Animals ; Ear, Inner ; growth & development ; Mice ; MicroRNAs

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

Fibroblast growth factor-21 (FGF-21) is an important metabolism regulator, however, whether FGF-21 has effects on cardiovascular remains unclear. In this study, H2O2-induced injury in H9c2 cells was used as a cell model, the anti-apoptosis potential and mechanism of FGF-21 against oxidative injury were evaluated by MTT assay, flow cytometry assay and real-time PCR. The results showed that FGF-21 could increase the cell survival of H2O2-induced injury in H9c2 cells and prevent H9c2 cells from oxidative stress-induced apoptosis. Furthermore, FGF-21 can elevate SOD activity and regulate Bcl-2/Bax expression in H9c2 cells. The results suggest that FGF-21 have protective effect against the H2O2-induced apoptosis in H9c2 cells.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fibroblast Growth Factors ; pharmacology ; Hydrogen Peroxide ; toxicity ; Malondialdehyde ; metabolism ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Protective Agents ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo study the anticancer effect in vitro and in vivo and mechanism of DHA compound.

METHODSCervical cancer cell line HeLa cells, glioma cell line U251 cells and mouse hepatoma H(22) tumor were used in this study. Transmission electron microscopy and fluorescence microscopy were used to observe the morphological changes of cell apoptosis. Western blot was used to detect the expression of caspase-3. RT-PCR was used to determine the effect on Bcl-2 and Bax mRNA transcription in U251.

RESULTSAntitumor effect was observed in vivo and in vitro. Typical morphological changes were seen in cancer cells. The level of caspase-3 was significantly increased and the content of Bcl-2 mRNA was decreased significantly, while the content of Bax mRNA was significantly increased in the U251 cells after treatment with DHA compound.

CONCLUSIONDHA compound can inhibit the growth of some types of tumors and the increase of caspase-3 and Bax mRNA and decrease of Bcl-2 mRNA may be involved in its mechanism of action.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Glioma ; pathology ; HeLa Cells ; Humans ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVESTo investigate CD3+CD56+ lymphocytes and their subsets in the peripheral blood of chronic hepatitis B patients and to explore the relationship between these cells and the pathogenesis of their diseases.

METHODSBlood samples from 53 chronic hepatitis B patients, 17 from HBV asymptomatic carriers (ASC) and 19 from healthy controls (HC) were collected. CD3+CD56+ lymphocytes were detected by flow cytometry (FCM), then the CD3+CD56+ lymphocytes were gathered to analyze their expressions of CD4, CD8, TCR Valpha24, TCRalpha/beta and TCRgamma/delta.

RESULTSThe number of CD3+CD56+ lymphocytes of chronic hepatitis B patients (7.4+/-4.6%) was more than those of ASC (4.5%+/-3.5%) and healthy controls (4.4%+/-3.7%). The expressions of TCR Valpha24 on CD3+CD56+ lymphocytes showed no significant differences among the three groups, but the expression of TCR Valpha24 on CD3-CD56+ lymphocytes of ASC ( 2.8%+/-1.4% ) was much more than that of the HC (1.7%+/-1.0%). For the subsets analysis, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of chronic hepatitis B (61.9%+/-16.8% and 68.1%+/-16.9%) were significantly higher than those of the HC (49.2%+/-15.6% and 56.4%+/-17.9%), while the TCRgamma/delta subsets of chronic hepatitis B and ASC (29.6%+/-15.4% and 30.5%+/-14.8%) were decreased significantly than those of the HC (41.4%+/-19.4%). On the other hand, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of severe chronic hepatitis B (69.0%+/-14.0% and 76.1%+/-12.9%) and CD8 subsets of moderate chronic hepatitis B patients (66.4%+/-14.9%) were significantly higher than those of the mild chronic hepatitis B patients (51.4%+/-16.2% and 62.1%+/-14.6%).

CONCLUSIONThe pathogenesis of chronic hepatitis B may positively relate to the high expression of CD8 on the CD3+CD56+ lymphocytes.

Adult ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol x L(-1) glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.
Adult ; Animals ; Bacterial Adhesion ; DNA, Bacterial ; analysis ; Denaturing Gradient Gel Electrophoresis ; Fusobacterium nucleatum ; physiology ; Humans ; Membranes, Artificial ; Mice ; Microbial Interactions ; physiology ; Polymerase Chain Reaction ; Protein Binding ; Saliva ; microbiology ; Streptococcus mutans ; physiology

OBJECTIVETo characterize the role of Toll-like receptor 4 (TLR4) in silica-induced production of tumor necrosis factor alpha (TNFalpha) from macrophage cell line.

METHODSThe human macrophage cell line THP-1 was incubated with silica suspension. Cell media were collected and TNFalpha levels in the supernatants measured with ELISA. To examine the involvement of TLR4 in silica-induced TNFalpha release, the neutralizing antibody (HTA125) against human TLR4 receptor was employed to pretreat THP-1 cells prior to silica treatment. Moreover, murine macrophages expressing wild type or mutated TLR4 were also treated with silica to verify the effect of TLR4 in silica-induced TNFalpha release.

RESULTSCompared with the control group [(3.18 +/- 0.41) pg/ml], the TNFalpha release in cells exposed to 100 microg/ml silica for 4 h and 8 h [(4.71 +/- 0.84), (6.22 +/- 0.58) pg/ml, respectively] increased 1.48 and 1.96 fold, respectively. Pretreatment of THP-1 cells with 20 microg/ml HTA125 antibody significantly blocked silica-induced TNFalpha release by 27%. Furthermore, the TNFalpha content released from cells expressing mutated TLR4 reduced by 30% in compared with that from the cells expressing wild type TLR4 after silica stimulation.

CONCLUSIONTLR4 mediates silica-induced TNFalpha release from macrophages.

Antibodies ; pharmacology ; Cell Line ; Humans ; Macrophages ; drug effects ; metabolism ; Silicon Dioxide ; toxicity ; Toll-Like Receptor 4 ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore treatments of severe post-kidney-transplant lung infection by integrative Chinese and Western medicine (ICWM), in order to elevate the curing rate as well as to lower the death rate.

METHODSBased on conventional ways of Western medical treatments of 18 cases of severe post-kidney-transplant lung infection, such as putting the patients in single individual ward, antibiotics to prevent infection, respiratory machines, blood filtration, nutritional support, steroids, and maintaining electrolytes balance, we applied integrated Chinese medicinal treatments, like altering conventional prescription "pneumonia III", and conducted clinical observation of effectiveness, and indexes including white blood cell (WBC), neutrophilic granulocyte, blood urea nitrogen (BUN), blood creatinine (Cr), etc.

RESULTSOf the 18 cases studied, 7 were already cured, 8 proved the treatment effective, 3 died. All clinical indexes had statistically significant changes compared with those of before treatment (P < 0.01).

CONCLUSIONICWM can increase curing rate and lower death rate.

Adult ; Aged ; Anti-Bacterial Agents ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunocompromised Host ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; Lung Diseases ; drug therapy ; immunology ; Male ; Middle Aged ; Opportunistic Infections ; drug therapy ; Respiratory Tract Infections ; drug therapy ; immunology

OBJECTIVETo investigate the expression characteristics of the gene of coiled-coil domain-containing protein 70 (Ccdc70) in the mouse testis and its potential role in spermatogenesis.

METHODSUsing expression profile microarray, we screened the mouse testis-specific gene Ccdc70, studied its expression characteristics in the mouse testis by RT-PCR, real-time PCR, Western blot and immunohistochemistry, followed by bioinformatic analysis of the Ccdc70 protein.

RESULTSThe Ccdc70 gene was expressed highly in the testis but lowly in the epididymis of the mice. The Ccdc70 protein was expressed mainly in the spermatocytes and round spermatids of the testis and in the epithelial cells of the epididymis. Bioinformatic analysis showed a structural domain in the Ccdc70 protein, which was highly conserved in mammalian evolution.

CONCLUSIONThe Ccdc70 gene is highly expressed in the mouse testis and mainly in the spermatocytes, round spermatids, and epididymal epithelial cells, which indicates that it is involved in the regulation of spermatogenesis and epididymal sperm maturation.

Animals ; Computational Biology ; Gene Expression Regulation, Developmental ; Male ; Mice ; Proteins ; genetics ; Spermatogenesis ; genetics ; Testis ; metabolism

Objective To explore surgical treatment of gliomas involving the motor eloquent area. Methods Twelve cases of gliomas involving precentral gyrus were underwent awake surgery procedures assis- ted with neuronavigation and brain functional mapping by cortical electrical stimulation.Results Eleven ca- ses acquired accurate location of both lesions and eloquent areas by neuronavigation and direct cortical stimula- tion.7 cases of motor cortices and 2 cases of motor speech centers were confirmed during the operation.Re- section,verified by postoperative MRI,was total in 8 cases (66.7%) and subtotal in 4 patients.Histological examination revealed an infiltrative glioma in all cases (8 low grade astrocytomas,2 high grade astrocytomas and 2 glioblastoma).Four patients had no postoperative deficit,while the other 8 patients were impaired, with,in all cases except one,complete recovery in 7 days to one month.Conclusion Direct cortical elec- trical stimulations and awake surgery offer a reliable,precise and safe method,allowing functional mapping es- pecially useful in case of infiltrative cerebral tumors in eloquent areas.This technique allows improvement in the quality of tumoral resection and concurrently a minimization of the risk of definitive postoperative neurologi- cal deficit.