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 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 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 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

AIM: To investigate the variation and significance of mRNA and protein concentration of myeloid cell leukemin-1 (Mcl-1) in apoptotic HepG2 cells induced by apoptin. METHODS: The apoptin expression vector pCDNA3.0-VP3 was transfected into HepG2 cells via liposome. Mcl-1 mRNA was analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction. The protein of apoptin, Mcl-1 and cytochrome C were detected by Western blotting. RESULTS: The VP3 gene was transfected into HepG2 cells successfully and expressed steadily. Compared to blank control, Mcl-1 mRNA and protein levels of VP3 positive cells were decreased (mRNA: 0.09%±0.00% vs 0.41%±0.14%, P<0.05; protein: 0.43%±0.01% vs 0.90%±0.04%, P<0.01). Released cytochrome C from mitochondrion was increased (0.98%±0.02% vs 0.62%±0.03%, P<0.01). CONCLUSION: In the course of the apoptosis of HepG2 cells induced by apoptin, the amount of Mcl-1 mRNA and protein is decreased, and released cytochrome C from mitochondrion is increased. The apoptosis induced by apoptin may be correlated with the down-regulation of Mcl-1 mRNA and protein.

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.

The development of the medical information platform of Henan province is cited as an example,to probe into the organization model of the platform,and the operating and organization model of third-party medical information providers,based on a description of the connotation and development status of the platform,and in consideration of the basic features,technology architecture and key technology requirements of such a platform.

Objective To explore the correlation of S100 protein with the prognosis of patients with primary retroperitoneal liposarcoma. Methods Analyzed the clinical data about 108 patients with primary retroperitoneal liposarcoma managed with surgery from January 2009 to June 2014. All patients were followed up. Patients were divided into S100-positive group(58 patients) and S100-negative group (50 patients) according to the immunohistochemical staining results. The overall survival time and all clinical data between two group were compared. Results All patients with primary retroperitoneal liposarcoma received radical surgical resection for the first time. The overall 5-year recurrence rate were 88.9%(96/108), and the median recurrence time was 32.7 months. The 1-year, 2-year, and 5-year recurrence rates of the S100- positive group were 25.9% (15/58), 53.4% (31/58), 96.6% (56/58), respectively, and the median recurrence time were 26.2 mouths. The 1-year, 2-year, and 5-year recurrence rates of the S100-negative group were 10.0%(5/50), 36%(18/50), 80.0%(40/50) and the median recurrence time were 40.0 mouths. Log-rank test showed that S100 protein expression was significantly associated with postoperative recurrence rates (c2=9.931, P=0.002) and survival time (c2=4.571, P = 0.033). The difference between gender, age, removal of the joint organs and tumor size showed no statistical significance on disease special survival (P>0.05). Cox regression analysis showed that S100 protein expression (OR=1.582, 95%CI:1.005-2.491) and histologic subtype (OR=1.531, 95%CI: 1.254-1.870) were independent risk factors of the prognosis of primary retroperitoneal liposarcoma patients. Conclusions S100 protein played a critical role in retroperitoneal liposarcoma carcinogenesis and its expression may be used as a potential survival predictor in patients with primary retroperitoneal liposarcoma.