OBJECTIVETo study the influence of HBV/P22 protein on the induced apoptosis of HepG2 cells.

METHODSIn vitro, two HepG2 strains were transfected with pcDNA3.1+ and pcDNA3.1+HBV/P22 respectively and the cells were exposed to Act D and TNF alpha for 6h and then the induced apoptosis was detected by flow cytometry (FCM) and TUNEL technique. Supernatant HBeAg was detected by Abbott reagent. The intracellular expression of HBV/P22 protein was measured by Western blot and immunochemistry. In vivo, three cell groups were inoculated into nude mice by subcutaneous injections. After two weeks, Act D and TNF alpha were injected into the tumors and then the induced apoptosis in the tissues was detected by TUNEL technique. The expression of HBV/P22 protein in the tumor tissues was detected by immunochemistry.

RESULTSIn vitro, in HepG2- pcDNA3.1+HBV/P22 cells, supernatant HBeAg was positive and intracellular HBV/P22 protein was positively expressed. The apoptosis proportion of HepG2-pCDNA3.1+HBV/P22 cells was markedly lower than HepG2 and HepG2-pcDNA3.1+ cells (P < 0.05). In vivo, HBV/P22 protein was expressed in the tumor tissues, and the apoptosis proportion in the group injected with HepG2-pcDNA3.1+HBV/P22 cells was markedly lower than those injected with HepG2 and HepG2-pcDNA3.1+cells (P < 0.05).

CONCLUSIONHBV/P22 protein could inhibit the induced apoptosis of HepG2 cells both in vitro and in vivo.

Animals ; Apoptosis ; Female ; Hep G2 Cells ; Hepatitis B Core Antigens ; genetics ; Hepatitis B e Antigens ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Transfection ; Viral Core Proteins ; genetics

OBJECTIVETo investigate the effects of the hepatitis B virus (HBV) P22e protein on the apoptosis of human hepatocellular carcinoma HepG2 cells.

METHODSHepG2 cells were transfected with recombinant plasmid pEGFP-HBVP22e and exposed to Act-D and tumor necrosis factor alpha (TNFalpha) treatment to induce cell apoptosis. Flow cytometry was performed to determine the proportion of cells containing sub-G1 DNA to represent the number of apoptotic cells. Laser scanning confocal microscopy was used to observe the nuclear alterations in the apoptotic cells.

RESULTSHepG2EGFP-C2HBVP22e cell strain showed a much delayed apoptosis as well as obviously lowered apoptotic rate in comparison with the HepG2 strain (P<0.01).

CONCLUSIONThe introduction and expression of extraneous gene HBVP22e significantly inhibits the apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Humans ; Transfection ; Viral Core Proteins ; metabolism

OBJECTIVETo test whether nuclear factor kappa B plays an important role in the apoptosis-inhibitory effect of hepatitis B virus (HBV) P22(e) protein.

METHODSHepG2 cells were transfected with recombination plasmid pEGFP-HBVP22(e). The Act-D and TNF alpha were used to induce apoptosis. NF-kappa B inhibitor ALLN were used to inhibit the signaling pathway. The activation of NF-kappa B was EMSA, and the nulear translocation of NF-kappa B was determined by immuno-staining.

RESULTSLaser scanning confocal microscopy and EMSA indicated that HBV P22(e) protein enhanced the nuclear translocation of NF-kappa B after apoptosis induction. ALLN treatment inhibited the nuclear translocation of NF-kappa B, and blocked the apoptosis-inhibiting effect of HBV P22(e) protein.

CONCLUSIONThis study indicates that HBV P22(e) protein inhibits apoptosis of hepatocyte via the NF-kappa B signaling pathway.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B virus ; genetics ; Humans ; Leupeptins ; pharmacology ; Liver Neoplasms ; metabolism ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Plasmids ; Signal Transduction ; drug effects ; Transfection ; Viral Core Proteins ; metabolism

OBJECTIVETo investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.

METHODThe effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.

RESULTAfter 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.

CONCLUSIONCur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.

Cell Cycle ; drug effects ; radiation effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Curcumin ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; radiation effects ; Humans ; RNA, Long Noncoding ; genetics ; Radiation Tolerance ; drug effects

As a self-protective mechanism of cells, autophagy of cells can maintain cell stability by degrading self-aging substances, and it can be highly induced. The ability of autophagy to degraded cells will decrease with age. The resorption phenomenon after lumbar disc herniation is one of the effective mechanisms in conservative treatment of lumbar disc herniation. The degenerative lesion of intervertebral disc is one of the main reasons of lumbar disc herniation. Cell autophagy is extensive participation in the degeneration of lumbar intervertebral disc, delaying the occurrence of degenerative disease. Futhermore, cell autophagy can potentially induce the occurrence of reabsorption. The study of cell autophagy has great significance to the degenerative disease of intervertebral disk and the reabsorption of lumbar disc herniation. And it is also of great significance for the clinical treatment of patients with lumbar disc herniation. For this reason, we should pay more attention to the study of cell autophagy in resorption.
Autophagy ; Humans ; Intervertebral Disc ; cytology ; pathology ; Intervertebral Disc Displacement ; pathology ; Lumbar Vertebrae ; pathology

Objective To analyze the internal and external factors of the portable integrated remote medical examination chest during military medical support to facilitate telemedicine service in the military hospital.Methods The self-developed chest had its advantages, disadvantages, opportunities and threatening factors analyzed qualitatively with SWOT method. Results The chest had advantages in meeting military and practical requirements as well as fulfilling military-civilian integration, disadvantages in deficiency of specialized mobile network, specific signs acquisition devices and mature operation mechanism, opportunities in national health program, military development and mobile internet as well as the threatening factors in non-unified interface standard,untimely policy issuing and unclear definition of responsibility,rights and interests.Conclusion Considerations have to be taken on internet application and policy institution to facilitate military telemedicine support.

OBJECTIVETo provide an cell model of immortalized lymphoblstoid B-cell lines for studying the biological characteristics of full-length hepatitis B virus (HBV) genome carrying the hot-spot mutations V60, G87, and L97.

METHODSV60, G87, and L97 mutation points were introduced into HBV p3.8 II plasmid containing 1.2 copy of HBV genome by means of site-directed mutagenesis. The HBV genome was amplified by PCR from p3.8 II and p3.8 II-V60, G87, L97 plasmid, and the PCR product was inserted into EBO-plpp eukaryotic expression vector. The recombinant vectors and the EBO-plpp vector were transfected into immortalized human lymphoblasts with lipofectamine 2000 and selected with hygromycin. Steady expression of the target genes was determined by RT-PCR, Western blotting and microparticle enzyme immunoassay.

RESULTSDNA sequence analysis indicated that the desired mutation was introduced into wild-type HBV DNA. HBsAg, HBeAg and HBcAg could be detected in EBO-HBV-transfected cell lysate or culture supernatant.

CONCLUSIONTransfectants that stably express HBV mutant antigen may provide a cell model to study the biological characteristics of HBV carrying hot-spot mutation in vitro.

B-Lymphocytes ; cytology ; virology ; Base Sequence ; Blotting, Western ; Cell Line, Transformed ; Cell Transformation, Viral ; DNA, Viral ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation, Viral ; Genetic Vectors ; Genome, Viral ; genetics ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo develop fluorescent amplified fragment length polymorphism (AFLP) method and to evaluate the its typing capability with pulsed-field gel electrophoresis (PFGE) in molecular typing of Vibrio cholerae.

METHODSForty-seven strains of V. cholerae, with different PFGE patterns, were selected as the reference group to optimize the selective primers of AFLP analysis. Eighty-three strains including 20 strains from one epidemic episode, isolated from different provinces during 1961 and 2005, were used to compare the typing abilities of AFLP and PFGE. LI-COR4300 DNA sequencing system was used for AFLP electrophoresis. The images were recorded by Saga(MX) software and transferred to BioNumerics for clustering analysis. A standard protocol for V. cholerae from PulseNet was used in PFGE.

RESULTSWhen comparison was made with different selective primers on AFLP based on the 47 strains, results showed that the optimized selective primer pair was EcoR I-G/Mse I-T, and the reproducibility of the tests was 99.2%. Eighty-three isolates showed 52 AFLP patterns and 44 PFGE patterns, with D values as 0.9545 (AFLP) and 0.9251 (PFGE) respectively.

CONCLUSIONThe protocol of fluorescent AFLP on V. cholerae typing was established. AFLP was higher than PFGE in discrimination of V. cholerae which could be used for molecular typing. When combined with PFGE, AFLP became a more insightful tool to identify genome difference of different isolates.

Amplified Fragment Length Polymorphism Analysis ; methods ; Electrophoresis, Gel, Pulsed-Field ; Genotype ; Phylogeny ; Vibrio cholerae ; classification ; genetics

OBJECTIVETo study the characteristics of epidemiology and molecular typing on Neisseria meningitidis serogroup C strains associated with outbreaks of Anhui province and sporadic cases in China, using pulsed field gel electrophoresis (PFGE).

METHODS212 Neisseria meningitidis serogroup C strains were isolated from invasive meningococcal cases, close contacts and healthy carriers, including 48 strains from Anhui province with 38 strains associated with serogroup C outbreaks. PFGE were performed by genomic DNA digestion with Nhe I restriction enzyme. The results of PFGE were analyzed by BioNumerics software (Version 4.0, Applied Maths BVBA, Belgium).

RESULTSA total number of 212 Neisseria meningitidis serogroup C isolates were typed by 43 patterns, named AH1 to AH43. In China, AH1 pattern was the major PFGE pattern with 69.3% (n = 147) of all strains, distributed in 11 provinces. Three types of PFGE patterns (AH1 to AH3) were found in 48 strains from Anhui province, in which, 93.8% (n = 45) belonged to AH1. 97.4% (n = 37) of 38 strains associated with serogroup C outbreaks in Anhui province showed AH1 pattern. A total of 53 serogroup C strains were isolated from invasive meningococcal cases with 67.9% (36/53) of AH pattern. 71.9% (87/121) of serogroup C strains isolated from contacts of invasive meningococcal cases was AH1 pattern and 63.2% (24/38) of the strains from healthy carriers showed AH1 pattern.

CONCLUSIONBy PFGE typing and analysis, AH1 pattern of Neisseria meningitidis serogroup C strains was proved to be the main clone which causing the outbreaks in Anhui province and might be responsible for the sporadic serogroup C meningococcal disease epidemics else where in the country.

Bacterial Typing Techniques ; China ; epidemiology ; DNA, Bacterial ; genetics ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Meningococcal Infections ; epidemiology ; Neisseria meningitidis, Serogroup C ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA

OBJECTIVETo investigate the situation of overactive bladder (OAB) in a community-based male population.

METHODSMale participants over 50 years old were randomly selected from multiple communities in Beijing. The evaluation of lower urinary tract symptoms (LUTS) including the International Prostate Symptom Score (IPSS), quality of life (QOL) score, prostate volume and post voiding residue (PVR) by abdominal ultrasonography, and maximum flow rate (Qmax). Definition of OAB was determined as the score of item number 4 in IPSS ≥ 2.

RESULTSOf 1656 male participants enrolled, a total of 1639 men met our study criteria. The mean age was (64 ± 10) years. The prevalence of OAB was 26.3% (431/1639), and was significantly related to age, IPSS, QOL score, prostate volume, PVR and Qmax (P < 0.01). The prevalence of OAB was closely associated with aging (P < 0.01) and the degree of LUTS (P < 0.01).

CONCLUSIONSThe prevalence of OAB increased with aging of the community-based male population. OAB would obviously affect the quality of life of the aging men.

Aged ; Aged, 80 and over ; Aging ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Prevalence ; Quality of Life ; Urinary Bladder, Overactive ; epidemiology