OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis C virus (HCV) recombinant NS5B-EGFP fusion protein and obtain a stable transfected HepG2 cell line.

METHODSThe coding region of NS5B gene of HCV was amplified by PCR and was digested by Xho I/Kpn I. This fragment was inserted into pEGFPN3 with T4 ligase and transformed E. coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into HepG2 cell by Lipofectin AMINE 2000. Cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level NS5B-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN3-ns5b was successfully constructed and the stable transfected HepG2 cell line expressing NS5B-EGFP fusion protein was obtained.

CONCLUSIONThe stable transfected HepG2 cell line could express NS5B-EGFP fusion protein, could be used for anti-HCV infection with ns5b gene as the target.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVETo evaluate the relationship between peroxisome proliferator-activated receptor-γ2 (PPARγ2) Pro12Ala polymorphism and type 2 diabetes mellitus (T2DM) in Chinese Han population.

METHODSPubMed, Chinese Biomedicine Database (CBM), China National Knowledge Infrastructure (CNKI) and Wanfang database were searched for all relevant articles investigating the association between PPARγ2 Pro12Ala polymorphism and T2DM that were available from January, 1990 to June, 2011. A total of 29 relevant articles were selected. A Meta-analysis was performed to estimate heterogeneity and the pooled odds ratio (OR) to evaluate the relationship between PPARγ2 Pro12Ala polymorphism and T2DM. The sensitivity analysis was also assessed.

RESULTSA total of 21 qualified articles including 3870 patients with T2DM and 3333 healthy controls were analyzed in the study. The frequencies of the allele Ala12 in T2DM and control groups were 4.13% (320/7740) and 4.56% (304/6666), respectively. There were not heterogeneity (χ(2) = 25.96, P = 0.17) among the 21 qualified articles. The pooled OR (95%CI) value of the frequencies of the PPARγ2 genotype (PA + AA)/PP calculated by fixed effects model was 0.96 (0.81 - 1.14) (P = 0.64). There was not heterogeneity among the remaining articles after excluding the article with the largest weight and the article with larger frequencies of the allele Ala12 respectively (χ(2) values were 24.23, 16.87 respectively, both P values > 0.05). The pooled OR (95%CI) value of the frequencies of the PPARγ2 genotype (PA + AA)/PP of the remaining articles were 1.01 (0.84 - 1.21) and 1.07 (0.89 - 1.28) after excluding the article with the largest weight and the article with larger frequencies of the allele Ala12 (both P values > 0.05).

CONCLUSIONPPARγ2 Pro12Ala polymorphism was not associated with type 2 diabetes mellitus in Chinese Han population.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Diabetes Mellitus, Type 2 ; genetics ; Gene Frequency ; Genotype ; Humans ; PPAR gamma ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo identify and characterize the epitope associated with the virus attachment protein (VAP) of hantaan virus.

METHODSThe monoclonal antibody 3G1 was used as the ligand to biospan from a phage-displayed 12-amino acid peptide library, then the positive phage clones were chosen and sequenced. The amino acid sequences of them were compared with that of hantaan virus G2 in homology. The characteristics of positive phage were studied by IFA and ELISA. A decapeptide combining to cell membrane was observed under laser scanning confocal microscope (LSCM).

RESULTSThe conservative motif PX(1-2) HX(0-2) H displaying on positive clones shared homologous amino acid sequence with G2 96YPWHTAKCHY105.

CONCLUSIONG2 96YPWHTAKCHY105 might play some roles in virus binding to host cell, and might be a possible key epitope of hantaan virus VAP.

Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; immunology ; metabolism ; Hantaan virus ; immunology ; Microscopy, Confocal ; Oligopeptides ; immunology ; metabolism ; Peptide Library ; Vero Cells ; Viral Proteins ; immunology ; metabolism

OBJECTIVETo study the clinicopathologic features and prognosis of primary lymphoma of breast.

METHODSForty cases of primary breast lymphoma, diagnosed according to the 2008 World Health Organization classification of hematopoietic and lymphoid tumors, were retrospectively studied. Immunohistochemistry was performed by SP method. The follow-up data were analyzed.

RESULTS(1) All the patients were females and the median age was 47 years. Unilateral and bilateral breast involvement were noted in 36 and 4 patients, respectively. The number of tumor were 31 cases (77.5%, 31/40) less than 3, and 9 cases (22.5%, 9/40) were 3 and more than 3. According to Ann Arbor staging system, 33 cases (82.5%) were in stage I to II and 7 cases (17.5%) in stage III to IV. The level of LDH in 9 cases (24.3%, 9/37) went up. For ECOG scores, 34 cases (85.0%) were 0 to 1 score and 6 cases (15.0%) were more than 2 scores. With respect to international prognostic index, 83.8% (31/37) were of score 0 to 2 and 16.2% (6/37) were of score 3 and more than 3. The axillary lymph nodes of 21 patients (53.8%, 21/39) were involved by the malignancy. (2) Histologically, 38 cases (95.0%, 38/40) were classified as B-cell lymphoma [including 27 cases (67.5%) of diffuse large B-cell lymphoma, 8 cases (20.0%) of mucosa-associated lymphoid tissue lymphoma, 2 cases of follicular lymphoma and 1 case of lymphoplasmacytic lymphoma]. The remaining cases included one case of peripheral T-cell lymphoma and one case of lymphoblastic lymphoma. Immunohistochemically, expression of CD20+/- CD79a were demonstrated in the 38 cases (95.0%) of B-cell lymphoma. The staining for CK was negative in all cases. In 33 cases, the positive rates of MUM-1, bcl-6 and bcl-2 were 57.6% (19/33), 30.3% (10/33) and 72.7% (24/33), respectively. Three cases were germinal center B cell phenotype and 21 cases were non-germinal center B cell phenotype. (3) Follow-up information was available in 37 patients (92.5%, 37/40). Twenty-three patients (62.2%, 23/37) were still alive and fourteen ones (37.8%, 14/37) died. For the 27 cases with diffuse large B-cell lymphoma, the five-year and disease-free survival rates were 48.0% and 36.0%, respectively.

CONCLUSIONSPrimary breast lymphoma is a rare disease entity. Diffuse large B-cell lymphoma is the commonest histologic type and the majority show a non-germinal center B cell phenotype. The level of LDH, number of tumor and international prognostic index are of prognostic significance.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; CD79 Antigens ; metabolism ; Cyclophosphamide ; therapeutic use ; Disease-Free Survival ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Interferon Regulatory Factors ; metabolism ; L-Lactate Dehydrogenase ; blood ; Lymphatic Metastasis ; Lymphoma ; drug therapy ; metabolism ; pathology ; surgery ; Lymphoma, B-Cell, Marginal Zone ; drug therapy ; metabolism ; pathology ; surgery ; Lymphoma, Follicular ; drug therapy ; metabolism ; pathology ; surgery ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; surgery ; Lymphoma, T-Cell, Peripheral ; drug therapy ; metabolism ; pathology ; surgery ; Mastectomy ; methods ; Middle Aged ; Neoplasm Staging ; Prednisone ; therapeutic use ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use ; Young Adult

BACKGROUNDMore and more Chinese drink hot water from water dispensers while many children were scalded due to this change. The present study aimed to propose a feasible strategy for prevention.

METHODSA retrospective study was conducted for all water dispensers related pediatric burns admitted to Changhai Hospital from January 2005 to December 2009.

RESULTSThe number of new cases and incidences of pediatric burns due to hot water from water dispensers was significantly increasing year after year. In the total 238 involved cases, 175 cases happened on males and 78.9% were at the age of 1 - 4 years. The burn areas were mainly located in upper extremities. All water dispensers in the surveyed families had no isolate protection devices and 85.2% of their locations were easy for children to reach. Nearly half of the children were in the same room with their guardians when injured. Total 196 burned children were playing the taps of water dispensers before injured, unfortunately, 80.6% of them have not been stopped until burned.

CONCLUSIONAs the kind of burns is quite serious and with bad outcome, some recommendations should be followed, such as buying water dispensers with protection devices, keeping children from touching them and so on.

Accidents, Home ; Adolescent ; Burns ; epidemiology ; etiology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Retrospective Studies ; Water

OBJECTIVETo prove the therapeutic effect of ginger-partitioned moxibustion on leukopenia induced by chemotherapy and effect on life quality of the patient with tumor after chemotherapy.

METHODSRandomized, controlled, multi-central cooperative method was used and the patients confirmed to the enrolled criteria were divided in-to two groups. The test group were treated with ginger-partitioned moxibustion at Dazhui (GV 14), Geshu (BL 17), Pishu (BL 20), etc.; and the control group with oral administration of Chinese patent medicine.

RESULTSOut of the 221 cases confirmed to program analysis, 113 cases were in the test group and 108 cases in the control group. After 10 days, the cured rate and the effective rate were 84.1% and 66.4% in the test group and 35.2% and 33.3% in the control group, respectively, with very significant differences between the two groups (both P < 0.01); fifteen days later, the therapeutic effects in the two groups were maintained. The two methods could improve clinical symptoms, with the test group being better than the control group. Any adverse response was not found in the two groups, and the injuries of functions of the heart, lung and kidney induced by chemotherapy had some improvement.

CONCLUSIONThe therapeutic effect of ginger-partitioned moxibustion on luekopenia induced by chemotherapy is reliable and is better than oral administration of Chinese patent medicine, with a better duplication.

Adult ; Aged ; Antineoplastic Agents ; adverse effects ; Female ; Ginger ; Humans ; Leukopenia ; therapy ; Male ; Middle Aged ; Moxibustion ; methods

BACKGROUNDDuring recent years, the incidence of serious infections caused by opportunistic fungi has increased dramatically due to alterations of the immune status of patients with hematological diseases, malignant tumors, transplantations and so forth. Unfortunately, the wide use of triazole antifungal agents to treat these infections has lead to the emergence of Aspergillus spp. resistant to triazoles. The present study was to assess the in vitro activities of five antifungal agents (voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin) against different kinds of Aspergillus spp. that are commonly encountered in the clinical setting.

METHODSThe agar-based Etest MIC method was employed. One hundred and seven strains of Aspergillus spp. (5 species) were collected and prepared according to Etest Technique Manuel. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48 hours at 35°C. MIC(50), MIC(90) and MIC range were acquired by Whonet 5.4 software.

RESULTSThe MIC(90) of caspofungin against A. fumigatus, A. flavus and A. nidulans was 0.094 µg/ml whereas the MIC(90) against A. niger was 0.19 µg/ml. For these four species, the MIC(90) of caspofungin was the lowest among the five antifungal agents. For A. terrus, the MIC(90) of posaconazole was the lowest. For A. fumigatus and A. flavus, the MIC(90) in order of increasing was caspofungin, posaconazole, voriconazole, itraconazole, and amphotericin B. The MIC of amphotericin B against A. terrus was higher than 32 µg/ml in all 7 strains tested.

CONCLUSIONSThe in vitro antifungal susceptibility test shows the new drug caspofungin, which is a kind of echinocandins, has good activity against the five species of Aspergillus spp. and all the triazoles tested have better in vitro activity than traditional amphotericin B.

Amphotericin B ; pharmacology ; Antifungal Agents ; pharmacology ; Aspergillus ; drug effects ; Echinocandins ; pharmacology ; Itraconazole ; pharmacology ; Lipopeptides ; Microbial Sensitivity Tests ; Pyrimidines ; pharmacology ; Triazoles ; pharmacology ; Voriconazole

Objective·To explore the association between neuropeptide Y (NPY) gene polymorphism and schizophrenia in Chinese Han population. Methods·Four single nucleotide polymorphism (SNP) loci (rs16148, rs1859290, rs16147, and rs16478 in NPY gene) were selected and genotyped by TaqMan genotyping assay in a case-control study with 678 schizophrenia cases (case group) and 685 healthy controls (control group). The allele, genotype and haplotype frequencies distribution of the SNPs between the groups were compared with SHEsis online software. Results·The distribution of genotype frequency of locus rs16478 showed a nominal statistically difference between the adult-onset schizophrenia (AOS group) and control group (χ2=6.66, P=0.036, P correction=0.144). Under recessive inherited model, the distribution of CC genotype frequency of locus rs1859290 showed statistically difference between the male schizophrenia cases and control group (P=0.012, OR=0.97, 95％CI 0.94-0.99, P correction=0.048). The distribution of haplotype CCTA (consisted of rs16148, rs1859290, rs16147, and rs16478) frequency showed statistically difference between the male AOS group and control group (8.1％ vs 13.2％, OR=0.57, P=0.010, P correction=0.040). Conclusion·The polymorphisms in NPY gene may be associated with schizophrenia in Chinese Han population.

Among the types of lung cancer, lung adenocarcinoma accounts for the majority, and its overall survival rate is poor. B-cell translocation gene 2 (BTG2) is a member of the antiproliferative gene family, belonging to the BTG/TOB family. Many studies have shown that BTG2 was abnormally expressed in many types of tumors, but its regulatory role in the radiosensitivity of lung adenocarcinoma remained unclear. In this study, we explored the expression level of BTG2 in patients with lung adenocarcinoma and its correlation with clinical prognosis through online database and tissue samples of lung adenocarcinoma patient. The results indicated that the expression level of BTG2 decreased significantly in lung adenocarcinoma patient with radiation resistance. Bioinformatics analysis confirmed that BTG2 could respond to radiotherapy in lung adenocarcinoma cell lines, and its low expression in lung adenocarcinoma patients was associated with poor prognosis (P < 0.05). The lentivirus overexpressing BTG2 (OE-BTG2) was transfected into human lung adenocarcinoma cell lines to increase the expression level of BTG2 including A549 and H1299. And the effect of BTG2 overexpression on the radiosensitivity of lung adenocarcinoma cell lines was detected by clone formation assay. Clone formation experiment confirmed that overexpression of BTG2 could significantly enhance the radiosensitivity of A549 and H1299 cell lines (P < 0.05). The expression levels of BTG2 and apoptosis related protein-Bax were detected by Western blotting (WB) and immunohistochemistry (IHC). The effect of BTG2 on radiation sensitivity of lung adenocarcinoma was further detected via nude mouse in vivo. WB experiment confirmed that BTG2 upregulation could significantly increase the apoptosis level of A549 and H1299 cells after radiation. Moreover, BTG2 overexpression can markedly enhance the radiosensitivity of lung adenocarcinoma (P < 0.05) and increase the protein level of Bax after radiation in vivo. In conclusion, BTG2 had low expression in lung adenocarcinoma patients and its low expression level was closely related to the poor clinical prognosis. Overexpression of BTG2 can increase the radiosensitivity of lung adenocarcinoma cell lines and promote the process of apoptosis after radiation, indicating a new target for overcoming the radiation resistance of lung adenocarcinoma.

Objective To analyze the surveillance data after schistosomiasis transmission interruption from 1985 to 2018, so as to provide scientific basis for the future prevention and control of schistosomiasis. Methods The data pertaining to snail survey, surveillance of schistosomiasis in humans and livestock, and health education were collected and analyzed in Songjiang District after schistosomiasis transmission interruption from 1985 to 2018. Results From 1985 to 2018, there were 8 townships, 68 villages and 1 462 settings with snails in Songjiang District, covering an area of 96.399 hm². There were 4 townships and 59 villages with snails newly identified in previously schistosomiasis non-endemic areas, with emerging snail habitats of 60.161 hm². A total of 26 148 snails were captured, and no infections were detected. The snail habitats were mainly distributed in the river course, ditches and farmlands, which accounted for 45.28%, 33.17% and 21.55% of total snail habitats, respectively. A total of 245 955 humans were subjected to the serological test, with a positive rate of 0.03%, and 37 humans were subjected to the parasitological test, with 3 positives detected. These three egg-positives were all imported chronic cases with schistosomiasis. A total of 9 109 livestock were detected using the serological test, and no infection was found. Most of the awareness rate of schistosomiasis control knowledge and the rate of correct behavior formation were over 80% in residents and students. Conclusions The achievements of schistosomiasis control are consolidated in Songjiang District; however, there are still risk factors of schistosomiasis transmission, including residual snails, snail diffusion and importation of external source of infection. The future surveillance of schistosomiasis should focus on snails and sources of infection.