2.Large Protein Production Through Split Intein-Mediated Trans-Splicing
Jing ZHANG ; Huan LIU ; Jing ZHOU ; Jian-Hua LIU ;
China Biotechnology 2006;0(12):-
Npu DnaE intein was used to produce some large proteins,which were difficult to obtain through conventional expression systems.A T7 expression system was described,by which the gene of T7 RNA polymerase is split into two pieces,and each piece fuses with Npu DnaE N-and C-terminal sequences respectively.Functional T7 RNA polymerase is created by mixing the two kinds of fusion constructs in vitro.The approach of split intein-mediated production of large proteins,in theory,readily generalizable to the purification of other large,cytotoxic or membrane proteins.
3.The relationship between cystatin C and glomerular filtration rate in creatinine-blind range elder patients.
Jian LIU ; Yonglun HANG ; Jing HOU
Chinese Journal of Practical Internal Medicine 2002;0(08):-
Objective To explore the relationship bewteen serum cystatin C (Cys-C )and glomerular filtration rate(GFR) in creatinine-blind range elderly patients. Methods From Jan.2002 to Dec.2003,in the Affiliated Hospital of Luzhou Medical College,the study was performed in 86 elderly patients with chronic renal disease.Serum Cys-C was determined by particle-enhanced immunoturbidimetry.Serum creatinine(Scr) was determined by auto-analyzer.Creatinine clearance rate(Ccr) was estimated by CockCroft-Cault formula.For statistical evaluation,linear regression analysis was used. Results Concentration of Cys-C increased in 56 patients(56.1%).When Ccr was higher than 59mL/min,concentration of Cys-C increased in 44.2% patients.Significant positive correlation was observed between Cys-C and Scr( r =0.89, P
4.Establishment of composite animal model of chronic periodontitis and IgA nephropathy, and the interrelation
Jing LI ; Jing ZHANG ; Jian LIU ; Xiaohong SANG ; Lina WANG
Chinese Journal of Nephrology 2013;29(11):842-848
Objective To analyse the relationship and mechanism between chronic periodontitis (CP) and IgA nephropathy (IgAN) by establishing animal model of chronic periodontitis and IgA nephropathy in SD rats.Methods Eighty health male SD rats were divided into four groups,control group (A,n=20),IgAN group (B,n=20),CP group (C,n=20),CP accompanied with IgAN group (D,n=20).CP model was established by ligating silk suture and besmeared pathogenic bacterium in rats dental cervix.Experimental IgAN model was established by lavage of bovine serum albumin (BSA) and injection of lipopolysaccharide (LPS) and carbon tetrachloride (CCl4).Ten rats were sacrificed in every group at the end of week 8 and 12.The blood,urine,kidney tissue samples were examined.The observation index included proteinuria,kidney and liver function,renal tissue pathology and periodontal tissue pathology.The data were statistically analysed.Results Animal models were established successfully.The levels of Scr and BUN in group A,B,C were not obvious difference,but that in group D was higher than the other third groups at 8 weeks (P < 0.05).The levels of Scr and BUN in group D and group B were significantly higher than that in group A,meanwhile that in group D was significantly higher than that in group B at 12 weeks(P < 0.05).The levels of 24 h urine protein in B,C,D groups were higher than that in group A at 8 weeks(P < 0.05),but at 12 week,that in group D was higher than that in group B and C (P < 0.05).At 8 weeks,glomeruli had little mesangial broadening and renal interstitium had mild hyperplasia of fibrous tissue in group B and D,and that in the group D significantly was heavier than that in group B.The focal varying degrees were glomerular mesangial proliferation hardening,glomerular mesangial broadening,focal segmental sclerosis,and diffuse or focal inflammatory cell infiltration in D group at the end of week 12.The score of PAS between group A and group C had no statistical significance.The scores of PAS in group B and D were higher than that in group A (P < 0.01),and that in group D was higher than that in group B (P < 0.01).Obvious inflammation of periodontal tissue was observed in group C and D,and modified sulcus bleeding index (MBI) were higher than that in group A and B,and MBI in group D was significantly higher than that in group C (P < 0.01).Conclusions The model can be used to research the change of biochemistry and pathology and observe the relationship between chronic periodontitis and IgAN.This study shows that there is relationship between chronic periodontitis and IgAN.Chronic periodontitis maybe make more serious pathology damage in kidney by inflammation mechanism.
5.Studies on purification and characterization of a protease from a marine bacterium, Pseudomons sp. 7~11
Qinghai LIU ; Gu JING ; Jing FENG ; Jian KONG ; Guirong MA ;
Chinese Journal of Marine Drugs 1994;0(03):-
A protease from the culture supernatant of marine bacteria Pseudomonas sp. 7~11 was discovered. One component of the protease was purified to homogeneity by ammonium sulfate precipitation, dialysis, ion exchange chromatography. The specific activity of the enzyme was raised from 19.46U?mg -1 to 13953115U?mg -1 ,which was 717.02 times that of the culture supernatant with a yield of 1.24%.The molecular mass of purified enzyme was estimated to be 3000Da about by SDS PAGE. The optimum temperature for the activity was observed to be 45℃ using casein as substrate.The optimum pH for activity the enzyme was 8.0 and it was stable between pH6~7 and below 50℃.The activity of the enzyme was inhibited by EDTA and IAA strongly. But was not inhibited by PMSF. the enzyme was inhibited Mn 2+ , Hg 2+ , Fe 2+ , Zn 2+ , Cu 2+ ,Pb 2+ and SDS. The inactivity of the enzyme with EDTA could be recovered partially by Mg 2+ .while Ca 2+ , Mg 2+ and (NH 4) 2SO 4 was activator for the activity of the enzyme. The activity of the enzyme was fairly stable in the presence of ethanol and urea, and was resistant to Tween 20.
6.Impact of overweight and obesity on flow mediated dilation in adult
Jing ZHAO ; Xi CHU ; Jing DONG ; Youqin LIU ; Jian DONG
Chinese Journal of Primary Medicine and Pharmacy 2017;24(11):1658-1661
Objective To explore the impact of overweight and obesity on flow mediated dilation (FMD) in adult people.Methods Intact clinical data of 183 healthy people, aged 30~65 in our hosipital were analyzed.According to BMI, all people were divided into obesity group (48 cases), overweight group(75 cases) and normal group (60 cases).FMD was detected by E tracing of ultrasonic.Weight, WC, BFP, baPWV, BP,FPG,CHO,HDL-C,LDL-C,TG and UC were observed at the same time.FMD of three groups were compared and the risk factors of FMD were analyzed.Results The level of FMD in the obesity group and overweight group was significantly higher than that in the normal group[(5.63±2.76)%,(6.45±2.54)% vs.(8.31±3.04)%](F=13.77,P<0.001);there was no statistically significant difference between the obesity group and overweight group (t=1.68,P>0.05).Age(β=0.078,95%CI: 1.004,1.164),BMI(β=0.465,95%CI 1.148,2.208),LDL-C(β=2.294,95%CI: 9.918,1.612) and UC(β=0.007,95%CI 0.986,1.000) were independent risk factors of decreased FMD(P<0.05).Conclusion Age, BMI, LDL-C and UC are independent risk factors of decreased FMD.
7.Influences of antenatal administration of taurine on cerebral neurogenesis of fetal rats with intrauterine growth restriction
Chen HUI ; Li JIAN ; Liu JING ; Liu LI ; Liu NA
Chinese Journal of Perinatal Medicine 2011;14(10):612-617
Objective To provide the experimental evidence for clinical application of taurine,rat model of intrauterine growth restriction (FGR) was made to investigate influence of prenatal administration of taurine on neurogenesis.Methods Fifteen pregnant rats were divided into control,FGR model and taurine groups,5 rats for each group.Rats in the control group were supplied with unlimited food and drink while the other two groups were fed by 40% food intake of the control group throughout pregnancy.Since gestational day 12,taurine (100 mg/kg) was added into diet of taurine group every day until term delivery.Brain tissues were obtained immediately after baby rats were born.Expression of proliferating cell nuclear antigen (PCNA),neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) of brain tissue was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry; meanwhile,numbers of PCNA-immunoreactive cells in subventricular zone,subgranular zone and cerebral cortex were compared with ANOVA test or q test.Results Levels of PCNA mRNA and GFAP mRNA expression in FGR group were significantly higher than those of control group (PCNA mRNA:1.002±0.011 vs 0.894 ± 0.040,P<0.01; GFAP-mRNA:1.012±0.013 vs 0.913±0.012,P<0.01).Compared to FGR model group,mRNA expressions of PCNA and GFAP in taurine groups were higher (1.090±0.029,P<0.01 ; 1.034±0.011,P>0.05).There was a significant decrease in the expression of NSE mRNA in FGR group compared with control group (0.796±0.036 vs 1.582±0.057,P<0.01),while the expression in taurine group (0.933±0.027) was significantly higher than that in FGR model group (P < 0.01).PCNA immunoreactive cells were mostly distributed in subventricular zone,followed by subgranular zone and cerebral cortex.Conclusions Prenatal application of taurine could enhance neurogenesis of FGR newborn rats and improve survival of neurons to ameliorate FGR brain damage.
8.Therapeutic effect of Turkish galls extract on IgA nephropathy model rats
Jing LI ; Yanjie LIU ; Xiaohong SANG ; Ting WANG ; Jian LIU
Chinese Journal of Pharmacology and Toxicology 2017;31(6):547-552
OBJECTIVE To investigate the effect of Turkish galls extract (TGE) on the expression of IgA in serum,urine and renal tissue of IgA nephropathy (IgAN) model rats.METHODS Fifty healthy male Sprague Dawley rats were randomly divided into normal control group,IgAN model group,and TGE 75,150 and 300 mg· kg-1 groups,10 rats per group.The model of IgAN rats was established with bovine serum albumin (BSA)+lipopolysaccharide (LPS)+carbon tetrachlorid (CCl4)for 12 weeks.From the 13th week,TGE was ig administrated once a day for 4 weeks.At the end of the 12th and 16th weeks,24 h urine protein was measured by BCA method.At the end of the 16th week,serum and urinary IgA levels were measured by enzyme linked immunosorbent assay(ELISA),serum creatinine(SCR) and blood urea nitrogen (BUN) were detected by an automatic biochemical analyzer,and the renal pathological changes were evaluated with an Oxford classification scoring system.The deposition of IgA immune complex in the kidney was observed by immunofluorescence assay.RESULTS At the end of 12th week,24 h urine protein increased in all IgAN groups (P<0.05),compared with normal control group.At the end of 16th week,24 h urine protein,IgA content in serum and urine,SCr and BUN content in serum,score in Oxford classification of renal tissue and deposition of IgA immune complex in the kidney in IgAN model group were all higher than in normal control group (P<0.05).Compared with IgAN model group,24 h urine protein,IgA content in serum and urine and SCr content in serum were decreased in all TGE groups (P<0.05),and BUN content in serum and deposition of IgA immune complex in the kidney decreased in TGE 150 and 300 mg·kg-1 groups (P<0.05).The score in Oxford classification of renal tissue was decreased in TGE 300 mg· kg-1 group only.CONCLUSION TGE has curative effect on IgAN model rats by reducing serum and urinary IgA and decreasing IgA immune complex deposition.
9.An experimental study of BALB/c mice immunized by vaccine gene pcDNA3- PAc against dental caries through different routes
Zhaohui YUE ; Jianguo LIU ; Jing WANG ; Jing ZHAO ; Wei WANG ; Jing JIANG ; Xiaoyan GUAN ; Jian ZHANG
Journal of Practical Stomatology 2010;26(2):145-148
Objective:To observe the anticaries effects of the vaccine gene pcDNA3- PAc against dental caries in BALB/c mice by different routes. Methods: BALB/c mice were immunized with the recombinant plasmid pcDNA3- PAc by the submandibular gland- target injection,the quadriceps femoris injection and the intranasal irrigation respectively. All the mice were immunized two times. The immune interval was two weeks. Saliva and serum samples were collected respectively at 0, 1, 2, 4 weeks after immunization. The specific antibodies were detected by ELISA. Results:The specific antibodies were up at one week after immunization in different routes. The peak time of the antibodies level appeared at 4 weeks.The level of salivary specific anti- PAc IgA induced by submandibular gland- target injection and that of serum IgG induced by thigh bone muscle injection was the highest, respectively. The differences of antibodies level between in experiment groups and negative control group or vacant comparison group were significant(P<0.01). Conclusion: The vaccine gene pcDNA3- PAc effectively induce local mucosal immune response and systemic immune response.