1.Study on LipofectinRM mediated murine SCF genetr ansduction into umbilical blood cells
Yubing CHEN ; Hongmei ZHANG ; Chunlei YU
Chinese Journal of Biochemical Pharmaceutics 2001;22(1):18-20
Purpose The aim is to study the transduction of m urine stem cell factor(SCF) into umbilical blood cells by LipofectinRMmediated and expression in them.Methods Murine SCF cDNA encoding extra cellu lar domain was isolated using PCR from plasmid pRC/CMV(containing SCF gene), and then recombined into the expression vector pcDNA3,and transferred into enrichme nt cultural umbilical blood cells. Semi-quantitative RT-PCR was used to examin e the expression on mRNA level.And the effects of supernatant of transfected umb ilical blood cells were investigated alone or in coordinate with GM-CSF by colo ny formation test of human bone marrow cells in semisolid culture.Results SCF mRNA was expressed in transfected umbilical blood cells.The su pernat ant of transfected umbilical blood cells could increase the number of CFU-GM, s ynergizing with GM-CSF.Conclusion The supernatant of umbil ical b lood cells transfected with vector containing mSCF gene can stimulate the colony formation of human bone marrow cells in combination with GM-CSF.
2.Effect of shenmai injection on myocardial eNOS mRAN expression in myocardial ischemic rats
Ruimin YU ; Yubing ZHOU ; Aihua XIONG ; Qinhe YANG
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To study the effect of shenmai injection (SMI), a Chinese medicine, on nitric oxide and the expression of endothelial nitric oxide synthase (eNOS) mRNA in myocardium of rats with experimental myocardial ischemia. METHODS: Rats were randomly divided into four groups: control, model (ischemia), SMI 1 and SMI 2 group. A rat model of acute myocardial ischemia was established by isoprenaline treatment and the lift of ST segment in ECG was used as the index of myocardial ischemia. The nitric oxide (NO) contents in serum and myocardium and the expression of eNOS mRNA in myocardium were measured. RESULTS: Compared with control group, ST segment of ECG was significantly elevated 20, 30, 40 min after myocardial ischemia in model group, the lift peak of ST segment occurred 20 min after myocardial ischemia, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium were significantly lowered in model group. Compared with the model group, in SMI 1 group and SMI 2 group, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium were significantly increased, the lift of ST segment were significantly reduced 20, 30, 40 min after myocardial ischemia. Compared with SMI 1 group, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium and the lift of ST segment were not statistically different in SMI 2 group. CONCLUSION: Shenmai injection can increase the expression of eNOS mRNA in myocardium and the content of NO, and protect against myocardial ischemia in rats.
3.Association of hs-CRP with abnormality of glucose metabolism in patients with essential hypertension
Guoqin LIU ; Ruixia YU ; Peikang DONG ; Yubing WANG
Chinese Journal of Primary Medicine and Pharmacy 2010;17(21):2911-2912
Objective To investigate the association of high-sensitivity C-reactive protein(hs-CRP) with the abnormality of glucose metabolism in patients with essential hypertension.Methods Based on the level of fast plasma glucose(FPG) and 2-hours-plasma glucose(2hPG),120 patients with essential hypertension were divided into 3 groups:Normal glucose tolerance group(NGT),impaired glucose tolerance group(IGT)and diabetes mellitus group (DM).The hs-CRP was measured.Results The level of hs-CRP gradually increased from NGT to DM groups(P<0.05).The level of hs-CRP was positively correlated with SBP,DBP,2hPG and FPG in patients with essential hypertension(r =0.48,0.37,0.46,0.31,P<0.05).Conclusion The abnormality of glucose metabolism in patients with essential hypertension was related with hs-CRP.
4.Systemic lupus erythematosus with secondary antiphospholipid syndrome: a retrospective clinicopathological analysis of 11 cases
Wenqing YU ; Yubing WEN ; Ruitong GAO ; Hang LI ; Xuemei LI
Chinese Journal of Nephrology 2010;26(12):875-879
Objective To analyze the clinicopathological characteris tics of systemic lupus erythematosus (SLE) with secondary antiphospholid syndrome (APS) . Methods Data of 11 cases of SLE with secondary APS (SLE with APS) admitted to Peking Union Medical College Hospital from January 2000 to March 2010 were retrospectively analyzed. Kidney biopsy was performed on all of these patients. Differences of clinicopathology and outcomes between SLE with and without APS were compared. Results Renal involvement was found in all the SLE with APS patients. The prominent clinical manifestations included hypertension (54.5%), nephrotic level of proteinuria (24 h proteinuria ≥3.5 g)(72.7%) and renal insufficiency (45.5%). Diastolic blood pressure, mean arterial pressure and glomerular filtration rate in SLE with APS were significantly higher than those in SLE without APS (all P<0.05). In 8 out of 11 cases (72.7%), APS nephropathy (APSN) in kidney biopsy was found, characterized by small vessel vaso-occlusive lesions. These included thrombotic microangiopathy (TMA), fibrous intimal hyperplasia (FIH), focal cortical atrophy (FCA) and tubular thyroidization. Among those, 5 cases (45.5%) had chronic APSN and 4 (36.4%) had acute APSN (one case had acute and chronic APSN at the same time). The incidences of APSN and acute APSN in the SLE with APS group were significantly higher than those in SLE without APS group (P<0.05). Conclusions The major renal manifestations of SLE with APS are hypertension, nephrotic level of proteinuria and renal insufficiency. Other than lupus nephritis, also a high incidence of APSN is found in SLE with APS patients.
5.Detection of apolipoprotein B-100 gene mutation in patients of primary hypercholesterolemia families.
Ruimin YU ; Yubing ZHOU ; Jiansheng FENG ; Qilu CAI ; Chunlan LIN
Chinese Journal of Practical Internal Medicine 2001;0(10):-
Objective To detect genetic mutation of apolipoprotein B-100(apoB-100)in patients with primary hypercholesterolemia.Methods One special segment of apoB-100 gene from nucleotide 10549 to 10895 was amplified by polymerase chain reaction(PCR).The PCR products were denatured and hybridized with specific aligonucleotide labeled with digoxigenin,and were analyzed by single strand conformation polymorphism(SSCP)to detect the apoB-100 gene mutation 3531CGC→CGT or other mutations.Results Overall 41 members of 11 primary hypercholesterolemia families were detected,but the above genetic mutation was not detected.Conclusion This genetic mutation is unlikely to exist or of significantly low incidence in Chinese population,and might not be the main cause of primary hypercholesterolemia in the 11 primary hypercholesterolemia families.
6.Type 2 diabetes in MKR transgenic mice cansed by skeletal muscle-specific double deficiency of IGF-1 and insulin receptors
Rong YU ; Xihua CHENG ; Wei HU ; Kaichun WEI ; Yubing GUO ; Junli LIU
Chinese Journal of Diabetes 2008;16(7):438-440
Objective To examine the circulating glucose and body weight in the transgenic MKR mouse model who expressed dominant-negative IGF-1 receptor and insulin receptor in skeletal muscle leading to systemic insulin resistance and diabetes. Methods MKR mice were genotyped by PCR analysis of tail DNA.And in these mice we examined the circulating glucose and body weight once a week from 1 to 16 weeks of age, and the circulating insulin and glucose tolerance at age of two-month-old by using C57 mice as controls. Results The descendents of MKR mice kept hereditary feature. And these mice had hyperglycaemia from 3 weeks of age,and an increasing body weight slowly(P<0.01).Twenty-fold significant hyperinsulinemia was observed in MKR mice,and they were glucose intolerant in 2-month-old male and female (P<0.01).Conclusions The MKR mouse is an excellent model of type 2 diabetes
7.Study on Bioequiavailability in Human Body between Domestic and Imported Roxithromycin
Yubing ZHU ; Dawei XIAO ; Jianjun ZOU ; Wei QIAN ; Yunfang HU ; Cuixia YU ; Rong GU
China Pharmacy 2005;0(16):-
OBJECTIVE:To study the bioequiavailability of domestic roxithromycin tablets and imported ones.METH?ODS:20male healthy volunteers took single dose of150mg roxithromycin tablet orally in a random crossover design,blood concentrations were determined by LC-MS.RESULTS:The main pharmacokinetic parameters of domestic and imported tablets were determined respectively as follows,AUC 0~72 were(72.81?23.85)(mg?n)/L and(72.63?20.86)(mg?h)/L,AUC 0~∞ were(74.41?24.45)(mg?h)/L and(74.42?24.45)(mg?h)/L,C max were(6.46?1.51)mg/L and(6.58?1.55)mg/L,t max were(1.9?0.5)h and(1.8?0.5)h,t 1/2 were(13.56?1.35)h and(14.18?1.50)h,the relative bioavailability of the homemade tablet to imported one was(99.8?11.2)%.CONCLUSIONS:Domestic and imported roxithromycin are bioequivalent.
8.Association between prognosis and renal vascular lesions in lupus nephritis patients
Yu TANG ; Hang LI ; Yubing WEN ; Lin DUAN ; Yan LI ; Xuewong LI
Chinese Journal of Nephrology 2008;24(7):449-455
Objective To examine the relationship between renal vascular lesions and clinical prognosis in lupus nephritis patients. Methods Renal biopsy specimens and clinical data of 451 patients with lupus nephritis in our hospital from January 1998 to February 2006 were enrolled in this study. According to renal vascular lesions, those patients were classified into six groups: no renal vascular lesion (NVL), immune complex deposits (ICD), lupus vasculopathy (LV),thrombotic microangiopathy (TMA), vasculitis (VAS) and arteriosclerosis (AS). The relationship of renal vascular lesions and the clinical features was retrospectively analyzed. Results (1) In 451 lupus nephritis patients, 247 eases (54.8%) had renal vascular lesion. The incidence of the ICD, LV, TMA, AS and VAS were 8.4%, 9.8%, 6.7%, 29.3% and 0.7%, respectively. (2) The incidence of vascular lesions in IV type lupus nephritis was 69.7%, which was higher than others.(3) Except ICD group, the incidences of renal insufficiency and hypertension were significantlyhigher in vascular lesion groups than those in NVL group (P<0.05). TMA group showed the most severe clinical manifestation, including anemia and depression of platelet. (4) The prognosis of TMAgroup was the worst. The prognosis among other groups was not significantly different.Conclusions Vascular lesions are common in lupus nephritis. TMA patients present not only the most severe clinical manifestation with the poorest prognosis. Other vascular lesions also indicate an elevation of blood pressure and Scr level, but without significant difference in prognosis.
9.Pharmacokinetics and bioavailability of two kinds of gliclazide sustained release tablets following a single and multiple dose in healthy volunteers
Jianjun ZOU ; Dawei XIAO ; Yubing ZHU ; Ling MO ; Cuixia YU ; Rong GU ; Yunfang HU ; Wei QIAN ; Sheng LOU
Chinese Journal of New Drugs and Clinical Remedies 2005;24(5):337-341
AIM: To compare the pharmacokinetics and relative bioavailability of the domestic and imported sustained-release tablets of gliclazide in healthy volunteers. METHODS:The study was performed by an four-period crossover design with singledose and multiple-dose administration. The plasmadrug concentrations of twenty male healthy volunteers were determined by liquid chromatography with mass spectrum detector method (LC-MS). RESULTS:The pharmacokinetic parameters after a single oral dose of the domestic and imported gliclazide tablets were (7.2+s 1.5) h and (6.9 +1.4) h for tmax, (13.4 ±1.2) h and (13.7 +1.3) h for t1/2, (2.4 +0.8) mg ·L-1and (2.3 ±0.6) mg· L-1 forcmax, (48 ±14)mg · h · L-1 and (48 +14) mg· h · L-1 forAUC0-60,(51+15) mg· h· L-1 and (50±14) mg· h· L-1for AUC0-∞, (22.4 ± 1.9 ) h and (22.8 ± 1.9 ) h for MRT, respectively. The steady state pharmacokinetic parameters after multiple doses of the domestic and imported gliclazide tablets were (6. 1 ± 1.4) h and (6.5+1.4) h for tmax, (4.6±0.9) mg· L-1 and (4.7±1.1) mg· L-1 for cmax, (0.23 ±0.08) mg ·L-1and (0.26±0.08) mg· L-1 forcmin, (1.6±0.3) mg·L-1 and (1.6±0.3) mg · L-1 for mean value of steady plasma-drug concentration (cav),(94±19) mg· h · L-1 and (95 ±20) mg · h · L-1forAUCss, (282 ±33)% and (283 ±43)% for degree of fluctuation DF ), respectively. The relative bioavailability of the domestic gliclazide tablet to the imported gliclazide tablet following a single and multiple dose were ( 102 ± 9) % and (99 ± 10 ) %, respectively. Main pharmacokinetic parameters between the two formulations in both single and multiples dose studies showed no statistical difference ( P >0.05 ). CONCLUSION: The result of two one side t-test shows that the two formulations are bioequivalent.
10.Specific expression of CPS-II in hyperammonemia-injured liver cells.
Chunli GUO ; Zujiang YU ; Chao HAN ; Qiongye WANG ; Yubing ZHOU ; Quancheng KAN
Chinese Journal of Hepatology 2015;23(5):358-362
OBJECTIVETo study the CPS-II mechanism underlying the pathological process of elevated blood ammonia leading to liver injury.
METHODSAn in vitro hyperammonemia hepatocyte cell model was constructed by exposure to various concentrations of NH4Cl. The subsequent changes to cellular morphology were observed by microscopy. to cell apoptosis were determined by flow cytometry, and to mRNA and protein expression of CPS-II were examined by real-time PCR and western blotting, respectively.
RESULTSExposure to NH₄Cl led to dose-dependent morphological damage, apoptosis and necrosis of the hepatocytes. The apoptosis rate was significantly higher for the high-dose group than for the control (no exposure) group (24.7% ± 2.39% vs. 4.1% ± 0.78%, q =8.06, P less than 0.05). Expression of the CPS-II mRNA was significantly elevated in response to NH₄Cl exposure (vs. the control group; F=191.881, P < 0.05).The CPS-II mRNA expression level increased with increasing NH₄Cl concentration (grey values: 1.040 ± 0.045, 1.641 ± 0.123, 2.285 ± 0.167 and 3.347 ± 0.124, respectively). The CPS-II protein expression level was also significantly enhanced in response to the NH₄Cl exposures (CPS-II protein and internal GAPDH grey value ratios: 0.099 ± 0.0130, 0.143 ± 0.025, 0.161 ± 0.036 and 0.223 ± 0.042, respectively; t=3.825, 3.968 and 6.908, P less than 0.05).
CONCLUSIONCPS-II mRNA and protein expression levels become elevated with increase in the NH₄Cl concentrations, suggesting that in addition to the urea cycle, CPS-II may play an important role in the ammonia metabolism under the condition of hyperammonemia.
Ammonia ; Apoptosis ; Hepatocytes ; Humans ; Hyperammonemia ; Liver ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Somatostatin