Objective A prokaryotic expression vector was constructed by inserting the coding sequence of mouse sperm specific lactate dehydrogenase C into pET-28a(+)and the recombinant mLDHC44 protein was purified by Ni+-NTA agrose.Methods The cDNA of mouse sperm specific lactate dehydrogenase C was obtained by RT-PCR,with total RNA of mouse testis tissues as templates.The coding sequence of mouse LDHC4 was amplified by PCR with specific primers.This recombinant vector was transformed into Escherichia coli BL21(DE3).The recombinant mLDHC4 protein was induced by isopropy-?-D-thiogalactoside(IPTG)and identified by sodium dodecyl sulfate polyacrylamide electrophoresis and LDH activity determination.After purified with Ni+-NTA agrose,the mLDHC4 protein was probed with antisera from the pVAX1-mLDHC4 vaccine(the eukaryotic expression vector of mouse sperm specific lactate dehydrogenase C)immunized BALB/c mice by Western blot analysis.Results After digested with BamH I-EcoR I,the recombinant plasmids produced right fragment which was about 1000bp.Sequencing showed that the sequence of the cloned fragment was in agreement with sequence in GenBank.This recombinant vector was named as pET-28a(+)-mLDHC4.With induction of IPTG,The recombinant protein with molecular weight of about 35 kD was expressed and the enzyme activity of this protein was high.After purified with Ni+-NTA agrose,this mLDHC4 protein formed a specific band by sodium dodecyl sulfate polyacrylamide electrophoresis and probed with antisera from immunized BALB/c mice and then formed a specific band in the nitrocellulose membrane.Conclusion The coding sequence of mouse lactate dehydrogenase subunit C had been cloned into the prokaryotic expression vector pET-28a(+)and the mLDHC4 protein could be expressed at a high level,the specificity of this protein was high and the activity was strong.

Objective To study the correlation between benign prostatic hyperplasia (BPH) and vesicoureteral reflux (VUR). Methods Intravenous radionuclide cystography (IVRC) was performed on 88 BPH patients and micturition cystourethrography on 30 to find if there was any VUR and the degree of reflux. Results 31 patients had VUR of which there were 6 grade Ⅰ,7 grade Ⅱ,6 grade Ⅲ,6 grade Ⅳ and 6 grade Ⅴ.9 were on the right,6 on the left and 16 bilateral. Conclusions VUR do occur in some BPH patients and IVRC is the better method for its diagnosis and evaluation.

With the rapid development of pharmacogenetics, more and more studies have shown evidence in the association between polymorphisms at the human leukocyte antigen (HLA) loci and severe adverse drug reactions (SADRs). Several HLA-B alleles proved to be associated with SADRs for drugs such as carbamazepine, allopurinol, lamotrigine, and lfucloxacillin. hTe USA Food and Drug Administration (FDA) has even recommended routine screening for HLA-B allele before the use of abacavir and carbamazepine. With the completion of human genome project and the Hapmapproject, several new pharmacogenetics approaches such as genome-wide association study (GWAS) have emerged. hTese newly developed methods will undoubtedly accelerate the identiifcation and clinical utilization of the pharmacogenetic biomakers. In addition, the immunogenetic mechanisms by which the HLA alleles cause SADRs are explored at the cellular and molecular level. hTis review focuses on the recent progresses in HLA alleles and ADRs regarding both the clinical translation and modern pharmacogenetic methods.

Objective By compared FAB classification of myelodysplastic syndrome with WHO classification of myelodysplastic syndrome,understanded the proposed WHO new classification of myelodysplastic syndrome as soon as possible.Methods Retraced and analysed 78 cases of myelodysplastic syndrome based on examination of the bone marrow.Restults(1)According to the FAB classification,divided myelodysplastic syndrome into 5 kinds:RA,RARS,RAEB,CMML,RAEB-T.(2)According to the WHO classification,its subtypes were adjusted again,and its concluded:RA,RARS,RCMD,RAEB-Ⅰ,RAEB-Ⅱ,5q-del.(3)There were 9 cases possess chromosome to revise particularly in the chromosome check 28 cases myelodysplastic syndrome. Conclusion WHO classification has clinical guiding significance for early diagnosis,therapy observation and prognosis decision.

Objective To summary the color Doppler imaging feature of localized Castleman disease.Methods From January 1997 to November 2011,32 localized Castleman diseases which were proved by pathology were analyzed.Results Round-like,hypoechoic,hypervascular lesions were showed in 23 hyalinevascular type lesions,2 of them with calcium,3 of them with structure liked lymph node hilum.Round-like,hypoechoic,hypervascular lesions or normal lymph node were showed in 5 plasma- type lesions and 4 mixed type lesions.Conclusions When round like,hypoechoic,hypervascular lesion is found by ultrasonography,Castleman should be considered.Calcium or lymph nod hilum-like structure is special finding in diagnosis of Castleman disease by ultrasonography.

AIM:To observe the effects of prostaglandin E2 on transforming growth factor-?1(TGF-?1)triggered human lung fetus fibroblast(HLF)transdifferentiation and connective tissue growth factor(CTGF),collagen type I(COLⅠ)expression.METHODS:HLFs were treated with TGF-?1,the cells underwent phenotypic change to myofibroblast.The marker of myofibroblast-?-smooth muscle actin(?-SMA)was detected by immunofluorescence.The ?-SMA content was measured by Western blotting.The changes in CTGF and COL Ⅰ at transcription levels were estimated by RT-PCR method.CTGF protein expression was evaluated by immunocytochemical.Cell culture medium hydroxyproline amount was measured by colormetric assay.RESULTS:PGE2 blocked TGF-?1 induced ?-SMA positive myofibroblast transformation(P

Objective To observe the effects of hydrotalcite on histological ulcer healing quality in patients with Helicobacter pylori (Hp) positive active gastric ulcer .Methods A total of 145 patients with H p positive active gastric ulcer were divided into two groups .The control group was treated with esomeprazole ,amoxicillin and furazolidone triple therapy .The treatment group was treated with above triple therapy and hydrotalcite .After four-week treatment ,gastroendoscopy was repeated .The sections of gastric biopsy specimens were stained with hematoxylin-eosin (HE) staining ,Van Gieson staining , Alcian blue-periodic acid stiff (AB-PAS ) staining , streptavidin-perosidase (SP ) immunohistochemistry staining and computer imaging analysis technology were applied to observe maturity type of mucosal structure at the margin of ulcer ,the content of collagen and neutral mucus ,and the changes of expression of epithelial growth factor receptor (EGFR) and basic fibrobast growth factor (bFGF) before and after treatment .Paired samples t test was performed for comparison before and after treatment .F test and Chi-square test were used for comparison between two groups .Results The percentage of maturity type of regenerated mucosal structure of treatment group was 62 .9% (39/62) ,however that of control group was 40 .6% (26/64) ,and the difference was statistically significant (χ2 =13 .09 , P=0 .03) .Compared with those before treatment , the content of collagen in granulation tissue and neutral mucus in regenerated mucosa increased in both groups after treatment , and the increase was more significant in treatment group ((55 .1 ± 10 .4)% and (45 .8 ± 7 .1)% ,respectively);and the differences were statistically significant (F= 12 .85 and 18 .17 ,both P<0 .01) .Compared with those before treatment ,the percentage of EGFR and bFGF positive cells of both two groups increased , and the increase was more significant in treatment group ((49 .5 ± 8 .4)% and (48 .8 ± 9 .4)% ,respectively) ,and the differences were statistically significant (F=12 .17 and 18 .73 ,both P<0 .01) .Conclusion Hydrotalcite combined with anti-H p triple therapy can improve the maturity degree of structure and function of regenerated mucosa at the margin of ulcer in patients with H p positive active gastric ulcer and then improve the healing quality of ulcer .

Objective To investigate the distribution and drug resistance of bacterial pathogens isolated from peritonsillar abscess .Methods Data on bacterial pathogens isolated from peritonsillar abscess in Wenzhou Central Hospital from January 2010 to December 2014 were retrospectively analyzed .Strains were identified with Vitek 32 identification system and the drug susceptibility test was performed with K-B method.Chi-square test for linear trend was performed to reveal the changes of distribution and drug resistance of the strains .Results A total of 2 864 bacterial strains were isolated in five years , in which 1 786 strains were Gram-negative bacilli (62.4%), and 1 078 (37.6%) strains were Gram-positive cocci. The positive rate of Gram-negative bacilli was on the rise during year 2010-2014 (χ2 =84.74, P<0.01), and the top three Gram-negative bacilli were Pseudomonas aeruginosa, Acinetobacter baumannii and Haemophilus influenzae, which accounted for 72.5%(1 295/1 786) of the total Gram-negative strains, and the positive rates of first two bacilli were on the rise (χ2 =83.75 and 24.74, P<0.01).Gram-positive cocci were mainly Staphylococcus aureus and Hemolytic streptococcus, which accounted for 83.2% ( 897/1 078) of the total Gram-positive strains.Resistance rates of Pseudomonas aeruginosa to ceftazidime, cefoperazone, piperacillin/tazobactam were on the rise (χ2 =16.17, 13.48 and 11.44, P<0.05), while resistance rates to gentamicin and amikacin were on the decline (χ2 =16.54 and 16.63, P <0.05). Resistance rates of Acinetobacter baumannii to ceftazidime, cefoperazone/sulbactam and piperacillin/tazobactam were on the rise (χ2 =12.52, 10.85 and 14.14, P<0.05).Resistance rates of Haemophilus influenzae to ampicillin were on the rise (χ2 =10.21, P<0.05), and the positive rate of β-lactamase producing strains was also on the rise (χ2 =10.38, P<0.05).Resistance rates of Staphylococcus aureus to cefazolin and methicillin were on the rise (χ2 =15.44 and 12.53, P<0.05), but no vancomycin resistant strain was found .Hemolytic streptococcus were sensitive to all commonly used antibiotics .Conclusions Peritonsillar abscess in Wenzhou Central Hospital is mainly induced by Gram-negative bacilli infection . Pseudomonas aeruginosa, Acinetobacter baumannii and Staphylococcus aureus are the top three bacterial pathogens , and are highly resistant to most antibiotics .

Objective To study the therapeutical effect of γ-aminobutyric acid on monocrotaline(MCT )induced pulmonary hy-pertension rats ,and to elucidate the expression of VEGF and MMP-9 .Methods Thirty SD rats were randomly divided into 3 groups :a normal control group(control group) ,a MCT-induced pulmonary hypertension group(model control group) ,and anγ-ami-nobutyric acid treatment group(treatment group) .The mean right ventricular pressure(mRVP)were detected ,the right ventricular hypertrophy index(RVHI)were measured ,WT% and WA% were evaluated ,and the expression of VEGF mRNA in the lung tissue and MMP-9 were detected wtih FQ-PCR and immunohistochemical staining method respectively .Results mRVP ,RVHl ,WT% , WA% and the expression of MMP-9 and VEGF mRNA of treatment group were lower than those in the model control group(P<0 .05) ,but higher than the control group(P<0 .05) .Conclusion GABA has a therapeutic effect on pulmonary hypertension rats through regulating the expression of VEGF mRNA and MMP-9 protein .

OBJECTIVE: To explore whether pre-administration of Ganoderma lucidum spore (GASP) can reduce incidence of neural tube defects (NTDs) induced by all-trans retinoic acid (ATRA) in pregnant mice. METHODS: Twenty pregnant mice were randomly divided into four groups: normal control group, solvent-treated group, ATRA-induced group, and GASP-treated plus ATRA-induced group. GASP solution, which was prepared with solvent (sodium carboxymethyl cellulose), was fed to the pregnant mice in the GASP-treated plus ATRA-induced group twice a day from embryo (E) 0 d to E10.5 d. The same dose of solvent was given to the pregnant mice in the solvent-treated group. At E7.75 d, ATRA (50 mg/kg) was given to the pregnant mice in both ATRA-induced group and GASP-treated plus ATRA-induced group for single time. Embryos were sampled from pregnant mice at E10.5 d. Then the incidence rate of NTDs in mouse embryo was calculated and the crown-rump length of mouse embryo was measured. The positive rate of nestin expression and the distribution of cell cycle of embryonic neural tube neuroepithelial cells were detected by histochemical staining technique and flow cytometry respectively. Reverse transcription-polymerase chain reaction method was used to detect the gene expressions of cyclin-dependent protein kinase 2 (Cdk2) and Cdk4 mRNAs. RESULTS: The incidence rates of NTDs in mouse embryos in the ATRA-induced group and the GASP-treated plus ATRA-induced group were 79.41% and 21.67% respectively, while the crown-rump length of mouse embryos in these two groups were (3.62+/-1.27) mm and (5.84+/-0.92) mm respectively. The positive rate of nestin expression in embryonic neural tube neuroepithelial cells of mouse embryo at E10.5 d in the ATRA-induced group was 32.44%, while that in the GASP-treated plus ATRA-induced group was 77.65%. The cell cycle of embryonic neural tube neuroepithelial cells was obviously arrested at G(0)/G(1) phase in the ATRA-induced group as compared with that in the GASP-treated plus ATRA-induced group. The Cdk4 mRNA was transcripted at a high level in embryonic neural tube in the GASP-treated plus ATRA-induced group, but the Cdk2 mRNA was not detected in this group. CONCLUSION: Pre-administration of GASP can reduce the incidence of NTDs induced by ATRA in pregnant mice.