2.Optimization on Fermentation and Purification of the Fibrinolytic Enzyme from Stenotrophomonas maltophilia
Gao-Xue WANG ; Chao-Jun LIANG ; Hai-Hong HUANG ; Jian-Fu WANG ; Ju-Lin YUAN ;
China Biotechnology 2006;0(09):-
The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia(DR-929) was investigated.The results showed the best fermentation are amidulin 2.0%,soya flour 1.0%,yeast extract 0.5%,NaCl 1.0%,CaCl2 0.02%,MgSO4 0.05%,inoculum of 36 hours,fermental time 4d,initial pH 8.0 or 9.0,temperature 25℃,volume of media 30ml,volume of inoculum 5% or 6%.The purification process includes the following steps: removing cells by the centrifugation,25%~70% saturation ammonium sulfate precipitation,HIC with Phenyl FF(high sub),IEC with Q-Sepharose FF,gel filtration chromatography with Superdex 75.SDS-PAGE electrophoresis was used to examine the purification effect,and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa.The purification factor and activity recovery of the fibrinolytic enzyme are 271.5 and 24.5%,respectively.
3.Acute toxicity of meso-2,3-dimercaptosuccinic acid on mice
Shi-Jun LIU ; Xue-Hai JU ; Ke-Ming SUN ; Zhi-Jie CHU ; Xian-Yong LIU ; Min ZHANG ; Jun-Qing GUO ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To explore acute toxicity of succimer on mice.Methods Twenty Kunming mice(10 males and 10 females) weighting approximately (21.2?2.3)g were acclimatized for 3 days prior to dosing,then were divided into control group and experiment group with 10 mice in each group according to body weight.Fasted for 12 hours,the mice in experiment group received intragastric administration of 160mg DMSA in deionized water in 24 hours,and the control group received the same volume of deionized water,and then they were observed for 7 days.Blood was collected into heparinized-tubes by removal of eyeball.All mice were sacrificed and brain,heart,liver and kidney were removed and washed with normal saline.The activity or amount of BUN,Scr,AST,ALT,SOD, GSH-PX and MDA were analyzed.Results (1)Given 160rag DMSA in 24 hours,gastrointestinal symptoms were main side effects.During the observation,experiment group lost weight due to the decrease of food-intake ,and some mice had slight hydroabdomen.(2)High dose of DMSA caused a significant inhibition of GSH-PX(P0.05).The hepatic cell was damaged accord- ing to the significant raise of MDA in liver(P0.05),which was related to acute toxicity on liver.Conclusion Succimer could inhibit the antioxidarrt systems and could do damage to liver and kidney.
4.Expression of EIIIA+ fibronectin in incised wound of rat's skin.
Meng HE ; Hai ZHAO ; Yi-wen SHEN ; Ai-min XUE ; Long CHEN ; Yun-ju GU ; Zi-qin ZHAO
Journal of Forensic Medicine 2014;30(6):409-412
OBJECTIVE:
To explore the relationship between the expression of EIIIA+ fibronectin in incised wound of rat's skin and injury time.
METHODS:
The wounding model was established by cutting the dorsal skin of 48 adult SD rats. The rats were sacrificed at the pre-set injury time as immediately, 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, and 8 h. The skin samples were taken at the margin of wound. The expression of the EIIIA? fibronectin was detected by immunohistochemistry and Western blotting and the relationship be- tween its expression and injury time was observed. Results The expression of EIIIA+ fibronectin was not observed immediately. The basal cell of skin began to show positive expression 0.5 h after injury. With the extension of injury time, positive staining became stronger. The value of relative optical density was gradually increased with prolonged injury time by the Western blotting analysis.
CONCLUSION
The expression of EIIIA+ fibronectin could be used for estimation of injury time in the early stage of skin injury.
Animals
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Fibronectins/metabolism*
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Immunohistochemistry
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Proteins/metabolism*
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Rats
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Rats, Sprague-Dawley
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Skin/metabolism*
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Staining and Labeling
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Time Factors
5.Effects of cervical mild hypothermia auxillary therapy on stress reaction in patients with cervical spinal cord injury
Xue-Mei PAN ; Hai-Zhang DENG ; Xue-Mei ZHANG ; Li-Ju JIN
Chinese Journal of Modern Nursing 2009;15(13):1201-1204
Objective To investigate the effcts of cervical mild hypothermia auxiliary therapy on stress reaction in patients with cervical spinal cord injury. Methods 84 cases cervical spinal cord injury patients were randomly divided into two groups:a cervical mild hypothermia group (42 cases)and a routine treatment group (42 cases), 64 healthy men who come for health examination were recruited and served as healthy control. Routine treatment were used in both treatment groups, in edition, the LM-HAT was just applied in mild hypothermia group. The changes of the concentrations of corticotropin releasing hormone (CRH), adrenocorticotropin hormone (ACTH), corticosteroid (Cor), arginine vasopessin (AVP) in serum and the symptoms of the stress dsease(hypertension, high blood sugar)were observed before treatment and 1 week after treatment. Results The concentrations of CRH, ACTH, Cor, AVP in serum and the symptoms of stress disease in mild hypothermia group and routine treatment group were improved significantly at week after treatment(P<0.05), but the therapeutic effects of mild hypothermia group were better than that of routine treatment group (P<0.05). Conclusions The cervical mild hypothermia auxiliary therapy could significantly inhibit the secretion of stress hormones, make the H-P-A axis stable and promote the stress disease recovery.
6.Formula Optimization in Renshen Jianxin Capsule Based on Uniform Design and Anti-myocardial Ischemia Effect.
Chua-hua YANG ; Yun-lun LI ; Hai-qiang JIANG ; Lei NIE ; Jiang-qing JU ; Shuai LI ; Xue-yi DING ; Shi-jun ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(9):1105-1108
OBJECTIVETo realize quadratic formula optimization of Renshen Jianxin Capsule (RJC) by screening Chinese herbs with major anti-myocardial ischemia effect in RJC and optimize their optimal dosages.
METHODSBy following "uniform design-pharmacodynamic experiment-mathematical modeling-formula optimization", authors employed U10(10(8)) uniform design in the experiment. Eight Chinese herbs contained in RJC were taken as observatory factors. Electrocardiograph (ECG) changes of myocardial ischemia induced by isoproterenol were taken as pharmacodynamic indices. The mathematical model between herbal factors and pharmacodynamic indices was established using stepwise regression analysis to screen Chinese herbs with major anti-myocardial ischemia effect. Their optimal dosages were optimized using the grid algorithm.
RESULTSThe regression equation was y =1. 7889 -0. 3247 Ginseng xSalvia Miltiorrhiza -0. 0663 Astragalus membranaceus xOriental Waterplantain tuber. Forecasting factors included were Ginseng, Salvia Miltiorrhiza, Astragalus membranaceus, and Oriental Waterplantain tuber. The optimal formula dosage calculated by the grid algorithm was Ginseng 1. 62 g, Astragalus membranaceus 4. 62 g, Salvia Miltiorrhiza 2. 43 g, and Oriental Waterplantain tuber 1. 66 g.
CONCLUSIONUniform design combined with stepwise regression analysis and grid algorithm were able to realize quadratic formula optimization of RJC.
Astragalus membranaceus ; Chemistry, Pharmaceutical ; standards ; Coronary Artery Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Electrocardiography ; Humans ; Isoproterenol ; Myocardial Ischemia ; drug therapy ; Panax ; Salvia miltiorrhiza
7.Effects of mycophenolate mofetil, valsartan and their combined therapy on preventing podocyte loss in early stage of diabetic nephropathy in rats.
Yan ZHANG ; Bing CHEN ; Xiang-hua HOU ; Guang-ju GUAN ; Gang LIU ; Hai-ying LIU ; Xue-gang LI
Chinese Medical Journal 2007;120(11):988-995
BACKGROUNDPodocyte has inflammatory role in the development of diabetic nephropathy (DN). Mycophenolate mofetil (MMF), an anti-inflammatory agent, can suppress macrophage infiltration and reduce renal injury in streptozotocin-induced diabetic rats. Angiotensin II receptor blocker (ARB), another renal protecting agent, can decrease podocyte loss in DN. In this study, we detected the expression levels of monocyte chemoattractant protein-1 (MCP-1) and nephrin to evaluate podocyte's role in inflammatory reaction in DN, observe and compare the effect of MMF alone and in combination with valsartan, on preventing podocyte loss in streptozotocin (STZ) induced diabetic rats.
METHODSDiabetic model was constructed in uninephrectomized male Wistar rats by single peritoneal injection of STZ (65 mg/kg). The successfully induced diabetic rats were randomly divided into four groups: diabetes without treatment group (DM), valsartan treated group (DMV), MMF treated group (DMM), and combined therapy group (DMVM). Normal rats of the same sibling were chosen as control (NC). At the end of the 8th week, serum biochemistry, 24-hour urinary protein (UP) and the ratio of kidney weight/body weight (RWK/B) were measured. The rats were sacrificed for the observation of renal histomorphology through light and electron microscope. Nephrin, desmin and MCP-1 levels were detected by semi-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1.
RESULTSCompared with group NC, serum glucose level, 24-hour UP and RWK/B in group DM were significantly higher (P < 0.01), and the nephrin mRNA level in DM group was significantly lower (P < 0.05). The nephrin mRNA expression levels in group DMV, DMM and DMVM were all higher than that of DM group (P < 0.05) and no significant differences were found among the three treatment groups (P > 0.05). Treatment with MMF, valsartan or their combination could significantly decrease the 24-hour UP and RWK/B, and suppress glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. In diabetic rats, the high expressions of desmin and MCP-1 in kidney were suppressed by valsartan, MMF or their combination.
CONCLUSIONSPodocytes are involved in the inflammatory reaction of diabetic rats. MMF could suppress MCP-1 and desmin expression, enhance nephrin expression, and attenuate proteinuria in diabetic rats. The combined therapy of valsartan and MMF did not show any superiority over monotherapies on renal protection. MMF may have renoprotective effect in early stages of diabetic nephropathy through preventing podocytes loss and anti-inflammatory activity.
Animals ; Chemokine CCL2 ; analysis ; Desmin ; analysis ; Diabetic Nephropathies ; drug therapy ; pathology ; Drug Therapy, Combination ; Immunohistochemistry ; Male ; Membrane Proteins ; analysis ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; therapeutic use ; Podocytes ; drug effects ; pathology ; Rats ; Rats, Wistar ; Tetrazoles ; administration & dosage ; therapeutic use ; Valine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Valsartan
8.Baseline study on a stroke-related cohort in the rural community of Shanghai.
Gui-qing WANG ; Jiu-yi HUANG ; Ji-ping GUO ; Yi-feng CAO ; Yong-ju YANG ; Xue-hai YU
Chinese Journal of Epidemiology 2006;27(1):12-14
OBJECTIVETo present data from a baseline investigation on stroke-related cohort population in rural area of Shanghai.
METHODSA cross-sectional study was carried out in a cluster sampling population aged 40 years and over. General information and data on common risk factors in the population were gathered and cerebral vascular hemodynamic indexes were checked. Hemodynamic score was estimated according to single indexes by unified methods. 5335 persons who had met the inclusion criteria were enlisted in the study. Exposure level of risk factors, prevalence of stroke, and hemodynamic indexes were analyzed and distributional characteristics were described.
RESULTSExposure rate of hypertension, heart disease, diabetes, family history of hypertension, overweight or obesity in males were 31.74%, 6.09%, 1.16%, 3.22%, 17.64%, 29.68% and were 32.76%, 9.22%, 1.55%, 3.84%, 19.22%, 29.44% in female respectively. Standard prevalence of stroke was 1167.3/100000, which in male was significantly higher than that in female (P < 0.05). The change of cerebral vascular hemodynamic indexes was significantly associated with age. Hemodynamic score in 21.3% of the subjects was below 75 points.
CONCLUSIONSAmong population of 40 years old and over in rural areas, hypertension was the principal risk factor regarding the rate of stroke. Prevalence of stroke in males was significantly higher than that in females. Abnormal rate of hemodynamic score was about 20% in this population.
Adult ; Age Factors ; Aged ; Aged, 80 and over ; Brain ; blood supply ; China ; epidemiology ; Cohort Studies ; Cross-Sectional Studies ; Female ; Hemodynamics ; Humans ; Hypertension ; complications ; epidemiology ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Rural Health ; Sampling Studies ; Sex Factors ; Stroke ; epidemiology ; physiopathology
9.Evaluation of aminoglycoside resistance phenotypes and genotyping of acetyltransferase in Escherichia coli.
Hai-shen KONG ; Xue-fen LI ; Ju-fang WANG ; Min-jin WU ; Xiao CHEN ; Qing YANG
Journal of Zhejiang University. Medical sciences 2006;35(1):83-86
OBJECTIVETo investigate the prevalence of aminoglycoside resistance and genotyping of acetyltransferase in Escherichia coli.
METHODSResistance phenotypes to 12 antibiotics of 44 Escherichia coli isolates were analyzed using agar dilution method and 3 aminoglycoside resistance genes aac(3)-I, II and aac(6')-I were determined by PCR method.
RESULTSIn 44 clinical isolates, the occurrence of ESBLs was 45.45%, resistance rates were discrepant for amikacin (18.18%), gentamicin (56.82%) and tobramycin (61.36%), the prevalence of phenotype TG (tobramycin and gentamicin) indicative of aac(3)-II production and TGA (tobramycin, gentamicin and amikacin) indicative of aac(6')-I production were 36.36% and 18.18%, respectively. The most common aminoglycoside resistance genotype of acetyltransferase was aac(3)-II (52.27%) and aac(6')-I was lower (29.55%), but no aac(3)-I was detected.
CONCLUSIONAt least 2 acetyltransferase genes exist in this area i.e. aac(3)-II and aac(6')-I.
Acyltransferases ; genetics ; Amikacin ; pharmacology ; Aminoglycosides ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Escherichia coli ; enzymology ; genetics ; Genotype ; Gentamicins ; pharmacology ; Phenotype ; Tobramycin ; pharmacology
10.A cohort study on the relationship between cerebrovascular hemodynamic changing and risk of stroke.
Ji-ping GUO ; Jiu-yi HUANG ; Yang-tai GUAN ; Hai-ying CHEN ; Yong-ju YANG ; Yin-ruo TAO ; Yi-feng CAO ; Ying XU ; Xue-hai YU
Chinese Journal of Preventive Medicine 2013;47(4):323-327
OBJECTIVETo study the role of cerebrovascular hemodynamic indexes (CVHI) changing in stroke and to provide reference for stroke prevention and risk factor study.
METHODSFrom 2003 to 2004, participants aged 40 years and above in two communities in Fengxian district were recruited by cluster sampling. Risk factors of stroke and CVHI were investigated and checked during baseline investigation. A total of 10 565 individuals completed the survey and met the inclusion criterion. After baseline investigation, the cohort was followed up for stroke occurrence. Relative risk (RR) of CVHI and common risk factors were estimated by cohort study design.
RESULTSAge of the cohort was (56.2 ± 11.4) years. 4444 (42.1%) were males and 6121 (57.9%) were females. Total follow-up duration was 67 885.7 person-years. A total of 195 stroke cases occurred and incidence density of stroke was 287.2 per 100 000 person-years. Stroke incidence in exposure groups of hypertension, heart disease and alcohol drinking was 3.47% (108/3118), 2.96% (21/710) and 2.50% (47/1882), respectively. The incidence in corresponding non-exposure group was 1.17% (87/7448), 1.77% (174/9855) and 1.70% (148/8683) respectively. There was significant difference between 2 groups (χ(2) value was 62.72, 4.56 and 4.94, respectively, P < 0.05). Stroke incidence in CVHI score < 25, 25 - 49, 50 - 74 and ≥ 75 groups was 9.12% (59/647), 5.68% (44/775), 2.52% (39/1545) and 0.72% (53/7403)(χ(2)trend = 273.57, P < 0.05), respectively. Incidence of stroke in 40 - 49, 50 - 59, 60 - 69, ≥ 70 years age group was 0.22% (8/3565), 1.28% (43/3357), 2.71% (50/1848) and 5.88% (94/1600) (χ(2)trend = 181.48, P < 0.05), respectively. Multiple Cox regression analysis indicated that RR (95%CI) value of hypertension and cigarette smoking was 1.40(1.02 - 1.92) and 1.59(1.19 - 2.12), respectively when comparing with non-exposure group. RR (95%CI) value in CVHI score < 25, 25 - 49 and 50 - 74 points group were 6.15 (4.08 - 9.26), 4.55 (2.98 - 6.96) and 2.68 (1.75 - 4.09), respectively when comparing with the score ≥ 75 points group. RR (95%CI) value in age 50 - 59, 60 - 69 and ≥ 70 years group was 4.61 (2.16 - 9.82), 7.81 (3.67 - 16.60) and 13.49(6.44 - 28.24), respectively when comparing with below 40 years group.
CONCLUSIONCVHI score is the strong independent predictive factor and hypertension, cigarette smoking and age are the independent risk factors of stroke.
Aged ; Brain ; physiopathology ; Cohort Studies ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Risk Factors ; Stroke ; epidemiology ; etiology ; physiopathology