Objective To establish the method of contact heat evoked potential(CHEP)and to explore the value of this evoked potential in pain testing of patients with cerebral infarction.Methods A total of 100 healthy volunteers and 30 patients were examined.The healthy volunteers were divided into 3 groups according to the length of their arms:(Group A:56.0～65.0 cm ;Group B :65.5～74.0 cm ;Group C :74.5～83.0 cm).A recently de- veloped heat-foil technique with a rapid temperature rising rate at 70℃/s was used to elicit pain and contact heat e- voked potentials.Contact heat was delivered via one circular thermode(diameter 27 mm,area 573 mm~2)and set at two intensity levels(49.5℃and 54.5℃)to three body sites:the thenar eminence,the dorsum of hand and proximal volar forearm.The subjects were asked to rate the pain with numerical rating scale after each stimulus and CHEP was recorded from Cz and Pz.The association between stimulus intensities and pain rating was explored,the main compo- nents of the evuked potential were watched.CHEP,sensory conduction velocity(SCV)and somatosensory evoked potentials(SEP)were performed in patients with hemi-anesthesia caused by cerebral infarction.Results The pain intensity ratings were 3.2?0.3 and 4.4?0.5 at thenar eminence,5.0?0.7 and 6.3?0.8 at the dorsum of hand and 5.3?0.6 and 7.2?0.5 at the proximal volar forearm when the temperature of 49.5℃and 54.5℃was applied, respectively;Three components,Cz/N550,Cz/P750 and Pz/P1000,were identified in the evoked potentials.Cz/ N550 and Cz/P750 appeared when the dorsum of hand and proximal volar forearm were stimulated.In contrast,Pz/ P1000 could be identified when nociceptors of thenar eminence and proximal volar fbrearm were excited.In the pa- tients with cerebral infarction,CHEP disappeared or became abnormal on one side,while SCV and SEP were normal on that side.Conclusion It was suggested that CHEP could be elicited reliably in the controls.CHEP is helpful in the assessment of analgesia in patients with cerebral infarction.

Objective To evoke cerebral potentials by stimulating nociceptive fibers with contact heat evoked potentials stimulator (CHEPS)and estimate the nerve conduction velocities of peripheral nerve fibers mediating these responses.Methods Subjects were set in supine position.A heat-foil technology with a rapid rising speed at 70 ℃/s was used to elicit pain and contact heat evoked potentials(CHEP).Contact heat was delivered via one circular thermode (diameter 27 mm,area 573 mm~2).Thermal stimuli were sent at two intensity levels (49.5 ℃ and 54.5 ℃) to three body sites:thenar eminence,the dorsum of hand and proximal volar forarm.Contact heat evoked potentials were recorded from Cz and Pz.A systemic effect between stimulus intensities and pain rating were observed,the main components of this evoked potential were observed.Nerve conduction velocity was calculated from latency difference of CHEP and center to center distance of distal and proximal stimulus arrays.Results The pain intensity rating was 3.2?0.3 and 4.4?0.5 when thenar eminence was stimulated at the temperature of 49.5 ℃ and 54.5 ℃ respectively;the rating was 6.3?0.8 and 7.2?0.5 when the dorsum of hand and proximal volar forarm were stimulated at the temperature of 54.5 ℃ respectively.Three components,Cz/N550,Cz/P750 and Pz/P1000,were found in the evoked potentials.Nerve conduction velocities of the fibers were (12.9?7.5) and (1.7?0.4) m/s respectively,which were corresponding to those of A8 fiber and C fiber.Conclusions CHEPs can be elicited reliably and stably.Velocities of peripheral nerve fibers demonstrate that A8 fiber and C fiber mediate the response.

Objective To determine the roles of MyD88 and Trif,critical adaptor proteins for TLR signaling,in production of donor-specific antibodies (DSA) and memory T cells in a presensitized mouse cardiac transplant model.Methods Skin grafts from Balb/c mice were transplanted into either wild type B6 mice or B6 Myd88 and Trif double knockout mice (Myd88/Trif DKO).The recipients were subsequently transplanted heterotopically with cardiac grafts from the same donors two weeks after skin transplantation.Plasma DSA levels and spleen phenotypical analysis were performed prior to heart transplant or at time of cardiac rejection by using flow cytometry.Results Recipients presensitized with skin grafts developed accelerated cardiac allograft rejection in the absence of Myd88 and Trif.However,plasma DSA,especially IgG2,was significantly decreased (P＜0.05) in Myd88/Trif DKO mice,compared to that in Wild Type mice at 2nd week after skin transplantation.The production of DSAs including all IgG subtypes was further reduced 3 days following heart transplantation in the Myd88/Trif DKO.In addition,MyD88/Trif DKO mice had impaired ability to generate memory T cells,as percentages of both CD44hi CD4+ and CD44hi CD8+ were significantly lower in the DKO than in Wild Type mice (P＜0.01,P＜0.05).Conclusion Simultaneous ablation of MyD88 and Trif in recipients significantly decreases the production of serum DSAs and spleen memory T cells following allogeneic skin and heart transplantation,supporting a crucial role of TLR signaling in adaptive immune responses in organ transplantation.

Objective To assess the right ventricular (RV) performance in patients with type 2 diabetes mellitus (2-DM) by two-dimensional speckle tracking imaging (2D-STI),and to explore the clinical value of RV longitudinal strain and strain rate.Methods Thirty-seven patients with 2-DM only and thirtyone patients coexisting diabetes and hypertension (DM + HTN) were studied.Thirty-nine healthy age matched persons served as control subjects.In each patient a conventional two-dimensional echocardiography was performed and also an echocardiographic study with strain/strain rate imaging was studied.Analysis of RV longitudinal systolic strain were obtained in the apical four-chamber view of the RV for the assessment of the RV free wall(F-PLSS),interventricular septum (S-PLSS) and the global RV wall (G-PLSS).The entire RV longitudinal peak systolic strain rate (G-SRs),peak early-diastolic strain rate (G-SRe) and peak late-diastolic strain rate (G-SRa) were performed in the apical four-chamber view.Results Compared with controls,F-PLSS,S-PLSS,G-PLSS,G-SRs and G-SRe were decreased in 2-DM and DM + HTN (all P ＜0.01),with lower values in DM + HTN (P ＜0.05).In the patients coexisting diabetes and hypertension,G-SRa was significantly lower than those in the control group (P ＜ 0.05).Compared with 2-DM,this difference was not significant in controls and DM + HTN (P ＞0.05).Conclusions No matte the patients of 2-DM with or without hypertension,the early changes of RV dysfunction can be found by 2D-STI.Patients coexisting diabetes and hypertension may have worse RV dysfunction.

Objective To evaluate arthroscopic PCL (posterior cruciate ligament) reconstruction with an implant fixaliun of hamstring tendon knot and bottle-neck technique.Methods Thirteen cases with PCL injury were treated with arthroscopic PCL reconstruction with an implant fixation of hamstring tendon knot and bottle-neck technique between April,2002 to June,2005.The patients were followed up fur at least seven months to evaluate the clinical effects by means of ROM of the joint,post drawer test,Lachman test,Lysholm and IKDC (International Knee Documents Committee) scores.Results The patients were followed up for an average of 13.4 months.No limitation of knee extension was found six months after operation.Eleven patients had normal knee flexion(＞140?), one had knee flexion of 120?and one 100?.In posterior drawer test,11 patients were gradeⅠand two gradeⅡ.In Lachman test,11 patients were rated as within gradeⅠand two gradeⅡ.By Lysholm knee function scale,the average knee scores were 90.47?4.13 and 78.82?2.46 respectively before and after operation,and the dif- ference was of statistical significance (t=2.416,P=0.029).By IKDC scale,six were grade A,five grade B and one grade C.Conclusion Arthroseopic PCL reconstruction with an implant fixation of hamstring tendon knot and bottle-neck technique is a reliable method to restore knee function and will have a prospect of wide application be- cause of its good tissue biocompatibility,less invasion,and less operative expenditure.

Objective To investigate the therapeutic effects of curcumin(Cur)on trinitrobenzene sulphonic acid(TNBS)induced colitis and to investigate cytokines change in colon mucosa,spleen and sera.Methods Colitis was induced in SD rats by intrarectal injection of TNBS(100 mg/kg).Experi- mental animals were divided into negative control group,TNBS group and Cur therapeutic group(Cur, 30 mg?kg~(-1)?d~(-1),intraperitoneal injection).Expression of cytokines mRNA in colon mucosa was observed by RT-PCR,intracellular cytokines of interferon(IFN)-?and interleukin(IL)-4 in splenocytes were detected by flow cytometry(FCM),concentrations of IFN-?and IL-4 in sera were determined by enzyme-link immunosorbent analysis(ELISA).Results After treatment with Cur,macroscopic scores (3.9?1.0 vs.2.2?0.7),myeloperoxidase(MPO)activity(15.0?2.6 vs.7.3?1.4),mRNA of IFN-?(1.02?0.07 vs.0.06?0.02,mRNAof IL-12(0.29?0.05 vs.0.11?0.01)and the ratio of IFN-?/IL-4(11.44?0.97 vs.0.38?0.10)in colon mucosa,proportion of IFN-?CD4~+(31.7?7.5 vs. 21.1?3.7)and the ratio of IFN-?/IL-4 CD4~+(19.9?5.1 vs.6.1?1.8)in splenocytes,concentra tions of IFN-?[(1528?159)pg/ml vs.(513?14)pg/ml] and the ratio of IFN-?/IL-4(19.5?4.1 vs. 4.2?0.6)in sera were all decreased(P＜0.05 or P＜0.01).Meanwhile,mRNA of IL-4(0.09?0.01 vs.0.15?0.04)and IL-10(0.28?0.08 vs.0.63?0.12)in colon mucosa,proportion of IL-4 CD4~+ (1.6?0.5 vs.3.4?1.1)in splenocytes and concentrations of IL-4 in sera[(81?15)pg/ml vs.(124?20) pg/ml] were all increased after the treatment of Cur(P＜0.05 or P＜0.01).Conclusions Cur showed ther- apeutic effects on TNBS-induced colitis,and the mechanism might be through regulating the balance of Th1/ Th2 in colon locally and systematically.

Objective To analyze the epidemiologic features of visceral leishmaniasis in different age groups in China from 2005 to 2012,and to provide a scientific basis for prevention and treatment of the disease.Methods The visceral leishmaniasis cases were obtained from China disease epidemic monitoring information system from 2005 to 2012.All the cases were divided into subgroups by demographic characteristics:infant (＜1 year old),childcare (≥1 and ＜4 years old),child (≥4 and ＜15 years old),adult (≥15 and ＜65 years old) and senior adult (above 65 years old).The epidemiologic features,such as gender,season of disease onset and area distribution of visceral leishmaniasis were analyzed.Results The total reported visceral leishmaniasis from 2005 to 2012 were 2 979,among which cases in infant,childcare,child,adult and senior adult were 24.9％,21.7％,20.0％,32.4％ and 1.0％,respectively.Infant cases were most reported in April; childcare cases in December which began to increase since September; child cases in October and adult and senior adult in March.The reported cases were mainly from Xinjiang,Gansu and Sichuan areas.However,age characteristics differed among regions,which were mainly infants,both childcare and children,and adults,respectively in this three areas.The duration from onset to diagnosis was shortest in infant group (11 d) and longer in adult group (15 d).Conclusions The number of visceral leishmaniasis cases in different age groups peaks in different months.Regional distribution is not even.Cases are most concentrated in Xinjiang,Gansu and Sichuan areas.Infants predominate in Xinjiang,childcare and children in Gansu and adults in Sichuan.

AIM: To determine the ultimate preservation time of murine cardiac grafts in 4℃ Ringer's solution. METHODS: Murine cardiac grafts were implanted to the abdominal vessels heterotopically 2, 4, 6, 8 and 10 hours after cold preservation. Graft survival rate and histological morphological changes, as well as the neutrilphil, T cell, macrophage infiltration, ICAM expression were determined by immunohistochemistry. RESULTS: The cardiac function-recovery rate and 1-week graft survival rate were 100% and 83.3% in 6-hour preservation group. Compared with non-preservation control group, no more apparent histological damages, cell infiltration and ICAM expression were found. CONCLUSION: The ultimate preservation time of murine cardiac graft in 4℃ Ringer's solution was 6 hours. [

Cell-free protein synthesis (CFPS) systems based on crude cell extracts have been used in protein expression in vitro. With the researchers' endeavor for decades, the CFPS system has been developed as an important research tool in many frontiers of fundamental and applied biology because of its clear genetic background and simplicity to control the reaction. The yield of CFPS systems derived from prokaryote or eukaryote has increased to several grams per liter with constantly decreasing cost. Nowadays grams of protein could be prepared using a large-scale cell-free system. Recently, the advantages on the expression of complicated, toxic and membrane proteins have shown the great potential of the CFPS systems. The rapid progress of this technology made us to believe that it will take an important place in biopharmaceutical industries undoubtedly.
Bioengineering ; trends ; Cell-Free System ; Drug Industry ; trends ; Protein Biosynthesis

Objective To investigate the relationships between the expression of VEGF,its receptor KDR/flk-1,cNOS mRNA and angiogenesis,cell proliferation,metastasis in human hepatocellular carcinoma(HCC). Methods Immunohistochemical analysis using antibodies against VEGF and its kinase insert domain receptor (KDR) was carried out.cNOS mRNA expression in HCC,liver cirrhosis and normal liver tissue was observed by in situ hybridization.CD34 immunostaining was used to measure the microvascular density(MVD)and proliferative index was evaluated by Ki-67 immunostaining. Results The expressions of VEGF and KDR of HCC were significantly related to MVD,proliferation and metastasis of HCC(P