Objective:To study the antigenicity of OMP extracted from Edwardsiella tarda.Methods:ELISA, Bactericidal test, Agglutinating test and Western blotting were used for testing the antigenic titers and immunogenicity of OMP.Results:In immunoblotting, by using ATCC15947 OMP antibody, the non pathgenic strains were negative, while all pathogenic strains except Et 122 gave positive results and had OMP bands of 33k, 35k, 38k, and 45k. OMPs of both ATCC 15947 and JEL4 could induce high antibody titers. Further more, the antibodies evoked by OMPs of ATCC 15947 of 33k or 35k could also protected mice to some degree when diluted.Conclusion:The 33k, 35k, 38k, and 45k of OMPs may be protective antigens, and the OMPs of Et could be a candidative component for vaccine.

Adolescent ; Adult ; Aged ; Bloodletting ; Child ; Child, Preschool ; Combined Modality Therapy ; Dermatitis ; therapy ; Female ; Humans ; Infant ; Insect Bites and Stings ; therapy ; Lymphangitis ; therapy ; Male ; Middle Aged ; Moxibustion ; Young Adult

Objective To study the mechanism of the resistance in Acinetobacter baumannii to common antibiotics.Methods Bacterial susceptibility test to ?-lactamatic antibiotics,quinolones,aminogiycosids were done by the Kirby-Bauer method and agar dilution method for 35 isolates.Penicillin-beta-lactamase,AmpCs,Metallo-beta-lactamase,ESBLs were detected by iodine-starch test,3-dimension test,microbiology sensitivity synergic test,disc agar diffusion method respectively.Outer membrane protein was analyzed by SDS-PAGE.Accumulation of ciprofloxacin was determined by direct Fluorescence method.The gene of Tem-1,aac-4 and gyrA were amplified by PCR while the gyrA was sequenced.Results 28 isolates were multi-resistant to common antibiotics in 35 isolates.16 isolates produced Penicillinase,10 isolates produced Cephalosporinase,2 isolates produced metal-beta-lactamase,3 isolates produced ESBLs.The analysis of outer membrane proteins showed that a protein of 29kD disappeared and 26kD protein enhanced in resistant isolates.The accumulation of ciprofloxacin in resistant isolates decreased.After treatment with NaN_3,the drug uptake increased to the normal level.Most of [QX(Y8]Tem-1 gene were positive except 2 drug resistant and 3 sensitive isolates.All of [QX(Y8]aac-4 were negative while gyrA were positive.DNA sequencing analysis revealed there had point mutation in the gyrA gene.Conclusion Beta-lactamase,active drug efflux,outer membrane protein permeability decreasement and gene mutation were the factors contributing to the antibiotics resistance of Abaumannii.

BACKGROUND: Recent studies indicate that after indirect co-culture of neonate rat myocardial cells and bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells can differentiate into myocardial cells successfully. OBJECTIVE: To induce human bone marrow mesenchymal stem cells (hBMSCs) to differentiate into endothelial cells using human umbilical vein endothelial cells by indirect co-culture. DESIGN, TIME AND SETTING: In vitro study of cell engineering was done at the Medical Research Center of Guangdong Provincial People’s Hospital between January and July 2007. MATERIALS: Small quantities of bone marrow were obtained from 11 children with congenital heart disease but without hematologic diseases through manubrium of sternum puncture in the congenital heart defect corrective surgery after the permission of family member. Umbilical cord of full-term normal delivery healthy newborn was provided by the Department of Obstetrics, the First Affiliated Hospital of Sun Yat-sen University. Fresh cattle jugular vein was provided by Guangdong Dali Meat Cattle Butchery. METHODS: The hBMSCs were isolated and purified using density gradient centrifugation method and were cultured in vitro. Human umbilical vein endothelial cells were obtained from newborn umbilical cord by enzyme digestion. Cell culture insert with semipermeable membrane combined with 6-well plate was used to do indirect co-culture induction. Human umbilical vein endothelial cells were expanded in the cell culture insert, passage 3 bone marrow mesenchymal stem cells were expanded in the 6-well plate outside the culture insert at the density of 1?105 cells/well, the initial ratio of the two kinds of cells was 1:5, then low-glucose DMEM culture solution containing 10% fetal bovine serum was added, cells were cultured for 14 days. Co-culture of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells was used as control. Introduced endothelioid cells were cultured and then seeded on the cell-free cattle jugular vein intravascular stent. MAIN OUTCOME MEASURES: Morphology changes of induced cells; introduced endothelioid cell surface antigen detected through immunocytochemical staining; the growth and adhesion condition of endothelioid cells on the intravascular stent observed under scanning electronic microsope. RESULTS: The morphologies of introduced endothelial cells were uniform, introduced endothelial cells presented a cobblestone-like appearance, they amplified fast and expressed endothelial cell-specific surface marker CD31 and vWF and the positive rate was over 99%. They also could form a continuous unicellular layer on the cell-free cattle jugular vein intravascular stent. CONCLUSION: Human umbilical vein endothelial cells can induce hBMSCs to differentiate into endothelial cells successfully and to adhere and grew on the cell-free cattle jugular vein intravascular stent through indirect co-culture method.

Objective To explore short term curative effects and adverse reactions of two different kinds of intraperitoneal hyperthermic chemotherapy (IPHC) drugs in the treatment of advanced ovarian cancer after cytoreductive surgery.Methods 76 patients with advanced ovarian cancer in the Second Affiliated Hospital of Zhengzhou University from July 2013 to November 2015 were divided into two groups:single-drugs group of 36 patients (IPHC with 5-fluorouracil after cytoreductive surgery combined with intravenous chemotherapy),combined treatment group of 40 patients(IPHC with 5-fluorouracil and carboplatin after cytoreductive surgery combined with intravenous chemotherapy).Short term curative effects,postoperative clinical indicators and adverse reactions of chemotherapy in two groups were compared and analyzed.Results The CA125 effective rates in single-drugs and combined treatment group were 86.11％ and 95％,and the difference showed statistically significant differences (P ＞ 0.05).The ascites remission rates in single-drugs and combined treatment group were 97.22％ and 97.5％,and the difference between two groups showed no statistically significant differences (P ＞ 0.05).Adverse drug reactions showed statistical difference in distribution of the bone marrow,liver damage and gastrointestinal toxicity.No statistical difference were found between the two groups in terms of distribution of renal damage and cardiovascular system damage.Conclusion IPHC after cytoreductive surgery in the treatment of advanced ovarian cancer is an effective means as adjuvant chemotherapy.The short-term curative effect of combined treatment group is obvious and adverse reactions can be tolerated.IPHC can be applied according to the patient's specific clinical situation.

Objective To explore a rapid method for classification of microorganisms.Methods The electrophorese fingerprinting, direct sequence of 16S rDNA and 16S-23S rDNA ISR after PCR, multiplex PCR for 16S rDNA and antibiotic resistance genes, were utilized to explore fast approaches of extracting total DNA from different clinical specimens.Results The specific 16S-23S rDNA ISR fingerprinting fragments were shown on the genus or species level in bacteria and fungi.So fingerprinting can be used to identify pathogenic microorganisms, to differentiate the evolution relations or to set the phylogenetic tree by comparing their DNA banding patterns with those of standard strains (NCCLS). Multiplex PCR was able to examine the special genes of genus or species, mecA gene, TEM, SHV and CTX gene in staphylococcus and ESBLs(E.coli or K.pneumoniae) at the same time.Conclusion The part of 16S rDNA sequencing and 16S-23S rDNA ISR genotypes by gel electrophoreses were useful for bacterial species identification in addition, it was clearly more rapid and accurate than culture technique, and the large numbers of strains can easily be examined.Multiplex PCR could provide a good method for identification of microorganisms and analysis of antibiotic resistance at the same time.

Objective To determinate the content of Danshensu in Xingkenin g Capsules by HPLC.Methods HPLC with Hypersil ODS2 column was used, MeOH- 1 % acetic acid( 12:88) as a mobile phase and detection wavelength at 280nm.Re sults The linear range of Danshensu was in the range of 0.256～ 1.278 ? g. Th e average recovery of Danshensu was 102.73 % with a RSD of 1.34 % .Conclusio n The method is simple with a good reproducibility and can be used for the qu ality control of Xingkening Capsules.

Objective To establish a method for the determination of patchouli alcohol in Huoxiang Qingwei Tablets by gas chromatography (GC).Methods HP- 5 column( 30 m? 0.32 mm? 0.25 ? m) was used and 5 % crosslinked phenyl polydimethyl siloxane was used as the mobile phase. The increase of column temperature was controlled by programming: the initial temperature was 150 ℃ and maintained for 23 min, and then rose at 8 ℃ /min up to 230 ℃ for 2 min. The injector temperature and the detector temperature both were 280 ℃ .The flow rate was 0.9 mL/min and the injection volume was 1 ? L, split ratio was 5∶ 1.Result A good linearity was obtained in the range of 0.057～ 1.140 mg/mL,r=0.9994. The average recovery was 99.38 % with a RSD of 1.82 % (n=5). Conclusion The method is simple and accurate and can be used for the quality control of Huoxiang Qingwei Tablets.

Aim Telomerase is highly expression in most tumor cells, and it is an ideal target for cancer molecular targeting therapy. It has been proved that wogonin effectively inhibits telomerase activity and tumor cell growth in vitro. The study was to explore the inhibitory effect of wogonin on the growth of tumor and telomerase activity of implanted human ovarian cancer cell line SKOV3 in nude mice. Methods Nude mice with implanted human ovarian cancer cells SKOV3 were randomly divided into five groups, viz. the high dose group of Wogonin(600 mg?kg-1),low dose group of Wogonin(300 mg?kg-1),normal control group, cisplatin therapy group(3 mg?kg-1), and combined therapy group(cisplatin plus wogonin).The weight of nude mice and the volume of tumor were regularly measured. DNA、RNA and protein were extracted from the tumor tissue. The length of telomere was examined by Southern blot. The expression of telomerase hTERT gene was detected by RT-PCR. The telomerase activity was examined by TRAP-PCR-silver staining. Results The wogonin significantly inhibit the growth of tumor when compared with controlled group.The inhibitory rate of high dose group and low dose group were 56.67% (P=0.002) and 38.10%(P=0.019), respectively. The inhibition rate of cisplatin therapy group was 50.83%(P=0.004). The suppress rate of combined group reached 66.9% and higher than any single therapy(P=0.002). The length of telomere in different concentration groups of wogonin was the same as that in the control group.Wogonin inhibited the expression of telomerase gene hTERT and telomerase activity. The inhibition is related to the dose of wogonin. Conclusion Wogonin suppresses the growth and telomerase activity of tumor. The inhibitory effect is related to the dose of wogonin. Combination of wogonin and cisplatin increase the inhibitory rate in nude mice tumor.

Objective: To investigate the effects of epigallocatechingallate (EGCG) on lipid metabolism related gene and long non-coding RNA (lncRNA) expression proifle by biochip technology, and to explore the possible relationship between the two elements.
Methods: HepG2 cell was cultured with EGCG at 25μmol/L for 24 hours, the total RNA was extracted and hybridized into the biochip of Human Transcriptome Array 2.0 for mRNA and lncRNA expression profile analysis. Bioinformatics technology was used to establish the possible relationship between lncRNA and the predicted target genes;the data obtained from biochip microarray was conifrmed by real time RT-PCR examination.
Results: The microarray revealed that EGCG treated HepG2 cell expressed 27 differential lipid metabolism genes and 11 of them involved in cholesterol metabolism. In addition, there 285 lncRNA expressions were up-or down-regulated. Bioinformatics technology indicated that the predicted target genes for lipid metabolism might be cis-or trans-regulated by lncRNA;the data from real-time RT-PCR was consistent with the data from biochip microarray.
Conclusion: Tea polyphenols improves lipid metabolism and lncRNA might be involved in the regulation of lipid metabolism related gene.