Serum types of enterovirus and clinical characteristics of 237 children with hand, foot and mouth disease in Shenzhen / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical and epidemiologic characteristics and the serum types of enterovirus of hand, foot and mouth disease (HFMD) in children.</p><p><b>METHODS</b>The RT-nPCR method was established with universal primers within 5' untranslated region of enterovirus and VP1 region of Coxsackievirus A16 (CAV16) and enterovirus 71 (EV 71). Enteroviruses were detected with RT-nPCR in 237 children with HFMD. Clinical and epidemiologic characteristics and serum types of enterovirus of the patients with HFMD were studied.</p><p><b>RESULTS</b>The patients'age ranged from 7 months to 11 years (mean 4.2 +/- 0.5 years). The majority (94.5%) were less than 6 years old. HFMD was mostly seen in spring and winter (67.9%). Oral mucosal pox or ulcer as well as hand and foot rashes were observed in all 237 patients. Fever occurred in 141 patients (59.5%). Of the 237 patients, 133 (56.1%) were RT-nPCR positive. Of the 133 cases, 38 were positive for EV71, 64 were positive for CAV16, and 31 were negative for both EV71 and CAV16. The patients infected by different types of enteroviruses had similar clinical characteristics. Gene colon and sequence analysis for 12 strains of enteroviruses PCR positive products presented as EV71 (n=5), CAV16 (n=5), ECHO13 (n=1), and CAV5 (n=1).</p><p><b>CONCLUSIONS</b>HFMD tends to occur in younger children less than 6 years old. The majority are affected in spring and winter. EV71 and CAV16 are common pathogens of HFMD. There is no relationship between clinical characteristics and serum types of enteroviruses in HFMD patients.</p>

Construction of replication-deficient recombinant adenoviral vector carrying HBsAg and HSP70 chimeric gene and its expression in vitro / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct a recombinant adenoviral vector carrying HBcAg-HSP70 chimeric gene by homologous recombination in bacteria and to detect its expression in vitro.</p><p><b>METHODS</b>Heat shock protein 70 gene from Mycobacterium tuberculosis were amplified by PCR and were cloned to adenoviral shuttle plasmid pAdTrack-CMV-HBsAg. Then the resultant pAdTrack-CMV-HBsAg-HSP70 was cotransfected into BJ5183 bacteria with the plasmid pAdeasy-1. The adenoviral plasmid carrying HBsAg-HSP70 gene (pAd-HBsAg-HSP70) was generated with homologous recombination in bacteria and the adenoviruses were produced in 293 cells. Several kinds of mammal cells (293 cells and Vero cells) were infected with adenoviruses and the expression of HBsAg-HSP70 was detected by RT-PCR and ELISA in vitro.</p><p><b>RESULTS</b>The adenoviral plasmids pAd-HBsAg-HSP70 were obtained by selection for kanamycin resistance and confirmed by restriction endonuclease Pac analyses. The recombinant adenoviruses Ad-HBsAg-HSP70 were packaged successfully in 293 cells. The titer of Ad-HBsAg-HSP70 was up to 2 x 10(12) pfu/L after the second passage of proliferation in 293 cells. HBsAg and HSP70 were expressed efficiently in mammal cells after infection.</p><p><b>CONCLUSION</b>The recombinant adenoviruses expressing HBsAg and HSP70 were constructed successfully which can be used further in study of gene therapy for HBV.</p>

Induction of specific cytotoxic T lymphocyte response against hepatitis B virus by hepatitis B virus S gene-modified dendritic cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore whether hepatitis B virus (HBV) S gene-modified dendritic cells (DCs) might induce a specific cytotoxic T lymphocyte (CTL) response.</p><p><b>METHODS</b>The recombinant adenoviruses carrying HBsAg genes were prepared and used to transfect DCs generated from cord blood. The efficacy of transfection was observed through the expression of enhanced green fluorescent protein (EGFP) in DCs and the expression of HBsAg was detected by ELISA. HBV S gene-modified DCs were co-cultured with T cells from cord blood and T cells stimulating activities were detected using mixed lymphocyte reaction (MLR). The CTL assay was carried out to assess the ability of CTL lines to lyse target cells of HepG(2)22.1.5 by measuring lactate dehydrogenase (LDH) release.</p><p><b>RESULTS</b>The results showed that HBV S genes were expressed in DCs with high efficacy by recombinant adenoviral vector. DCs had a normal shape after transfection. The result of MLR showed that HBV S gene-modified DCs could effectively stimulate naive T cells to proliferate. The induced specific CTL lines could lyse target cells of HepG(2)22.1.5.</p><p><b>CONCLUSIONS</b>HBV S gene-modified DCs enhanced the function to induce a specific CTL effect, showing its promise for developing anti-viral vaccine in future.</p>

Expression of HBV preS2/S gene in mammalian cells transferred with adenoviral vector / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study HBV preS2/S gene expression effects in mammalian cells transferred with recombinant adenoviral vector.</p><p><b>METHODS</b>The replication-deficient recombinant adenoviral vector (Ad-HBs) carrying HBV preS2/S gene were constructed by homologous recombination in bacteria. The 293 cells, Vero cells, HepG2 cells and mesenchymal stem cells (MSCs) were infected with adenoviruses. The expressions of enhanced green fluorescent protein (EGFP) were observed with fluorescence microscope and the expressions of HBsAg were detected by RT-PCR and ELISA in vitro.</p><p><b>RESULTS</b>More than 90% of 293 cells, Vero cells, HepG2 cells or MSCs expressed EGFP after transfection at the MOI of 20 and the titers of HBsAg were more than 3.229 (A value) in culture supernatant.</p><p><b>CONCLUSION</b>The HBV preS2/S gene was not only expressed efficiently in immortalized cells, but also expressed efficiently in stem cells with the recombinant adenoviruses vector.</p>

Detection of glutathione S-transferase and lung resistance-related proteins in acute leukemia and its clinical significance / 中南大学学报(医学版)

OBJECTIVE@#To explore the relationship among intracellular glutathione S-transferase activity (GST), the expression of lung resistance-related proteins (LRP) in acute leukemia, and its clinical effects.@*METHODS@#The GST activity of bone marrow mononuclear cells and LRP expression in 57 acute leukemia patients were detected by the spectrophotometry assay and immuno-cytochemistry (SABC), respectively.@*RESULTS@#The GST activity of bone marrow mononuclear cells in the acute leukemia group was significantly higher than that of the control group (P < 0.01). The GST activity of mononuclear cells in acute leukemia was positively correlated with the percentage of blast in the bone marrow (r = 0.30, P < 0.05). The GST activity of mononuclear cells in the untreated acute leukemia group was obviously higher than that of the complete remission group (P <0.01). The GST activity in the refractory or relapsed acute leukemia group was significantly higher than that of the complete remission group and untreated leukemia group (P <0.05). In post-chemotherapy 13 of 17 the LRP-positive patients were the non-remission, 12 of the 20 LRP-negative patients were the complete remission. The curative rate of the LRP-positive group was the significantly lower than the LRP-negative group (P < 0.05). The GST activities of non-remission patients in the LRP-positive and LRP-negative group obviously increased.@*CONCLUSION@#The increase of GST activity in the bone marrow mononuclear cells is related to the clinical curative effects and the proliferation of blast in acute leukemia. Detection of LRP and GST activities in acute leukemia may have a reference value in judging the leukemia with drug resistance and estimating the prognosis.

Detection of SEN virus (subtype D/H) infection in Shenzhen / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of newly identified single-chain DNA virus (SENV) infection in Shenzhen.</p><p><b>METHODS</b>Nested polymerase chain reaction (nPCR) was established using primers from ORF1 region of SENV genome. Six hundred and one sera samples from different populations were detected for SENV DNA (D and H subtype) by nPCR. Products of PCR were cloned into T-vector and sequenced.</p><p><b>RESULTS</b>The positive rates of SENV DNA in different populations were as followed: 27.8% in patients with hepatitis B, 22.2% in patients with hepatitis C, 26.9% in hemodialysis patients and 39.3% in IDUs. Among blood donors, the positive rates of SENV DNA were 28.1% in unqualified blood donors, 31.3% in blood donors with an elevated ALT levels and 15.1% in qualified blood donors. The infection rates of SENV in unqualified blood donors and blood donors with an elevated ALT levels were obviously higher than in qualified blood donors (chi(2) = 8.29, P < 0.01 and chi(2) = 6.03, P < 0.01). There was a 6.8% difference of nucleotide between SENV-D standard subtype and 6 isolates with 13.5% difference of nucleotide between SENV-H standard subtype and 4 isolates from Shenzhen.</p><p><b>CONCLUSION</b>Results suggested that SENV infection was common in high-risk groups in Shenzhen.</p>

Change of peripheral blood CD5~+B cells in systemic lupus erythematosus patients / 中华风湿病学杂志

Objective To explore change of peripheral blood CD5+B cells in the systemic lupus ery- thematosus(SLE)patients and the association of this change with the disease activity,Methods The percent- age of CD5~+B ceils in peripheral blood in 57 patients with SLE and 35 healthy controls was determined by flow eytometry.Levels of anti-dsDNA,complement C3 and C4,anti-nuclear antibody(ANA),anticardiolipin anti- body(ACL),were also measured.Results The percentage of CD5~+B cell in SLE patients(2.1?0.4)% was high- er than that in normal controls [(1.5?0.4)%](P＜0.05).In active disease group,the percentage of CD5~+B cells [(2.5?0.5)%] was significantly higher than that in inactive SLE patients [(1.4?0.5)% ](P＜0.01).The percent- age of CD5~+B cell associated with anti-dsDNA,ANA,ALA positively,with C3 negatively and had no associa- tion with C4.Conclusion The percentage of CD5~+B cells in the SLE is significantly high and has correlation and some relationship with the SLE.