Amifostine ; pharmacology ; Animals ; Benzene ; toxicity ; Blood ; drug effects ; Blood Cell Count ; Male ; Mice ; Random Allocation

To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

OBJECTIVESTo investigate the expression of Survivin in patients with extrahepatic cholangiocarcinoma (EHCC) and its relationship with clinicopathological features of EHCC, and the correlation between the expression of Survivin and lymph node micrometastasis, tumor markers, and the prognosis of EHCC.

METHODSThe expression of Survivin protein in paraffin-embedded specimens of 59 patients with EHCC and their 20 para-carcinoma tissues were evaluated by S-P method of immunohistochemical staining. The correlation between the expression of Survivin and the lymph node micrometastasis, clinicopathological features of EHCC and the prognosis of EHCC were analyzed.

RESULTSThe positive expression rate of Survivin protein was 67.8% (40/59) in paraffin-embedded specimens of 59 patients with EHCC and was 20.0% (4/20) in para-carcinoma tissues, and difference between carcinoma tissues and para-carcinoma tissues was significant (P<0.01). Histological differentiation in EHCC had a negative correlation with the expression of Survivin protein, while the expression of Survivin protein in EHCC had a positive correlation with TNM of EHCC, lymphatic vessel infiltration, lymph node metastasis and perineural invasion (P<0.05). The serum CA19-9 levels in the positive group with expression of Survivin protein was (290,300+/-55 500) U/L and was obviously higher than that in the negative group [(113,300+/-31,400) U/L, P<0.05]. The mean survival time of the patients with negative expression of Survivin protein was higher than that of the patients with positive expression (43.5 vs. 21.1 months, P<0.01). Screened to significance univariate, the multivariate analysis through Cox proportional hazard model analysis showed that lymph node metastasis, residual tumor margins, and expression of Survivin protein were independent prognosis factors of the patients with EHCC (P<0.05, P<0.01, P<0.01).

CONCLUSIONSThe expression of Survivin protein in EHCC has a negative correlation with histological differentiation, while has a positive correlation with lymphatic vessel infiltration and serum CA19-9 concentrations. The expression of Survivin protein maybe an independent prognosis factor of the patients with EHCC.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Extrahepatic ; Cholangiocarcinoma ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis

Objective To learn the epidemiological characteristics of HIV/AIDS cases aged 15-24 years in Shaoxing for making targeted prevention and control measures among young people. Methods To analyze the HIV/AIDS cases' distribution, route of transmission and reported route according to the cases which were reported 2003-2016 and aged 15-24 years. Results There were 325 cases had been reported. The epidemic had spread outward to each county, and transient population was accounted for 64.00%, and the ratio of male to female was 2.46:1. Career was widely dispersed, and farmers/migrant workers accounted for 31.08% , and students accounted for 10.15% . Homosexual transmission accounted for 41.29% , and heterosexual transmission accounted for 51.08% . The cases that were locality census register/high school education or above/unmarried adolescent, and professional for business services and student's homosexual transmission ratio was higher than that of heterosexual transmission. Voluntary counseling and testing was the major detection route which accounted for 16.9%. Conclusion The results showed that the male who aged 15-24 years was the most susceptible population. It is necessary to carry out effective measures to prevent and control HIV/AIDS transmission among young people, especially MSM and out-of-school youth.

OBJECTIVETo construct a system of I-SceI and induce a site-specific DNA double-strand break (DSB) in the genome of HepG2 for using this system in future exploration of the potential mechanisms of HBV integration by DSB repair.

METHODSThe eukaryotic expression plasmid pEGFP2 was constructed and transfected into human hepatoma cell line HepG2. The positive neomycin-resistant transfected cell clones were generated by G418 selection. Then the positive cells containing an 18-bp I-SceI endonuclease site were transfected transiently with pCMV(3NLS) I-SceI, an I-SceI expression plasmid. At 24 h post-transfection with pCMV (3NLS) I-SceI, gamma-H2AX, as an early cellular marker of DSB, was detected using immunocytochemistry and Western blot analysis.

RESULTSRestriction analysis and DNA sequencing verified that the plasmid pEGFP2 was successfully constructed. gamma-H2AX increased significantly in cells transfected with the I-SceI system.

CONCLUSIONSGenomic DSB can be induced into HepG2 by introducing an I-SceI system. The cell model could provide us with a practical tool for further study to see if DSB is a potential target for HBV integration.

Carcinoma, Hepatocellular ; genetics ; DNA Breaks, Double-Stranded ; DNA Repair ; Flap Endonucleases ; genetics ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; Plasmids

BACKGROUNDEndostatin is a potent inhibitor of tumor angiogenesis. In the preliminary studies, we developed a mutant endostatin containing Arg-Gly-Asp-Arg-Gly-Asp (RGDRGD) sequences. In this study, we compared the antitumor effects of mutant endostatin and Bcl-2 antisense oligonucleotides both in combination and individually.

METHODSThe artificially synthesized Bcl-2 ASODN (antisense oligonucleotides) included a translation-initiation site and was transfected into the bladder cancer cells by Lipofectamine. Cell growth was investigated by the tumor cell growth chart, MTT assay, caspase-3 activity detection assay, AO/EB fluorescein stain, and the annexin V-FITC apoptosis detection assay. In the in vivo study, UM-UC-3 bladder cancer cells were subcutaneously implanted into nude mice and the growth of tumor was examined. The ultrastructure of the tumor tissues in the treated and control groups were observed.

RESULTSThe cell growth chart showed that the cell population of the treated combination group decreased by 52.04% compared to the control group. The inhibition rate of the treated combination group was (79.66 ± 6.79)%, whereas those of the individual ASODN and ES groups were (53.39 ± 3.22)% and (50.22 ± 5.46)% respectively. In the caspase-3 activity detection using AO/EB fluorescein stain and annexin V-FITC apoptosis detection assay, the co-inhibitory effect was higher than the individual inhibitory effects (P < 0.05). There were significant differences in the inhibition of the solid tumor growth in the in vivo study.

CONCLUSIONSOur findings indicated that Bcl-2 antisense oligonucleotides enhance the antitumor effects of mutant endostatin both in vitro and in vivo. We noted the synergistic effects of Bcl-2 antisense oligonucleotides combined with mutant endostatin.

Angiogenesis Inhibitors ; administration & dosage ; Animals ; Cell Line, Tumor ; Drug Synergism ; Endostatins ; administration & dosage ; Mice ; Thionucleotides ; administration & dosage ; Urinary Bladder Neoplasms ; pathology

OBJECTIVETo study diagnosis and therapeutic efficacy of transient oeteoporods of the hip (TOH).

METHODSFrom January 2005 to February 2010, 5 patients with TOH were treated with traditional methods. All the patients were male, with an average age of 38.6 years (ranged, 27 to 46 years). The clinical manifestation, physical examination and imageology characteristic was investigated. The therapeutic efficacy was evaluated by Harris hip score.

RESULTSAll the patients were followed up, the duration ranged from 12 to 36 months (averaged, 24 months). The Harris hip score before treatment were 63.1, 86.0, 74.9, 63.6 and 64.8 respectively, while after 6 months treatment, the scores improved to 90.5, 94.5, 89.7, 93.9 and 87.8 respectively. Moreover, 6 months later, the abnormal signal disappeared in MR imaging and X-ray.

CONCLUSIONTransient osteoporosis of the hip is a self-resolving condition and a self-limited disease, the expectant treatment is useful for it.

Adult ; Female ; Hip Joint ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Osteoporosis ; diagnosis ; therapy ; Retrospective Studies

Aged ; Humans ; Hypercalcemia ; complications ; Liver Neoplasms ; complications ; Lymphoma ; complications ; Male

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Diabetes Mellitus ; epidemiology ; etiology ; Female ; Glucose Intolerance ; epidemiology ; Glucose Tolerance Test ; Humans ; Hypertension ; complications ; Male ; Mass Screening ; Middle Aged ; Obesity ; complications ; Prevalence ; Rural Health ; Sampling Studies ; Surveys and Questionnaires ; Urban Health

OBJECTIVESTo evaluate the correlation between signal/cutoff (S/CO) ratios of anti-HCV EIA and their true positivity for determining the predictive value of S/CO ratios.

METHODSOne hundred and fifty-nine samples of blood from donors positive for anti-HCV at the initial screening were collected from Beijing, Guangzhou, Hangzhou, Kunming and Urumchi. All the samples were retested by Ortho and 6 Chinese domestic anti-HCV EIA kits in duplicate, and detected for HCV RNA (NAT) using Chiron Procleix HIV/HCV system (transcription mediated amplification, TMA). The HCV RNA negative samples were further tested for anti-HCV by Chiron RIBA 3.0. Either NAT or RIBA positive samples were interpreted as the true positive.

RESULTSAll 7 anti-HCV EIA kits had a significant correlation between S/CO ratios and true positivity. The S/CO ratio of Ortho > or = 3.8 predicted the true positivity in 96.1% of the samples tested. The S/CO ratios of BGI-GBI, GWK, SABC, KHB, InTec, and Wantai were > or = 7.0, > or = 10.0, > or = 6.0, > or = 10.0, > or = 8.6, > or = 14.0 and predicted 96.1%, 96.1%, 97.3%, 96.0%, 96.1%, 96.0% of the true positivity, respectively.

CONCLUSIONSThe S/CO ratios of anti-HCV EIA kits are associated with the true positivity. S/CO ratios of Ortho, BGI-GBI, GWK, SABC, KHB, InTec and Wantai predicting > or = 95% true positivity are > or = 3.8, > or = 7.0, > or = 10.0, > or = 6.0, > or = 1 0.0, > or = 8.6 and > or = 14.0, respectively.

Blood Donors ; Hepacivirus ; isolation & purification ; Humans ; Immunoenzyme Techniques ; methods ; Predictive Value of Tests ; Reagent Kits, Diagnostic ; Sensitivity and Specificity