Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

This study aimed to evaluate the in vivo efficacy of combined ABZ-interferon (IFN)-α treatment for CE in mice.After 5 months of secondary infection with protoscoleces,mice were randomly allocated into four groups:ABZ-treated group,IFNαtreated group,ABZ+IFN-α group and untreated control group.Drugs in diverse treated groups were respectively administered for 2 months,of which,sera were respectively collected in 0 d,7 d,14 d,28 d,36 d,48 d,and 60 d.Mice were then euthanized and associated indications were investigated to evaluate the therapeutic efficacy.Results showed that ABZ+IFN-α induced a significant reduction of the number,size as well as weight of cysts,compared with that in ABZ (P＜0.05) or untreated group (P＜0.01) respectively.This effect was associated with ultrastructural modification of the cyst in ABZ+IFN-α group.Interestingly,significant decrease of IL (interleukin)-10 in serum and in vitro production by spleen cells with ABZ+ IFN-α treatment was observed in comparison with untreated control (P＜0.01).Serum IgE,IgG and subsets were respectively decreased in ABZ+IFN-α treatment,compared with that in control group (P＜0.01).Our findings demonstrated that combination of ABZ with IFN-α may contribute to an efficient therapeutic regimen of human and animal CE.

OBJECTIVETo analyze the epidemiological characteristics of HIV infected immigrant marriage women in rural areas in Zhejiang province.

METHODSA retrospective investigation was conducted among 118 HIV infected immigrant marriage women, who were detected in active or passive surveillance in rural areas in Zhejiang province. The investigation information included the basic demographic characteristics, risky behavior, the status of HIV infection among their spouses and children. EpiData 3.0 and SPSS 13.0 software were used for database set up and bivariate and multiple logistic regression analysis.

RESULTSBy the end of 2009, a total of 118 HIV/AIDS were reported among immigrant marriage women. Of which, 53.4% (63/118) of the subjects were from Yunnan province and 33.1% (39/118) were minority nationality, and 30.6% (39/108) of their spouses were diagnosed HIV infection when these targeted women were detected. In discordant couples follow up with female partner being HIV positive, the spouse seroincidence rate was 2.09 per 100 person years (3/143.5 person years). Subjects who had sexual behavior beyond marriage accounted for 83.9% (99/118) of total, and 69.5% (82/118) subjects had former boyfriend or former husband in former inhabited places. In the 36 concordant-positive couples, 63.9% (23/36) reported never using condom in in-marriage sexual behavior before being detected HIV positive. Bivariate analysis results revealed that the significant difference for the HIV positive testing rate for a couple among different time duration that an immigrated woman moved to Zhejiang province due to marriage was observed (χ(2) = 8.708, P = 0.003). While the number and proportion of HIV positive couples for those immigrated women who lived in Zhejiang province for less than 2 years, 2 - 5 years, more than 5 years were 12 (20.7%), 9 (31.0%), 11 (57.9%), respectively. Multiple logistic regression analysis showed that the long time length when an immigrated woman was detected positivity after she moved to Zhejiang province was the risk predictor of their husband being infected (OR = 2.223; 95%CI: 1.288 - 3.836).

CONCLUSIONIt is showed that premarital unsafely sexual behavior, in-marriage sexual behavior without condom, delayed detection of HIV are risk factors for HIV infection and transmission.

Adolescent ; Adult ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; Humans ; Marriage ; Middle Aged ; Retrospective Studies ; Risk-Taking ; Rural Population ; Sexual Behavior ; Transients and Migrants ; statistics & numerical data ; Young Adult

Cell Culture Techniques ; methods ; Fixatives ; Humans ; Immunohistochemistry ; methods ; Tissue Fixation ; methods

OBJECTIVEFrequency, type and clinical implications on protease and reverse transcriptase drug resistance mutations were investigated and phylogenetic analysis in antiretroviral drug-naïve AIDS patients was carried out in Henan province.

METHODS45 plasma samples were separated from the anticoagulatory whole blood, from which reverse transcription-polymerase chain reaction technique was used to amplify the partial pol gene. The sequences were analysed for genotypic antiretroviral resistance and phylogenetic relation through landing the websites http://hivdb.stanford.edu and http://hiv-web.lanl.gov, under BioEdit and DNAClub software.

RESULTSPartial pol sequences of 36 samples were successfully amplified. The major mutation rate of resistance to protease was 8.3% (3/36), including types D30A, V32A, G73C and V82A. Minor mutation rate of resistance was 100%, including types of L63PS (36/36), I93L (35/36), V77IL (34/36), A71IVT (10/36) and D60E (2/36). The mutation rate of resistance to reverse transcriptase was 38.9% (14/36). Mutation-scoring and clinical implication clewed drug resistance rates were 5.6% (2/36) and 22.2% (8/36) to protease inhibitors and reverse transcriptase inhibitors respectively, while 1 sample was potentially low-level resistant to all of the protease inhibitors and 3 samples to part of the reverse transcriptase inhibitors. Phylogenetic analysis revealed that the pol gene of 36 samples were highly homologous and having a near relative to B.US.83.RF ACC M17451. 36 samples seemed to have the same infection source while their resistance mutations were not due to drug-resistant virus infection but to the evolving of virus in vivo.

CONCLUSIONMost of the antiretroviral drug-naïve AIDS patients in Henan province were sensitive to the currently available antiviral medicine, but antiviral treatment must be in accordance with the strict procedure and to keep better adherence, to avoid the epidemics caused by drug-resistant virus.

Acquired Immunodeficiency Syndrome ; genetics ; Adult ; Anti-HIV Agents ; pharmacology ; China ; Drug Resistance, Viral ; genetics ; Female ; Genes, pol ; genetics ; Genotype ; HIV Protease ; genetics ; HIV Protease Inhibitors ; pharmacology ; Humans ; Male ; Mutation ; Phylogeny ; RNA-Directed DNA Polymerase ; genetics ; Reverse Transcriptase Inhibitors ; pharmacology

BACKGROUND AND OBJECTIVENasopharyngeal carcinoma (NPC) is known for its propensity for distant metastases. Lung metastasis is one of the most important causes of death for patients with NPC. Solitary metastatic lung tumor from NPC is a distinctive group associated with a better survival. This study was to find a more effective treatment modality and prognostic factors for the group.

METHODSClinical data of 105 cases of solitary metastatic lung tumor from NPC were retrospectively analyzed. Survival rate was calculated by the Kaplan-Meier method. The difference of survival between the patients treated by different modalities was evaluated by the log-rank test. The Cox univariate and multivariate analyses of gender, age, pathologic type, stage, adjuvant chemotherapy, evaluation of treatment for NPC, disease-free interval, size of metastatic tumor, pulmonary hilar and/or mediastinal lymph node metastasis, treatment modalities, recurrent distant metastases and/or relapse of NPC were conducted.

RESULTSThe local control rate was 53.8% in chemotherapy group, 88.0% in radiotherapy ± chemotherapy group, and 96.4% in operation ± chemotherapy group (P < 0.01). The most promising progression-free survival (PFS) and overall survival (OS) were obtained with operation ± chemotherapy and followed by radiotherapy ± chemotherapy. Both of them showed much better efficacy than chemotherapy (P < 0.001). The Cox multivariate analysis showed that recurrent distant metastases and/or relapse of NPC affected the survival (OR = 2.087, 95% CI = 1.277-3.410, P = 0.003). The T stage of NPC, size of metastatic tumor, hilar and/or mediastinal lymph node metastasis, and the treatment modality were independent prognostic factors.

CONCLUSIONSOperation ± chemotherapy and radiotherapy ± chemotherapy are better treatment of solitary metastatic lung tumor from NPC, which could improve the local control and prolong the PFS and OS. Chemotherapy is recommended for patients with higher T stage of NPC, size of metastatic tumor ≥ 3 cm, pulmonary hilar and/or mediastinal lymph node metastasis.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; secondary ; therapy ; Combined Modality Therapy ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; secondary ; therapy ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Pneumonectomy ; methods ; Radiotherapy, High-Energy ; Remission Induction ; Retrospective Studies ; Survival Rate ; Young Adult

OBJECTIVETo study the effect of oleic acid (OA) on expression of aquaglyceroporin genes, AQP3 and AQP9, in hepatocyte steatosis and to investigate the underlying molecular mechanisms using an in vitro system.

METHODSHepG2 cells were treated with OA at different concentration to establish in vitro models of nonalcoholic hepatocyte steatosis. The corresponding extents of hepatic steatosis modeling were assessed by oil red O staining and optical density (OD) measurements of the intracellular fat content. The model lines were then treated with inhibitors of the PI3K/Akt and p38 MAPK signaling pathway factors and effects on AQP3/9 expression was measured by real time RT-PCR and western blotting.

RESULTSThe fat concentration, indicative of hepatic steatosis, increased in conjunction with increased concentrations of OA (0 less than 250 less than 500 mumol/L). OA exposure also down-regulated AQP3 mRNA and up-regulated AQP9 mRNA levels in a concentration-dependent manner. The most robust changes in expression occurred in response to the 500 mumol/L concentration of OA for both AQP3 (0.47+/-0.18; t = 4.5450, P less than 0.05) and AQP9 (1.57+/-0.21; t = 3.0306, P less than 0.05). Treatment with OA + PI3K pathway inhibitor (LY294004) significantly decreased AQP9 mRNA expression (4.55+/-0.62) as compared to the control group (1.00+/-0.10; t = 9.7909, P less than 0.01), that 500 mumol/L OA group (2.43+/-0.53; t = 4.5018, P less than 0.05), and the LY294002 group (1.90+/-0.16; t = 7.1683, P less than 0.01). Treatment with p38 MAPK pathway inhibitor (SB230580) significantly increased the OA-suppressed level of AQP3 mRNA to the level detected in the control group (1.27+/-0.11; t = 5.7455, P less than 0.01) and decreased the OA-stimulated AQP9 mRNA (0.38+/-0.09; t = 6.5727, P less than 0.01). No significant changes in mRNA expression of AQP3/9 were observed with inhibition of the ERK1/2 and JNK signal transduction pathways. The OA-induced changes in protein expression levels of AQR3 and AQP9 followed a similar trend of the genes. Finally, OA suppressed the level of phosphorylated Akt (from 0.21+/-0.02 to 0.13+/-0.03; t = 3.8431, P less than 0.05) but elevated the level of phosphorylated p38 (from 0.58+/-0.06 to 1.02+/-0.10; t = 12.5289, P less than 0.01). Again, OA treatment produced no significant affect on ERK1/2 and JNK phosphorylation.

CONCLUSIONOA down-regulates AQP3 expression by stimulating the p38 MAPK signaling pathway, and up-regulates the AQP9 by blocking the PI3K/Akt pathway and activating the p38 MAPK signaling pathway.

Aquaporin 3 ; metabolism ; Aquaporins ; metabolism ; Fatty Liver ; metabolism ; pathology ; Gene Expression Regulation ; drug effects ; Hep G2 Cells ; Humans ; Oleic Acid ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVE: To study the correlation of splicing mutations at the 5' end of the DMD gene with their phenotypes. METHODS: DMD gene mutations were analyzed using Multiplex Ligation Probe Amplification (MLPA) and Sanger sequencing. Co-segregation analysis was performed for the pedigrees of the probands. Influence of mutations on protein function was predicted by bioinformatic analysis. RESULTS: Three novel splicing mutations were identified in three patients with different phenotypes. Patient 1 carried a c.31+3insT mutation and presented primarily with dilated cardiomyopathy (XLDC). There was no clinical signs of skeletal myopathy. Bioinformatic analysis predicted that the mutation may inactivate the splicing donor of intron 1 and lead to premature termination of protein translation. Patient 2 carried a c.264_264+4delTGTAA mutation, which led to loss of splicing donor site for intron 4, and manifested Becker muscular dystrophy (BMD). The mutation was predicted to result in skipping of exon 4. The defective protein may still retain most of its function. Patient 3 had Duchenne muscular dystrophy (DMD) and carried a c.832-3C>T mutation which was predicted to decrease the activity of splicing acceptor of intron 8, resulting in usage of alternative acceptor site or retain of intron 8. All related transcripts may cause premature termination of protein translation and complete loss of protein function. The three mutations were all inherited from the mothers of the patients. CONCLUSION Three novel splicing mutations were discovered at the 5' end of DMD gene in three patients with different disease phenotypes. Our study may facilitate understanding of the influence of splicing mutations at the 5' end of the DMD gene on dystrophin function and the correlation between genotypes and phenotypes.
Dystrophin ; genetics ; Humans ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Phenotype ; RNA Splicing

Objective To analyze the relationship between strain subtypes and transmission of HIV infection on marriage-based immigrant women,their spouses and children in rural area of Zhejiang province.Methods Marriage-based immigrant women with HIV infection,their HIV infected spouses and children in rural area in Zhejiang province,were selected as study objects.Analysis on genetic sequence and epidemiologic information was carried out.Subgenomic gag was amplified by nest-PCR analysis on the whole blood samples.Genetic subtype characterization and the source of HIV strains were analyzed.Relationships on sequences were also examined by phylogenetic tree analysis.Results Genetic sequences of 72 samples from HIV infected marriage-based immigrant women were obtained.The genetic subtypes comprised 21 CRF01_AE (29.2％),12CRF07 BC (16.7％),31 CRF08 BC (43.1％),6 B (8.3％),2 C (2.8％).HIV strains from 45 cases (62.5％) were similar to the prevalent HIV strains in the province where former census of marriagebased immigrant women were registered.In total,there were 26 (70.3％) cases from Yunnan province.84.7％ of the infected women had heterosexual behaviors before settling down in Zhejiang province.Genetic sequences of 17 pairs showed the same subtype between the couples and data from phylogenetic tree analysis supported the assumption of transmission linkage in the family.Conclusion The HIV subtype strains detected in those HIV infected marriage-based immigrant women in the rural area of Zhejiang province characterized with diversity,showing CRF08_BC and CRF01_AE were the main HIV strain subtypes.HIV infection originated mainly fiom Yunnan province and nearby regions.Heterosexual behaviors of the marriage-based immigrant women in the original region where they had their residence registration,seemed to be the primary high risk factors for these women.Surveillance and intervention programs on these marriage-based immigrant women and their family members should be improved.

OBJECTIVETo explore etiologic fraction (EF) and interaction of serum hepatitis B surface antigen (HBsAg) carriage and other risk factors for primary hepatocellular carcinoma (PHC) in Wenzhou, Zhejiang, China.

METHODS1:1 matched case-control study was carried out in Wenzhou, with 180 cases of PHC and 180 controls. EF and interactions of serum positive HBsAg [HBsAg(+)] and other risk factors for PHC were analyzed by Mantel-Haenszel stratified method and conditional multiple logistic regression.

RESULTSSerum HBsAg(+), poor economic status during the past five years, preferring intake of pickled vegetables, history of PHC in their first-degree relatives, and negative life events all were risk factors for PHC, with EFs of 0.728, 0.245, 0.224, 0.084, and 0.234, respectively. There existed interactions of HBsAg(+) with other risk factors, including poor economic status during the past five years, preferring intake of pickled vegetables, history of PHC in their first-degree relatives, and negative life events, with etiologic fractions attributable to interaction [EF (A x B)] of 0.770, 0.630, 0.848, and 0.627, and indices of interaction of 0.789, 0.638, 0.852, and 0.634, respectively.

CONCLUSIONSMain risk factor for PHC in Wenzhou, Zhejiang, China could include HBsAg(+), poor economic status during the past five years, preferring intake of pickled vegetables, history of PHC in their first-degree relatives, and negative life events. HBsAg(+) plus any of the following factors, such as poor economic status during the past five years, preferring intake of pickled vegetables, history of PHC in their first-degree relatives, and negative life events, could increase the risk of PHC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; etiology ; Female ; Hepatitis B Surface Antigens ; blood ; Humans ; Liver Neoplasms ; etiology ; Logistic Models ; Male ; Middle Aged