1.Inhibition of protein phosphatases induces apoptosis in neuroblastoma cells
Ruohan XIA ; Shengyuan LIU ; Jianzhi WANG
Chinese Journal of Neurology 2000;0(04):-
Objective To investigate the relationship between protein phosphatase inhibition, tau hyperphosphorylation and neuronal death seen in Alzheimer disease (AD). Methods Co culture of protein phosphatase inhibitor okadaic acid (OA) and neuroblastoma cells (SH SY5Y), by agarose gel electrophoresis to detect DNA fragmentation, and in situ hybridization by TdT mediated biotin labeled dNTP nick end labeling (TUNEL) to further detect the cell apoptosis. Results Incubation of SY5Y cells with 10 nmol/L OA for 24 or 48 hours led to the appearance of DNA fragmentation and a remarkable increase of positive cells from 2 16%?0 94% to 18 05%?3 57% ( P
2.Effect of ovariectomization on Alzheimer-like phosphorylation of Tau in hippocampus of rats
Qi ZHANG ; Maoqiong HU ; Xiaoyan DAI ; Yi LIU ; Jianzhi WANG
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To explore the effect of ovariectomization on Alzheimer-like phosphorylation of Tau in hippocampus of Sorague-Dawlery rats. METHODS: An animal model was developed using ovariectimized (OVX) rats, and the phosphorylation of Tau protein was measured by Western blot. RESULTS: The levels of phosphorylated Tau at PHF-1epitope were elevated in ovariectimized rat brain hippocampus 4 weeks and 8 weeks after ovariectomization, when compared with sham-OVX rats (P0.05). CONCLUSION: Ovariectomization may induce Alzheimer-like hyperphosphorylation of Tau protein in brain hippocampus of rats. [
3.Application of mandible and zygomatic arch measurement and related analysis in medical aesthetics
Xiaoheng ZENG ; Jianzhi LIU ; Sixin OUYANG ; Jianguo TAN ; Shenghua CHEN
Chinese Journal of Medical Aesthetics and Cosmetology 2001;0(03):-
Objective To provide the data of mandible and zygomatic arch, including the space distance of mandible angles, depth and length of zygomatic arch, by direct measurement and related analysis at home position of corpse in Chinese Han for medical aesthetics. Methods The vernier callipers were used to directly measure the every indexes of mandible and zygomatic arch in the formalin-fixed male and female adult corpses (50 each) and to proceed the related analysis. Results The results showed that male mandible thickness was (12.60?1.26) mm, mandible height (33.60?3.35) mm, mandible length (77.60?7.80) mm , width of mandible ramus (34.80?3.50) mm, height of mandible ramus (67.20?6.70) mm, depth of zygomatic arch (9.54?1.19) mm, length of zygomatic arch (40.60?4.97)mm, and thickness of mandible angle (12.20?0.67) mm in male; Mandible thickness was (10.70?0.98) mm, mandible height (29.40?2.64) mm, ramus of mandible height (74.00?7.07) mm, width of mandible (31.10?3.04) mm, height of mandible ramus (64.00?5.53) mm, depth of zygomatic arch (8.67?1.09) mm, l ength of zygomatic arch (36.50?4.59) mm, and thickness of mandible angle (13.80?1.28) mm in female. Conclusions The breadth between bilateral angle of mandible could come to reckoning the depth of zygomatic arch, the length of zygomatic arch and other indexes of mandibles. Those data are useful in aesthetics and preparation of the compound.
4.The possible mechanism of insulin reducing amyloid beta peptide in the brain cortex of diabetic rats
Zhongsen QU ; Yongbo ZHAO ; Wenwen LIU ; Jianzhi WANG
Chinese Journal of Neurology 1999;0(06):-
Objective To investigate the reduction of amyloid beta peptide (A?) production induced by insulin in the cortex of diabetic rat. Methods 15 male SD rats were randomly divided into control group, diabetes group, and diabetes plus insulin group. The diabetic rats were induced by streptozotocin. The activity of glycogen synthase kinase-3 (GSK-3) was measured by 32P liquid scintillography for incorporated radioactivity in control group, DM group, DM+insulin group, the production of A? was determined by sandwich ELISA in each group, and the expression of APP was determined by Western-blot. Results In control group, the activity of GSK-3 (1.04?0.11), the production of A? (A?_ 40 (40.92?5.34) pg/?l, A?_ 42 (29.64?3.19) pg/?l)and the levels of full-length APP(1.05?0.08) was low, but in DM group, the activity of GSK-3 (2.02?0.12) and the production of A?(A?_ 40 (67.53?11.69) pg/?l, A?_ 42 (45.02?4.10) pg/?l) increased (P
5.Disturbance of GSK-3 and PP-2A Induced tau Hyperphosphorylation in Diabetic Rat Brain
Zhongsen QU ; Yongbo ZHAO ; Wenwen LIU ; Jianzhi WANG
Progress in Biochemistry and Biophysics 2006;0(08):-
The changes of glycogen synthase kinase-3(GSK-3)and protein phosphatase-2A(PP-2A),and the role of them in the regulation of the abnormally hyperphosphorylated some sites of tau in the cortex of diabetic rat were investigated.The diabetic rat model was induced by streptozotocin.The activities of GSK-3,PP-2A were measured by liquid scintillation for incorporated radioactivity in control,DM,DM + Li2CO3 groups.The level of hyperphosphorylated tau and the expression of 2PP-2A was measured respectively by Western blot.It is suggested that GSK-3 activity increases,PP-2A activity and expression decrease,and hyperphosphorylated tau be produced at Ser198/Ser199/Ser202 and Ser396/Ser404 in DM rats cortex.After the DM rat were treated with Li2CO3,the inhibition of GSK-3 activity and the improvement of PP-2A activity were found,and hyperphosphorylation of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 were deduced.These studies firstly suggested that an increase of GSK-3 activity might inhibit PP-2A activity,and which produce hyperphosphorylated of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 in DM rat cortex in common.
6.Selection of peptide specifically binding to bladder carcinoma by using phage display in vivo
Junqian LUO ; Fan ZHANG ; Xiaofeng YANG ; Fang LUO ; Jiehao LIU ; Jianzhi PANG ; Sanhua YAN ; Xiaolei ZHANG
Chinese Journal of Immunology 2015;(4):509-513
Objective:To screen the peptide binding to human bladder carcinoma cells specifically by using phage display technology in vivo.Methods: Nude mice were inoculated with bladder carcinoma cells BIU87 for establishing tumor-bearing mice model.The Ph.D.-C7CTM Peptide Library was injected intravenously via tail vein.Then we screened Phage containing exogenous peptides binding to bladder transitional carcinoma cells specifically.The phage peptide homed to the tumor tissues was obtained after 3 rounds screening in vivo.The phage clones affinity to BIU87 were identified by immunohistochemistry and ELISA.The positive peptide was synthetized by chemical methods after sequencing the positive monoclonal phage DNA.The tumor cell specificity of target peptide was identified by confocal laser scanning microscope and flow cytometry.Results:After 3 rounds screening in vivo,enrichment rate of phage was 4.334×102 times.Immunohistochemistry results showed that the dyeing of the tumor tissue had a rising trend following each round of phage screening,while liver had a lot of non-specific binding phage because the phages were metabolized through liver and kid-ney.The 30 phage clones were identified by ELISA and 10 clones had a strong affinity on BIU87 among 24 positive clones.Three amino acid sequences of positive phage clones were obtained.The highest rate of repeat sequences CSSPIGRHC(8/10) named NYZL1 and the FITC-C6-NYZL1 peptide was synthesized.Our results showed that it could bind to bladder carcinoma cells BIU87 specifically.Conclusion:We obtained the small molecular peptide NYZL1 binding to human bladder carcinoma specifically by means of phage display in vivo,which provide a theoretical basis for bladder carcinoma early diagnosis and targeted therapy.
7.Experimental study on the characters of the FITC-CSNRDARRC molecular probe and its targeted ability for orthotopic transplanted tumor of bladder cancer in vivo
Sanhua YAN ; Jianzhi PANG ; Xiaofeng YANG ; Jiehao LIU ; Ziqiang ZHANG ; Fan ZHANG ; Junqian LUO
Chinese Journal of Urology 2015;36(9):699-704
Objective To investigate the characters and targeted ability of FITC-CSNRDARRC molecular probe in labeling orthotopic transplantation tumor of bladder cancer in vivo.Methods From July 2013 to June 2014,the stability and characters of FITC-CSNRDARRC molecular probe were detected by spectrophotometer and molecular imaging and the optimum concentration and imaging time window were determined.30 BALB-C nude mice were randomly divided into experimental group (n =20) and control group(n =10).In control group,5 of them (group A) were ligated bilateral ureter,others(group B) were not.We established orthotropic transplanted bladder tumor (BIU-87) model by operation.And 0.2 ml probes (220 μmol/L) was then injected intravenously in all mice after 2 weeks.We obtained images and analyzed average gray value of the heart,lung,liver,spleen,bilateral kidney and orthotropic transplantation bladder tumor by using optical probe molecule fluorescence imaging system after 30 min,1 h,2 h,4 h and 12 h,respectively.Results After injected the FITC-CSNRDARRC molecular probes intravenously at 220 μmol/L,the fluorescence signal of tumor tissue strengthened gradually.The optimal imaging time window was 4 hours after injection.The illumination and temperature had little effect on the fluorescence signal.With the time passing after injection,the intensity of florescence signal progressively increasing,which reached the peak at4 h.The average gray value of tumor tissue at 1 h,2 h,3 h,4 h,5 h,6 h,8 h,12 h were 74.22,76.2,80.11,89.38,83.29,85.1,81.22,83.01,respectively.The fluorescence signal of normal tissue weakened gradually with the passage of time.Only liver and gall bladder could notice the fluorescence signal 4 hours after injection in group A.However,the relatively strong fluorescence signal could be found in liver and gall bladder in group B.Conclusions The characters of fluorescence probe are affected by its concentration.Its optimal concentration of labeling tumor is 220 μmol/L.The optical imaging time window was about 4 h after intravenous injection.The FITC-CSNRDARRC molecular probe can specifically bound to orthotopic transplanted tumor of bladder cancer in vivo.
8.Effects of EGF and bFGF on expression of microtubule-associated protein tau and MAP-2 mRNA of mononuclear cells derived from human umbilical cord blood
Wenhai YAN ; Mengde CAO ; Jianzhi WANG ; Jirong LIU ; Guangming GONG ; Yan XU ; Xuefei HAN ; Ying XING
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To explore the regulatory effects of cytokines such as EGF, bFGF on expression of neural-specific molecules in mononuclear cells (MNCs) cultured in H-DMEM medium. METHODS: The umbilical cord blood samples were collected from health puerperal natural delivery. The mononuclear cells were isolated by centrifugation over Lymphoprep and planted in T-75 flasks containing H-DMEM medium with or without addition of EGF, bFGF or EGF plus bFGF at a final concentration of 20 ?g/L, respectively. Phenotypic changes were monitored by inverse phase-contrast microscopy. Tau and MAP2 mRNA were determined by reverse-transcript polymerase chain reaction (RT-PCR). Tau and MAP2-positive cell were determined by immunocytochemistry. RESULTS: The expression of tau protein mRNA was negative in uncultured cells, but MAP2 mRNA was positive. In cultured cells, tau protein mRNA expressed positively, MAP2 mRNA expression was upregulated by EGF+bFGF, EGF or bFGF compared with control group (no cytokines). The upregulatory capability of EGF+bFGF to MAP2 mRNA expression was stronger than that of EGF or bFGF alone. The same upregulatory tendency was noted in tau mRNA expression. In the group of control, bFGF, EGF, EGF+bFGF, the rate of MAP2-positive cells was 14.4%, 19.6%, 25.6%, 33.5%, respectively. Tau protein-positive cells were 13.5%, 15.3%, 21.4%, 29.8%, respectively. Under inverse microscopy, the freshly isolated MNCs were small and round, after culturing, the cells became larger with some big, long cytodenrites in the EGF+bFGF group, with 1 or 2 threadlike cytodenrites in the EGF group, or with some short multi-dendron like-astrocyte in the bFGF group and control group, but the number of astrocyte-like cells in the control group was less than that in bFGF group. CONCLUSION: MNCs derived from human umbilical cord blood cells express some neural specific molecules and are upregulated by cytokines, especially EGF and bFGF, which have the synergetic action. [
9.Analysis and follow-up of dTP value in patients with coronary heart disease complicating diabetes
Zhijun WANG ; Jun TIAN ; Jianzhi ZHOU ; Zengli DIAO ; Yuling HUANG ; Tienan LIU ; Haitao LI
Chongqing Medicine 2017;46(22):3063-3065,3069
Objective To analyze the dTP value in the patients with coronary heart disease (CHD) complicating diabetes mellitus (DM) and its relationship with major adverse cardiovascular events (MACE) and rehospitalization.Methods Two hundreds and seventy CHD patients were selected as the research subjects,including 136 cases of non-MD and 134 cases of DM.Their clinical condition was recorded.The indicators such as height,body mass,blood pressure and heart rate were measured.ECG,echocardiography,coronary angiography and other examiantions were carried out.The various indicators were detected.11-dh-TXB2 and 6-k-PGF1a levels were detected in the two groups and then dTP value was calculated.The 1-year follow-up was performed,MACE and rehospitalization were recorded.Epdate software was used for building a database and SPSS 17.0 software was applied for conducting the statistical analysis.Results The dTP level in the f non-DM and DM patients were 1.8 ± 0.6 and 2.0 ± 0.7 respectively,the difference was statistically significant (P< 0.05).For the non-DM CHD group,hs-CRP,systolic blood pressure,diastolic pressure,lesions number and severe lesions number were correlated with dTP level(P<0.05).For the complicating DM CHD group,hs CRP,blood glucose,CHO level,lesions number and severe lesions number were correlated with dTP level(P<0.05).After 1-year follow-up,MACE had 33 cases (24.3%) in the non-DM group and 44 cases (32.8%) in the DM group respectively,the difference was not statistically significant (P>0.05).The rehospitalized cases had 12 cases (8.8%) in the non-DM group and 24 cases (17.9 %).in the DM group respectively,the difference was statistically significant (P< 0.05).The dTP levels of MACE occurrence and non-MACE occurrence were 2.3 ± 0.8 and 1.8 ± 0.6 respectively,the difference was statistically significant (P<0.05).The dTP levels of rehospitalized patients and non-rehospitalized patients were 2.4 ± 1.0 and 1.9 ±-0.6 respectively,the difference was statistically significant(P<0.05).Conclusion The dTP level in the patients with CHD complicating DM is significantly increased,suggesting that platelet is obviously activated,moreover higher dTP level increases the risk of MACE and rehospitalization.So the anti-platelet therapy should be strengthened.
10.The analysis and follow-up study of Clopidogrel resistance of Coronary heart disease combine with diabetes patients
Zhijun WANG ; Zeyu WANG ; Shuo WANG ; Jianzhi ZHOU ; Ning LIU ; Zengli DIAO ; Yuling HUANG ; Tienan LIU ; Haitao LI
The Journal of Practical Medicine 2017;33(3):447-450
Objective To analyze Clopidogrel Resistance (CR) and influencing factors of coronary heart disease (CHD) with diabetes (DM) patients and evaluatc the relationship of CR and major adverse cardiovascular events (MACE) and readmission of CHD with DM patients.Methods 270 CHD patients were enrolled.Clinical conditions of CR were measured by adenosine diphosphate (ADP) induced maximum platelet aggregation rate (MPAR).After 1-year follow-up,MACE events and rehospitalization were recorded.Results CR of NDM and DM patients were 45 (33.1%) and 78 cases (58.2%) respectively,and the difference was significant (P < 0.001).Factors of CR of CHD DM patients included heart rate,TG level,the number of severe coronary artery disease.MACE events of CS and CR patients were 35 (23.8%) and 47 patients (38.2%) respectively,and the difference was significant (P =0.010).The readmitted patients of CS and CR groups were 15 cases (10.2%) and 27 patients (22.0%) respectively,and the differcnce was significant (P =0.008).The MACE of CR and CS patients in DM group were 32 (41.0%) and 12 cases (21.4%) respectively,and thc difference was significant (P < 0.05).The Readmitted cases of CR and CS patients in DM group were 19 (24.4%) and 5 (8.9%) respectively,and the diffcrcnce was significant (P < 0.05).Conclusions CR of CHD DM patients increased significantly.The influencing factors of CR of CHDDM are including heart rate,TG level,the number of severe coronary artery disease.MACE events and rehospitalization rate were significantly increased in CHD patients with DM AR.Therefore,it should be further strengthened the anti-platelet therapy for CHD patients with DM.