Lactobacillus casei was selected as an antigen delivery vehicle for the development of oral vaccine to express recombinant LTB and porcine parvovirus (PPV) VP2 protein. The fusion protein gene encoding PPV VP2 protein and LTB, was cloned into the surface expression vector pPG, and then the recombinant expression vector pPG-VP2-LTB was electrotransformed into Lactobacillus casei 393, generating recombinant strain pPG-VP2-LTB/L. casei 393. After induced by 2% Lactose in MRS broth, an about 78 kD protein was detected in the recombinant Lactobacillus casei by SDS-PAGE. The result of Western blot indicated that the protein possessed the antigenic specificity same as the native virus protein. The result of the whole bacteria cell ELISA indicated that the LTB protein was expressed at the same time. The results of indirect immunofluorescence test and immuno-gold electron microscopy showed that the interest protein was expressed on the surface of L. casei 393. The results provide potential for the development of lactic acid bacteria oral vaccine of PPV, which used LTB as mucosal adjuvant.

This study was conducted to explore a proper training model of interns' clinical thinking ability under the construction of a new four-year system of medical laboratory technology courses, combined with the establishment of innovative standard whole process practice mode. Multi-teaching methods of clinical thinking, such as explanation of laboratory sheet, interactive teaching based on micro digital system, interdisciplinary multiple information system, combined PBL teaching and intern report, were applied and evaluated in the laboratory. Integrated application of these methods remarkably improved the intern's com-prehensive professional quality and their practice performance. All methods received high evaluation from both the interns and teachers.

OBJECTIVETo investigate effect of tumor necrosis factor-alpha (TNF-alpha) on the Src-suppressed C kinase substrate (SSeCKS) in C6 glioma cells.

METHODSCultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-alpha (2 ng/mL) for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-alpha (0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL) for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS's subcellular localization.

RESULTSTNF-alpha induced rapid phosphorylations of protein kinase C (PKC) substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-alpha induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220.

CONCLUSIONTNF-alpha activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.

A Kinase Anchor Proteins ; Animals ; Astrocytes ; metabolism ; Brain Neoplasms ; metabolism ; Cell Cycle Proteins ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation ; Glioma ; metabolism ; Immunohistochemistry ; Protein Kinase C ; metabolism ; Protein Transport ; physiology ; Random Allocation ; Rats ; Signal Transduction ; physiology ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; administration & dosage ; physiology

Objective To evaluate whether the patients with Parkinson′s Disease (PD) having higher occurrence rate of frac-ture .Methods CMB ,CNKI ,PubMed ,Embase ,Web of Science ,Medline ,Embase and Cochrane Library were retrieved ,meanwhile which was assisted by the manual retrieval .The retrieval time was until February 2017 .The cohort studies on the occurrence rate of fracture in PD patients were collected .Then the included studies were analyzed after the data extraction and treatment evaluation . Results A total of 1160 articles were retrieved ,finally 11 cohort studies were included ,involving 988723 subjects .The analysis showed that the fracture occurrence risk in PD patients was significantly higher than that in the control group (RR=2 .09 ,95% CI:1 .91-2 .28) ,in which the occurrence rate of hip fracture was significantly higher than that in the control group (RR=2 .33 ,95%CI:1 .79-3 .02) ,while the occurrence rate of spinal fracture had no statistical difference (RR=1 .33 ,95% CI:0 .78-2 .27) ,and the fracture occurrence risk in male and female patients of PD group was significantly higher than that in the control group (RR=2 .40 , 1 .69 ,95% CI:2 .21-2 .60 ,1 .62-1 .76) .Conclusion The fracture occurrence risk in PD patients is significantly higher than that in the control group ,due to existence of certain geographic bias and publication bias risk ,it is needed more high quality clinical stud-ies to accurately evaluate whether the fracture risk in PD patients being much higher .

OBJECTIVE: To establish a 15-plex rapid STR multiplex amplification system. METHODS: Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives. RESULTS: Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure. CONCLUSION The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Alleles ; Chromosome Mapping ; DNA/genetics* ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Humans ; Microsatellite Repeats/genetics* ; Polymerase Chain Reaction/methods* ; Racial Groups/genetics* ; Sensitivity and Specificity ; Tandem Repeat Sequences

The present study investigated the contribution of endogenous heme oxygenase (HO)/carbon monoxide (CO) system to hypertension pathogenesis of rats. Zinc deuteroporphyrin 2,4-bisglycol (ZnDPBG),an inhibitor of heme oxygenase (HO), was used to inhibit HO activity in vivo. It was found that the blood pressure of rats with HO inhibition was significantly elevated, and plasma levels of adrenaline, noradrenaline,endothelin, nitrate and nitrite were significantly increased. HO activity and HbCO formation within vascular smooth muscle tissues were significantly inhibited after administration of ZnDPBG. Furthermore, administration of exogenous CO into HO inhibiting rats led to MABP decrease, but injection of HO substrate, heme-Llysinate, had no effect on HO inhibition-induced hypertension. In spontaneously hypertensive rats, injection of exogenous CO resulted in a significant decrease of MABP, and heme-L-lystnate had a similar effect with exogenous CO. These data show that HO/CO system has an anti-hypertension biological action, suggesting that endogenous CO plays an important role in hypertension pathogenesis.

A sepsis model induced by cecal ligation and puncture was used to study the role of endogenous carbon monoxide in hypotension pathogenesis of rats during septic shock. After administration of zinc deuteroporphyrin 2,4-bisglycol (ZnDPBG),an inhibitor of heme oxygenase (HO),blood pressure (BP),HO activity and carbon monoxide (CO) release from vascular muscle tissue were measured. The results showed that BP of sepsis rats,including systolic and diastolic arterial BP,decreased significantly while HO activity and CO content were significantly increased. In contrast,after administration of ZnDPBG,BP of sepsis rats was significantly increased while the HO activity and CO production were significantly decreased. These findings suggest that HO activity and CO release within vascular musculature are increased during septic shock;inhibition of HO may elevate BP of rats during septic shock through a decrease of endogenous CO production. It is concluded that endogenous CO derived from vascular muscle cells plays an important role in regulating vascular tone,and the up-regulation of HO activity followed by subsequent CO production contributes to hypotension pathogenesis during septic shock.

Objective To elucidate the potential mechanisms by which mesothelin(MSLN)contributes to chemotherapy resistance in high-grade serous ovarian cancer(HGSOC).Methods A Meta-analysis utilizing public ovarian cancer databases was performed to evaluate the correlation between MSLN expression levels and overall survival(OS)in ovarian cancer patients.Pathway enrichment analysis was employed to identify key signaling pathways regulated by MSLN and their roles in chemotherapy resistance.Additionally,the TCGA-HGSOC database was analyzed to examine genomic features associated with MSLN-mediated chemotherapy resistance.To validate the biological function of MSLN in chemotherapy resistance,an intraperitoneal metastasis model was established using MSLN-knockdown ID8 ovarian cancer cells in mice.Results Elevated MSLN expression was significantly associated with poor patient prognosis(HR:1.42,95%CI:1.16-1.74).Differential gene expression and pathway enrichment analyses revealed that high MSLN expression upregulates resistance-associated genes and pathways involved in drug metabolism and DNA-binding signaling.Genomic association analysis showed a negative correlation between high MSLN expression and chromosomal instability features,specifically CX3,CX11,and CX13 scores.In vivo studies demonstrated that MSLN knockdown enhanced the tumor-suppressive effects of cisplatin.Conclusion High MSLN expression represents a potential biomarker for poor prognosis and chemotherapy resistance in HGSOC patients,suggesting MSLN as a promising target for therapeutic intervention.

OBJECTIVETo survey the prevalence of enterotoxigenic Escherichia coli (ETEC) and Laribacter hongkongensis (LH) and their drug resistance in diarrhea patients in Guangzhou.

METHODSWe detected 646 fecal cases collected between Sep 2008 and Oct 2009 from the out-patient and emergency departments in a hospital. EC enriched culture medium was used for enrichment. MAC- and CMAC-specific culture media were used to isolate ETEC and LH from the specimens. The biochemical agents API20NE and API20E were employed for biochemical identification, and PCR was used for genetic identification. K-B disk diffusion method was used for antimicrobial susceptibility testing.

RESULTSNo LH was detected in the total 646 patients, and 38 patients were positive for ETEC, with a detection rate of 6%. Antibiotics resistance test showed that 38 strains of ETEC had a high resistance rate to penicillin, tetracycline and sulfa, but remained sensitive to cephalosporins.

CONCLUSIONSLH may have a low prevalence in Guangzhou. The incidence of diarrhea caused by ETEC tends to decrease as compared with that a decade ago, and further multi-center survey is needed for confirmation. Consumption of aquatic products may be one of the major risk factors for ETEC infection. Cephalosporins can be used for ETEC-induced diarrhea.

Adolescent ; Adult ; Bacterial Infections ; epidemiology ; microbiology ; Cephalosporins ; pharmacology ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; microbiology ; Drug Resistance, Bacterial ; Enterotoxigenic Escherichia coli ; drug effects ; isolation & purification ; Escherichia coli Infections ; epidemiology ; microbiology ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Neisseriaceae ; drug effects ; isolation & purification ; Prevalence ; Young Adult

OBJECTIVETo express and purify polyphosphate kinase (PPK) from Proteus mirabilis and prepare the polyclonal antibody against PPK.

METHODSThe antigenicity and hydrophobicity of PPK were analyzed using software. The N-terminal conservative sequence containing 309 amino acids was selected as the target peptide, and its corresponding gene sequence with modification based on prokaryotic cells-preferred codon was synthesized and inserted into plasmid pET28b(+). The constructed recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced with IPTG. The expressed fusion protein was purified using Ni-affinity chromatography. The purified protein was injected along with adjuvant in rabbits to prepare the polyclonal antibodies against PPK.

RESULTS AND CONCLUSIONPPK fusion protein expressed by E. coli was purified successfully using Ni-affinity chromatography. ELISA result demonstrated that the harvested rabbit anti-sera against PPK had a high titer of 1:512 000, and Western blotting showed a good specificity of the antibody, which can be used further study of the role of PPK in the pathogenesis of Proteus mirabilis infection.