Objective To investigate the effect of telmisartan and irbesartan on PPARα transcriptional activity, and to clarify their molecular mechanisms in improving glucose and lipid metabolism. Methods The structural expression vectors, including pCMV-PPARα, pGL3-PPRE and the internal control vector pRL-TK, were transiently eo-transfected into COS-7 cells using SuperFect, the cells were eontinously cultured with various concentrations of telmisartan and irbesartan, and then the PPRE controlled luciferase activity was determined by using a dual-luciferase reporter gene assay system. PPARα mRNA and protein expression levels were detected by RT-PCR and Western blot after 3T3-L1 adipoeytes were treated with various concentrations of telmisartan or irbesartan. Results (1) Both telmisartan and irbesartan stimulated PPARα transcriptional activity in concentration-and time-dependent manners in cultured COS-7 cells with the maximal effect at 60 h, with the results increased by 3.8 and 2.6 folds respectively at the concentration of 100 μmol/L compared with control group (both P<0.01). (2) The PPARγ antagonist GW9662 did not inhibit fenofibrate, telmisartan and irbesartan-stimulated PPARα transcriptional activities. (3) Both telmisartan and irbesartan increased PPARα mRNA and protein expression levels in a dose-dependent manner in 3T3-L1 adipocytes. Conclusion Angiotensin type 1 receptor blockers, telmisartan and irbesartan, can both increase PPARα transcriptional activity, which may contribute to their metabolic effects.

Objective To explorer the effect of angiotensin type 1 receptor blockers (ARB) on activating peroxisome proliferator-activated receptor (PPAR) ? and ?.MethodsLuciferase gene reporters of PPAR? and PPAR? were constructed in COS-7 cells.The cells were then cultured with various concentrations (0,0.01,0.1,1,10 and 100 ?mol/L) of valsartan,losartan,irbesartan or telmisartan for 12,24,36,48 and 60 hours;or co-cultured with PPAR? antagonist GW9662 (10,30?mol/L) for 1 hour,and then cultured with various concentrations (0,1 and 10?mol/L) of irbesartan or telmisartan for 12,24,36,48 and 60 hours.The peroxisome proliferator-response element (PPRE) luciferase activity was determined by Dual-Luciferase Reporter Gene Assay system.3T3-L1 cells were induced to differentiate into adipocytes,and then co-cultured with 10?mol/L of valsartan,losartan,irbesartan or telmisartan for 24 hours,and the expressions of PPAR? and PPAR? mRNA and protein were detected by RT-PCR and Western blotting respectively.ResultsStimulation of valsartan and losartan didn't increase the transcriptional activities of PPAR? and PPAR? in COS-7 cells,while of irbesartan and telmisartan significantly increased the transcriptional activity of PPAR? and PPAR? in COS-7 cells.After co-cultured with 100 ?mol/L of irbesartan or telmisartan for 48 hours,the PPAR? transcriptional activities reached their peak values (43.3?13.0 and 47.8?11.8 respectively),and were significantly higher than that of control group (4.3?0.5,P

Objective To study the protection effect of acupuncture at acupoint ST 36 in rat with cold-restrainted stress-induced ulcer,and to observe the expression of NOS in hypothalamus and adrenal. Methods Ulcer Index in rats was used to study the protection effect of acupuncture, and and RT-PCR was employed to study the expression of NOS in hypothalamus and adrenal. Images were analyzed with semi-quantitative method. Results The ulcer Index was significantly decreased in rats with stress-induced ulcer by acupuncture as indicated by a comparison with those without acupuncture. The expression of NOS1 in hypothalamus was increased by acupuncture protect stress-induced ulcer. The increased expression of NOS2 was involved in stress-induced ulcer, and acupuncture decreased its expression. The expression of NOS3 in hypothalamus had the similar reaction to NOS2, but the decreased effect of acupuncture was limited. The expression of NOS1 and NOS3 in adrenal were increased by cold stress, only the expression of NOS1 could be repressed with acupuncture. There was no NOS2 expression in adrenal in stress-induced ulcer rats. Conclusion The protection effect in stress-induced ulcer exerted by acupuncture at acupoint ST 36 were mediated by increase of the physiological expression of NOS1 in hypothalamus as well as decrease of the expression of NOS2,NOS3 in hypothalamus and repress of the expression of NOS1 in adsenal.

0.05)]. Conclusion The elevated serum HA is linked to a potential marker in kidney transplantation.

Objective To evaluate the clinical value of the level of plasma procalcitonin,blood lactic acid an1 endotoxin in patients of severe pneumonia complicated with sepsis.Methods The 40 cases of severe pneumonia complicated with sepsis(observation group)were analyzed retrospectively,they were divided into survival group included 20 cases and the death group included 20 cases.Meanwhile the 20 cases of healthy persons were selected as control group.The worst score of Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) within 24 hours after admission were record.The level of plasma procalcitonin,blood lactic acid and endotoxin were compared between three groups.In addition do a correlation study between the above indexes and the score of APACHE Ⅱ.Results The level of plasma Procalcitonin,blood lactic acid and endotoxin of observation group increased significantly compared with the control group [(0.02±0.01 ng/ml,0.87 ± 0.27 mmol/L,4.15±1.63 pg/ml) vs (18.29±11.02 ng/ml,6.55 ± 3.02 mmol/L and 15.5±10.38 pg/ml),t=10.48,11.79,6.75,all P＜0.05].The level of plasma procalcitonin,blood lactic acid and endotoxin of the death group increased significantly compared with the survival group [(9.52±2.93 ng/ml,4.26±1.78 mmol/L,7.62±3.04 pg/ml) vs (27.06±8.88 ng/ml,8.84± 2.14 mmol/L and 23.39± 9.00 pg/ml),t=8.39,7.35,7.42,all P＜0.05].In the all patients of severe pneumonia complicated with sepsis,there was positive correlation among plasma procalcitonin,blood lactic acid,endotoxin and the score of APACHE Ⅱ (r=0.919,P=0.001;r=0.914,P=0.002;r=0.909,P=0.004).Conclusion The level of plasma procalcitonin,blood lactic acid and endotoxin are very important indexes in assessment of the severity and the prognosis of severe pneumonia complicated with sepsis,that has important value in clinical application.

ObjectiveTo study the special effect of oxytocin and vasopressin mRNA expression by acupuncturing acupoint ST36.Methods78 SD male rats were divided into four groups, acupuncturing ST36 group, acupuncturing para-acupoint ST36 group, bondage group and control group. RT-PCR was used to observe the oxytocin and vasopressin mRNA expression in 2,4,6,8 hours after acupuncture, semi-quantitative result was obtained by using image analysis system.ResultsOxytocin mRNA expression in hypothalamus in 2,4 hours was the statistically significant different in acupuncturing para-acupoint ST36 group contrast with other three groups, while there was not statistically different between all groups on oxytocin mRNA expression in 6,8 hours. Vasopressin mRNA expression in hypothalamus in 2,4,6,8 hours was statistically depressed in acupuncturing acupoint ST36 contrast with other three groups.ConclusionOxytocin mRNA expression in hypothalamus increases for short time after acupuncturing para-acupoint ST36. Vasopressin mRNA expression in hypothalamus decreases for long time after acupuncturing acupoint ST36.

ObjectiveTo investigate effects of electroacupuncture(EA) of Zusanli(ST36) on changes of substance P(SP) mRNA expression in rat brain stem.Methods78 SD rats were evenly randomized into Zuanli-EA group, non-acupoint group, bondage groups(at the end of 2h,4h,6h,8h after 30 minute EA stimulation in each group) and control group. RT-PCR and image processing were used to study the change of SP mRNA level in brain stem.ResultsIn control groups,there was SP expression. An increase of SP mRNA expression was seen in Zusanli group and non-acupoint group comparing with control group at the end of 2h after 30min EA stimulation(P<0.05).There was difference between above two groups at the end of 6h and 8h(P<0.05).ConclusionEA stimulation elicited an accelerated expression of SP gene. Zusanli-EA stimulation can increased this expression, which may constitute one of the mechanisms for gastrointestinal motility and eletroacupuncture analgesia.

Objective To amplify the recombinant adenovirus vector carrying rat angiotensin Ⅱ type 2 receptor (AT2R) gene using human embryonic kidney (HEK) 293A cell lines and to construct a pancreatic islet βcell model overexpressing AT2R by transfecting the adenovirus vector into rat insulinoma (INS-1) cell lines.Methods Recombinant adenovirus vector Ad-G-AT2R-EGFP and control vector Ad-CMV-EGFP were amplified with HEK 293A cells and the titer of the adenovirus was detected .After both adenovirus vectors were transfected into INS-1 cells,AT2R and angiotensin Ⅱtype 1 receptor(AT1R) gene expressions were tested using real-time PCR, Western blotting, immunofluorescence staining and confocal laser-scanning microscopy .Results The titer of amplified Ad-G-AT2R-EGFP and Ad-CMV-EGFP was re-spectively 9 ×109 pfu/ml and 8 ×109 pfu/ml.Transfection of Ad-G-AT2R-EGFP into INS-1 cells induced an increase in AT2R mRNA expression in a dose-dependent manner , and significantly increased AT2R mRNA and protein expression compared with Ad-CMV-EGFP-or mock-transfection.Conclusion The recombinant adenoviral vector carrying AT2R gene is successfully amplified and an INS-1 cell model overexpressing AT2R is constructed by transient transfection , which can contribute to further study of the role of AT2R in pancreatic islet βcells.

BACKGROUND:In joint surgery, the commonly used autologous chondrocyte transplantation often used to repair cartilage defects, and poor integration is one of the reasons that leading to failure repairing. Chondrocytes migration capability is proven to have correlation with integration and some pathways, such as Src-phosphorylated phospholipase Cγ1-extracellular regulated kinase 1/2 has been confirmed to have correlation with the migration ability of chondrocytes, but the correlation with the integration is stil unknown. OBJECTIVE:To determine the chondrocyte signaling pathways involved in autologous chondrocyte migration and their effects on cartilage integration in autologous chondrocyte implantation. METHODS:Articular chondrocytes were isolated from immature pig knee joints. The cells were divided into four groups:Src group, phosphorylated phospholipase Cγ1 group, extracellular regulated kinase 1/2 group and control group, then the Boyden chambers were used to quantify the chondrocyte migration. The chondrocytes/cartilage ring integration model was developed and cultured for 28 days, and then histology, biochemistry, biomechanics, western blot analysis and celltracking analysis were performed to observe the differences between the control group and the suppression groups. RESULTS AND CONCLUSION:The migration ability of chondrocytes was significantly decreased after pretreated with inhibitors. After the chondrocytes/cartilage ring co-cultured for 28 days, Western blot analysis showed that the pathway inhibitors has been presented in the entire culture cycle. The number and length of chondrocytes migrated into the integration area, col agen secretion level, matrix and mechanical strength in the control group were higher than those in three suppression groups. The results suggest that chondrocyte migration ability can affect the cartilage integration capability through Src-phosphorylated phospholipase Cγ1-extracellular regulating kinase 1/2 signal transduction pathway.

BACKGROUND:With the development of tissue engineering, autologous chondrocyte implantation is often used to repair cartilage defects. And poor integration is one of the common reasons that lead to failure repairing. Many models in vitro are used for related studies.
OBJECTIVE:To develop an interface integrated model of tissue engineered cartilage repair in vitro and to evaluate the effect.
METHODS:Cartilage integration model in vitro was established in pigs. Total y 21 cartilaginous rings were obtained and divided into agarose gel group (n=18) and control group (n=3). In agarose gel group, cartilage rings were covered with agarose gel. Chondrocytes were separated and implanted into the ring. The leakage of cells around the cartilage rings was observed. The sections were stained for histological observation at 1, 2, 4 weeks. The average area of neochondrocytes was measured and compared.
RESULTS AND CONCLUSION:The results from the control group were not processed, because there was no chondrocyte aggregate formation in the center of the explant ring due to earlier chondrocyte leakage outside the explant. While no chondrocytes were found outside the explant ring in the agarose gel group. Tissue sections of the agarose gel group were stained by hematoxylin and eosin, alcian blue, Safranin-O and col agen type II immunohistochemistry at 1, 2, 4 weeks. Neochondrocytes proliferated within cartilage ring, and produced extracellular matrix. After 2 weeks of incubation, these inserted chondrocytes were significantly increased. There was no statistical y significant increment between 2 weeks and 4 weeks (P>0.05), although the area was further increased by 4 weeks. This model provides a convenient simulation of the cartilage integration process in vitro and has a potential application in studies of cartilage integration and cartilage tissue engineering.