1. Mechanism of delayed encephalopathy in acute carbon monoxide poisoning rats and mouse nerve growth factor in early intervention
Chinese Pharmaceutical Journal 2013;48(24):2123-2127
OBJECTIVE: To observe the mouse nerve growth factor on early intervention in rats with acute carbon monoxide intoxication delayed encephalopathy control effects, and preliminarily discuss the pathogenesis of delayed encephalopathy. METHODS: Ninety-two weight only 180-230 g, male Wistar rats were randomly divided into two experimental groups (n=40), and a healthy control group (n=12), namely the CO poisoning group (COP) of the group; mouse nerve growth factor (NGF plays group); healthy controls (NC group). Experimental group was according to 150 mL·kg-1 intraperitoneal injections of CO in the preparation of animal model of acute carbon monoxide poisoning, in the control group rats were injected with equal volume of air. NGF group rats were injected within 30 min after poisoning muscle injection of mouse nerve growth factor, once a day, every time 18 μg·kg-1, total 7 days; Control group and the COP were given intramuscular isodose physiological saline. NGF group rats were treated with 18 μg·kg-1·d-1 NGF (total 7 times); NC group and COP group treated with the same dose saline. 90 min, 7 day, 14 day and 21 day groups of rats serum myelin basic protein (MBP) levels were monitored. The routine preparation of brain tissue frozen pathology slice, line HE and MBP immunohistochemical staining. Animal intelligence is assessed by Morris water maze experiment, in the 120 s in rats to determine correctly through the platform number less than 3 times DEACMP. RESULTS: All rats showed typical toxic after CO poisoning. Experimental group rats died 29, including COP death cases in the group, the model NGF group of 15, 14, mortality rates were 37.50%, 35.00% respectively. Morris water maze experiment results showed that the COP in the group 8 rats were judged to be delayed encephalopathy; model NGF group has 5 only be judged to be delayed encephalopathy; COP group rats serum MBP content increased significantly, model NGF group close to normal.Differences were obvious in poisoning after 7 days, 14 days.Pathological examination showed the COP group delayed encephalopathy rats in poisoning 90 min-21 d in hippocampus, subcortical neurons damage, such as myelin basic protein depigmentation pathological changes.The above pathological changes can be observed in each group, the COP group rats lesion severity is heavy, model NGF group was lighter. CONCLUSION: Mouse nerve growth factor can significantly reduce the incidence of delayed encephalopathy in rats, but can't reduce the mortality. Myelin basic protein (MBP) in the pathogenesis of delayed encephalopathy may play an important role.
2.Construction of serine protease gene mutant and study on pathogenicity of serine protease gene of Entero-coccus faecalis
Li-Xian WU ; Wen HUANG ; Sun XIANG ; Ping XIAO ;
Chinese Journal of Infectious Diseases 2000;0(02):-
Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.
3.Construction of a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum and expression in Escherichia coli BL21 (DE3)
Li, ZHANG ; Wen-gui, LI ; Jin-ping, XIANG
Chinese Journal of Endemiology 2013;32(6):608-611
Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.
4.Effect of Daidzein intravitreal injection on optic nerve injury in rats
Zhong-Jun, TANG ; Zhen-Ping, HUANG ; Wen-Jing, YANG ; Yong-Xiang, ZOU ; Ji-Ping, CAI
International Eye Science 2014;(8):1378-1381
AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages.
METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups.
RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.
5.Screening of High Daptomycin-producing Strain by He-Ne Laser Irradiation and Streptomycin Resistance Screening Method
Wen-Yu LU ; Jian-Ping WEN ; Jing-Hua FAN ; Bo-Xiang CAO ; Bing SUN ;
Microbiology 1992;0(03):-
The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.
6.Construction of Hexose Transporter-like HXT1 Deletion Mutant in Pichia pastoris
Wen-Wen ZHANG ; Ping ZHANG ; Yao-Ji XUAN ; Xiang-Shan ZHOU ; Yuan-Xing ZHANG ;
Microbiology 2008;0(09):-
Glucose was transported by the large number of hexose transporters in yeast cells. There were 18 hexose transporter genes had been identified in Saccharomyces cerevisiae. However,as an excellent expression system,there was no information of these genes had been reported in Pichia pastoris. Based on high homologous recombination efficiency in yeast,we chose G418 resistance for screening,200 bp were cloned from the up and down sequences of HXT1 ORF respectively,then ligated to the 5′ and 3′ end of G418 resis-tance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates,finally we obtained one HXT1 gene deletion mutant named GS115?HXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.
7.Survey of defluoridation water improvement project in Shanxi province from 2005 to 2009
San-xiang, WANG ; Jun, LI ; Zheng-hui, WANG ; Qing-zhen, JIA ; Xiang-dong, ZHANG ; Xiao-tian, CHENG ; Xin-ping, WEN
Chinese Journal of Endemiology 2011;30(1):56-59
Objective To learn the present status of defluoridation water improvement project in Shanxi province in order to provide scientific basis for speeding up the prevention and control of endemic fluorosis.Methods According to "The National Technical Scheme for Endemic Disease Control" from 2005 to 2009, the investigation points were selected in the counties that implemented the measures of water improvement and defluoridation,the status of drinking water defluoridation Project was investigated, and the water fluoride levels were determined by fluoride selective ion electrode. Results The primary status was surveyed in 1658 water improvement and defluoridation projects in 51 counties. The resource of drinking water for water improvement and defluoridation projects was mostly ground water[accounting for 93.12% (1544/1658)]. Among 1658 water improvement and defluoridation projects 1405 projects worked well(accounting for 84.74%) and 190 projects intermittently worked (accounting for 11.46%). Sixty three projects abandoned (accounting for 3.80%), in Datong basin the abandoned projects accounted for 36.36% (12/33). Water fluoride content of 1595 water improvement and defluoridation projects had been determined, among them water fluoride content of 874 projects were above 1.0 mg/L (accounting for 54.80%). The situations of exceeded national standard in the five basins was different(H = 33.22,P < 0.01). The rate of over national standard of fluoride levels in drinking water was 88.37%(38/43) in Datong basin. Therefore, in Datong basin water improvement should be strengthened. Conclusions In Shanxi province the water improvement and defluoridation projects are basically running normally. However, the qualified rate is lower for the water improvement and defluoridation projects. The water improvement status varies dramatically among areas.The situation is still grim in Shanxi province. Water improvement and defluoridation needs to be strengthened to improve the effect of prevention and control of the disease.
8.Studies on the Capacity of Phosphate Dissolving and the Identification of Strain BL-11
Chun-Mei DU ; Shu-Chao JIN ; Wei WANG ; Wen-Xiang PING ; Hui YANG ;
Microbiology 1992;0(02):-
The morphology,cultural characteristics,physiological and biochemical properties of phosphate solubilizing strain BL-11 were studied.Combine with its 16S rDNA sequence analysis result strain BL-11 was identified as Brevibacillus laterosporus.Results showed that the phosphate solubilizing efficiency of strain BL-11was up to 10.91% when with Ca_ 3 (PO_ 3 )_ 2 as the sole phosphorus source in the medium,while the efficiency of that was up to 1.56% when with sands as the sole phosphorus source.An optimal media of BL-11 strain for solubilizing phosphate was obtained by orthogonal test.It was composed of sugar 20g/L,(NH_ 4 )_ 2 HCO_ 3 0.3g/L,MgSO_ 4 .7H_ 2 O 0.5g/L,NaCl 0.3{g/L},KCl 0.5g/L,FeSO_ 4 0.03g/L,MnSO_ 4 .H_ 2 O 0.03{g/L.}_ While other conditions were consisted of initial pH7.0-8.0,180 r/min,and 30℃.
9.Isolation,Screening and Identification of Antagonistic Microorganisms Against Magnaporthe grisea
Min-Xia WANG ; Ying-Wen XU ; Ping-Zhong CAI ; Yue-Wu XIANG ; Guang-Jun REN ;
Microbiology 1992;0(03):-
Three hundred and twenty-one bacteria strains were obtained from rice leaves,stem,root tissue and paddy field soil,of which the number of strains which can inhibit mycelium of Magnaporthe grisea growth markedly was fifty-seven through fermentation in 2.0 mL Eppendorf tube,and among these fifty-seven strains,five strains were strongly antagonistic to Magnaporthe grisea.These five strains was identified for their morphologic,physiological and biochemical characteristics,and the results showed that one strain(No.156)was bacillus subtilis,two strains(No.171 and No.177)were Bacillus pumillus and two strains(No.192 and No.279)were Bacillus ploymyxa.
10.Surveillance of bacterial resistance from a hospital in Chongqing from 2004 to 2005
Chao ZHUO ; Bei JIA ; Wen-Xiang HUANG ; Xing-Ping ZHENG ; Chong-Zhi LI ;
Chinese Journal of Infection and Chemotherapy 2007;0(05):-
Objective To investigate the antimicrobial resistance of clinical isolates from a hospital in Chongqing during one year according to CHINET project.Methods Disc diffusion test (K-B method) was employed to study the antimicrobial resistance. WHONET5 was used for data analysis.Results In one year period from 2004 to 2005,690 non-duplicate isolates were collect- ed.Enterobacter isolates showed the lowest resistance rate to imipenem and meropenem.About 37.5% of E.coli and 31.4% of K.pneumoniae isolates produced ESBLs,respectively.All ESBLs-producing strains were susceptible to imipenem and mer- openem.About 37.2%,39.4% and 48.9% of P.aeruginosa isolates were resistant to imipenem,meropenem and ceftazidime, respectively.Pandrug-resistant (PDR) P.aeruginosa was isolated from our hospital.All strains of A.baumannii were sus- ceptible to imipenem and meropenem.About 37.7% of A.baumannii were resistant to cefoperazone-sulbactam.Twenty-nine strains showed the same resistant pattern among non-susceptible strains of A.baumannii,mainly derived from 2 clones by PFGE analysis.Conclusions The surveillance results suggest that prevalent strain resistant to cefoperazone-sulbactam may pres- ent in some ICUs.Resistance rate to cefoperazone-sulbactam increased significantly.