activate hepatic stellate cells.

The role of autophagy in Hif-1α modulated activation of hepatic stellate cells was illustrated in current work. Autophagy markers were determined in livers of Schistosoma japonicum infected mice and hypoxia or LPS treated human hepatic stellate cell, LX-2 cells. The action of Hif-1 to autophagy was defined as increase of autophagy markers was significantly suppressed in Hif-1α siRNA transfected cells upon hypoxia or LPS stimulation. The function of autophagy in activation of LX-2 cells was assessed as increase of activation markers was blocked using autophagy inhibitors under hypoxia and LPS stimulation. Conclusively, Hif-1α regulates activation of hepatic stellate cell by modulating autophagy.

Hypoxia-inducible factor-1alpha and MAPK co-regulate activation of hepatic stellate cells upon hypoxia stimulation.

BACKGROUND: Hepatic stellate cell (HSC) plays a key role in pathogenesis of liver fibrosis. During liver injury, hypoxia in local micro-environment is inevitable. Hif-1α is the key transcriptional regulation factor that induces cell's adaptive responses to hypoxia. Recently, it was reported that MAPK is involved in regulation of Hif-1α activity. AIMS: To explore whether Hif-1α regulates HSC activation upon hypoxia, and whether MAPK affects Hif-1α-regulated signaling cascades, thus providing new targets for preventing liver fibrosis. METHODS: Hif-1α expression in livers of Schistosomajaponicum infected BALB/c mice was detected with western blot and immunohistochemistry. A rat cell line of HSC, HSC-T6, was cultured in 1% oxygen. HSC activation, including F-actin reorganization, increase of vimentin and α-SMA, was detected with western blot or immunocytochemistry. Cells were transfected with specific siRNA to Hif-1α, expression of activation markers, transcription of fibrosis-promoting cytokines, secretion of collagen I were detected with western blot, Real Time PCR and ELISA. Lysate from HSC-T6 cells pretreated with PD98059, a specific MEK1 pharmacological inhibitor, was subjected to detect Hif-1α ubiquitination and nuclear translocation with western blot and immunoprecipitation. RESULTS AND CONCLUSIONS: Hif-1α apparently increased in liver tissues of Schistosomajaponicum infected mice. 1% O2 induced F-actin reorganization, increase of Hif-1α, vimentin and α-SMA in HSC-T6 cells. Hif-1α Knockdown inhibited HSC-T6 activation, transcription of IL-6, TGF-ß and CTGF and secretion of collagen I from HSC-T6 cells upon hypoxia. Inhibition of MAPK phosphorylation enhanced Hif-1α ubiquitination, and inhibited Hif-1α translocation into nucleus. Conclusively, Hif-1α and MAPK participate in HSC activation upon hypoxia.

Epidemiological distribution and genotype characterization of hepatitis C virus and HIV co-infection in Wuhan, China, where the prevalence of HIV is low.

Little is known about the epidemiological characteristics of HIV/hepatitis C virus (HCV) co-infection in cities in China with low HIV prevalence. This study evaluated the level of exposure to different risk factors associated with HCV transmission and characterized the distribution of HCV genotypes in 356 HIV-1-positive patients in Wuhan, central China. HIV transmission routes were distributed as follows: heterosexual contact, male-to-male sexual contact, intravenous drug use, blood transfusion, and unknown route. HCV antibodies were detected by a third-generation enzyme-linked immunosorbent assay. HCV-positive plasmas were subjected to RNA extraction, RT-PCR amplification, and sequencing. Phylogenetic analysis characterized HCV subtypes and the evolutionary origin of circulating HCV strains. Ninety-two of 356 (25.8%) patients infected with HIV were anti-HCV-positive. Among co-infected patients, the predominant risk for HCV transmission was intravenous drug use (87.3%). Six HCV subtypes (1a, 1b, 2a, 3a, 3b, and 6a) were detected. HCV genotype 6a was most prevalent, occurring in 39.3% of all patients, followed by genotypes 1b (24.7%), 3b (18.0%), and 3a (9.8%). The least frequent genotypes were 1a (4.9%) and 2a (3.3%). Intravenous drug use was strongly associated with genotype 6a, and infection by blood or blood product transfusion was strongly associated with genotype 1b. Genotype 2a was detected only among those infected by male-to-male sexual contact. The distribution of HCV subtypes suggests that the city plays a crucial role as a hub of HCV transmission in China. Exposure to multiple risk factors associated with HCV transmission was common among patients co-infected with HIV and HCV.

Hair selenium levels in hepatic steatosis patients.

The purpose of this study was to assess hair selenium levels of liver patients suffering from hepatic simple steatosis and non-alcoholic steatohepatitis (NASH) in central areas of China. Selenium was measured by an atomic absorption spectrophotometer equipped with the hydride generation system. The levels of selenium in healthy individuals ranged between 0.3 and 0.9 µg/g, and mean hair selenium levels in the male population and female population were 0.59 ± 0.18 and 0.57 ± 0.15 µg/g, respectively. These concentrations did not vary significantly (P > 0.05) in relation to the gender. One hundred-eighteen individuals of both sexes aged between 15 and 60 years with hepatic simple steatosis and NASH were selected for this study. The mean and standard deviation of hair selenium concentrations observed in male and female patients with hepatic simple steatosis were 0.54 ± 0.16 and 0.50 ± 0.15 µg/g, respectively, while the mean and standard deviation of hair selenium concentrations observed in male and female patients with NASH were 0.40 ± 0.14 and 0.41 ± 0.12 µg/g. Analysis of t test showed a significant difference between NASH (P < 0.001) patients in hair selenium concentrations when compared with controls.

Identification of novel splice variants and exons of human endothelial cell-specific chemotaxic regulator (ECSCR) by bioinformatics analysis.

Recent discovery of biological function of endothelial cell-specific chemotaxic regulator (ECSCR), previously known as endothelial cell-specific molecule 2 (ECSM2), in modulating endothelial cell migration, apoptosis, and angiogenesis, has made it an attractive molecule in vascular research. Thus, identification of splice variants of ECSCR could provide new strategies for better understanding its roles in health and disease. In this study, we performed a series of blast searches on the human EST database with known ECSCR cDNA sequence (Variant 1), and identified additional three splice variants (Variants 2-4). When examining the ECSCR gene in the human genome assemblies, we found a large unknown region between Exons 9 and 11. By PCR amplification and sequencing, we partially mapped Exon 10 within this previously unknown region of the ECSCR gene. Taken together, in addition to previously reported human ECSCR, we identified three novel full-length splice variants potentially encoding different protein isoforms. We further defined a total of twelve exons and nearly all exon-intron boundaries of the gene, of which only eight are annotated in current public databases. Our work provides new information on gene structure and alternative splicing of the human ECSCR, which may imply its functional complexity. This undoubtedly opens new opportunities for future investigation of the biological and pathological significance of these ECSCR splice variants.

[Impacts of HIV-1 resistance mutations associated with nucleoside reverse transcriptase inhibitors on viral fitness].

Nucleoside reverse transcriptase inhibitors which act as a major component of highly active antiretroviral therapy regimens are widely used in treatment of Acquired Immune Deficiency Syndrome. However, the emergence of drug-resistant variants of HIV-1 severely limits the effectiveness of these drugs. Many drug resistance mutations confer a fitness cost, which can be partially overcome by compensatory mutations or other molecular mechanisms. This review focuses on the impacts of resistance mutations emerging during treatment with nucleoside reverse transcriptase inhibitors on viral fitness, and inter actions between these mutations.

Structural and functional characterization of two alternative splicing variants of mouse Endothelial Cell-Specific Chemotaxis Regulator (ECSCR).

Endothelial cells (ECs) that line the lumen of blood vessels are important players in blood vessel formation, and EC migration is a key component of the angiogenic process. Thus, identification of genes that are specifically or preferentially expressed in vascular ECs and in-depth understanding of their biological functions may lead to discovery of new therapeutic targets. We have previously reported molecular characterization of human endothelial cell-specific molecule 2 (ECSM2)/endothelial cell-specific chemotaxis regulator (ECSCR). In the present study, we cloned two mouse full-length cDNAs by RT-PCR, which encode two putative ECSCR isoform precursors with considerable homology to the human ECSCR. Nucleotide sequence and exon-intron junction analyses suggested that they are alternative splicing variants (ECSCR isoform-1 and -2), differing from each other in the first and second exons. Quantitative RT-PCR results revealed that isoform-2 is the predominant form, which was most abundant in heart, lung, and muscles, and moderately abundant in uterus and testis. In contrast, the expression of isoform-1 seemed to be more enriched in testis. To further explore their potential cellular functions, we expressed GFP- and FLAG-tagged ECSCR isoforms, respectively, in an ECSCR deficient cell line (HEK293). Interestingly, the actual sizes of either ECSCR-GFP or -FLAG fusion proteins detected by immunoblotting are much larger than their predicted sizes, suggesting that both isoforms are glycoproteins. Fluorescence microscopy revealed that both ECSCR isoforms are localized at the cell surface, which is consistent with the structural prediction. Finally, we performed cell migration assays using mouse endothelial MS1 cells overexpressing GFP alone, isoform-1-GFP, and isoform-2-GFP, respectively. Our results showed that both isoforms significantly inhibited vascular epidermal growth factor (VEGF)-induced cell migration. Taken together, we have provided several lines of experimental evidence that two mouse ECSCR splicing variants/isoform precursors exist. They are differentially expressed in a variety of tissue types and likely involved in modulation of vascular EC migration. We have also defined the gene structure of mouse ECSCR using bioinformatics tools, which provides new information towards a better understanding of alternative splicing of ECSCR.

Genetic characterization of Human astrovirus infection in Wuhan, People's Republic of China, 2007-2008.

Human astrovirus (HAstV) was an important cause of viral gastroenteritis in infants in Wuhan city based on our previous study. The aim of the study was to investigate the nature of HAstV infection in Wuhan, People's Republic of China, especially in adults. Stool specimens were collected from 361 children and 301 adults with diarrhea from July 2007 to June 2008 and were tested for HAstV RNA by RT-PCR. The 348-bp PCR product of positive samples was further sequenced and analyzed for multiple sequence alignment and phylogenetic tree. HAstV RNA was detected in 2.33% (7/301) adults, which was significantly lower than that in children (13.57%, 49/361). HAstV-positive patients were either older than 50 years of age or younger than 3. Genetic analysis showed that the HAstV strain in adults was the same as that in children in 2007-2008. Contrarily, HAstV strains prevalent in 2007-2008 showed genetic characteristics different from those in 2004-2005 and belonged to two new groups of HAstV-1b. Thus, our data characterized HAstV infection in Wuhan 2007-2008, suggesting that HAstV infection also played an important role in adults in Wuhan, especial in patients of >50 years, and should be included for routine diagnosis in the population with diarrheal illness.

Endothelial cell-specific molecule 2 (ECSM2) localizes to cell-cell junctions and modulates bFGF-directed cell migration via the ERK-FAK pathway.

BACKGROUND: Despite its first discovery by in silico cloning of novel endothelial cell-specific genes a decade ago, the biological functions of endothelial cell-specific molecule 2 (ECSM2) have only recently begun to be understood. Limited data suggest its involvement in cell migration and apoptosis. However, the underlying signaling mechanisms and novel functions of ECSM2 remain to be explored. METHODOLOGY/PRINCIPAL FINDINGS: A rabbit anti-ECSM2 monoclonal antibody (RabMAb) was generated and used to characterize the endogenous ECSM2 protein. Immunoblotting, immunoprecipitation, deglycosylation, immunostaining and confocal microscopy validated that endogenous ECSM2 is a plasma membrane glycoprotein preferentially expressed in vascular endothelial cells (ECs). Expression patterns of heterologously expressed and endogenous ECSM2 identified that ECSM2 was particularly concentrated at cell-cell contacts. Cell aggregation and transwell assays showed that ECSM2 promoted cell-cell adhesion and attenuated basic fibroblast growth factor (bFGF)-driven EC migration. Gain or loss of function assays by overexpression or knockdown of ECSM2 in ECs demonstrated that ECSM2 modulated bFGF-directed EC motility via the FGF receptor (FGFR)-extracellular regulated kinase (ERK)-focal adhesion kinase (FAK) pathway. The counterbalance between FAK tyrosine phosphorylation (activation) and ERK-dependent serine phosphorylation of FAK was critically involved. A model of how ECSM2 signals to impact bFGF/FGFR-driven EC migration was proposed. CONCLUSIONS/SIGNIFICANCE: ECSM2 is likely a novel EC junctional protein. It can promote cell-cell adhesion and inhibit bFGF-mediated cell migration. Mechanistically, ECSM2 attenuates EC motility through the FGFR-ERK-FAK pathway. The findings suggest that ECSM2 could be a key player in coordinating receptor tyrosine kinase (RTK)-, integrin-, and EC junctional component-mediated signaling and may have important implications in disorders related to endothelial dysfunction and impaired EC junction signaling.

Changes in PGRMC1, a potential progesterone receptor, in human myometrium during pregnancy and labour at term and preterm.

This study assesses the role of progesterone receptor membrane component 1 (PGRMC1) in actions of progesterone (P4) on human myometrium during pregnancy and labour. Myometrial tissues were obtained from non-pregnant patients during hysterectomy or pregnant women undergoing C-section at term and preterm, before and during labour. PGRMC1 expression in myometrial tissues and in a human myometrial cell line (HM9) was assessed by western blots and RT-PCR. The subcellular localization of PGRMC1 in HM9 was performed by immunofluorescence staining. Isometric contractions of myometrial tissues were obtained in response to P4 with and without addition of specific antibodies against PGRMC1. Endogenous and over-expressed PGRMC1 proteins are detected by western blots in myometrial tissues, HM9 and 293 cells, respectively. PGRMC1 is localized to the plasma membrane, cytoplasm and nuclear membranes. PGRMC1 is lower in myometrium of women at term either not in labour (P = 0.004) or in labour (P = 0.005) compared with tissues from women in preterm non-labour. PGRMC1 levels are also decreased (P = 0.02) in myometrial tissues from women during preterm labour compared with preterm non-labour. P4 rapidly inhibits contractions of myometrial tissues compared with control (P < 0.05) in vitro. Pretreatment of myometrial strips with PGRMC1 antibody, suppresses the P4-induced relaxation (P < 0.05). PGRMC1 may mediate the non-genomic action of P4 and the relaxation effect on human myometrium during pregnancy. A decrease in PGRMC1 during term or preterm labour might contribute to the 'functional withdrawal' of P4 action and shift the balance to a state of heightened uterine contractility.

Prospective study of colonization and infection because of Pseudomonas aeruginosa in mechanically ventilated patients at a neonatal intensive care unit in China.

BACKGROUND: Ventilator-associated pneumonia (VAP) is an important nosocomial infection at neonatal intensive care units (NICU), frequently caused by Pseudomonas aeruginosa. A 6-month prospective study from January 2009 through June 2009 was performed to investigate the respective contribution of endogenous and exogenous transmission of P aeruginosa in the respiratory colonization or/and infection in the mechanically ventilated patients at a NICU to identify routes of lung infection with P aeruginosa and to assess risk factors for colonization or respiratory infection with P aeruginosa. METHODS: Samples from oropharyngeal swab, tracheobronchial aspirates, gastric aspirate, and rectal swab were obtained in each patient after intubation and then twice a week. Surveillance cultures for the presence of P aeruginosa from environmental surfaces of the ICU were taken once every 5 days during the study period. Pulsed-field gel electrophoresis was used to characterize the clonal relatedness of the strains by SpeI-digested genomic DNA. RESULTS: Eighteen patients (78.3%) had colonization of the upper respiratory tract. Sixteen (69.6%) patients with colonization of the respiratory tract were infected from other patients or environmental surfaces, which was considered exogenous, and, among strains causing pulmonary infection, there were 4 (50%) patients with exogenous infection. Eight of these developed VAP after a mean of 9 ± 3.4 days. The incidence of P aeruginosa VAP on the unit was 6.2%. The respiratory tract was the earliest site of colonization in all patients of VAP. Low birth weight, duration of mechanical ventilation, previous ampicillin group use, and previous second-generation cephalosporins use were independently associated with patient-related acquisition of P aeruginosa. CONCLUSION: Our results confirm that the upper respiratory tract acts as an important reservoir of P aeruginosa colonization and infection in the mechanically ventilated patients and emphasize the importance of exogenous acquisition of P aeruginosa. A combination of early identification and eradication of airways colonization by P aeruginosa plus infection control measures may be the basis to prevent pulmonary infection.

Downregulation of human telomerase reverse transcriptase through anti-C-myc siRNA in human colon cancer Colo 320 cells.

The c-Myc and human telomerase reverse transcriptase gene (hTERT) gene are frequently deregulated and overexpressed in malignancy. hTERT activity is induced by c-Myc and strategies designed to inhibit c-Myc expression in cancer cells may have considerable therapeutic value. We designed and used a short hairpin RNA to inhibit c-Myc expression in Colo 320 cells and validated its effect on cell proliferation. In this study, four c-Myc-shRNA expression vectors were constructed and introduced into Colo 320 cells. The effects of c-Myc silencing on tumor cell growth was assessed by soft agar assay and DNA synthesis and MTT experiments. The expressions of c-Myc and hTERT were also assessed by real-time reverse transcription-polymerase chain reaction and Western blot analysis. Upon transient transfection with plasmid encoding shRNA, it was found that expression of c-Myc and hTERT decreased in shRNA-transfected cells. The downregulation of c-Myc and hTERT inhibited cell growth, shortened telomere lengths, and suppressed telomerase activity. In conclusion, our findings demonstrate that shRNA of c-Myc can inhibit the DNA replication in Colo 320 cells effectively and reduce telomere length and telomerase activity; therefore, it could be used as a new potential anticancer tool for therapy of human colon cancer.

Relationship between genomic types of Escherichia coli and clinical diseases.

In this study, by analysis of genome structures of E. coli, the relationships between the genomic types of E. coli and the associated diseases were investigated. Samples of sputum, urine and other excretions from patients with different infective diseases were collected. And 62 E. coli strains were isolated from these samples. Intact bacterial genomic DNA was cleaved with I-CeuI, separated by pulsed field gel electrophoresis and then typed on the basis of cleavage map. The results showed that 7 I-CeuI sites were found in all the genome structures of the 62 E. coli, indicating that there were 7 rrn operons in the genomes. The size of genome ranged from 4,500 kb to 5,000 kb. According to the genome structures, 62 E. coli strains were divided into 30 genome types. It was concluded that genome structures of E. coli isolated from the patients with different infective diseases varied to some extent, suggesting that some genome types of E. coli were closely related to some infective diseases.

Identification of new subtype of astrovirus type 3 from an infant with diarrhea in Wuhan, China.

Human astrovirus is one of the important causes for viral gastroenteritis in young children. In previous study where we examined the molecular epidemiology of human astrovirus (HAstV) infection in infants in Wuhan City, we isolated and identified a new subtype (WH1859) of HAstV genotype 3 from an infant with diarrhea. The sequence analysis of this strain showed that the complete region of ORF2 of WH1859 contains 2385-bp of nucleotides that encode 795 amino acids. Because WH1859 strain has the identity of less than 95% with the distance of more than 0.05 to the reference strains of HAstV-3, WH1859 represents a distinct subtype within HAstV-3 strains. Further studies are needed to determine the role of this new subtype strain of HAstV in viral gastroenteritis among young children.

Reduced antigenicity of naturally occurring hepatitis B surface antigen variants with substitutions at the amino acid residue 126.

BACKGROUND: Substitutions at amino acid residue 126 of hepatitis B surface antigen (HBsAg) occur frequently in hepatitis B virus (HBV) isolates from patients with chronic HBV infection. These substitutions occur naturally, but their significance for viral persistence is unclear and requires further investigation. METHODS: We amplified coding regions of HBsAg by PCR using sera from 1 patient chronically infected with HBV. Three representative clones of HBsAg with amino acid residues 126Ile (I), 126Thr (T) and 126Ser (S) were selected from sequenced clones. HBsAg 126Ala (A) mutants of subtype C/adr and D/adw were generated by site-directed mutagenesis. The HBsAg expression vectors were constructed and transiently transfected into HepG2 cells. The binding reactivity of HBsAg to anti-HBs antibodies was tested by chemiluminescent microparticle immunoassay and by immunofluorescence staining with polyclonal and monoclonal anti-HBs antibodies. RESULTS: Diverse HBsAg variants with substitutions at amino acid residue 126 co-existed in a chronically infected HBV patient. HBsAg with the substitution 126S showed a significantly low antigenicity, while the binding reactivity to anti-HBs of other HBsAg with 126I, 126T and 126A were comparable. CONCLUSION: HBsAg with the 126S substitution has a reduced antigenicity, which may contribute to immune escape in chronic HBV infection.

Relationship between Genomic Types of Escherichia coli and Clinical Diseases / 华中科技大学学报（医学）（英德文版）

In this study, by analysis of genome structures of E. coli, the relationships Between the genomic types of E. coli and the associated diseases were investigated. Samples of sputum, urine and other excretions from patients with different infective diseases were collected. And 62 E. coli strains were isolated from these samples. Intact bacterial genomic DNA was cleaved with I-CeuI, separated by pulsed field gel electrophoresis and then typed on the basis of cleavage map. The results showed that 7 I-CeuI sites were found in all the genome structures of the 62 E. coli, indicating that there were 7 rrn operons in the genomes. The size of genome ranged from 4500 kb to 5000 kb. According to thegenome structures, 62 E. coli strains were divided into 30 genome types. It was concluded that genome structures of E. coli isolated from the patients with different infective diseases varied to some extent, suggesting that some genome types of E. coli were closely related to some infective diseases.

Study on the blood-borne virus co-infection and T lymphocyte subset among intravenous drug users.

AIM: To investigate the features of various blood-borne virus infections and co-infection in intravenous drug users (IDUs), and to examine the correlation of T lymphocyte subsets with virus co-infection. METHODS: Four hundred and six IDUs without any clinical manifestation of hepatitis and 102 healthy persons were enrolled in this study. HBV-DNA and HCV-RNA were detected by fluorescence quantitative PCR. HBsAg, HBeAg, anti-HBc, anti-HCV, HDV-Ag, anti-HGV, anti-HIV, and HCMV-IgM were assayed by enzyme-linked immunosorbent assay (ELISA) and immunochromatographic tests. The levels of Th1 and Th2 cytokines were measured by ELISA and radioactive immune assay (RIA). The T lymphocyte subpopulation was detected by using fluorescence immunoassay. The similar indices taken from the healthy persons served as controls. RESULTS: The viral infection rate among IDUs was 36.45% for HBV, 69.7% for HCV, 47.3% for HIV, 2.22% for HDV, 1.97% for HGV, and 3.45% for HCMV. The co-infection rate of blood-borne virus was detected in 255 of 406 (62.81%) IDUs. More than 80% (161/192) of subjects infected with HIV were co-infected with the other viruses, such as HBV, HCV. In contrast, among the controls, the infection rate was 17.65% for HBV and 0% for the other viruses. Our investigation showed that there was a profound decrease in the proportion of CD4/CD8 and the percentage of CD3 and CD4, but not in the percentage of CD8. The levels of PHA-induced cytokines (IFN-gamma and IL-4) and serum IL-2 were obviously decreased in IDUs. On the other hand, the level of serum IL-4 was increased. The level of IFN-gamma and the percentage of CD4 were continuously decreased when the IDUs were infected with HIV or HIV co-infection. IDUs with HIV and HBV co-infection was 15.1% (29/192). Of those 29 IDU with HIV and HBV co-infection, 51.72% (15/29) and 37.93% (11/29) were HBV-DNA-positive and HBeAg-positive, respectively. But, among IDUs without HIV infection, only 1.68% (2/119) of cases were HBV-DNA-positive. CONCLUSION: HCV, HBV and HIV infections are common in this population of IDU, leading to a high incidence of impaired Th1 cytokine levels and CD4 lymphocyte. IDUs with HIV and HBV/HCV co-infection have lower expression of Th1 cytokine with enhancement of the Th2 response. HIV may be causing HBV replication by decreasing Th1 function.