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.

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

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

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.

Objective To explore the effect of different doses of irbesartan on osteopontin expression and fibrosis in diabetic rat kidney. Methods Sixty-three g-week old male Wistar rat were randomly divided into control group (Ctrl group,n=7),diabetes group (DM group,n=14),30 mg·kg-1d-1 hydralazine administrated group (DM+Hyd group,n=12),25 mg·kg-1·d-1 irbesartan administrated group (DM+Irb25 group,n=10),50 mg·kg-1 ·d-1 irbesartan administrated group(DM+Irb50 group,n=9) and 200 mg·kg-1·d-1 irbesartan administrated group (DM+Irb200 group,n=11).Four weeks after modeling,rats were administered with the corresponding dose of irbesartan.After 12 weeks,urinary albumin excretion rate (UAER),endogenous creatinine clearance rate (Ccr) were measured; morphology and collagen deposition in rat kidney were observed by PAS and Masson staining respectively; Ang Ⅱ content in kidney was measured by ELISA; renal tissue TGF-β1 and OPN mRNA expression were detected by real-time PCR. Results UAER and Ccr in the intervention groups of irbesartan were significantly decreased compared with DM group (P＜0.05).UAER and Ccr in DM+Irb200 group were significantly lower than those in DM+Irb25 group and DM + Irb50 group (P＜0.05).Glomerular hypertrophy,mesangial matrix expansion,tubular lesions and deposition of collagen fiber were siginficant in diabetic rats compared with Ctrl,and prevented after administration with different doses of irbesartan.Ang Ⅱ protein level and TGF-β1,OPN mRNA expression in renal tissue of diabetic rats were significantly higher than those in Ctrl group.Ang Ⅱ,TGF-β1,and OPN mRNA expression was significantly reduced after administration with different doses of irbesartan,and with the increase of irbesartan,the above indicators were decreased P＜0.05).Renal local Ang Ⅱ level was positively correlated with OPN mRNA expression (r=0.74,P＜0.01). Conclusion Irbesartan reduces renal TGF-β1,OPN mRNA expression by decreasing kidney local Ang Ⅱ in dose-dependent manner,and eventually reduces tubulointerstitial fibrosis,which plays a role in kidney protection.

Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Objective To study the neuroprotective effects of erythropoietin on intracerebral hemorrhage (ICH). Method The rat models of 1CH were produced by injecting autologous blood into caudate necleus by using stereotatic techique. One hundred ten male wistar rats were randomly divided into 4 groups, namely normal group,sham operation group, 1CU group, and EPO treatment group. The immunohistochemistry and TUNEL were used to detect expressions of Bc 1-2 and Bax,and apoptosis cells. LSD- t and Pearson correlation were used to analyzing data. Results The positive cells of TUNEL Bcl-2 and Bax in ICH group and EPO group obviously increased over 6 hours,and reached peak 72 hours later,and decreased over 120 hours,and the positive cells in different intervals significantly decreased in ICH group and EPO group compared with those in sham operation group (P < 0.01). The positive cells of TUNEL and Bax in EPO group in different intervals significantly decreased compared with those in ICH group (P < 0.01). The Bcl-2 positive cells in EPO group in different intervals significantly increased compared with those in ICH group (P < 0.01). The Bax protein expression, Bax/Bcl-2 and apoptosis presented positive correlation (P < 0.01). Conclusions Apoptosis may induce some brain injury after ICH,and EPO can decrease the number of apoptotic cells after ICH by up-regulating Bcl-2 and down-regulating Bax.

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.

?AIM:To investigate the correlation between dry eye and different degrees of diabetic retinopathy ( DR ) in type 2 diabetic patients.?METHODS: In the cross-sectional study, 340 patients (340 eyes) with type 2 diabetes were enrolled. Tear film function tests including tear meniscus height, tear film breakup time ( BUT ) , fluorescein staining, Schirmer Ⅰtest were performed followed by surveying questionnaires about dry eye. Retinal status was evaluated by retinal color photography and indirect ophthalmoscopy exam with dilated pupils to evaluate DR and whether companied by macular edema.?RESULTS:The prevalence of dry eye was 49. 41%. The mean duration of diabetes in patients with dry eye was 11.15±7.07a, while 6.92±5.45a without dry eye(P<0.01). Dry eye had the positive correlation to the development of DR. The incidence of dry eye in people with mild nonproliferative diabetic retinopathy ( NPDR ) , moderate NPDR, severe NPDR and proliferative diabetic retinopathy (PDR) was 1. 097 times, 1. 724 times, 2. 86 times and 5. 43 times respectively, compared with people without DR. The occurrence of dry eye in people with macular edema increased by 3. 697 times compared with people without macular edema.?CONCLUSION: Dry eye was more prevalent in people with type 2 diabetes. The incidence of dry eye increased gradually with the occurrence and development of diabetic retinopathy.

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