Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.

Objective To investigate the age-associated changes of ultrastructure,mRNA and protein expressions of H+-K+-ATPase in elderly gastric parietal cell. Methods Fifty patients with relative normal stomach without gastroduodenal diseases were enrolled,including younger group (aged 20-59 years,n=19) and elderly group (aged≥60 years,n=31).Furthermore,the elderly group was divided into 3 subgroups:60-69 years old (n =11 ),70-79 years old (n=10 ),above 80 years old (n =10).The ultrastructure of gastric parietal cell was observed under electron microscope.The expression of H+-K+-ATPase α subunit mRNA and H+-K+-ATPase β subunit protein were assessed by quantitative real-time PCR and Western-blot,respectively.The ageing-associated changes of all these data were respectively compared. Results No significant difference was showed in the morphology of gastric parietal cell and acid-secretion-associated organelles among all the groups.The average ratio Am to Ac (Am means the area of mitochondria,Ac means the area of cytoplasm) of gastric parietal cell and the average At to Ac ratio (At means the area of secretory canaliculi and tubulovesicular system )between younger group and elderly group had no significant difference[(48.4±7.5) ％ vs.(50.6±7.6) ％,t=-0.775,P=0.444; (13.8±4.1) ％ vs.(12.2±4.7) ％,t=0.984,P=0.332].Meanwhile,there were no distinctions in the expression of H+-K+ -ATPase α subunit mRNA and H+-K+-ATPase protein among all elderly subgroups(F=1.522,2.32,P=0.24,0.114).However,the mRNA expression of H+-K+-ATPase a subunit was higher in the elderly group than in the younger group(t=-3.682,P=0.001).Furthermore,the expression of H+ -K+ -ATPase protein in the elderly group was increased as compared with younger group(t=-3.389,P=0.004). Conclusions Acidsecretion-associated organelles of human gastric parietal cell have no degeneration and the expression of H + -K+-ATPase is in trend of increase with aging,indicating that healthy elderly people have the basis of ultrastructure and molecular biology to maintain well function of acid secretion.

BACKGROUND: Idiopathic scoliosis is a common spinal deformity in teenagers, which is managed mainly by orthomorphia. However, due to great trauma, long operative duration and large blood loss, a great amount of blood transfusion is needed during the surgery. Allogeneic blood transfusion should be reduced in order to release blood insufficient, decline blood transfusion expense, as well as avoid transfusion diseases. OBJECTIVE: To investigate the value of controlled hypotension combined with autotransfusion in idiopathic scoliosis orthomorphia.METHODS: Intraoperative controlled hypotension was performed during posterior orthomorphia surgery on all the 46 cases of idiopathic scoliosis, 17 cases in which were served as the control group, who underwent allogeneic blood transfusion without autotransfusion, while the other 29 cases were served as the experimental group, who underwent autotransfusion that including reinfusion of preoperative deposited autologous blood and intra-operative salvaged autologous blood. The blood loss volume and transfusion status in two groups were observed. RESULTS AND CONCLUSION: Blood loss volume in the control group was 400-1 000 (867±161) mL, and that in the experimental group was 350-1400 (842±376) mL, There was no marked difference between the two groups (P > 0.05). The volume of allogeneic blood transfusion in the control group was 500-1 800 (845±332) mL, which was greater than that in the experimental group [0-1 300(423±237) mL] (P < 0.01). The results suggested that controlled hypotension reduces intraoperative bleeding, and postoperative autotransfusion minimizes the need of allogeneic blood transfusion.

Objective To investigate the efficacy of microsurgery treatment with small craniotomy lateral fissure approach for hyperten-sive basal ganglia intracranial hemorrhage. Methods From December 2013 to February 2015, 30 patients with hypertensive basal ganglia intracranial hemorrhage ( blood loss within 30~60 mL) were analyzed respectively. Summarized the methods and techniques of this microsur-gery treatment and analyzed its advantages and disadvantages compared with the traditional operation. Results The hematomas were excluded more than 90% which were proved by reexamination after opreation. There was one case of temporal lobe infarction,but there was no re-bleed case and death case. The recovery of consciousness is good, and the limb function and language function were improved by early rehabilitation treatment. Conclusion Microsurgery treatment with small craniotomy lateral fissure approach for hypertensive basal ganglia intracerebral hemorrhage has the advantages of minimal invasion,complete hematoma removal and low complication and disability rates, which is worthy of promotion and application.

Animals ; Cytochrome P-450 CYP1A2 ; Cytochrome P-450 CYP2C19 ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochromes ; metabolism ; Liver Cirrhosis ; enzymology ; Rats

Background The sex hormones plays an important role in the incidence of dry eye,especially for the regulation of function.However,the effects of sex hormones on lacrimal gland epithelial cells are below understand.Objective This study was to investgate the effects of estradiol and testosterone on the apoptosis of lacrimal gland cells induced by H2O2.Methods The lacrimal gland tissue was obtained from 2- or 3-month-old clean male New Zealand rabbits and the lacrimal gland epithelial cells were cultured in vitro using esplant culture method.The cells were identified by pan cytokeratin antibodies with immunocytochemistry.lacrimal gland epithelial cells were incubated in the 96 well plate at the density of 5 × l04 cells/ml for 44 hours.Estradiol or testosterone with the concentrations of 1 × 10-5,1 × 10-6,1 × 10-7,1 × 10-8 mol/L were added into the medium for 24 hours respectively and 1× 10-4 mol/L H2O2 treated the cells for 1 hour to induce the apoptosis in experimental groups.The cells treated by only 1 × 10-4 mol/L H2O2 were used as apoptotic control group,and the cells cultured by regular method were used as blank control group.The cell viability in different groups was detected using MTT at 570 nm ( A570 ),and the apoptotic rates of the cells were assayed using Annexin V/PI double staining.This use and maintain of experimental animals followed the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The cultured cells showed the irregular polygon in shape,and about 80％ cells was positive response for cytokeratin.MTT assay showed that the lower A570 values were detected in the H2O2-induced group,various concentrations of estradiol or testosterone groups compared with blank control group (P＜0.01 ).The A570 values in 1 × 10-5,1 × 10-6,1 × 10-7 mol/L estradiol groups or 1 × 10-6 mol/L testosterone group were significantly higher than ones of H2 O2-induced group (P＜0.01 ).Compared with corresponding concentrations of testosterone groups,the A570values in various concentrations of estradiol groups were elevated( P＜0.01 ).The apoptosis rates at the early and later phase were significantly declined in both estradiol group and testosterone group in comparison with H2 O2-induced group (P ＜ 0.01,P＜ 0.05 ),and those in estradiol group were lower than the testosterone group( P＜0.01,P＜0.05 ).Conclusions Estradiol and testosterone suppress the apoptosis of lacrimal gland cells induced by H2O2,and the stronger effect is found in estrogen.The inhibition of estrogen on lacrimal gland cell apoptosis show a dose-dependent manner to some extent.

Objective To evaluate the risk of plague occurrence via surveying and analyzing indoor rat density and flea index in natural villages having previous plague experience. Methods During August to September 2007, 30 natural villages experiencing previous plague were selected based on the surveillance data, and then all households were coded with numbers and 20 households in each village were randomly selected via computer. Cages and sticky papers were set in 600 selected households to capture rats and fleas. Rat density, flea prevalence, flea index and median were estimated. Results One hundred thirty-three Rattus flavipectus and 33 Suncus murinus were caught and averaged rat density was 2.8 rats per one hundred cage. nights (166/6000), the median was 5 rats each village. One hundred and one mice infected fleas, flea prevalence on rats was 60.8% (101/166), 296 Xenopsylla cheopis and 48 Leptopsylla segnis were collected. Rat flea index was 2.1 fleas per rat (344/166). A total of 315 dissociated flea was caught, average dissociated flea index was 0.026 fleas per sticky paper (315/11888). The median was 5.5 dissociated fleas per village. Of dissociated fleas, Ctenocephalides felis felis (205) and Xenopsylla cheopis (103) accounted for 97.8% (308/315). The proportion for species of the rat flea and the dissociated flea was different(Fisher test: P < 0.01). The rat flea was significantly associated with the rat density(r = 0.68, P < 0.01), but the dissociated flea was significantly associated with neither the rat density(r = -yield than fried wheat batter(χ2 = 5.59, P < 0.05). Conclusions In these villages having previous plague experience of Lianghe County, Rattusflavipectus was dominant species of indoor rats, Xenopsylla cheopis and Ctenocephalides felis felis were dominant species of rat flea and dissociated flea, respectively. Mengsong, Bangdu, and Tangjiatun village had potential risk of plague emergence.

To synthesize and evaluate antimicrobial activity of novel heterocyclic compounds, the corresponding title aroyl ureas have been synthesized by the reaction of 2-amino-5-(3-pyridyl)-1, 3, 4-thiadiazole with aroyl isocyanates. Their antimicrobial activities in vitro were tested by disk diffusion methods and broth microdilution according to M-27A protocol recommended by NCCLS. Twelve new compounds were obtained, and their structures were confirmed by MS, IR, 1H NMR and elemental analysis. The biological screening tests showed that most of the compounds have some antifungal activities in vitro. Aroyl ureas incorporating pyridyl thiadiazole ring may be developed as novel antifungal candidate drugs and are worthwhile to be further studied.
Antifungal Agents ; chemical synthesis ; chemistry ; pharmacology ; Candida albicans ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Phenylurea Compounds ; chemical synthesis ; chemistry ; pharmacology ; Thiadiazoles ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo investigate the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

Objective To explore the predictors on the abundance of Rattus (R.) tanezumi in households of commensal rodent plague foci.Methods Thirty natural villages that experienced previous plague cases in Lianghe county,Yunnan province,were selected followed by random selection of 20 households in each village through computer technique.Live traps were set in households to capture small mammals which were then identified to species in the field according to their morphological features.Data on potential factors for abundance of R.tanezumi were collected through questionnaires and field observation and were coded and computerized using EpiData software and further analyzed by hurdle regression model under R software.Results A total of 166 rodents (133 R.tanezumi and 33 Suucus murinus) were captured.Results from final multilevel hurdle regression model showed that the likelihood of R.tanezumi captures increased by 1.67-to 2.76-fold in households belonged to Dai ethnic families that stored foodstuff in metal pails,often raising dogs,and having adjacent houses.The number of R.tanezumi captures increased by 2.18-fold in the villages where over 80％ of the households would raise chickens.In the villages with communal latrine,the likelihood and the number of R.tanezumi capture increased 1.93-fold and 2.38-fold,respectively.While the likelihood of R.tanezumi captures would reduce by 45％-61％ in those households where there were cats and cattle being raised and maize grown in the village.The number of R.tanezumi captures would reduce by 63％ in the households where there were outside toilets.Conclusion The abundance of R.tanczumi seemed to be closely related to the ecological environment factors.Programs on plague control and prevention should relate to ecological factors that influencing the abundance of R.tanezumi.