1.Effects of siRNA targeting ADAMTS2 gene on hepatic stellate cells
Zhiqiang SHI ; Meng LUO ; Qing XU ; Wei CHEN ; Hong ZHOU
Chinese Journal of Hepatobiliary Surgery 2012;18(3):205-210
Objective To explore the preventive and therapeutic role of silencing type Ⅰ rat platelet-binding protein motifs depolymerization protein-like metalloproteinase 2(ADAMTS2)by siRNA on experimental liver fibrosis in vitro.By studying the mechanism of siRNA silencing of ADAMTS2,we also aim to evaluate the feasibility of ADAMTS2 as a target for anti-liver fibrosis therapy.Methods Three pairs of siRNAs targeting ADAMTS2 mRNA 2237,2597 and 690 targets were designed and synthesized by utilizing RNA design software.The most effective siRNA was chosen to transfect HSC-T6 cell line to test the tendency of hepatic stellate cell(HSC)activation and ex pression of ADAMTS2,COL1α1,COL(I),α-SMA,TGF-β1,MMP-2 and TIMP-3.These were quantified using real time-PCR,Western blotting,and MTT assays.Results Of the same dosage and time of injection,siRNA 2237 inhibited ADAMTS2 gene expression significantly more than other siRNAs.siRNA-ADAMTS2 2237 markedly inhibited ADAMTS2 gene and protein expression of HSCT6 with more than 80% efficiency.Conversely,siRNA-ADAMTS2 2237 markedly reduced the gene and protein expressions of COL(I),α-SMA and TGF-β1 on HSC-T6 and inhibited the proliferation of HSC.Conclusions siRNA-ADAMTS2 2237 could effectively knockdown the gene and protein expression of ADAMTS2 in HSC-T6 cell lines.Silencing ADAMTS2 by siRNA significantly inhibited the activation,proliferation of HSC and the gene and protein expressions of COL(I),α SMA,and TGF-β1,and it may have a potential anti-fibrotic effect.ADAMTS2 might be an efficient target for anti-fibrotic therapy.
2.Changes in the flux of hexosamine biosynthesis pathway in skeletal muscle of rats with insusulin resistance induced by high fat diet
Jinhua GU ; Jianhua SHI ; Qing ZHU ; Hong ZHOU ; Jiliang XU
Chinese Journal of Endocrinology and Metabolism 2010;26(1):43-46
Objective To investigate the effect of hexosamine biosynthesis pathway on the development of insulin resistance induced by high fat diet.Methods Normal male SD rats were randomly divided into three groups:control(fed with normal chow),high fat(fed with high fat diet for 13 weeks),and rosiglitazone (intragastric administration with rosiglitazone for 5 weeks)groups.After 13 weeks,all the rats were sacrificed,serum and muscle triglycerides(TG),serum total cholesterol(TC),and serum and muscle free fatty acids(FFA) were measured.Insulin sensitivity wss evaluated by insulin sensitivity index(ISI)and glucose infused rat(GIR) with the hyperinsulinemic englycemic clamp technique.The flux of HBP in skeletal muscle was detected with the expression level of glutamine-fructose-6-phosphate transaminase(GFAT)mRNA(RT-PCR),the content of UDPGlcNAc(HPLC)and the level of O-GlcNAc glycosylation in skeletal muscle proteins(Western blot). Results Compared with control group,senlm TG,TC,FFA and muscle TG,FFA levels of high fat group increased(aII P<0.01).both ISI and GIR decreased(both P<0.01),and the leveIs of GFAT mRNA(0.51±0.05 vs 0.18±0.02),UDP-GlcNAc[(6.18±0.86 vs 2.42±0.36)nmol/g],and O-GIcNAc glycosylation of skeletal muscle proteins in high fat group were raised(all P<0.01).In rosiglitazone group,serum and muscle TG.FFA welc deceased(all P<0.01).insulin sensitivity was increased(P<0.05)and the flux of HBP[GFAT mRNA 0.27±0.03,UDP-GIcNAc(2.62±0.32)nmol/g]was reduced(all P<0.05)as compared with high fat group. Conclusions High fat diet-induced insulin resistance in rats is correlated with the increased flux of HBP in skeletaI muscle.which is decreased by rosiglitazone.
4.Effect of alanyl-glutamine dipeptide on glutathione concentration and antioxidation in burned rats
Yingqin SHI ; Wenjie ZHANG ; Qing WANG ; Xiaopeng ZHANG ; Jianli ZHOU ;
Parenteral & Enteral Nutrition 1997;0(01):-
Objectives:In the study, effect of Ala Gln on glutathione concentration and antioxidation were investigated in burned rats . Methods:Thirty male SD rats were randomly divided into three groups,A:control group,B:TPN group,C:Ala Gln PN group.The nutrient solution was administered through central venous catheter for 7 days.Glutathione concentration and levels of xanthine oxidase(XOD) and superoxide dismutase(SOD) in the serum were determined. Results:Compared with control group and TPN group,the levels of serum glutathione and SOD in group C were significantly increased( P
5.Keshan disease in Hubei Province from 2007 to 2014: analysis of surveillance data
Suhua ZHOU ; Guanghai WU ; Biao MOU ; Qing SHI
Chinese Journal of Endemiology 2015;34(6):437-439
Objective To study the disease trend of Keshan in Hubei Province and provide a basis for scientifically making control and prevention strategy.Methods According to Chinese Surveillance Plan of Keshan Disease,physical examination and electrocardiogram were done for surveillance subjects and suspected patients were filmed chest X-ray in Keshan disease areas.Results Totally 4 822 people were monitored from 2007-2014.The numbers of females (2 589,53.69%) were more than those of the males (2 233,46.31%).The group of people under the age of 15 accounted for 52.47% (2 530/4 822) which was higher than that of the group over the age of 55 (19.14%,923/4 822).The rate of electrocardiographic abnormality was the highest in 2011 (61.33%,268/ 437) and the lowest in 2014 (5.61%,23/410).In the eight years,only potential type of Keshan disease patients were detected,and the incidences from 2007 to 2014 were all lower than 3% and had a decreasing trend yearly.No Keshan patient was detected in 2012-2014.Ninety-one suspected cases were taken chest X-ray.Most of the suspected cases (63.74%,58/91) had normal cardiothoracic ratio.The cardiothoracic ratio increased,excepting one,other 32 were all potential type Keshan disease patients.Conclusions The disease trend of Keshan disease in Hubei Province is weak.Nevertheless,surveillance and health education are indispensable,in case the disease relapses.
6.Clinical features of 29 cases of children with extracranial cerebral embolism
Qing HAN ; Yu SHI ; Kai ZHOU ; Wenwei TANG ; Deyu ZHAO
Chinese Journal of Applied Clinical Pediatrics 2016;31(24):1889-1893
Objective Through the analysis of cases,to improve the cognition of clinicians on extracranial vas-cular embolism diseases(VED)in children.Methods The clinical information included incidence trend,thrombophi-lia,clinical features,treatment and prognosis of 29 children with VED,who were admitted from January 201 1 to Decem-ber 201 5 in Nanjing Children′s Hospital Affiliated to Nanjing Medical University,and retrospectively analyzed.Results The annual rate of VED increased from 0 to 1 .49 per 1 0 000 hospital admissions from January 201 1 to December 201 5.In 29 cases,there were 20 male and 9 female,with an average age of 6.03 years old (2 h -1 3.83 years old).In-fants accounted for >20%(6 /29 cases).The average age of the female,non infected group,rheumatic disease group and venous thrombosis group was higher than that of the male,the infection group,the non rheumatic disease group and the arterial embolization group,but there was no significant difference(all P >0.05).Thrombophilia were as follows:in-fection disease,heart disease,kidney disease syndrome,rheumatoid disease,tumor,and twins embolization syndrome. Risk factors included central venous line,immobility,dehydration,glucocorticoids use,etc.Heart disease was the leading cause of non thrombotic embolism,while other thrombophilic diseases resulted in thrombotic embolism.Pulmonary em-bolism was mainly seen in patients with severe pneumonia,especially mycoplasma pneumoniae pneumonia (MPP ).The older the age,the more typical clinical manifestations,and significantly elevated D dimer,fibrin/fibrinogen degradation products tips VED.Treatment depends on the degree of risk.Thirty -seven point five percent(3 /8 cases)of children with arterial embolism were less than 1 year old,and the mortality 25.00% (2 /8 cases)was significantly higher than that of venous thrombosis 1 7.65%(3 /1 7 cases)and 1 1 .76% (2 /1 7 cases).Conclusion Heart disease mainly cause arterial embolism and infection,while nephritic syndrome and rheumatic disease mainly cause venous embolism.Pulmona-ry embolism is mainly seen in children with severe pneumonia,especially MPP.The risk of arterial embolization is higher than that of venous embolism.
7.Surveillance and Analysis of Nosocomial Infection among Jizheng Patients
Jiyang ZHOU ; Yi SHI ; Xuelei LI ; Qing HE
Chinese Journal of Nosocomiology 2006;0(06):-
OBJECTIVE To realize the status and the impact factors of nosocomial infection among Jizheng patients.METHODS To investigate 562 former inpatients suffered from Jizheng in hospital from Jan 2004 to Dec 2005.RESULTS The incidence rates of nosocomial infection that calculated by the patients and the cases were 14.59% and 16.55%,respectively.Infection mostly occurred on the 22nd day in hospital on average.The main infection sites were lower respiratory tract(48.78%),urinary tract(15.85%),and alimentary tract(12.20%).The most risk factor was aging.CONCLUSIONS The high incidence of nosocomial infection is due to various factors.We should take measures respectively aiming at different factors to reduce the occurrence of nosocomial infection.
8.Application of 3-dimensional speckle tracking imaging in assessing the left ventricular systolic function of patients with uremia after a single hemodialysis
Bingyi ZHANG ; Ruiqiang GUO ; Qing ZHOU ; Ming SHI ; Liming ZHOU ; Hongning SONG ; Tuantuan TAN
Chinese Journal of Ultrasonography 2014;23(8):651-655
Objective To assess the impact of a single hemodialysis on the left ventricular systolic function of uremia patients with 3-dimensional ultrasound speckle tracking imaging (3D-STI).Methods Thirty-seven clinically stable outpatients with uremia undergoing hemodialysis were studied.The echocardiographic examinations were obtained in 30 minutes before and after homodialysis.Twenty nine normal subjects with age and sex matched were selected as control groups.Conventional ultrasound was recorded and then left ventricular mass index(LVMI) was calculated.The peak systolic mitral annular velocity S' was recorded by tissue Doppler imaging.3D-STI imaging were recorded from standard left ventricular apical 4-chamber views before and after a single hemodialysis.Left ventricular end diastolic volume (LVEDV),left ventricular end systolic volume (LVESV),left ventricular ejection fraction (LVEF),and left ventricular global longitudinal peak systolic strain (LVGLS),left ventricular global radial peak systolic strain (LVGRS),left ventricular global circumferential peak systolic strain (LVGCS),left ventricular global area peak systolic strain (LVGAS) were measured.The indicators of conventional ultrasound and 3D-STI were compared between the pre-hemodialysis group and the normal control group,and also between pre-and post-hemodialysis groups.Results ①Compared with the control group,the left ventricular end-diastolic diameter (LVDD),interventrieular septal thickness at diastole (IVSD),left ventricular posterior wall thickness at diastole(LVWPD),left atrial diameter(LAD) in the group before a single hemodialysis were significantly higher (P <0.05 for all).Meanwhile,the S' was significantly lower (P >0.05) and LVMI was significantly higher in the pre-hemodialysis group than the control group(P < 0.05).The LVDD,LAD,left atrial volume index(LAVI) were significantly lower in the post-hemodialysis group than that in the pre-hemodialysis group(P <0.05 for all).②Results of 3D-STI showed that the LVEF,LVGLS,LVGCS,LVGRS,LVGAS was significantly lower,however LVEDV,LVESV were higher in the pre-hemodialysis group compared with the control group (P < 0.05 for all).After a single hemodialysis,LVEDV,LVESV,LVGLS were significantly lower than before(P <0.05 for all),but LVEF,LVGRS,LVGCS,LVGAS were not significantly changed (P > 0.05 for all).Conclusions After a single hemodialysis,the whole systolic function of the left ventricular was not changed.but the left ventricular global longitudinal peak systolic strain was significantly lower.The parameter of LVGLS was sensitive to the change of preload and was volume-load dependent.The preload of the patients should be sufficiently considered when the LVGLS are measured in the patients with uremia undergoing hemodialysis.
9.Inhibitory effect of genistein on the proliferation of the anterior pituitary cells of rats
Qing-Hong ZHANG ; Yu-Zhen HU ; Shi-Sheng ZHOU ; Fu-Zhou WANG
Acta Physiologica Sinica 2001;53(1):51-54
The effects of tyrosine kinase inhibitor genistein on the proliferation of rat anterior pituitary cells and mouse AtT-20 cells were studied using techniques of cell culture, ~3H-TdR incorporation, flow cytometric analysis and electron microscope. Genistein significantly inhibited the proliferation of rat anterior pituitary cells and mouse AtT-20 cells. Genistein (50 and 100 μmol/L ) blocked the proliferation of AtT-20 cells at G0/G1 and G2/M phases and evoked an apoptotic peak of these cells with an apoptotic ratio of 19.9% and 36.4%. The apoptotic cells were also observed under the electron microscope. In consequence, genistein, as a tyrosine kinase inhibitor, can significantly inhibit the proliferation of pituitary cells possibly by inducing apoptosis, and the tyrosine kinase activity may play a key role in the proliferation and differentiation of pituitary cells.
10.The expression and function of retinoic acid-inducible gene Ⅰ in monocyte-derived dendritic cells in patients with hepatitis B virus infection
Gangde ZHAO ; Qing XIE ; Hui WANG ; Baoyan AN ; Huijuan ZHOU ; Nina JIA ; Lanyi LIN ; Cuicui SHI ; Qing GUO ; Hong YU
Chinese Journal of Infectious Diseases 2009;27(12):727-732
Objective To investigate the expression and function of retinoic acid-inducible gene Ⅰ(RIG-Ⅰ) in monocyte-derived dendritic cells (MoDC) at different stages of hepatitis B virus(HBV)infection and to explore the role of RIG-Ⅰ in the disease progression after HBV infection. Methods Peripheral blood samples were collected from 28 hepatitis B virus-infected persons, including 21 cases of chronic hepatitis B (CHB) and 7 of acute hepatitis B (AHB). Eighteen healthy subjects were recruited as controls. Purified CD14~+ monocytes were isolated by CD14 microbeads. MoDCs were induced from CD14~+ monocytes with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 for 7 days, and then were infected with vesicular stomatitis virus (VSV) to stimulate RIG-Ⅰ expression. The mRNA expression levels of RIG-Ⅰ, interferon (IFN )-promoter stimulating factor-1 (IPS-1) and IFN-β at 16 hours and 24 hours after infection with VSV were measured by real-time quantitative polymerase chain reaction (PCR). Data with normal distribution were tested by analysis of variance. Continuous variables between groups were compared using Mann-Whitney U test. Comparison among multiple groups was done by Kruskal-Wallis test. Results The expression levels of RIG-Ⅰ in MoDCs from CHB patients were significantly lower than those in AHB patients and healthy controls at 16 hours (2.44±2.03, 19. 54±3. 15, 21. 48±8. 39, respectively; F=7.451,P=0.002) and 24 hours (2. 68±2. 93, 10. 31 ±3. 88, 14. 01 ±5. 04, respectively, F = 7. 908, P = 0. 001)following VSV stimulation. The IPS-1 levels in both CHB patients and AHB patients were higher than those in healthy controls at 16 hours (2. 05±l. 08, 1. 99±1. 56, 0. 60±0. 31, respectively) F=7.246,P =0.003) and 24 hours (2. 27±2. 16, 3.24 ± 1.21, 1. 08±0. 73, respectively; F= 13. 598, P = 0. 001).Furthermore, the IFN-β expression levels were significantly lower in CHB patients compared to AHB patients and healthy controls at 16 hours and 24 hours after VSV stimulation. Conclusions The expressions of RIG-Ⅰ and IPS-1 in MoDC are abnormal in HBV infected persons, which indicates that RIG-Ⅰ signaling pathway might be blocked by HBV. The impaired function of MoDC may play a role in HBV infection and chronicity.