Objective To investigate the effects of morphine dependence and withdrawal on neurosteroids and amino acid transmitters of rat amygdala. Methods Morphine dependence was induced by pretreatment with increasing doses of morphine for 7 days. Withdrawal was precipitated by naloxone (2mg/kg). Withdrawal syndromes were observed and scored. After decapitation, amygdala was dissected out. Nomadic and conjugated neurosteroids were extracted using liquid-liquid extraction and solid phase extraction. Concentrations of neurosteroids including dehydroepiandrosterone (DHEA), pregnenolone (PREG), allopregnanolone (AP), dehydroepiandrosterone sulfate (DHEAS) and pregnenolone sulfate (PREGS) were detected with HPLC-MS. Concentrations of glycine (GLY), glutamate (GLU) and gamma-aminobutyric acid (GABA) were quantitated by HPLC-ECD with pre-column OPA derivatization. Results Compared with saline control, the DHEA level in rat amygdala of morphine dependent group decreased by 33% (P＜0.01). Compared with naloxone control, the PREG and AP levels in rat amygdala of morphine withdrawal group increased by 45% (P＜0.05) and 42% (P＜0.05) respectively; the GABA level decreased by 18% (P＜0.01). Compared with morphine dependent group, the PREG and PREGS levels in rat amygdala of morphine withdrawal group increased by 60% and 40% respectively (P＜0.05); the glycine level decreased by 14% (P＜0.05). Conclusion The DHEA in rat amygdala may play a role in the development of morphine dependence but not involved in the manifestation of withdrawal symptoms. Other neurosteroids (including PREG, AP and PREGS) in rat amygdala seem to be involved in withdrawal but not in dependence. The synthesis and release of inhibitory amino acids in amygdala were depressed when withdrawal was precipitated by naloxone. The results suggest that different changes of neurosteroids and amino acids exist in stages of morphine dependence and withdrawal.

Objective:To study the efficacy of prostaglandin E1 in the patients with diabetic nephropathy and its inhibition on inflammatory factors. Methods:Totally 86 cases of diabetic nephropathy from June 2013 to June 2015 in our hospital were selected,and according to the random number,they were randomly divided into the observation group and the control group. The control group was treated with the conventional dietary restriction and therapy regimen including lowering blood pressure and blood sugar. The patients in the observation group were given prostaglandin E1 additionally. After the treatment,24h urine protein quantity,serum creatinine and blood urea nitrogen and the other basic indicators of renal function and the contents of sICAM-1 and hs-CRP were detected. Results:After the treatment,the contents of 24h urine protein,serum creatinine,blood urea nitrogen, sICAM-1 and hs-CRP in the two groups were notably decreased when compared with those before the treatment,and the decrease in the observation groups was more significant than that in the control group,and the difference was statistically significant(P <0. 05). During the course of treatment,no obvious adverse reactions appeared in the two groups. Conclusion:Prostaglandin E1 in the adjuvant treatment of diabetic nephropathy patients shows better therapeutic efficacy and inhibition on inflammatory factors with higher security,which should be promoted in clinics.

Objective: To investigate the effect of morphine dependence and withdrawal on the levels of neurosteroids in hippocampus of male rat.Methods: Rats were given (ip) increasing doses of morphine to form morphine physical dependence, withdrawal syndromes were precipitated by naloxone. The conditioned place preference (CPP) was used to establish morphine psychological dependence. The concentrations of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), pregnenolone (PREG), pregnenolone sulfate (PREGS), and allopregnanolone (AP) in rat hippocampus and plasma were quantified by liquid chromatography-mass spectrometry. Results:The rat model of morphine physical and psychological dependence were successfully established by ip increasing doses of morphine for 7 days and 5mg?kg~ -1 morphine for 10 days respectively. Compared with saline control group, morphine physical dependence increased DHEA and PREG contents in rat hippocampus (0.88?0.19/0.67?0.17,t=2.52,10.94?2.02/7.53?2.64,t=3.24,P

Aim To detect the effect of morphine dependence and withdrawal on the levels of neurosteroids and amino acid neurotransmitters in nucleus accumbens in rat morphine dependent model. Methods Nucleus accumbens was dissected out from morphine dependent and naloxone precipitated withdrawal rats. The contents of neurosteroids including dehydroepiandrosterone, pregnenolone, allopregnanolone, dehydroepiandrosterone sulfate and pregnenolone sulfate were detected with liquid chromatography-negative atmospheric pressure with ionization mass spectrometry(LC-MS). The contents of glycine, glutamate and ?-aminobutyric acid were quantitated by HPLC-ECD with precolumn derivatization. Results Compared with saline group,in nucleus accumbens of morphine withdrawal rats, the level of dehydroepiandrosterone sulfate (P

Objective: To study the regulatory effects of antifibrotic cytokines, interleukin 10 (IL-10), hepatocyte growth factor (HGF), and interferon-gamma (IFN-γ) on activity of TGF-β1 gene promoter, so as to assess the antifibrotic mechanism of cytokines. Methods: Sequence - 1328-+812 of TGF-β1 gene, which contains the - 509 C>T polymorphism, was selected as putative promoter. The recombinant constructions containing - 1328-+ 812 of TGF-β1 gene and CAT reporter gene (phTGF2. 14T, phTGF2. 14C) were constructed and transfected into HepG2 cells with liposomal transfection method, then the transfected HepG2 cells were treated with IL-10(4 ng/ml), HGF(10 ng/ml) or IFN-γ(20 ng/ml). Reporter gene activity was analyzed by ELISA. Results: Reporter gene activity in cells transfected with phTGF2. 14C was significantly higher than those transfected with phTGF2. 14T (P<0.01). IFN-γ significantly inhibited the reporter gene activity in HepG2 cells transfected with phTGF2. 14C or phTGF2. 14T(P<0.05); HGF significantly increased the reporter gene activity in cells transfected with phTGF2. 14C (P<0.05). IL-10 had no effects on the activities of cells transfected with phTGF2. 14C or phTGF2.14T. Conclusion: C allele at - 509 can increase the promoter activity of TGF-β1 gene in HepG2 cells. The antifibrotic effect of IFN-γ might be related to its inhibitory effect on the putative promoter activity of TGF-β 1 gene; the antifibrotic effects of HGF and IL-10 may not be through regulation of TGF-beta1 gene transcription.

Gene Library ; Humans ; Nucleic Acid Hybridization ; methods ; Pulmonary Fibrosis ; diagnosis

Objective The research was to achieve Shanghai resident satisfaction of primary health service after new health reform.Methods During self-compiled questionnaire survey,the research adapted stratified random sampling to select 9 community health centers in Shanghai urban,rural and suburb areas.Results Shanghai resident satisfaction rate of new health reform was 85.2％.Satisfaction rate of basic medical security system was 85.0％.Satisfaction rate of basic medical service was 96.5％.Satisfaction rate of basic public health service was 95.1％.Age,region,job,education and income were main factors to satisfaction of primary health service.Conclusion From an overall point of view,Shanghai resident satisfaction of primary health service after new health reform was high.However,the level of basic medical security and convenience of reimbursement were not high.Medical expenses of primary health care institution had not been effectively controlled.Basic medicine could not fully meet the medication needs of primary health care institutions.

Objective To investigate the effects of Simo decoction on ghrelin and calcitonin gene-related peptide (CGRP) in diabetic rats.Methods A total of 48 SD rats were divided into healthy control group,diabetes group (DM),DM treated with low dose Simo decoction group (DMTL),DM treated with medium dose Simo decoction group (DM-TM),DM treated with high dose Simo decoction group (DM-TH) and DM prevented with medium dose Simo decoction group (DMPM) ; there were eight rats in each group.The rats of DM group and other intervention groups were intraperitoneally injected with 1％ streptozotoein (STZ) to establish the model.The rats of healthy control group were injected with solvent.At eight weeks after the model successfully established,the rats of each intervention group were gavaged with low dose of Simo decoction (0.15 ml · kg-1 · d-1 ),medium dose of Simo decoction (1.5 ml · kg-1 · d-1 ) and high dose of Simo decoction (3.0 ml · kg-1 ·d-1 ) for two weeks; the rats of control group and DM groups were gavaged with equal volume of 0.9％NaCl.The rats of DM-PM group were gavaged with Simo decoction at 1.5 ml · kg-1 · d-1 for 10 weeks after the model successfully established.Rats were sacrificed at the end of drug intervention and gastric antrum tissues were dissociated for detecting the expression of ghrelin and CGRP by real time polymerase chain reaction (PCR).The data were analyzed by one way ANOVA and t test.Results There was no significant difference in ghrelin expression at mRNA level in gastric antrum tissues between diabetic rats and control group.However after intervened by low dose and medium dose of Simo decoction,the ghrelin expression increased (1.45 ± 0.34,1.87 ± 0.68 compared with 0.87 ±0.28,Dunnett's T3 test,P＜0.05).There was no effects of high dose Simo decoction on the expression of ghrelin in diabetic rats.There was no significant difference in ghrelin between DM-TM and DM-PM.The expression of CGRP at mRNA level in gastric antrum tissues of diabetic rats was significantly higher than that of control group (4.61 ± 1.67 compared with 1.06 ± 0.40,t =5.843,P＜0.01).There was no effects of low dose Simo decoction on CGRP expression of diabetic rats.The expression of CGRP of diabetic rats was down-regulated by medium and high dose of Simo decoction intervention.There was no significant difference in CGRP expression between DM-TM and DM-PM.Conclusions Simo decoction can promote ghrelin expression in gastric antrum of diabetic rats.The expression of CGRP in gastric antrum of diabetic rats significantly increased,but Simo decoction may down-regulate the expression of CGRP in gastric antrum of diabetic rats.

Objective To discuss the diet balance by the enteral nutritional support for patients after hematopoietic stem cell transplantation. Methods We divided 47 patients after hematopoietic stem cell transplantation into the balanced-diet group and the ordinary-diet group. The balanced-diet group received diet according to "diet exchange share" method and took food by the proportional share of 5 kinds of food. The ordinary-diet group took food according to the habits of patients.The effect of nutritional support was compared between the two groups. Results The body weight,white blood cell,hemoglobulin,platelet,total protein and proprotein in the balanced-diet group at the 90th day were different from those at the 30th day after transplantation(t value were -8.5,-4.7,-4.2,-10.2,-3.9,-14.1 and -80.9),P ＜ 0.01.In the ordinary-diet group only body weight,platelet and proprotein at the 90th were statistically different from those at the 30th day after transplantation (t value were -4.5,-4.6 and -14.9), P ＜ 0.01. Conclusion Balanced diet contributed to the rehabilitation and nutritional support after hematopoietic stem cell transplantation.

Ubiquitination on target proteins is the signal of cellular protein degradation.Ubiquitin ligase E3 is one of the key enzymes in ubiquitination,it recognizes a specific substrate protein and recruits an ubiquitin conjugating enzyme E2,mediating the ubquitin transfer from the E2 to the substrate protein.Ubiquitin ligase E3 can be divided into two subfamilies according to their different structure characters and function mechanisms,the HECT(homologous to E6AP C terminus) family and the RING-finger family.Members of the HECT E3 share the common HECT catalytic domain,which can bind to an E2 and load the ubiquitin on themselves before catalyzing the transfer of ubiquitin to the target proteins.While the RING-finger E3 all contain an similar E2 binding domain and a unique substrate binding part,mediating direct ubiquitin transfer from the E2 to the substrate.The most recent progresses in the stuctural and functional studies of these two E3 famlies were summarized.