Objective To evaluate the role of calcium/calmodulin-dependent kinase Ⅱ alpha (CaMK Ⅱα) in the central nucleus of the amygdale (CeA) in fentanyl-induced hyperalgesia in rats and the relationship with miniature excitatory postsynaptic currents (mEPSCs).Methods Thirty-two male Sprague-Dawley rats,weighing 50-80 g,in which the CeA was successfully cannulated,were randomly divided into 4 groups (n=8 each) using a random number table:control 1 group (group C1),fentanylinduced hyperalgesia 1 group (group FIH1),KN92 group,and KN93 group.Normal saline was injected subcutaneously,and dimethyl sulfoxide (DMSO) was given into the amygdale in group C1.In group FIH1,fentanyl was injected subcutaneously (60 μg/kg per time,4 times in total,15-min interval,cumulative dose of 240 μg/kg) to establish the model of hyperalgesia.In KN92 and KN93 groups,KN92 and KN93 10 nmol were given into the CeA after establishing the model.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal threshold (TWT) were measured at 6 and 7 h after fentanyl or normal saline injection.Another 12 Sprague-Dawley rats were selected and randomly divided into either control 2 group (group C2) or fentanyl-induced hyperalgesia 2 group (group FIH2) using a random number table with 6 rats in each group.The brains were removed and sliced 12 h later,and the frequency and amplitude of mEPSCs were recorded.KN93 10 nmol was then added to the artificial cerebral spinal fluid,and the frequency and amplitude of mEPSCs were recorded by whole cell patch-clamp technique.Results Compared with group C 1,the MWT and TWT were significantly decreased at 6 h after fentanyl or normal saline injection in FIH1,KN92 and KN93 groups,and at 7 h after fentanyl or normal saline injection in FIH and KN92 groups (P＜0.05).Compared with group FIH1,the MWT and TWT were significantly increased at 7 h after fentanyl or normal saline injection in group KN93 (P＜0.05),and no significant change was found in group KN92 (P＞0.05).Compared with group C2,the frequency and amplitude of mEPSCs were significantly increased before administration of KN93 (P ＜ 0.05),and no significant change was found in the frequency and amplitude of mEPSCs after administration of KN93 in group FIH2 (P＞0.05).Compared with the value before KN93 administration,no significant change was found in the frequency and amplitude of mEPSCs after administration of KN93 in group C2 (P＞0.05),and the frequency and amplitude of mEPSCs were significantly decreased after administration of KN93 in group FIH2 (P＜ 0.05).Conclusion Activation of CaMK Ⅱ α in the CeA enhances synaptic excitation in neurons,which is involved in fentanyl-induced hyperalgesia in rats.

Objective To explore the role of extracellular signal-regulated kinase (ERK)in central nucleus of amygdala (CeA)in the mechanism of fentanyl-induced hyperalgesia (OIH)in rats. Methods Step 1:12 healthy male Sprague-Dawley rats,weighing 60-100 g,were randomly divided into OIH and Control group.The mechanical paw withdrawal threshold (PWT)and the thermal paw withdrawal latency (PWL)were tested at pre-and post-OIH induction.Then the level of p-ERK in the CeA was analyzed by Western blotting.Step 2:After successful induction of OIH and catheterization in CeA,another 30 SD male rats were randomly divided into 5 groups (n = 6 each):Group OIH;Group OIH+U0124;Group OIH+U0126(0.1 5 nmol);Group OIH+U0126(0.45 nmol)and Group OIH+U0126 (1.5 nmol),then 0.3 μl of DMSO,U0124 (1.5 nmol),U0126 (0.1 5 nmol,0.45 nmol,1.5 nmol)was given through the catheter separately.PWT and PWL were tested before cathe-terization,at pre-OIH induction,post-OIH induction and 0.5 h after CeA drug administration.After the last test of pain threshold,the rats were sacrificed and CeA tissues were sampled for analyzing the expression of p-ERK by western blot.Results In step 1 compared with control group,PWT and PWL of OIH group were sharply decreased post-OIH induction (P <0.05),concomitant increase of the expression of p-ERK in CeA in OIH group was also observed.In step 2,both PWT and PWL were sharply decreased post-OIH induction (P <0.05).Intra-CeA U0126 injection,but not U0124, reversed both behavioral hyperalgesia and molecular activation of ERK in CeA in a dose-dependent manner (P <0.05).Conclusion ERK plays a pivotal role in the maintenance of fentanyl-induced hy-peralgesia.Targeting inhibition of ERK activation in CeA can alleviate fentanyl-induced hyperalgesia.

OBJECTIVETo provide evidence for Chinese medical treatment of children with EB virus infection by exploring its clinical efficacy from multiple angles.

METHODSTotally 81 children patients were randomly assigned to the treatment group (46 cases) and the control group (35 cases). Patients in the treatment group took Chinese medical decoction, while those in the control received intravenous dripping of Ganciclovir and oral administration of pidotimod. The treatment period for the two groups was 2 weeks. Patients were followed-up till the 12th week. Clinical symptoms such as fever, lymphadenopathy and hepatosplenomegaly, as well as lab indices such as abnormal lymphocyte percentage, EB virus antibody, virus DNA load, T cell subsets, immunoglobulin, and so on were observed before and after treatment, at week 4 and 12 of follow-ups.

RESULTS(1) The total effective rate at week 2 was 95.6% in the treatment group, higher than that of the control group (94.3%), but there was no statistical difference between the two groups. (2) The time for defervescence, duration of pharyngeal hyperemia, duration of swollen tonsils was shorter in the treatment group than in the control group (P<0.05). The subsidence of lymphadenopathy, hepatomegaly, and abnormal lymphocytes was better in the treatment group than in the control group (P < 0.05). (3) The positive cases of peripheral blood hetero-lymphocyte was significantly reduced after treatment, at week 4 and 12 of follow-ups both in the treatment group and the control group (P < 0.01). The expression of IgA and IgM decreased after treatment in the two groups when compared with before treatment in the same group (P < 0.05, P < 0.01). IgG in the treatment group also obviously decreased after treatment, at week 4 and 12 of follow-ups (P < 0.05, P < 0.01), while it decreased only after treatment in the control group (P < 0.05). Activities of AST and ALT in the treatment group and the AST activity in the control group were markedly improved when compared with those before treatment (P < 0.05). Compared with the control group, the abnormal lymphocyte positive case number obviously decreased in the treatment group after treatment, at week 4 and 12 of follow-ups (P < 0.05). (4) After treatment, at week 4 and 12 of follow-ups, CD3+ and CD8+ significantly decreased; CD4+, CD4/CD8, and B cells significantly increased in the two groups, when compared with before treatment (P < 0.05). NK cells significantly increased more in the treatment group after treatment, at week 4 and 12 of follow-ups, higher than before treatment as well as the control group (P < 0.05). (5) EB viral DNA and EB viral CA-IgM negative conversion case numbers significantly increased in the two groups after treatment, at week 4 and 12 of follow-ups (P < 0.05). Compared with the control group, EB viral DNA and EB viral CA-IgM negative conversion case numbers significantly increased in the treatment group after treatment and at week 4 of follow-ups (P < 0.05).

CONCLUSIONSTreatment of EB virus infection by Chinese medical treatment was effective. It could promote the recovery of EB viral infection, and reduce the risk of vicious disease after EB viral infection.

Adolescent ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Epstein-Barr Virus Infections ; drug therapy ; immunology ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Male ; Phytotherapy ; T-Lymphocyte Subsets ; immunology

OBJECTIVE To analyze the profile of the pathogens and their drug resistance isolated from children with lower respiratory tract infection in Wenzhou area from Jan 2004 to Dec 2006.METHODS Lower respiratory tract secretions were obtained from children with lower respiratory tract infection for bacterial culture.The K-B method was applied for the antibiotic susceptibility test.RESULTS Total 1605 strains were isolated.The isolating rates of Gram-positive cocci,Gram-negative bacilli and fungi were 24.9%,61.2% and 14.0%,respectively.60.5%,and 54.6% of the isolates of Escherichia coli and Klebsiella pneumoniae produced extended-spectrum beta-lactamases(ESBLs).The rate of Penicillin-resistant Streptococcus pnenmoniae(PRSPN)was 30.6%.20.5% isolates of Staphylococcus aureus were MRSA.All isolates of Enterobacteriaceae and Gram-positive cocci were susceptible to imipenem and vancomycin in vitro.CONCLUSIONS Gram-negative bacilli are still the primary pathogens resulting in lower respiratory tract infection in children.Fungi and muti-drug-resistant bacteria are on the rise trend.

Objective To investigate the effect of portal hypertension (PH) on the activity of local renin angiotensin system(LRAS). Method The expression of local renin and local angiotensinogen mRNA in liver,splenic artery and vein of PH patients was detected with RT-PCR. Result Expression of local renin mRNA in the liver in control group was 0.19?0.12, significantly lower than that of splenic artery (0.45?0.12) and splenic vein (0.39?0.12) respectively(P

Objective To investigate the anti-oxidant effect and the possible mechanisms ofpicrodide II in cerebral ischemia/reperfusion injuries in rats. Methods A total of 90 adult, healthy, mmale Wistar rats were used to established the middle cerebral artery occlusion reperfusion (MCAO/R) models by intraluminal monofilament suture on the left external-internal carotid artery. The treatment group and the positive control group were respectively injected with 1.0％ picroside II (10 mg/kg, 250 μl) and salvianic acid A sodium (10 mg/kg, 250 μl) via the tail vein, and the negative control group and sham-surgery group were injected with 0.1mol/L phosphate buffer saline (PBS) 250 μl. The neurological deficit scores were evaluated with Bederson's test. The cerebral infarction volume was observed with tetrazolium (TTC) staining. The apoptosis positive cells were counted by terminal deoxynucleotidyl transferase dUTP nick-end labeling and the expressions of inducible nitric oxide synthase (iNOS) and superoxide dismutase (SOD) were detected with immunohistochemical assay.The concentration of iNOS and SOD proteins in brain tissue was detected by enzyme linked immunosorbent assay.Results Neurological behavioral malfunction appeared in all the rats with MCAO/R. The infarction focuses emerged in the ischemic hemisphere following the MCAO/R injuries. The number of apoptotic cells and the expression of iNOS increased while the SOD reduced after MCAO/R. After the treatment of picrodide Ⅱ, the nervous behavioral function (1.28±0.38)improved, the infarction volume(68.73±4.46)％ reduced, the number of apoptosis positive cells(6.10± 1.26), the expressions and the concentrations in brain tissue of iNOS(4.67+0.51)decressed while those of SOD (0.53 ±0.14) increased significantly compared with the negative control groups(t=3.16、 2.51、 4.15、3.12、 3.25, P＜0.05). Conclusion PicrodideⅡ might play a neuroprotective effect by inhibiting the neuronal apoptosis and the expressions of iNOS and SOD after cerebral ischemia/reperfusion injuries.

Child ; Graft Survival ; HLA Antigens ; genetics ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Polymerase Chain Reaction ; Tissue Donors ; Transplantation Chimera ; genetics ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; immunology

Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.

Objective To analyze the cost-effectiveness of integrated pharmaceutical care (IPC) in community-based patients with chronic obstructive pulmonary disease (COPD).Methods The COPD patients were divided randomly into intervention group (group A,n =79) and control group (group B,n =71).Group A received the IPC intervention measures,including serial lectures of pharmaceutical knowledge,home supervision and web-based communications for while group B group had none.Results As compared with group B,group A obviously improved on the knowledge of pharmaceutical treatment effect and side effect (95.1％,67.4％ vs.63.1％,21.9％,x2 =12.445,P=0.000 vs.x2 =55.557,P=0.000).There were notable differences between two groups [ ( 1.77 ± 1.23 ) vs.( 2.42 ± 1.66 ) s,t =2.583,P=0.011],pulmonary rales [ (0.01 vs.0.01)s,Z =2.370,P=0.018],6-min walk distance (6 MWD) [ (457 ± 67 ) vs.(425 ± 72) m,t =2.760,P =0.007 ],vital capacity ( VC ) [ ( 1.60 ± 0.25 ) vs.( 1.49 ± 0.23 ) L,t =2.718,P =0.007 ) ] andquality-of-life items ( 48 ± 10 vs.52 ± 11,t =2.624,P =0.010) after intervention.Analyze of cost-effectiveness show that groupA was superior to group B [ total effect total cost(TE/TC =764.6/4936,15.49％ )vs.(TE/TC =1509.4/4708),32.06％,x2 =6.183,P ＜ 0.01 ].Conclusion IPC can improve the disease condition and achieve excellent cost-effectiveness for community-based COPD patients.

The Interleukin-2 gene cDNA was cloned into the Pichia pastoris expression vector pPICZB,which is under the control of the alcohol oxidase promoter AOX1. The linearized recombinant plasmid of BoIL2-pPICZB,digested by Sac I ,was transformed into X-33 strains by electroporation. The multi-copy insert transformants were screened by Zeocin-resistance and induced by 1% methanol. The intracellular expression products were tested by SDS-PAGE analysis and Western blotting. Purified recombinant BoIL2 was gained by metal-chelating affinity chromatographic (MCAC). Assay with murine CTLL-2 cells showed that the recombinant BoIL2 exhibited the biological activity.