Effects of Rapamycin and mammalian target of rapamycin - small interfering RNA (mTOR siRNA)on proli-feration,apoptosis and extracellular matrix in premature rats lung fibroblasts exposed to hyperoxia / 中华实用儿科临床杂志

Objective To investigate the effects of Rapamycin and mammalian target of rapamycin - small in-terfering RNA (mTOR siRNA)on the proliferation,apoptosis and collagen Ⅰ(COLⅠ),collagen Ⅲ(COLⅢ)and fi-bronectin(FN)in premature rats lung fibroblasts exposed to hyperoxia. Methods 900 mL/ L volume fraction of oxygen was used to establish hyperoxia - damaged cell models,and the premature rats lung fibroblasts were divided into air control group,hyperoxia group,hyperoxia + rapamycin group and mammalian target of rapamycin - small interfering RNA transfection group. Cell proliferation was assessed by using 3 -(4,5 - Dimethylthiazol - 2 - yl)- 2,5 - dipheny-ltetrazolium bromide assay. Apoptosis were detected by Annexin V - FITC and propidium lodide (PI)double staining. The expressions of COLⅠ,COLⅢ and fibronectin was assessed by using enzyme linked immunosorbent assay and Bcl - 2,P53 and pro - fibrotic factors of connective tissue growth factor(CTGF)and transforming growth factor β(TGF - β)by using Western blot. Results Compared with the air control group,the proliferation of lung fibroblasts decreased and the apoptosis increased in the hyperoxia group,while the contents of COLⅠ(28. 30 ± 0. 53 vs. 17. 43 ±0. 37),COLⅢ(27. 86 ± 1. 02 vs. 17. 43 ± 0. 37)and fibronectin(32. 87 ± 0. 42 vs. 21. 57 ± 0. 47),P53(0. 810 ± 0. 119 vs. 0. 160 ± 0. 018),TGF - β(0. 580 ± 0. 108 vs. 0. 210 ± 0. 008)and CTGF(0. 590 ± 0. 017 vs. 0. 220 ± 0. 007)were also increased but the expression of Bcl - 2(0. 150 ± 0. 004 vs. 0. 600 ± 0. 130)protein was decreased, and the differences were all statistically significant (all P < 0. 01). Compared with the hyperoxia group,the proliferation of lung fibroblasts was increased in the hyperoxia + rapamycin group,but the apoptosis was decreased,the contents of COLⅠ(23. 17 ± 0. 60 vs. 28. 30 ± 0. 53),COLⅢ(17. 09 ± 0. 58 vs. 27. 86 ± 1. 02)and fibronectin(28. 11 ± 0. 68 vs. 32. 87 ± 0. 42),P53(0. 430 ± 0. 008 vs. 0. 810 ± 0. 119),TGF - β(0. 380 ± 0. 008 vs. 0. 580 ± 0. 108)and CTGF (0. 040 ± 0. 006 vs. 0. 590 ± 0. 017)were decreased while the expression of Bcl - 2(0. 290 ± 0. 009 vs. 0. 150 ± 0. 004) protein was increased,and the differences were all statistically significant (all P < 0. 01). In the mTOR siRNA transfec-tion group,compared with the hyperoxia + rapamycin group,the proliferation of lung fibroblasts was increased,but the apoptosis was decreased;the contents of COLⅠ(15. 71 ± 0. 34 vs. 23. 17 ± 0. 60),COLⅢ (13. 85 ± 1. 36 vs. 17. 09 ± 0. 58)and fibronectin(20. 18 ± 0. 28 vs. 28. 11 ± 0. 68),P53(0. 300 ± 0. 006 vs. 0. 430 ± 0. 008),TGF - β(0. 150 ± 0. 002 vs. 0. 380 ± 0. 008)and CTGF(0. 140 ± 0. 004 vs. 0. 040 ± 0. 006)were decreased while the expression of Bcl - 2 (0. 460 ± 0. 012 vs. 10. 290 ± 0. 009)protein was increased,and the differences were all statistically significant (all P < 0. 01). Conclusion Rapamycin and mTOR siRNA can protect lung injury caused by hyperoxia and have a certain inhibitory effect on pulmonary fibrosis,and mTOR siRNA effect is more obvious,so the mechanism may be through the inhibition of mTOR signaling pathway.

Role of mammalian target of rapamycin protein in oxidative stress-induced lung injury and pulmonary fibrosis in immature rats / 中国新生儿科杂志

Objective To study the changes of transforming growth factor-β (TGF-β),connective tissue growth factor (CTGF) and collagen 1 (COL1) in newborn rat's lung tissue and the expressions of 4EBP1 (eukaryotic promoter) and P7OS6K (mammalian target of rapamyein pathway downstream target protein) after rapamycin and hyperoxia intervention,and to study the influence of mammalian target of rapamyein (mTOR) pathway on hyperoxic lung injury and the possible intervention methods.Method A total of 48 21-day-old neonatal rats were assigned into 8 groups (n =6),including air control group,3 d group (3 days after hyperoxic exposure),7 d group (7 days after hyperoxic exposure),14 d group (14 days after hyperoxic exposure),air + RAPA group (air + rapamycin),3 d + RAPA group (3 days after hyperoxic exposure + rapamycin),7 d + RAPA group (7 days after hyperoxic exposure + rapamycin) and 14 d + RAPA group (14 days after hyperoxic exposure + rapamycin).In the hyperoxic group,newborn rats were exposed to 90％ oxygen for 3,7,14 days respectively.The rats in the hyperoxia + rapamycin intervention groups received intraperitoneal injection of rapamycin and inhaled high concentrations of oxygen for 3,7,14 days respectively.Air ± rapamycin group received intraperitoneal injection of rapamycin for 3 days.To study the pathological changes of lung tissures after hyperoxia and rapamycin intervention,we used ELISA to detect the changes of TGF-β,CTGF and COL1 and Western blot to detect the variations of mTORC1,P70S6K and 4EBP1 expression.Result TGF-β,CTGF,COL1 levels at 3 days,7 days and 14 days after hyperoxic exposure (TGF-[β:33.7±2.8 vs.58.6 ±3.1 vs.98.8 ±1.5 ng/mg,CTGF:50.1 ±1.8 vs.68.7 ± 2.2 vs.94.4 ±2.5 ng/mg,COL1:471.9 ±5.7 vs.529.7 ±7.0 vs.556.4 ±8.5 ng/mg) were significantly higher than the air control group (TGF-β:25.5 ± 1.9 ng/mg,CTGF:41.7 ± 1.4 ng/mg,COL1:414.4 ± 8.9 ng/mg) (P ＜ 0.01).While the levels in rapamycin intervention group were significantly lower than all the hyperoxia + rapamycin intervention groups (P ＜ 0.01).The lung tissue pathological grades in 3 d + RAPA group and 7 d + RAPAgroup were significantly lower than those in the 3 d group and 7 d group (3.5 ± 0.8 vs.6.3 ± 2.3 and 9.7 ± 2.0 vs.14.0 ± 2.4) (P ＜ 0.01).The mTORC1,P70S6K,4EBP1 expressions in 3 d + RAPA group were lower than 3 d group (mTORC1:0.26 ± 0.04 vs.0.29±0.08,P70S6K:0.29±0.01 vs.0.31 ±0.08,4EBP1:0.31 ±0.06 vs.0.33 ±0.06) (P＜0.05),while the expressions in 7 d + RAPA and 14 d + RAPA groups were significantly lower than 3 d + RAPAgroup (P ＜0.01).Conclusion mTOR signal pathway may be involved in the repairing process of hyperoxic-induced lung fibrosis.Rapamycin can reduce the levels of TGF-β,CTGF and COL1 and inhibit the expressions of mTOR pathway downstream target protein P70S6K and 4EBP1,thus reduce lung injury atearly stage.

The value of color doppler flow imaging in diagnosis of liver cirrhosis upper gastrointestinal bleeding about portal vein hemodynamic changes / 中国医学装备

Objective:To investigate the value of color doppler flow imaging (CDFI) in the diagnosis of liver cirrhosis upper gastrointestinal bleeding in portal vein hemodynamic changes. Methods: 96 cases of patients with liver cirrhosis were selected who were diagnosed in our hospital, according to the history whether patients had a gastrointestinal bleeding or not. They were divided into bleeding group(45 cases) and no bleeding group(51 cases). At the same time, we chose the hospital physical examination center of 42 cases of healthy volunteers as a control group, using color doppler flow imaging portal venous blood flow mechanics parameters, including diameter, average blood flow velocity and blood flow of portal vein(PV) and splenic vein(SV) and compare the data of the three groups.Results: Compared with control group, the patients with liver cirrhosis, the blood vessel diameter have increased whether bleeding or not. The average blood flow velocity is slower and PVF is larger, and the differences between them are statistically significant(t=3.579,t=3.670,t=4.750,t=3.951,t=6.116,t=5.371;P<0.05). Conclusion: The clinical application of color doppler flow imaging(CDFI) in patients with cirrhosis portal hemodynamic change is not only simple noninvasive, and there is important diagnostic value in the detecting parameters.

Cell signaling pathways and bronchopulmonary dysplasia / 国际儿科学杂志

The bronchopulmonary dysplasia(BPD),one of the most common complications in prema-ture infants,has become one of the most difficult problems in neonatal intensive care unit. The molecular mecha-nism of BPD is extremely complicated,of which the pathogenesis process requires the participation of many sig-nal transduction pathways. This article summarizes the probable relationship of mitogen activated protein kinases signal pathways,nuclear factor-κB pathways,transforming growth factor beta pathways,Wnt pathways,mTOR pathways with BPD.

Effects of liraglutide on the expression of genes related to cholesterol metabolism in ApoE-/-mice with hypoadiponectinemia / 中华内分泌代谢杂志

Objective To investigate the effects of liraglutide on gene expression related to cholesterol metabolism in ApoE-/-mice with adiponectin deficiency. Methods Thirty six ApoE-/-mice fed with the high-fat diet were subdivided into four groups. One group was given 100 μl(1×109PFU) of adenoviral pAd-U6-GFP(GFP group, n=6). The second group received 100 μl of adenoviral pAd-U6-Acrp30(ADI group, n=10). The third group was given 100 μl of adenoviral pAd-U6-Acrp30 and liraglutide(HEA group, n=10) and the fourth group was given only 100 μl sterile saline(HF group, n=10). Insulin sensitivity and glucose metabolism were assessed by the hyperinsulinemic-euglycemic clamp technique using 3-[3H] glucose as a tracer. Plasma adiponectin level was evaluated using a commercially available ELISA kit. The mRNA expressions of genes involved in cholesterol metabolism were measured by quantitative realtime PCR. Results Fasting blood glucose(FBG), free fatty acids(FFA), total cholesterol, triglyceride, low density lipoprotein cholesterol, adiponectin, and fasting plasma insulin(FINS) in ADI mice were significantly higher than those in the other groups(P＜0.01), while high density lipoprotein cholesterol was significantly lower(P＜0.05). During the clamp, glucose infusion rate(GIR) in ADI group was significantly lower than the other groups(P＜0.01), and hepatic glucose production(HGP) significantly higher in ADI group(P＜0.01). The mRNA expressions of INSIG2 and LDLR in ADI group were significantly down-regulated in HEA group(P＜0.01 or P＜0.05), while HMGCR and SREBP-2 were significantly up-regulated in HEA group(P＜0.01 or P＜0.05). Conclusions Liraglutide regulates a number of genes involved in cholesterol metabolism and ameliorates hypercholesterolemia by elevating plasma adiponectin level.

Comparison between MR relaxometry techniques and dual-energy X-ray absorptiometry in evaluation of osteoporotic postmenopausal rats / 中华放射学杂志

Objective To evaluate MR relaxometry techniques and dual-energy X-ray absorptiometry (DXA) for the diagnosis of osteoporotic diseases in rats. Methods Thirty 3-month-old female rats were randomly divided (using completely randomized grouping method) into two groups (each contained 15 rats). Animals in group A without osteoporotic castration were included as normal controls, whereas osteoporotic castration was created in each animal in group B. Three parameters (BMC, BMD, Hbmdl)was measured for both groups by DXA at two time points, one immediately before the castration and another at the 12 th week after the castration. Then animals from the control group and the osteoporotic group went through the following three diagnostic procedures using a 1.5 T MR system: (1) A fast multi echo gradient echo (MEGRE) pulse train sequence with different inter-echo intervals (1000, 500, 400, 300, 200, 100) to obtain the T_2~* value. (2) A multi-echo fast spin echo sequence to obtain the T_2map. (3) A conventional spin-echo (CSE) sequence to obtain the T_1map. The statistical difference between group A and group B was tested by t-test to analyze parameters. And, the most significant parameter for diagnosis ofosteoporotic diseases was picked out from all parameters by Fisher Sequential diseriminant analysis. At the end of experiments, animals were killed and histopathological examination was performed on the femurs of animals from both control and osteoporotic groups. Results (1) Histopathological examination confirmed the presence of osteoperosis in all animals in group B. (2) BMD was picked out from 3 DXA parameters (BMC,BMD,Hbmdl) by fisher stepwise discriminant analysis, and its discriminant rates was 87.6%. (3) All 2-sample t-test results(t=6.20, 4.79, 5.18, 5.22, 5.59, 4.37, 6.14, 5.12, 5.09, 4.99, 5.57, 4.84, 4.07, 2.98, 6.75 individually) for MR relaxometry parameters(T_2~* 1000,R_2~* 1000,T_2~* 500,R-2~* 500,T_2~* 400,R_2~* 400,T_2~* 300,R_2~* 300,T_2~* 200, R_2~* 200, T_2~* 100, R_2~* 100, T_2map, R_2map, T_1map) showed statistically significant differences between groups A and B (P=0.01 for T_2~* map, P=0.00 for all other parameters) except the R_2map(P=0.07). (4) Using fisher stepwise discriminafion method in the analysis of 14 parameters of MR relaxometry techniques and 3 parameters of dual X-ray absorptiometry(T_2~* 1000,T_2~* 500,T_2~* 400,T_2~* 300,T_2~* 200,T_2~* 100,T_2map, R_2~* 1000, R_2~* 500, R_2~* 400, R_2~* 300, R_2~* 2OO, R_2~* 100,T_1map,BMC,BMD,Hbmdl), we found that the most significant difference was from the T_2map and T_1map. Conclusions The MR relaxometry parameter-T_2map in the present study is shown to be appropriate parameter for the diagnosis of osteoperotie diseases, and stability of magnetic field plays an important role in this process. It would be the optimal method to make a diagnosis of osteoporotic diseases with both MR relaxometry and DXA technological means.

Brain activation of acupuncture stimulation at Zusanli acupoint on heroin addicts / 中华行为医学与脑科学杂志

0bjective To observe the different brain activation of acupuncture and electroacupuncture on healthy subjects and healthy subjects.Methods The different brain activation involved in heroin addiction between healthy subjects and addicts was detected by fMRI.Acupuncture point used in present study Was Zusanli (ST 36).Results Different brain activations between healthy subjects and addicts during electroacupuncture were hypothalamus(X0,Y2,Z9,t=7.36,P<0.01),anterior cingulate(X5,Y49,Z8,t=4.11,P<0.01),tempo-ral gyrus(X61,Y12,Z8,t=3.05,P<0.01).The difference of activated regions during conventional acupuncture between healthy subjects and heroin addicts was thalamus(X2,Y16,Z12,t=2.87,P<0.01),parahippocampus (X17,Y52,Z3,t=3.14,P<0.01),and hypothalamus(X0,Y2,Z9,t=6.98,P<0.01).Conclusion Regions with significant activation detected by fMRI are different during acupuncture in heroin addicts and in the healthy subjects.Notably,the hypothalamus activation is more robust in the addicts than in the healthy subjects during ac-upuncture stimulation.

A Study of Right Hemisphere Function of the Nonverbal Learning Disabled Children / 中国临床心理学杂志

Objective: To study the right hemisphere function state of NLD children. Methods: Adapting tachistoscopic vision and Benton Revised Visual Retention Test (VRT), three groups involving 20 children each were studied. Results: Under the tachistoscopic vision, NLD children achived poorly in recognition of nonverbal stimulus; and they do worse also in VRT test, with more errors of omission and distortion. Conclusion: Compared with normal children, the function of NLD children's right hemisphere is relatively weaker.

Triglyceride level changes and potential mechanism on alcoholic fatty liver cell model induced in vitro:with or without diammonium glycyrrhizinate / 重庆医科大学学报

Objective:To study the triglyceride level changes with or without Diammounium Glycyrrhizinate(DIG)and the potential mechanism on alcoholic fatty liver cell model induced in vitro.Methods:MTT assay was used to screen the optimal concentration of ethanol and DIG injection.Ultra-microstructure was observed under transmission electron microscope.Intracellular triglyceride concentration was measured by hol-automatic biochemistry.Expressions of peroxisome proliferator-activated receptor-gamma (PPAR?),sterol regulatory element binding protein-1(SREBP-1)and SREBP cleavage activating protein(SCAP)were determined by immunocyto-chemistry(ICC).Results:1.The screened optimal DIG concentration was 2.273?g/ml.2.Masses of lipid droplets in cytoplasm were observed under transmission electron microscope.3.Hol-automatic biochemistry measured intracellular triglyceride concentration to be(1.84?0.16)mmol/L and(1.05?0.12)mmol/L respectively in Ld30 group and DIG group,and the difference was statistically significant(P