1.Quantitative cardiotoxicity assessment of gambogic acid using multiple cellular phenotype analysis
Chinese Journal of Pharmacology and Toxicology 2017;31(1):73-79
OBJECTIVE To evaluate the cardiotoxicity of a widen-spectrum antineoplastic drug, gambogic acid, through quantitative multiple cellular phenotypic characterization. METHODS H9c2 cell line was used as a model with doxorubicin (Dox) and amiodarone (Ami) as positive controls, hypaconi?tine as negative control and 0.1% DMSO as normal control. An optimized protocol was established to identify the morphology and function of cell nuclei. The effect of drugs on cell viability, nuclear area (Hoechst33342), mitochondria mass (MitoTracker Deep Red) and cytoplasmic calcium ion mobilization (Rhod2 AM)was studied. EC50 and Z′values were calculated to evaluate the degree of toxicology and to estimate the precision and false-positive rate, respectively. RESULTS Dose-response analysis indicated that EC50 of Dox on cell viability, nuclear area, mitochondrial mass was 0.72, 0.014 and 1.21μmol · L-1, respectively. On the other hand, EC50 of Ami on the parameters of cell viability, nuclear area and mitochon?drial mass was 14.83, 6.72 and 4.54μmol·L-1, respectively with Z′value above 0.5. Hypaconitine decreased the SER ridge of mitochondria. Gambogic acid caused significant mortality of H9c2 cells and induced nuclear shrinkage as Ami did. The EC50 values of cell viability and nuclear area were 0.24 and 1.16 μmol · L- 1. Meanwhile,gambogic acid disturbed the mitochondrial function as indicated by the increased mitochondrial area (EC50=0.44 μmol · L-1), abnormal SER Ridge(EC50=0.99 μmol · L-1) and decreased mitochondrial mass(EC50=1.21 μmol · L- 1). Cellular calcium mobilization was lower than normal (EC50=0.41 μmol · L-1). CONCLUSION The EC50 values of positive controls calculated from our assessment are similar those reported in literature. A multi-parameter and simultaneous evaluation enables a comprehensive analysis of the morphology of nuclei and mitochondria of cardiomyocytes and a preliminary assessment of the mechanisms of toxicity.
2.Etiology of recurrent diabetes ketoacidosis in children with type 1 diabetes
jie, YAN ; chun-xiu, GONG ; yan-mei, SANG ; chen, ZHU
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To analyze the etiology of diabetes ketoacidosis(DKA) in children with type 1 diabetes.Methods Totally 850 person-time of type 1 diabetes children in recent 20 years in our hospital were selected as studied subjects. Two hundred and twenty-five person-time of them were hospitalized because of DKA.Fifty-six cases (131 person-time) were due to recurrent DKA.These patients were classified into 2 groups according to onset time: group 1(diagnosed from 1982 to 1991) and group 2(diagnosed from 1992 to 2001).Results The analysis of recurrent DKA suggested that 71.8 % of them was due to infection, 20.4 % of them did not obey diabetic diet and 9.2 % of them discontinued insulin injection. The etiology of DKA showed no difference in two groups. The number of recurrent DKA in two groups was significantly different (P
3.Correspondence analysis of chinese medical constitution features in different ages population.
Jie DI ; Yan-bo ZHU ; Qi WANG ; Yang-yang WANG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(5):627-630
OBJECTIVETo explore the distribution features of Chinese medical constitutions in different ages population, thus providing scientific evidence of constitution process theory.
METHODSRecruited were 21 948 cases from a survey of Chinese medical constitutions and health conditions in 9 provinces or municipalities across China (including Jiangsu, Anhui, Gansu, Qinghai, Fujian, Beijing, Jilin, Jiangxi, and Henan) from Dec 2005 to Jan 2007. The body constitution type of individual was diagnosed using discriminant analysis on the basis of Chinese medical constitution questionnaire. By using correspondence analysis, the correlation between the general population, genders, ages, and Chinese medical constitution types was studied.
RESULTSConstitutions of yin-deficiency type, wetness-heat type, qi-depression type, and special diathesis type often occurred in the population ranging 15 -24 years old. Gentleness type mostly occurred in the population ranging 25 -44 years old. During this time period, phlegm-wetness type and wetness-heat type were liable to occur in males, while blood-stasis type was liable to occur in females. Qi-deficiency type and yang-deficiency type were most often seen in those older than 45 years. Phlegm-wetness type and blood-stasis type body constitution were also liable to occur in those older than 45 years.
CONCLUSIONSThe distribution features were different in different Chinese medical constitutions. Different constitution types exist in different genders. Different constitution types exist different ages population.
Asian Continental Ancestry Group ; Body Constitution ; China ; Discriminant Analysis ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Qi ; Surveys and Questionnaires ; Yang Deficiency ; Yin Deficiency
4.Effect of Sangge Mixture on Cerebral Circulation and Metabolism in Rabbits
Fuwen WANG ; Jie LI ; Yan ZHU ; Shulan XU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(03):-
The effect of Sangge Mixture (SM) on cerebral circulation and metabolism in rabbits was observed.The results showed that SM increased the blood flow of brain and cerebral cortex in a dose-dependent manner and had no effect on the blood flow of femoral arteries. Meanwhile, SM markedly increase the oxygen consumption and glucose intake in rabbit brain. It is suggested that SM can improve cerebral circulation, with a certain degree of selectivity.
5.Quality standard of Zhike Pingchuan Capsules
Ping CHEN ; Ying-Long ZHU ; Qiang WEI ; Bian-Jie YAN ;
Chinese Traditional Patent Medicine 1992;0(03):-
AIM: To establish the quality standard for Zhike Pingchuan Capsule (Radix Panacis Quinquefolii, Semen Pruni Armeniacae, Radix Scutellariae, Radix Glycyrrhizae, Bulbus Fritillariae Cirrhosae, etc.). METHODS: Radix Panacis Quinquefolii, Radix Glycyrrhizae, Bulbus Fritillariae Cirrhosae in Zhike Pingchuan Capsule were identified by TLC and ginsenoside Rb_1 was determined by HPLC. The analysis was carried out on C_~18 column by HPLC. The mobile phase was CH_3CN-H_2O(34∶66). The flow rate was 1.2 mL?min~-1 and the detection wavelength was at 203 nm. The column temperature was at 40.0 ℃ and sensitivity was 0.02 AUFS. RESULTS:The average recovery was 97.20% and RSD was 2.25% and the linear range of ginsenoside Rb_1 was in 0.612-~6.120 ?g. CONCLUSION:This method is simple, rapid with a good reproducibility. This method can be used for the quality control of Zhike Pingchuan Capsule.
6.Effects of hepatocyte growth factors on contractile function of disused gastrocnemius atrophy in rats
Yan HUANG ; Bin YU ; Dang WANG ; Jie ZHU ;
Chinese Journal of Orthopaedic Trauma 2004;0(06):-
Objective To explore the possible effects of hepatocyte growth factors (HGF) on prevention and therapy of disused gastrocnemius atrophy in rats by observing their effects on the wet weight and contractile function of the gastrocnemius of fixed hind limbs. Methods Twenty-four Wister male rats were randomly divided into a blank group, a contrast group and an experimental group. Their right hind limbs were fixed for four weeks with plastic piaster from metatarsus to 1 cm up the knee. Their knee joint was fixed in 100?flexion and their ankle joint in 60?plantar flexion. Rats in the contrast group were injected with 0. 2 mL of normal saline, and rats in the ex- perimental group were injected with HGF (10 mg/kg, 10 mg/mL). Their whole gastrocnemius of the right hind limb was dissected four weeks later to measure its contractile function and wet weight. Results Compared with the blank group, single maximal contraction, maximal tetanus, and wet weight of the gastrocnemius in the contrast and experimental groups decreased significantly (P<0. 05), and fields of electric stimulation also decreased signifi- cantly (P<0. 05). Compared with the contrast group, single maximal contraction, maximal tetanus, and wet weight of the gastrocnemius in the experimental group were significantly superior (P<0.05), and fields of electric stimulation significantly increased. Conclusion Since after a local injection of HGF, the contractile function and wet weight can be significantly increased, HGF may have a preventive effect on disused atrophy.
7.Effect of hepatocyte growth factor on hepatocytes after partial hepatectomy in cirrhotic rats
Hong ZHU ; Yongzhang GUO ; Li LI ; Jie ZHANG ; Zhibin YAN
Chinese Journal of General Surgery 1993;0(01):-
Objective To study the effect of hepatocyte growth factor (HGF) on hepatocytes after partial hepatectomy in chronic liver injury rats.Methods 30% hepatectomy was performed in rats with cirrhosis induced by CCL 4 and alcohol,and subcutaneously injection of cow HGF into intraabdomen was done with different dosages.Results Small dose (0.16mg?100g -1d -1)of HGF did not stimulate liver regeneration or enhance liver function after partial hepatectomy in cirrhotic rats.Large dose (1.3mg?100g -1d -1) of HGF stimulated liver regeneration and enhanced liver function after partial hepatectomy in cirrhotic rats(P
8.Effects of STAT3 aberrant activation on malignant transformation of human umbilical cord mesenchymal stem cells when exposed to human breast cancer microenvironment
Ling WANG ; Jing ZHU ; Jie TIAN ; Bin TAN ; Sha YAN
Basic & Clinical Medicine 2015;(10):1314-1319
Objective_To investigate whether hUCMSCs undergo malignant transformation when exposed to MCF-7B breast cancer microenvironment and whether the abnormal activation and over expression of STAT3 play an impor-tant role in this transformation.Methods_The experiment was divided into three groups:blank group ( hUCMSCs were separately cultured),experimental group(hUCMSCs were indirectly co-cultured with MCF-7B breast cancer cells),positive control group(MCF-7B breast cancer cells were separately cultured).Morphology of cells was detec-ted by invertedmicroscope.Cell cycle was detected by flow cytometry.The mRNA expression of STAT3, c-Myc and Bcl-xL was tested by real-time PCR.The protein expression and location of p-STAT3,c-Myc and Bcl-xL were detected by immunofluorescence.The protein expressions of p-STAT3,STAT3,c-Myc and Bcl-xL were also meas-ured by Western blot.Results_The experimental group cells showed typical morphology of the tumor cells.The cells proportion of experiment group in G1 phase was significantly lower than that of the blank group(P<0.05),but which in S and G2 phase were significantly higher than those of the blank group(P<0.05).The mRNA expression levels of STAT3 ,c-Myc and Bcl-xL in experimental group was significantly higher than those in blank group ( P<0.05).p-STAT3,STAT3,c-Myc and Bcl-xL protein were significantly higher than those of the blank group(P<0.05),and they were mainly located in the nuclei.The protein expression of STATS also showed significant changes in experimental group.Conclusions_hUCMSCs trends to malignant transformations when exposed to MCF-7B breast cancer microenvironment.The abnormal activation and over expression of STAT3 are of important factors leading to the malignant transformation of hUCMSCs.
9.Attenuated store-operated Ca2+ entry in mesenteric artery endothelial cells of aged rat
Xia WANG ; Jie LI ; Yan ZHU ; Juan DU ; Bing SHEN
Chinese Journal of Geriatrics 2015;34(4):434-437
Objective To observe attenuated store-operated Ca2+ entry in superior mesenteric artery endothelial cells of aged rat.Methods The pressure myograph was applied to measure the vasodilation of superior mesenteric artery induced by bradykinin (BK) in aged versus non-elder rats.Primarily cultured MAECs were treated by thapsigargin (TG) and BK to deplete intracellular Ca2+ stores for comparing SOCE.SOCE of MAECs was detected by calcium imaging technology and was compared between aged and non-elder rats.The expression levels of Orai 1 and stromal interaction factor 1 (STIM 1) were observed by immunofluorescence method.Results Compared with the nonelder rats (70.0 ± 4.1 %),BK-induced vasodilation in aged rats (27.0 ± 4.9)% was declined by (60.7±4.3) %.SOCE induced by BK and TG in primarily cultured MAECs was attenuated by 37.6% and 39.2% in aged rats (1.71±0.18 and 1.06±0.03) respectively as compared with non-elder rats (2.72±0.39 and 1.75±0.06).The expressions of SOCE's components Orai 1 and STIM 1 in MAECs were decreased obviously in aged rats than in non elder rats.Conclusions SOCE of MAECs and SOCE-induced vasodilation are significantly attenuated in aged rats.The decreased expression level of Orai 1 and STIM 1 may contribute to this alteration.
10.ox-LDL enhances the mRNA expressions of MMP-9 and LOX-1 in human umbilical endothelial cells and how LOX-1 acts as in inflammation course
Hezhong ZHU ; Xiaoyu SHEN ; Jiajuan CHEN ; Qingmin PAN ; Jie YAN
Journal of Chinese Physician 2012;14(7):920-922
Objective To observe how ox-LDL impacts the mRNA expressions of MMP-9 and LOX-1 of human umbilical endothelial cells (HUEVC) and how LOX-1 acts as in inflamation course.Methods HUVEC were incubated in vitro.mRNA Expressions of MMP-9 and LOX-1 were determined by reverse transcription polymerase chain reaction (RT-PCR).Results Compared to the control group(0.252±0.032;0.279 ±0.041 ),ox-LDL significantly increased the mRNA expressions of MMP-9 and LOX-1 (25 ng/group 0.486 ± 0.012,0.586 ± 0.02;50 ng/L group 0.668 ± 0.011,0.739 ± 0.014; 100 ng/group 0.817 ±0.030,0.872 ±0.003,P <0.01 ).Those expressions were increased by ox-LDL( 1.020 ±0.039)in a concentration-and time-dependent manner.MMP-9 mRNA(0.872 ±0.046) was reduced when LOX-1 was inhibited by polyinosinic acid ( P < 0.01 ).Conclusions The mRNA expressions of MMP-9 and LOX-1 were induced by ox-LDL in HUVEC.Inhibition of LOX-1 may decrease the expression of MMP-9.Those data demonstrate that LOX-1 is involved in the process of ox-LDL-induced MMP-9 expression.