α4ß7high CD4+ T cells are prone to be infected by HIV-1 and associated with HIV-1 disease progression.

INTRODUCTION: To investigate the characteristics of ß7high CD4+ T cells during HIV-1 infection and the relationship between ß7high CD4+ T cells and HIV-1 disease progress. METHODS: This study enrolled 124 HIV-1-infected patients, including 80 treatment naïve patients (TNs), 41 patients who underwent antiretroviral therapy (ARTs), and three long-term no progression patients (LTNPs). Nineteen matched healthy subjects were included as controls (HCs). The characteristics and frequency of ß7high CD4+ T cells were analyzed using flow cytometry. An in vitro culture experiment was used to study HIV-1 infection of ß7high CD4+ T cells. Real-time polymerase chain reaction was performed to quantify HIV-1 DNA and CA-RNA levels. RESULTS: The frequency of ß7high CD4+ T in the peripheral blood was significantly decreased and negatively correlated with disease progression during chronic HIV-1 infection. A large proportion of ß7high CD4+ T cells showed Th17 phenotype. Furthermore, ß7high CD4+ T cells were preferentially infected by HIV-1 in vitro and in vivo. There were no significant differences of HIV-1 DNA, and CA-RNA levels between ß7high CD4+ T and ß7low CD4+ T subsets in HIV-1 infected individuals after antiviral treatment. CONCLUSION: The ß7high CD4+ T cells were negatively correlated with disease progression during chronic HIV-1 infection. ß7high CD4+ T cells are susceptible to infection with HIV-1 and HIV-1 latent cells.

[Construction and expression of the Echinococcus granulosus recombinant BCG-EgG1Y162].

OBJECTIVE: To construct and express Echinococcus granulosus recombinant bacille Calmette-Guerin (BCG) strain rBCG-EgG1Y162. METHODS: The encoding gene of the antigen EgG1Y162 of E. granulosus was recombined with E. coli-Mycobacterium shuttle expression plasmid vector pMV361 by genetic engineering technique, and transformed into E. coli for amplification. The recombinant plasmid rpMV-EgG1Y162 was identified by PCR, double digestion with restriction enzymes, and sequence analysis. The confirmed rpMV-EgG1Y162 was transformed into BCG strain via electroporation technique to construct the recombinant rBCG-EgG1Y162. After identification by PCR and double digestion with restriction enzymes, the recombinant strain was cultured for about 2 weeks. In order to induce the expression of target protein, the rBCG was placed in 45 degrees C for 30 min. SDS-PAGE and Western blotting were used to analyze the expressive protein. RESULTS: The product of recombinant plasmid rpMV-EgG1Y162 was approximately 360 bp by PCR amplification and double digestion with restriction enzymes, consistent with the expected fragment length. Sequencing results showed that the inserted sequence was correct. The rBCG-EgG1Y162 grew well and the identification of PCR and enzyme digestion revealed accuracy. The results of SDS-PAGE and Western blotting showed that the relative molecular weight (M(r)) of the protein was about 71 000. CONCLUSION: The E. granulosus rBCG-EgG1Y162 strain is constructed and expressed.

[Cloning and sequence analysis of the egG1Y162 gene of Echinococcus granulosus].

A pair of primers (egG1Y162) were designed according to the nucleotide sequence of Echinococcus multilocularis emY162 antigen gene. Using genomic DNA and cDNA from protoscoleces and adult worms of E. granulosus as templates, PCR was performed with the primers to obtain fragments of egG1Y162 gene. PUCm-T/egG1Y162 recombinant plasmids and PUCm-T/egY162 cDNA recombinant plasmids were constructed and identified by PCR, digestion with restriction enzyme and sequencing. The egG1Y162 antigen gene was amplified in protoscoleces and adult worms of E. granulosus. The size of the egG1Y162 gene was 1 648 bp and cDNA was 459 bp, and GenBank accession numbers were AB458258 and AB458259, respectively.

Role of immune tolerance in BALB/c mice with anaphylactic shock after Echinococcus granulosus infection.

This study tested the hypothesis that immune tolerance mediated by regulatory T (Treg) cells is protective against cystic echinococcosis (CE)-induced anaphylactic shock. BALB/c mice were inoculated with protoscoleces of Echinococcus granulosus. After 3 months, the presence of cysts in the peritoneal cavity was confirmed after which a subset of mice was sensitized using a cyst fluid suspension to induce anaphylactic shock. While IgE levels were significantly higher in both groups inoculated with E. granulosus as compared to the healthy control group (both P < 0.01), sensitized mice had higher IgE levels as compared with those with E. granulosus alone (P < 0.05). Mice inoculated with E. granulosus alone and sensitized mice both had significantly higher histamine levels as compared to the healthy controls. The proportion of CD4(+)CD25(+)Foxp3(+) Treg cells relative to CD4(+) cells was significantly higher in mice inoculated with E. granulosus alone (P < 0.0167); significantly higher interleukin-10 (IL-10) and tumor growth factor-ß (TGF-ß1) levels were also noted in this group (all P < 0.01). In contrast, IL-13 and IL-17A levels were significantly higher in the sensitized mice (both P < 0.05). Taken together, these data suggest that the biphasic changes in Treg cell and cytokine levels may be associated with anaphylactic shock induced by CE.

Correlation analysis between plasma D-dimer levels and orthopedic trauma severity.

BACKGROUND: The correlation between the plasma D-dimer level and deep vein thrombosis has not been conclusive in various studies. The aim of this research was to study the relationship between plasma D-dimer levels and the severity of orthopedic trauma by retrospective examination of orthopedic trauma cases. METHODS: Clinically acute trauma and non-acute trauma patients were selected and their plasma D-dimer levels were measured. Plasma D-dimer levels in patients of these two groups were compared. The relationship between the plasma D-dimer level and the severity of the trauma was also studied. RESULTS: There were 548 cases in the acute trauma group and 501 cases in the non-acute trauma group. The levels of plasma D-dimer were significantly higher in the acute trauma group than in the non-acute trauma group (P < 0.01). In the acute trauma group, the correlation between the D-dimer level and the number of fractures was a positive linear correlation (r = 0.9532). CONCLUSIONS: Elevated plasma D-dimer is common in trauma patients. The D-dimer level and the number of fractures in the trauma patients are closely correlated. D-dimer is not only an indicator for the diagnosis of deep vein thrombosis and pulmonary embolus, but also an indicator of the severity of trauma in acute trauma patients.

Transmembrane protein 166 expression in esophageal squamous cell carcinoma in Xinjiang, China.

OBJECTIVE: Transmembrane protein 166 (TMEM166) expression in esophageal squamous cell carcinoma (ESCC) and remote normal esophageal tissues was examined to assess any role in tumour biology. METHODS: TMEM166 mRNA expression in 36 cases with ESCC (36 tumour samples, 36 remote normal esophageal tissue samples) was detected by RT-PCR. TMEM166 protein expression was analysed in paraffin-embedded tissue samples from the same cases by immunohistochemistry. RESULTS: Semi-quantitative analysis showed TMEM166 mRNA expression in ESCCs to be significantly lower than in remote normal esophageal tissues (0.759 ± 0.713 vs. 2.622 ± 1.690, P=0.014). TMEM166 protein expression was also significantly reduced (69.4% vs. 94.4%, P<0.01). CONCLUSION: TMEM166 mRNA and protein expression demonstrated significant reduction in ESCCs compared with remote esophageal tissues, albeit with no correlation with tumour size, differentiation, stage, and lymph node metastasis, suggesting a role in regulating autophagic and apoptotic processes in the ESCC.

[Regulation on phenotypic and functional maturation of dendritic cells by apolipoprotein A-I].

AIM: to investigate the effects of apolipoprotein A-I (ApoA-I) on the peripherial blood dendritic cell (PBDC) and monocyte derived DC (MDDC) in vitro. METHODS: isolate PBDC or monocyte by cell isolation kit, monocyte were induced to MDDC by treated with GM-CSF plus IL-4 for 6 days, and then collect PBDC and MDDC treated them with apoA-I, LPS or TNF-α for 24 hours. Then check the cell surface marker and phagocytic capacity by flow cytometry. ELISA was used to detect the levels of cytokine secretion. T cells were stained with CFSE and T cell proliferation was assessed by flow cytometry. RESULTS: collect the PBDC and MDDC with high purity. In the presence of ApoA-I, the surface markers on MDDC, such as CD40, CD86 and MHC-II, were up-regulated which were detected by flow cytometry. CD83 expression on both PBDC and MDDC was remarkably increased. ApoA-I DC demonstrated decreased the phagocytic capacity. ApoA-I also stimulated MDDC to produce IL-12 and TNF-α. Furthermore, ApoA-I can induce considerable Th cell proliferation. CONCLUSION: ApoA-I can induce the maturation and activation of MDDC and PBDC, including the cytokine secretion, specific antigen presentation and T cell proliferation and decreasing the phagocytic capacity. Therefore, ApoA-I may attribute to the immune response in AS process.