1.Effect of Rapamycin on Proliferation and Cell Cycle of Human Bladder Carcinoma Cell Line 5637
Guangyi SHAN ; Cheng FU ; Ang CHEN ; Bin HU ; Huan BI
Journal of China Medical University 2015;(3):230-233
Objective To evaluate the effect of rapamycin on proliferation and cell cycle of human bladder cancer cell line 5637. Methods Total?ly 5637 cells were maintained in RPMI 1640 containing 10%fetal bovine serum,100 U/mL penicillin and 100μg/mL streptomycin for adherent cul?ture. Cells were grown at 37℃in a 5%CO2 incubator. The 5637 cells were treated with various concentrations of rapamycin solution(50 nmol/L, 100 nmol/L,200 nmol/L,and 400 nmol/L). The control group was daily treated with single RPMI 1640 without medication. The growth repression rate for 5637 cells was evaluated by MTT. Flow cytometry was used to investigate the effect of rapamycin of different concentrations on cell cycle of 5637 cells. Results Compared to the control group,rapamycin can inhibit the proliferation of 5637 cells in a concentration and time dependent manner. Rapamycin inhibited 5637 cells at G0/G1 thus inhibiting cell proliferation but not inducing apoptosis. Conclusion Rapamycin inhibited growth of 5637 bladder carcinoma cells and arrested cell cycle at G0/G1,indicating that mammalian target of rapamycin may play an important role in proliferation of 5637 cells and act as a new tumor therapeutic target in patients with bladder cancer.
3.Effects of Salvianolate on Myosin Heavy Chain in Cardiomyocytes of Congestive Heart Failure Rats.
Cheng CHEN ; Xiang-gu ZOU ; Shan-dong QIU ; Hui CHEN ; Yong-zhong CHEN ; Xiu-ming LIN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(7):871-876
OBJECTIVETo explore the effect of Salvianolate on myosin heavy chain (MHC) in cardiomyocytes of congestive heart failure (CHF) rats.
METHODSSixty male SD rats were divided into 6 groups according to random digit table, i.e., the normal control group (NCG), the model group, the Captopril group (CAG), the low dose Salvianolate group (LSG), the high dose Salvianolate group (HSG), the Captopril and high dose Salvianolate group (CSG), 10 in each group. CHF rat model was established with peritoneal injection of adriamycin in all rats except those in the NCG. Equal volume of normal saline was peritoneally injected to rats in the NCG, once per week for 6 successive weeks. Corresponding medication was started from the 5th week of injecting adriamycin. Rats in the CAG were administered with Captopril solution at the daily dose of 10 mg/kg by gastrogavage. Rats in the LSG and the HSG were administered with Salvianolate solution at the daily dose of 24.219 mg/kg and 48.438 mg/kg respectively by gastrogavage. Salvianolate was dissolved in 2 mL 5% glucose solution and administered by peritoneal injection. Rats in the CSG were peritoneally injected with high dose Salvianolate solution and administered with Captopril solution by gastrogavage. Two mL normal saline was peritoneally injected to rats in the model group, once per day for 8 successive weeks. Eight weeks later, the cardiac function and myocardial hypertrophy indices were detected by biological signal collecting and processing system. mRNA expression levels of alpha-MHC and beta-MHC in cardiac muscle were detected by fluorescence quantitative PCR. Expressions of protein kinase C (PKC) in cardiac muscle were detected by Western blot.
RESULTSCompared with the normal control group, heart mass index (HMI) and left ventricular mass index (LVMI) obviously increased in the model group (P < 0.01). Compared with the model group, HMI and LVMI decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). It was more obviously lowered in the CSG group than in the CAG group (P < 0.05). Compared with the NCG, the mRNA expression level of alpha-MHC in cardiac muscle decreased, the mRNA expression level of p-MHC and the expression of PKC in cardiac muscle increased in the model group (P < 0.01). Compared with the model group, the mRNA expression level of alpha-MHC in cardiac muscle was increased, and the mRNA expression level of beta-MHC and the expression of PKC in cardiac muscle were decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). There was statistical difference between the CSG group and the CAG group (P < 0.05).
CONCLUSIONSSalvianolate could up-regulate the mRNA expression level of alpha-MHC, and down-regulate the mRNA expression level of beta-MHC in cardiac muscle. Its mechanism might be related to decreasing the expression of PKC.
Animals ; Captopril ; Doxorubicin ; Drugs, Chinese Herbal ; Heart Failure ; metabolism ; Male ; Myocardium ; Myocytes, Cardiac ; drug effects ; metabolism ; Myosin Heavy Chains ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley
4.The influence of the aqueous extract of ephedra sinica treating in brain damage after subarachnoid hemorrhage in rats
Zhengheng TANG ; Shihao GAO ; Tunan CHEN ; Fei LI ; Dongyi CHEN ; Youan SHAN ; Hua FENG ; Yuan CHENG
Chongqing Medicine 2015;(25):3481-3484
Objective To observe the therapeutic effect and mechanism of the aqueous extract of ephedra sinica on brain damage after subarachnoid hemorrhage(SAH)in rats.Methods Totally 50 rats of Sprague-Dawley were randomly divided into control group,model group and three groups treated with different concentrations(4,12,36 mg/kg).The changes of the cerebral water content,malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and hydroxy radical of brain tissue were recorded,and he-matoxylin-eosin(HE)staining was used to test the subarachnoid haemorrhagia and oedema,and immunohistochemistry and western blot were carried out to assay the expression of complement C3 in brains of different animal in different group 3d after operation. Results On the postoperative 3 days,compared with the model group,the content of MDA、GSH-Px activity and hydroxyl radical of 12、36 mg/kg treatment groups significantly reduced(P <0.01 ),the content of MDA of 4 mg/kg treatment group not change (P >0.05),but GSH-Px activity and inhibition of hydroxyl radical significantly reduced(P <0.01),and the cerebral water content of 12,36 mg/kg groups were obviously lower compared with model group.The expression of complement C3 was significantly lower on 36 mg/kg treatment group and edema reduced.Conclusion The aqueous extract of ephedra could significantly inhibit the activity of complement C3,prevent the production of MDA、GSH-Px and hydroxyl radical,reduce the severity of cerebral edema and the in-flammatory response,which has a better therapeutic effect SAH animals.
5.Clinical analysis of 36 cases of sinus-straddling hematoma after craniocerebral injury.
Shan-lang YIN ; Shan-cheng CHEN ; Yang ZHENG ; Ze-liang YE
Journal of Southern Medical University 2006;26(1):130-1p following 130
OBJECTIVETo investigate the relation between sinus-straddling hematoma (SSH) and venous sinus injury and explore the approaches for surgical management.
METHODSThirty-six cases of stride sinus hematoma were reviewed to observe the incidence rate of sinus injury complicated with (SSH) and explore its surgical management.
RESULTSThe incidence rate of venous sinus injury following (SSH) was 80.56% (29/36), and appropriate surgical management yielded good therapeutic effect in these patients.
CONCLUSIONIntracranial stride sinus hematoma is often accompanied by venous sinus injury, and adequate preoperative risk evaluation may improve the success rate of the operation.
Adolescent ; Adult ; Brain Injuries ; complications ; China ; epidemiology ; Cranial Sinuses ; injuries ; Female ; Hematoma ; epidemiology ; etiology ; Humans ; Incidence ; Intracranial Hemorrhage, Traumatic ; epidemiology ; etiology ; Male ; Middle Aged
6.Spatial characteristics of non-iodized salt at household level based on geographic information system in Chongqing
Cheng-guo, WU ; Ya-lin, CHEN ; Xing-jian, LUO ; Xin-shu, LI ; Shan-shan, LI ; Bang-zhong, XIAO
Chinese Journal of Endemiology 2012;31(6):635-639
Objective To analyze the spatial distribution characteristics of non-iodized salt at household level based on geographic information system (GIS) in Chongqing.Methods The database of non-iodized salt at county level from 2001 to 2010 was established in Chongqing.By using GIS technology,the spatial distribution and spatial autocorrelation were analyzed by ArcGIS 9.3 software.Results The rate of non-iodized salt was fluctuated between 2.35%-5.78% during 2001-2006 and the rate was reduced to less than 2.00% after 2007.The result of spatial autocorrelation analysis on non-iodized salt from 2001 to 2006 indicated that Moran's Ⅰindex was 0.145578,0.078801,0.108033,0.091957,0.127749,0.214302,respectively(Z value was 3.066275,1.977321,2.541619,2.309972,2.900446,3.874203,respectively,all P < 0.05).The spatial distribution of non-iodized salt had marked spatial cluster through Chongqing region from 2001 to 2006.The result of local spatial autocorrelation analysis from 2001 to 2006 indicated that Fengdu and Fuling were two high-risk areas(all P < 0.05).Dianjiang,Yubei,Jiangbei,Wulong and Banan were also confirmed as high-risk areas in 2001,2005 and 2006(all P < 0.05).The results also indicated that the distribution of non-iodized salt in the seven high-risk areas was positively correlated.The result of spatial autocorrelation analysis on non-iodized salt from 2007 to 2010 indicated that Moran's Ⅰ index was 0.018361,0.016186,0.040769,-0.059691,respectively (Z value was 1.093310,0.787361,1.071811,-0.583820,respectively,all P > 0.05).The spatial distribution of non-iodized salt was at random on the whole from 2007 to 2010.However,there were four local high-risk areas.The distribution in Fengdu and Dianjiang was positively correlated,while that in Jiangjin and Shizhu was negatively correlated.Conclusions The distribution of non-iodized salt at households level in Chongqing is changed from spatial distribution before 2006 to random distribution after 2007,but there are high value areas,which should be taken as the focus of monitoring.
7.Study of the detection and significance of intracellular bacterial communities in patient with catheter-associated urinary tract infections
Zhen DU ; Ludong QIAO ; Wei YAN ; Cheng TIAN ; Qing CAI ; Shan CHEN
Chinese Journal of Urology 2017;38(1):51-54
Objective To detection the urine of bacteria hyphae and intracellular bacterial communities in patients with indwelling urinary catheter and discuss intracellular bacterial comnmunities in the pathogenesis of catheter-related urinary tract infection.Methods From May 2014 to February 2016,95 cases with D-J stent indwelling were enrolled in this study,including 38 male patients and 57 female patients.The mean age was (43 ±21)years old,ranging from 25 to 83 years old.We recorded those patient g clinical symptoms,middle urine culture results.If the middle urine culture was positive,further pathology test and scanning electron microscopy for bacteria hyphae and intracellular bacterial communities would be considered.Results The middle urine culture showed positive in 21 cases (22%,21/95);The classification of bacteria included E.coli in 11 cases,dung enterococcus in 2 cases,klebsiella pneumonia in 4 cases,pseudomonas aeruginosa in 3 cases,epidermis staphylococcus aureus in 1 case.Among those 21 patients,9 cases had the symptoms of fever and shiver.Urine pathology testing found hyphae in 6 cases (6%,6/95).all others were E.coli infection.For scanning electron microscope,6 cases were found rodshaped bacteria and hyphae.3 cases were found intracellular bacterial communities.Conclusions The presence of intracellular bacterial communities made urothelial itself the source of endogenous bacteria of urinary tract infection.Catheter-related urinary tract infections in patients with recurrence maybe basically homology bacteria.
8.Role of JLP on the epithelial to mesenchymal transition in renal tubular epithelial cells
Shan LIU ; Huiming WANG ; Guohua DING ; Qi YAN ; Dou FU ; Cheng YANG ; Zhaowei CHEN
Chinese Journal of Nephrology 2016;32(8):612-616
Objective To observe the effect of JLP on transdifferentiation of human renal proximal tubular epithelial cells (HK-2),and to investigate the role of p38 MAPK signaling pathway in this process.Methods The knock-down plasmids of JLP were constructed.HK-2 cells were randomly divided into four groups:negative control cells (Ctrl-shRNA group),knock-down jlp cells (jlpshRNA group),negative control cells with FGF-2 treatment (FGF-2 group) and knock-down jlp cells with FGF-2 treatment(jlp-shRNA +FGF-2 group).The expressions of JLP,E-cadherin,TGF-β1,α-SMA,p-p38 MAPK protein were detected by Western blotting.After the induction of FGF-2 for 24 hours,the expressions of α-SMA,COL-Ⅰ,FN were detected by immunocytochemistry.Results Compared with Ctrl-shRNA group,the expression of JLP protein was significantly down-regulated in FGF-2 group.Compared with FGF-2 group,the expressions of TGF-β1,α-SMA,p-p38 MAPK protein were significantly up-regulated,while E-cadherin protein was significantly down-regulated (P < 0.05).Compared with FGF-2 group,the expressions of α-SMA,COL-Ⅰ,FN immunostaining increased markedly in jlp-shRNA+FGF-2 group.Conclusion Scaffolding protein JLP is critical in preventing EMT in the course of fibrosis through the inhibition of p-p38 activation in HK-2 cells.
9.Screening and Identification of an Independent-glutamic Acid Strain Producing Poly (?-glutamic acid)
Qing-Shan SHI ; Cheng-Bin LI ; Chun-Hua WANG ; You-Sheng OUYANG ; Yi-Ben CHEN ;
Microbiology 1992;0(02):-
17 strains of bacterium that produced a large amount of ?-PGA when it was grown aerobically in a culture medium containing ammonium salt and sugar as sources of nitrogen and carbon respectively,were isolated from bean products.With the following identifications of colony morphology,physiological and biochemistry experiments,and genetics,the strain PGA-O-7 was classified as a Bacillus subtilis.The PGA production 2.8 (mg/mL) was obtained when it was grown in a medium containing 3% ammonium sulfate and 4% glucose at 30℃ for 72h with sharking.
10.Tissue distribution of exendin-4 in rats
Guo AI ; Zhihang CHEN ; Chengqi SHAN ; Jinjing CHE ; Yunan HOU ; Yuanguo CHENG
Chinese Journal of Pharmacology and Toxicology 2008;22(2):95-101
AIM To investigate the tissue distribution of exendin-4 after administration in healthy rats. METHODS Exendin-4 was radioiodinated by the Iodo-GenTMmethod. Tissue distribution of [125I]exendin-4 was investigated after sc administration of [125I]exendin-4 at 3 μg·kg-1 in rats. Both total radioactivity and trichloroacetic acid (TCA) precipitated radioactivity were used to calculate the levels of [125I]exendin-4 in rats plasma and tissue samples after sc administration. RESULTS The tissue distribution of [125I]exendin-4 after sc injection showed substantial disposition in kidneys, lungs, bladder and pancreas. The rank order of normalized tissue distribution was kidneys>lungs>bladder>pancreas>intestine>plasma>adrenals>jejunum>lymph>liver>spleen>heart>marrow>thymus>testicles>brain>muscle>adipose. CONCLUSION [125I]Exendin-4 underwent a rapid and wide distribution in the tissues throughout the whole body within the time course examined. TCA precipitated radioactivity in kidneys was the highest, however, only trace amounts of [125I]exendin-4 was detected in the brain.