Objective To investigate and analyze the effect of serum micro inflammatory and nutritional indexes of psychological intervention combined with enalapril maleate tablets on the treatment of the patients with maintenance hemodialysis. Methods 100 patients with chronic renal failure in Jiande the first people's hospital from February 2015 to August 2016 were selected as the subjects, and randomly divided into the control group and the experimental group. The control group and the experimental group were given enalapril maleate tablets, at the same time, the experimental group were received psychological intervention. The changes of serum micro inflammation indexes and nutritional indexes in the the two groups were compared and analyzed. Results The HsCRP, TNF alpha levels in the experimental group decreased significantly than those in the control group, the differences had statistical significant (P<0.05). The levels of prealbumin and plasma albumin in the experimental group were significantly higher than those in the control group (P<0.05). After treatment, the SAS score in the experimental group was (42.3±5.2) points, and the SDS score was (40.1±5.9) points. The SAS score in the control group was (60.9±9.2) points, and the SDS score was (59.1±7.4) points. The scores of depression and anxiety in the experimental group were significantly lower than those in the control group (P<0.05). Conclusion Psychological intervention combined with enalapril maleate tablets used in maintenance hemodialysis patients, which can significantly improve patients' serum inflammation index, restore the nutritional status of patients, eliminate the negative emotion of patients, with further clinical promotion and application significance.

The effects of advanced glycation end-products (AGEs) on number and activity of vascular endothelial cells (VEC) from hone marrow stem cells (BMSC) of mice were investigated. 100 μ/ml AGEs markedly inhibited differentiation of BMSC into VEC with decreased vascular endothelial growth factor receptor-2 positive cells, hut 20 μg/ml AGEs had no effect.

Research on novel pullulanase has major significance on the domestic industrialization of pullulanase and the breakdown of foreign monopoly. A thermophilic bacteria LM 18-11 producing thermostable pullulanase was isolated from Lunma hot springs of Yunnan province. It was identified as Anoxybacillus sp. by 16S rDNA phylogenetic analysis. Full-length pullulanase gene was cloned from Anoxybacillus sp. LM18-11. The optimum temperature of the pullulanase was between 55 and 60 degrees C with a half-life as long as 48 h at 60 degrees C; and its optimum pH was between 5.6 and 6.4. V(max) and K(m) of the pullulanase was measured as 750 U/mg and 1.47 mg/mL, which is the highest specific activity reported so far. The pullulanase crystals structure showed a typical alpha-amylase family structure. The N-terminal has a special substrate binding domain. Activity and substrate binding were decreased when the domain was deleted, the V(max) and K(m) were 324 U/mg and 1.95 mg/mL, respectively. The pullulanase was highly heterologous expressed in Bacillus subtilis by P43 promoter. The extracellular enzyme activity was 42 U/mL, which increased more than 40 times compared to the initial strain. This pullulanase has good application prospects.
Anoxybacillus ; classification ; enzymology ; China ; Glycoside Hydrolases ; metabolism ; Hydrogen-Ion Concentration ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Temperature

Objective To investigate the effect of intrathecal morphine preconditioning (ITMP) on the excitability of substantia gelatinosa (SG) neurons in the dorsal horn of the spinal cord in a rat model of myocardial ischemia-reperfusion (I/R).Methods Thirty-six adult male Sprague-Dawley rats,weighing 200-300 g,in which intrathecal catheters were successfully placed without complications,were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (group S),group I/R,and group ITMP.Myocardial I/R injury was produced by occlusion of the left anterior descending branch of the coronary artery for 30 min followed by 120 min reperfusion.In group ITMP,the rats received intrathecal morphine 3 μg/kg (10 μl) by three cycles of 5 min infusions interspersed with 5 min infusion-free periods starting from 30 min before ischemia,and the equal volume of normal saline was given instead of morphine in group I/R.At 10 min of reperfusion,6 rats randomly selected in each group were sacrificed,and the T2-6 segments of the spinal cords were acutely isolated to prepare spinal cord slices.The resting potential,threshold of action potential (APT),peak of action potential (APP) and action potential duration in SG neurons in the dorsal horn of spinal cord slices were determined using the whole-cell patch-clamp technique,and the number of action potentials evoked by currents of 40,60,80 and 100 pA was recorded.At 120 min of reperfusion,6 rats randomly selected in each group were sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size (IS) and area at risk (AAR),and IS/AAR ratio was calculated.The expression of c-fos in the T2-5 dorsal horns of the spinal cords was detected by Western blot.Results Compared with group S,the IS/AAR ratio was significantly increased,the expression of c-fos was up-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was increased,APT was decreased,and APP was increased in group I/R (P＜0.05).Compared with group I/R,the IS/AAR ratio was significantly decreased,the expression of c-fos was down-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was decreased,APT was increased,and APP was decreased in group ITMP (P＜0.05).Conclusion The mechanism by which ITMP attenuates myocardial I/R injury is related to decrease in the excitability of SG neurons in the dorsal horn of the spinal cord and reduction of responses to nociceptive stimuli in rats.

Aim To explore the effects of acupuncture and sinomenine on the withdrawal symptoms,on the spontaneous discharge and synaptophysin expression of amygdala of heroin-addicted rats.Methods 75 SD rats were randomly and equally divided into 5 groups consisted of control,heroin-addicted,acupuncture,sinomenine and combined treatment group.The model of heroin-addicted rats were established with heroin subcutaneous injection,which in acupuncture,sinomenine and combined group were subsequently punctured at 3 acupoint(Neiguan,Baihui and Zusanli),intramuscular injected with sinomenine and treated with combined methods(acupuncture and sinomenine)respectively.The withdrawal symptoms of rats in different group were evaluated,and then the spontaneous discharges and synaptophysin expression of amygdala were detected with electrophysiological and immunohistochemical technology respectively.Results The heroin-addicted rats were successfully established,being suggested by typical withdrawal symptoms induced by naloxone(P<0.01 vs control),which were alleviated in the 3 treatment groups(acupuncture,sinomenine and combined ) with varying degree(P<0.01 vs heroin group).The electrophysiological results suggest that the forms of low-frequency and single irregular discharges in amygdala were increase,bunchy/cluster and complex discharges were decreased(P<0.05 vs control),which were reversed apparently in sinomenine and combined treatment groups(P<0.05 vs heroin group),closed to control.The immunohistochemical results show that expression of synaptophysin in amygdala was increased obviously(P<0.01 vs control),which was decreased in the 3 treatment groups(acupuncture,sinomenine and combined),P<0.01 vs addicted group.Conclusion Treatments of acupuncture and sinomenine administration can alleviate withdrawal symptoms of heroin-addicted,and rectify the abnormal spontaneous discharges and expression of synaptophysin in amygdala of heroin-addicted rats for certain degree,especially with combined treatments.

Objective To investigate the effect of intrathecal morphine preconditioning on the expression of nerve growth factor (NGF) in the dorsal root ganglia (DRG) in a rat model of myocardial ischemia-reperfusion (I/R).Methods Thirty healthy adult male Sprague-Dawley rats in which intrathecal catheters were successfully placed without complications,weighing 250-350 g,were randomly divided into 5 groups (n =6 each) using a random number table:sham operation group (S group),I/R group,intrathecal morphine preconditioning group (ITMP group),μ receptor antagonist CTOP + intrathecal morphine preconditioning group (CTOP + ITMP group),and CTOP control group (CTOP group).Myocardial ischemia was induced by 30 min of occlusion of the anterior descending branch of the left coronary artery followed by 120 min of reperfusion in all the groups except S group.Intrathecal morphine preconditioning was produced by 3 cycles of 5 min intrathecal injection of morphine 3 μg/kg (10 μl) at 5 min intervals within 30 min before ischemia in ITMP group.In CTOP+ITMP and CTOP groups,1 μg/μ1 CTOP 10 μl was injected intrathecally at 10 min before morphine preconditioning and 40 min before ischemia,respectively.At 120 min of reperfusion,the rats were sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size,and DRGs were removed for determination of the expression of NGF by using immunohistochemistry and Western blot.Results Compared with S group,the myocardial infarct size was significantly increased,and the expression of NGF in DRGs was significantly up-regulated in I/R group (P＜0.05).Compared with I/R group,the myocardial infarct size was significantly decreased,and the expression of NGF in DRGs was significantly down-regulated in ITMP group (P＜ 0.05),and no significant change was found in the parameters mentioned above in CTOP group (P＞0.05).Compared with ITMP group,the myocardial infarct size was significantly increased,and the expression of NGF in DRGs was significantly up-regulated in CTOP+ITMP and CTOP groups (P＜0.05).Conclusion The mechanism by which intrathecal morphine preconditioning reduces myocardial I/R injury is related to activation of spinal μ receptors,inhibition of NGF expression in DRGs,and reduction of responses to noxious stimulation in the rats.

Objective To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by morphine preconditioning in the rats with chronic heart failure in vitro.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,aged 6-7 weeks,in which doxorubicin 2 mg/kg was injected via the tail vein once a week for 6 consecutive weeks to induce chronic heart failure,were studied.At the end of 8th week,30 rats with chronic heart failure were randomly divided into 5 groups (n =6 each) using a random number table:sham operation group (group Sham),I/R group,morphine preconditioning group (group MPC),SB203580 (p38MAPK inhibitor) + morphine preconditioning group (group SBM),and SB203580 group (group SB).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the anterior descending branch of the left coronary artery followed by 120 min of reperfusion to establish the model of myocardial I/R injury.After equilibration,the hearts were subjected to 3 cycles of 5 min perfusion with K-H solution containing morphine 1 μmol/L at 5-min intervals before ischemia in group MPC.In group SBM,the hearts were perfused with K-H solution containing SB203580 (5 μmol/L) for 45 min starting from l0 min before morphine preconditioning until 5 min of ischemia.In group SB,morphine preconditioning was not performed,and the hearts were only perfused with K-H solution containing SB203580 (5 μmol/L) starting from 40 min before ischemia until 5 min of ischemia.At 15 min of equilibration (baseline),5 and 10 min of reperfusion,the coronary effluent was collected to detect the activity of lactate dehydrogenase (LDH) using the chemical colorimetry.At 10 min of reperfusion,the expression of phosphor-p38MAPK (p-p38MAPK) in the myocardium was determined by Western blot in Sham,I/R and MPC groups.At 120 min of reperfusion,the area at risk (AAR),total areas of right and left ventricles (LV+RV),and infarct size (IS) were measured,and the IS/AAR ratio was calculated.Results Compared with group Sham,the LDH activity in coronary effluent during reperfusion and IS/AAR ratio were significantly increased in the other groups,and the expression of p-p38MAPK was significantly up-regulated in I/R and MPC groups (P＜0.05).Compared with group I/R,the LDH activity in coronary effluent during reperfusion was significantly decreased,the expression of p-p38MAPK was significantly up-regulated,and the IS and IS/AAR ratio were significantly decreased in group MPC (P＜0.05),and no significant change was found in the LDH activity in coronary effluent,IS and IS/AAR ratio in SBM and SB groups (P＞0.05).Compared with group MPC,the LDH activity in coronary effluent during reperfusion was significantly increased,and the IS and IS/AAR ratio were significantly increased in group SBM (P＜0.05).Conclusion The mechanism by which morphine preconditioning reduces myocardial I/R injury is related to activation of p38MAPK signaling pathway in the rats with chronic heart failure in vitro.

ObjectiveThe heroin-dependent animal model of rats was used to investigate the effects ofheroin on regulation of pain perception in the dorsal and ventral hippocampus of the heroin-dependent rats. Meth odsHeroin was injected subcutaneously twice a lay for 9 days according to the principle of daily increasing dose in the Sprague-Dawley rats. From the 10th day,rats were given heroin at dose of 27 mg · kg-1 once a day until the14th day, then the unit discharges of the dorsal and ventral hippocampus of rats were observed respectively afternoxious electric stimulation of the rat-tail by the extracellular single-unit recording with glass microelectrodes. ResultsWhen given noxious stimulation, most of the neurons in the dorsal hippocampus in the heroin-dependent ratswere unaffected(59.09％ ) ,whereas in the control rats ,the ratio of the neurons of the dorsal hippocampus affectedby noxious stimulation was about 66.67％, respectively(P ＜ 0.05 ). However,in the ventral hippocampus, the ratioof the neurons activated,inhibitory or unaffected was 20. 69％ ,41.38％ and 37.93％ from the control and was40.74％ ,33. 33％ and 25.93％ from the heroin group respectively with no significant difference between two groups (P ＞ 0.05 ) . ConclusionHeroin changed the regulation of pain perception in the hippocampus,primarily the dorsal hippocampus of rats.

Objective To investigate the effect of long hairpin RNA ( lhRNA) expression vector targeting HBV X gene ( HBx) on replication of hepatitis B virus ( HBV) and gene expression.Methods Four kinds of small interference RNAs ( siRNAs) were synthesized and lhRNA expression vectors targeting HBx were constructed.Four siRNA oligonucleotides and two lhRNA expression vectors were transfected into HepG2.2.15 cells.HBsAg, HBV DNA in culture supernatants and HBx mRNA in HepG2.2.15 cells were detected by time-resolved immunofluorometric assay, real-time quantitative PCR, and reverse transcription PCR, respectively.Negative sequence group or empty vector group was taken as the control.Independent-samples t test was performed to evaluate the inhibition effect on replication of HBV and gene expression. Results Compared with the negative control, HBsAg, HBV DNA level in culture supernatants and HBx mRNA in HepG2.2.15 cells were significantly decreased after siRNA-1 and siRNA-4 transfected at high concentrations (60 nmol/L or 90 nmol/L) (P<0.05), especially the HBsAg and HBV DNA levels in the siRNA-1 transfection group, which were significantly decreased at 24, 48 and 72 h after transfection ( P<0.05 or P <0.01 ) . Two lhRNA expression vectors ( pMD-HBxlh1 and pMD-HBxlh4 ) were successfully constructed and transfected into HepG2.2.15 cells, HBsAg and HBV DNA level in transfected cells was significantly lower than those in negative control (P<0.05).Conclusion The novel siRNA-1 is confirmed to target HBx gene and lhRNA expression vector targeting HBx can effectively inhibit the replication of HBV and expression of HBV gene.

Objective To analyze the reasons of focal liver lesions that difficult to detect by conventional ultrasound ultrasound-CT/MR fusion imaging.Methods 101 lesions which were confirmed by pathology or clinical diagnosis standards were recruited in the research.All of them were difficult to detect by conventional ultrasound but CT/MR display clearly.Ultrasound-CT/MR fusion imaging was used to observe the size,location and internal echo of the lesions,as well as the background of the surrounding liver parenchyma.Results All cases were successfully registrated,the registration time were 2-6min [(4.1 ±0.6)min].For these 101 lesions,93.1％(94/101) of which the diameter ≤20 mm,56.4％ (57/101) were located in hepatic segments near the diaphragm (such as S2,S4,S7,S8),78.2％ (79/101) were internal isoecho,and 79.2％(80/101) in the background of liver cirrhosis.Conclusions The important reasons that focal liver lesions detected difficult by conventional ultrasound includes:lesion size,location,internal echo and the hepatic background.