Objective To investigate the impact of citreoviridin(CIT) on mRNA expression of mitochondrial respiratory chain synthesis related transcriptional regulation gene,mitochondrial membrane(MMP) potential and reactive oxygen species (ROS) in cardiomyocytes of rat.Methods Viability of rat primary cardiomyocytes treated with different concentrations of CIT (0,1,2,3,4,5,6,7,8,9,10 μmol/L) for 24 h was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method.Based on the MTT curve,median inhibitory concentration(IC50) was calculated using SPSS 13.0.High-,medium-and low-dose groups of CIT(1.650,1.234,0.715 μmol/L) were defined corresponding to 99％,95％ and 90％ of cardiomyocyte viability,respectively.CIT was not added in as the control group.After 24 hours,the mRNA expression levels of peroxisome-proliferator-activated receptor γcoactivator(PGC-1α),nuclear respiratory factor 1 (Nrf1) and nuclear respiratory factor 2(Nrf2) in cardiomyocytes were detected by reverse transcriptase polymerase chain reaction(RT-PCR).Changes of MMP and intracellular ROS were determined by a fluorescence microplate reader using 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1) and 2,7-dichlorofluorescein diacetate (DCFH2-DA) as fluorescent probes.Results Compared with 0 μmol/L CIT group [(89.4 ± 17.6)％],viabilities of rat primary cardiomyocytes treated with 2-10 μmol/L CIT groups[(80.2 ± 20.2)％,(74.4 ± 18.7)％,(63.2 ± 8.9)％,(51.5 ± 18.8)％,(39.0 ± 15.7)％,(22.6 ± 10.5)％,(19.9 ± 4.9)％,(20.7 ± 4.8)％,(18.5 ± 3.3)％] decreased significantly(all P ＜ 0.05).The IC50 value of cardiomyocytes after24 h treatment with CIT was 4.6 μmol/L The PGC-1α mRNA expressions ofhigh-,medium-and low-dose groups(0.431 ± 0.041,0.619 ± 0.031,0.653 ± 0.037) were significantly lower compared to that of the control group(0.776 ± 0.081,all P ＜ 0.05).The Nrf1 mRNA expression of high-dose group(0.358 ± 0.05) was significantly lower compared to that of the control group(0.580 ± 0.098,P ＜ 0.05).Nrf2 mRNA expressions of the high-and medium-dose groups(0.352 ± 0.041,0.472 ± 0.011) were significantly lower than that of the control group (0.667 ± 0.091,all P＜ 0.05).Compared with the control groups[(100.00 ± 0.00)％,(100.00 ± 0.00)％],the MMP levels of high-,medium-and low-dose groups[(55.3 ± 3.3)％,(69.8 ± 4.7)％,(81.8 ± 2.7)％] were significantly lower and the ROS levels[(606.0 ± 46.3)％,(275.0 ± 53.5)％,(158.9 ±29.5)％] were significantly higher(all P ＜ 0.05).Conclusions CIT inhibits the biosynthesis of mitochondria in primary cardiomyocytes and induces oxidative stress.Myocardial injury is caused by cardiomyocyte apoptosis through mitochondrial pathway,which leads to myocardial injury.

Objective To investigate the effect of menthol compound decoction on the growth and acid metabolism of cariogenic bacteria and to find out the anticariogenic mechanism of Menthol compound decoction .Methods Streptococcus mutans ,Streptococ‐cus sanguinis and Porphyromonas gingivalis were chosen as the experimental bacteria .Firstly ,slip diffusion method was used to ex‐amine the inhibitory effect of menthol compound decoction on cariogenic bacteria .Secondly ,observing the effect of menthol com‐pound decoction of different concentration on acid metabolism .Results Menthol compound decoction could inhibit the growth of Streptococcus sanguinis and Porphyromonas gingivalis .And it also inhibited the acid production of cariogenic bacteria ,but the con‐centration of menthol compound decoction were different .Conclusion Menthol compound decoction may have an inhibitory effect on the growth and acid metabolism of cariogenic bacteria .

Objective　To observe the characteristics and statistics in the bi-variable distribution of thyroid volume (ml) and urine iodine level (μg/L) of children aged 8,9,10 years in China,1999.Methods　Classing analysis.Results　①Data of 12982 cases collected from all country are analyzed with cluster method and three clusters are delimitated as follows;②in first cluster,according to the calculation of median,the volume of thyroid gland is small (2.2ml) and the level of urinary iodine is high (369.1μg/L);③in second cluster,the volume of thyroid gland is middle size (3.4ml) but the level of urinary iodine is significantly lower (88.0μg/L) than that in the first cluster;④in third cluster,the thyroid volume is highest (4.1ml) and the level of urine iodine is high (363.4μg/L),too;⑤the meanings of the classifying results are discussed in detail.Conclusions　The results of the cluster analysis reported here and the typical analyses of the previous papers suggested that no low iodine problems be found out but the correlation between the increasing of thyroid volume and the higher level of urine iodine be observed by us in the studies.

Objective　To discover and discuss the charac ter and statistics of frequency distribution of thyroid volume (TV) in Chinese c hildren aged 8,9,10 years for setting up an effective strategy to control the endemic goiter in China.Methods　The TV is measured by B-ultrasonography.The da ta of a great sample collected from all country are treated with computer centra lly in Chinese Research Center for Endemic Disease Control.Results　①In 1999,the statistics of TV(ml) of children aged 8,9,10 years in China are 3.6 of mean,3.4 of median,3.0 of mode,5.9 of 95% point and 7.9 of 99% point.②The values of 95% point in frequency distribution of TV in 8,9,10 years of boy in order are 4.8,5.6 and 6.3,and in girl they are 5. 2,5.8 and 6.5.③The disparity in medians of TV of children is very significant between the studied provinces;the lowest is Shanghai,the mediums are Anhui and Sh anxi,and the higher ones are Chongqing,Guizhou and Xinjiang,and the medians of the TV in these three categories are 1.2,3.7 and 4.0 in order.④Differential effe cts of palpation method in detection of TV are discussed carefully in this paper .Conclusions　The cause inducing to the significant diffe rences of children TV in several regions in China is not known yet,but it is not the nutritional problem of iodine certainly.

?ln recent years, the number of patients with diabetes increase rapidly. Diabetic retinopathy ( DR ) , one of the complications of diabetes, is also the important aspect of current and future prevention of blindness in our country. Now, more and more scholar have noticed the important role of immune inflammation in the pathogenesis of diabetic retinopathy. ln this article, we reviewed the role of interleukin-6 ( lL-6 ) in diabetic retinopathy.

Objective To study the prevalence of dilated cardiomyopathy and the quasi-latent Keshan disease in villages of non-endemic areas of six Keshan disease endemic provinces in China,and to provide reference values for proposing a elimination standard of keshan disease.Methods County,township and village was selected as the study area by using multi-stage sampling in non-Keshan disease areas of Sichuan,Shanxi,Henan,Shandong and in Keshan disease areas of Chongqing and Yunnan.In each county two townships were selected and in each township one village was chosen.The residents of the villages sampled were surveyed by questionnaire,physical examination,electrocardiogram (ECG) and cardiac echocardiography.Suspected dilated cardiomyopathy patients had chest X-ray.Dilated cardiomyopathy patients were diagnosed according to the criteria proposed by 2006 World Health Organization/International Society and Federation of Cardiology (WHO/ISFC).Results The number of investigated villages was 126 and 54 139 people were surveyed by questionnaire and clinical examination.Ten patients with dilated cardiomyopathy were found,the prevalence was 18.47/100 000(10/54 139),and its 95％ confidence interval was 18.11/100 000-18.84/100 000.A total of 197 patients with quasi-latent Keshan disease were found,the prevalence was 363.88/100 000 (197/54 139),and its 95％ confidence interval was 362.27/100 000-365.49/ 100 000.The prevalence of male quasi-latent Keshan disease was 353.34/100 000(83/23 490) and of female was 372.07/100 000(114/30 639).The number of subjects with normal and abnormal ECG was 45 222 and 8917,respectively,and the rate of abnormal ECG was 16.47％.The highest rate of abnormal ECG was 38.28％ (1585/4141) in Chongqing.The lowest rate of abnormal ECG was 8.10％ (1175/14 507) in Yunnan.The highest detection rate of T wave and ST segment changes was 4.67％ (2528/54 139).In abnormal ECG indices,the detection rate of Henan,Shandong and Chongqing was higher,and all of them were higher than 10.0％.Conclusions We suggest that the reference baselines of dilated cardiomyopathy and quasi-latent Keshan disease in Keshan disease areas of the six provinces in the south of China be 18.47/100 000 and 363.88/100 000,respectively.

Objective To observe the expression of matrix metalloproteinases(MMP)-1,MMP-3 and iNOS in cartilage of experimental rabbit osteoarthritis at different time intervals after anterior cruciate ligament transection(ACLT)operation.The aim of this study is to provide the theoritical evidence for using ACLT rabbit model in Osteoarthritis(OA)research.Methods Unilateral ACLT was performed on 27 randomly selected while rabbits and underwent unilateral arthrotomy was performed on the other 9 white rabbits as the control group.Nine randomly selected white rabbits in experimental group were killed and 3 white rabbits in the control group at 4th,8th and 12th week respectively.Cartilage degradation of femoral condyles was evaluated macr-oscopically,mRNA expression level and protein expression level of MMP-1,MMP-3 and iNOS was measured by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry respectively.Results Forepart OA cartilage degradation was observed at the 4th week and became more severe at the 8th week after ACLF.Afterpart cartilage degradation was evident at the 12th week after ACLT while cartilage still remained normal in the control group,mRNA expression level and protein expression level of MMP-1.MMP-3 and iNOS were increased at the 4th week and became higher gradually at the 8th,12th week after ACLT compared with the control group.Expression distribution of MMP-1,MMP-3 and iNOS bad different patterns respectively.Conclusion It is suggested that the process of OA cartilage degradation can be simulated by ACLT model and MMP-1,MMP-3 and iNOS may be good markers in therapeutical research of OA.

OBJECTIVETo construct the folic acid deficient model in zebrafish and observe the abnormal cardiac phenotypes, to find the optimal period for supplementing folic acid that can most effectively prevent the heart malformation induced by folic acid deficiency, and to investigate the possible mechanisms by which folic acid deficiency induces malformations of heart.

METHODThe folic acid deficient zebrafish model was constructed by using both the folic acid antagonist methotrexate (MTX) and knocking-down dhfr (dihydrofolate reductase gene). Exogenous tetrahydrofolic acid rescue experiment was performed. Folic acid was given to folic acid deficient groups in different periods. The percent of cardiac malformation, the cardiac phenotypes, the heart rate and the ventricular shortening fraction (VSF) were recorded. The out flow tract (OFT) was observed by using fluorescein micro-angiography. Whole-mount in situ hybridization and real-time PCR were performed to detect vmhc, amhc, tbx5 and nppa expressions.

RESULTAbout (78.00 ± 3.74)% embryos in MTX treated group and (68.00 ± 6.32)% embryos in dhfr knocking-down group had heart malformations, including the abnormal cardiac shapes, the hypogenesis of OFT and the reduced heart rate and VSF. Giving exogenous tetrahydrofolic acid rescued the above abnormalities. Given the folic acid on 8 - 12 hours post-fertilization (hpf), both the MTX treated group (20.20% ± 3.77%) and dhfr knocking-down group (43.40% ± 4.51%) showed the most significantly reduced percent of cardiac malformation and the most obviously improved cardiac development. In folic acid deficient group, the expressions of tbx5 and nppa were reduced while the expressions of vmhc and amhc appeared normal. After being given folic acid to MTX treated group and dhfr knocking-down group, the expressions of tbx5 and nppa were increased.

CONCLUSIONSThe synthesis of tetrahydrofolic acid was decreased in our folic acid deficient model. Giving folic acid in the middle period, which is the early developmental stage, can best prevent the abnormal developments of hearts induced by folic acid deficiency. Folic acid deficiency did not disrupt the differentiations of myosins in ventricle and atrium. The cardiac malformations caused by folic acid deficiency were related with the reduced expressions of tbx5 and nppa.

Animals ; Atrial Natriuretic Factor ; metabolism ; Cell Differentiation ; drug effects ; Folic Acid ; metabolism ; Folic Acid Deficiency ; genetics ; metabolism ; Gene Knockdown Techniques ; Heart ; drug effects ; embryology ; growth & development ; T-Box Domain Proteins ; metabolism ; Zebrafish ; embryology ; genetics

Objective To study the effect of atorvastatin on the expressions of intercellular adhesion molecule-1 (ICAM-1) and transforming growth factor-β1 (TGF-β1) in the spinal cord of rats with experimental autoimmune encephalomyelitis (EAE).Methods Eighty healthy female Wistar rats were randomly divided into 4 groups:normal control group (n=20),EAE model group (n=20),low-dose treatment group (n=20) and high-dose treatment group (n=20).Each group was equally divided into 2 sub-groups:14-d group and 21-d group according to the day that they were sacrificed.EAE rat models were established by immunizing the rats with fresh guinea-pig spinal cord homogenate plus complete Freund's Adjuvat (CFA).Rats of the treatment groups were fed with atorvastatin at dasage of 2 mg/(kg·d)and 10 mg/ (kg·d),respectively; and rats of the normal control group and EAE model group were fed with physiological saline.The severity of EAE was scored according to the signs and symptoms,and NIHSS was performed.Pathological changes were observed with the aid of HE staining.The levels of ICAM-1 and TGF-β1 were detected by immunohistochemistry.Results The rats in the high-dose treatment group had significantly lower incidence of disease,decreased CNS inflammation focus,lower level of ICAM-1 and higher level of TGF-β1 as compared with rats of the EAE model group and low-dose treatment group (P＜0.05).Conclusion Atorvastatin could ameliorate EAE of rats in a dose-dependent manner,whose effect might be related to the decreased expression of ICAM-1 and increased expression of TGF-β1.

OBJECTIVETo study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment.

METHODSNinety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-β(1) in lung tissue for each mouse were determined, and the protein level of TGF-β(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected.

RESULTSThe survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 ± 11.65, 44.52 ± 9.48, 43.27 ± 6.01 and 40.82 ± 5.90 mg/g respectively, which were significantly lower than that (74.27 ± 3.68) of PQ group (P < 0.01). The TGF-β(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 ± 7.27, 27.75 ± 5.84 and 21.31 ± 6.82 ng/ml respectively, which were significantly lower than that (52.52 ± 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-β(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01).

CONCLUSIONPF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs.

Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Paraquat ; poisoning ; Pulmonary Fibrosis ; chemically induced ; drug therapy ; pathology ; Pyridones ; therapeutic use ; Transforming Growth Factor beta ; metabolism