Objective Study on the pattern of changes of TGF-?1 and type I receptor occurred in the experimental fluid percussion brain injury model for the purpose of providing the scientific basis for molecular pathological diagnosis, forensic identification, clinical treatment as well as further ascertaining the molecular mechanism of brain injury. Methods Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of injury groups were subjected to moderate lateral fluid percussion brain injury (0.2 MPa). The injury groups were then subdivided into 30min, 1h, 3h, 6h, 12h,1d, 3d and 7d sub-groups according to the time elapsed after injury. Immunohistochemistry SP method was used for studing the immunoreactivity of both TGF-?1 and T?R I factors. Results (1) In the brain of normal control and sham operation control groups, the low expression levels of TGF-?1 and T?R I were observed; (2) The gradual increase of TGF-?1 and T?R I immunoreactivity could be observed 1 to 3d after injury both in cortex and brain stem, and sustained at the high level up to 7d; (3) In hippocampus, the gradual increase occur during 12h to Id after brain injury, and sustained the high level at 3d, then declined at 7d. Conclusion The results suggested that brain injury induced the gene eypressions of the TGF-?1/ T?R I . The TGF-?1/ T?R I may contribute to maintance of nerve cell survival and the repair of damaged neural tissues after CNS injury and the patterns of their level change were quite regular and can be used for timing of injury in forensic medicine aspect.

BACKGROUND: Effect of nerve growth factor(NGF)on angiogenesis of regenerative perpheral nerve is still researched up to now.OBJECTIVE:To evaluate the dose-response relationship and mechanism of NGF on angiogenesis of regenerative sciatic nerve in rats.DESIGN:Randomized controlled animal study.SEFFING:Department of Neurology,the 324 Hospital of Chinese PLA;the Third Laboratory,Institute of Battle Surgery,Daping Hospital of the Third Military Medical University of Chinese PLA.MATERIALS:A total of 32 healthy adult Wistar rats weighing 200-250 g and of both genders were selected in this study.Silica gel conduit was provided by Shanghai Xinya Medical Rubber-goods Factory and NGF was provided by Sigma Company,USA.METHODS:The experiment was carried out in the Third Laboratory,Institute of Battle Surgery,Daping Hospital Affiliated to the Third Military Medical University of Chinese PLA from August 2003 to February 2005.Experimental grouping:All rats were randomly divided into 4 groups,including saline group,50,100 and 500 ng NGF groups with 8 rats in each group.Experimental intervention:Sciatic nerves were cut to establish defect of 10 mm in length,and then the defect was bridged with silica gel conduit in the left latter limb.After the bridging,rats in the saline group were perfused with 5 μL saline,and rats in the NGF groups were perfused with 2.5,5 and 25μL NGF(20 mg/L),respectively.Experimental evaluation:On the 30th day,immunohistochemical technique was used to detect the expressions of CD34,vWf,vascular endothelial growth factor(VEGF)and trkA in newborn vascular endothelial cells of regenerative sciatic nerve;meanwhile,their forms were performed with quantitative analysis.MAIN OUTCOME MEASURES:Pathological and immunohistochemical changes of nerve regeneration of rats in each group.RESULTS:All 32 rats were involved in the final analysis,①Results of pathological sections of nerve regeneration in rats:Distal end of sciatic nerve was completely bridged at 30 days after operation and thickness of sciatic nerve in silica gel conduit lined in 500 ng NGF group＞50 and 100 ng NGF group＞saline group.Numbers of fiber and regularity of arrangement of sciatic nerve were superior in NGF groups to those in saline group,In addition,angiogenesis was obviously observed at 30 days after operation.②Results of immunohistochemical staining:There were significant differences among three NGF groups as compared with saline group in the expressions of four antigens (P＜0.05).Expressions of four antigens were increased in 500 ng NGF group as compared with those in 100 and 50 ng NGF groups(CD34:94.2±6.4,74.2±10.9,77.0±11,0;vWf:116.2±20.0,72.0±13.1,68.0±9.7;TrkA:105.4±10.6,57.8±11.5,58.8±6.5;VEGF:89.0±3.0,48 6±7.4,35.2±2.9;P＜0.05 or P＜0.01).CONCLUSION: NGF can promote angiogenesis of the regenerative peripheral nerves and show a dose-response relationship,while its mechanism may be related to promoting expressions of TrkA and VEGF.

Background Corneal blindness is one of the major blinding eye diseases in China.With the development and progress of tissue engineering technology,tissue-engineered corneas offers a new approach to the treatment of corneal diseases.To select and cultivate ideal seed cells is a foundation of construction of tissueengineered corneas.Objective This study was to evaluate the efficiency of stripe off the Descemet membrane with endothelium plus enzymic digestion in the isolation of corneal endothelial cells and analyze the bionomics of rabbit corneal endothelial cells (CECs) in vitro.Methods Descemet membrane was stripped from fresh cornea of New Zealand rabbit under the dissection microscope.Descemet membrane with endothelium was incubated in trypsin and EDTA solution at 37 ℃ and then purified for CECs subculture in vitro.The morphology of the cultured cells was observed under the inverted microscope and marked by CM-Dil dye solution.Then the shape of the cells was observed by hematoxylin and eosin staining and the cells were identified for the expression of neuron specific enolase (NSE) using immunochemistry.The viability of the cells were evaluated by trypan blue staining.The surface structure of the cells were examined under the scanning electron microscope.Intercellular zonula occludens-1 (ZO-1) was identified by immunofluorecsence staining.Results A large number of purified CECs were obtained from Descemet membrane with endothelium through enzymic digestion.Cultured cells grew well and formed monolayer 5-7 days later with the cobblestone stone-like arrangement.The survival rate of the cells was 95％.CECs presented with the red annular fluorescence for CM-Dil with the labeling rate ＞90％.NSE was positively expressed in the cytoplasm.Polygon CECs were seen by hematoxylin and eosin staining and showed the brown staining.Abundant microvilli on the cellular surface and interconnected foot process were seen under the scanning electron microscope.ZO-1 showed the green fluorescence.Conclusions The method of striping off the corneal Descemet membrane with endothelium plus enzymic digestion can obtain abundant CECs.Cultured cells have good biological properties.This study may offer a feasible application in the engineering of corneal transplant membrane.

OBJECTIVE:To systematically evaluate the efficacy and safety of modified infusion(2-4 h infusion or continuous 24 h infusion)versus traditional infusion(0.5-1 h infusion)of meropenem in the treatment of severe infectious,and to provide evi-dence-based reference for clinic treatment. METHODS:Retrieved from Medline,CJFD,VIP database and Wanfang database, modified infusion(test group)versus traditional infusion(control group)of meropenem in the treatment of severe infections were collected,and Mata-analysis was performed by using Rev Man 5.0 statistical software after extracting data and evaluating quality. RESULTS:A total of 13 studies were included,involving 1 012 patients. Results of Meta-analysis showed the effective rate [RR=1.25,95%CI(1.10,1.43),P<0.001] and bacterial eradication rate [RR=1.25,95%CI(1.05,1.48),P=0.01] in test groups were sig-nificantly higher than those of control group,and there were no significant differences in the mortality rate [RR=0.74,95%CI (0.46,1.18),P=0.21] and incidence of adverse reactions [RR=0.81,95%CI(0.48,1.39),P=0.45]. CONCLUSIONS:Compared with traditional infusion of meropenem,extended or continuous infusion can improve efficacy in the treatment of severe infections, with similar safety. Due to methodology limit of included studies,large-scale and high quality RCT are required for further valida-tion of the conclusions.

Objective To investigate the value of X-ray Knife in the stereotaxis radiotherapy of lung cancer.Methods 52 cases with pulmonary carcinoma were treated with X-ray knife,before and after that,general radiotherapy was applied.After whole radiotherapy,2 or 3 months later,all patients were followed by CT scan.The changes of tumor and clinical appearances were compared before and after therapy.The curative effect was estimated according to clinical appearances and CT findings.Results 6 patients were complete response(11.5%) ,29 patients were partial response(55.8%) and 13 patients were no changes or reduced less than 50%.The total responsive rate was 92.3%.Conclusion The treatment of lung cancer with X-ray Knife can give tumor accuracy high dosage irradiation.The short term therapeutic effectiveness is significant and safe on clinical.

OBJECTIVE:To systematically review the effect of organic cation transporter 2 [(OCT2)808G>T] gene polymor-phism on the metformin hydrochloride pharmacokinetics in vivo,and to provide evidence-based reference for clinical medication. METHODS:Retrieved from PubMed,EMBase,Foreign Medical Journey Service,CJFD,VIP database and Wanfang database,re-lated studies about the effect of (OCT2)808G>T gene polymorphism on the metformin hydrochloride pharmacokinetics in vivo were collected,and Meta-analysis was performed by using Rev Man 5.1 statistics software. RESULTS:A total of 5 retrospective studies were included,involving 172 patients. The result of gene type was type GT,type TT and type GG. Results of Meta-analysis showed,compared with type GT volunteers,type TT could prolong the half-time period of metformin hydrochloride;compared with type TT,type GG could increase the peak concentration. However,(OCT2)808G>T gene polymorphism had no effects on the renal clearance rate,creatinine clearance rate and area under the drug-time curve. CONCLUSIONS:(OCT2)808G>T gene poly-morphism has certain effect on the half-time period and peak concentration of metformin hydrochloride in vivo of health volunteer, and has no effect on the renal clearance rate,creatinine clearance rate and area under the drug-time curve. Due to the limit of re-search methodological quality,large-scale and high quality studies are required for further validation of the conclusions.

OBJECTIVE:To systematically review the effect of ticagrelor versus prasugrel on platelet reactivity,and provide evi-dence-based reference for clinical treatment. METHODS:Retrieved from PubMed,CJFD and Wanfang Database,randomized con-trolled trials(RCT)about the effect of ticagrelor versus prasugrel on platelet reactivity were collected. Meta-analysis was performed by using Rev Man software after data extract and quality evaluation by Cochrane 5.1.0. RESULTS:Totally 17 RCTs were enrolled,involv-ing 2 757 patients. Results of Meta-analysis showed,regardless of Verity Now(VN)detection method [MD=15.43,95%CI(-0.39, 31.25),P=0.06] or vasodilator stimulus phosphoprotein(VASP)detection method [MD=-3.04,95%CI(-8.98,2.90),P=0.32], ticagrelor and prasugrel had the same effects on platelet reactivity under loading dose,the differences were not statistically significant;regardless of VN detection method [MD=-48.94,95%CI(-58.04,-39.84),P<0.001] or VASP detection method [MD=-14.32, 95%CI(-20.45,-8.20),P<0.001],the effects of ticagrelor were better than prasugrel on platelet reactivity under maintenance dose,the differences were statistically significant. CONCLUSIONS:At the loading dose,there was no difference between ticagrelor and prasugrel,but ticagrelor has more benefits than prasugrel under maintenance dose.

Objective To compare the image quality of diffusion-weighted MR neurography (DW-MRN) of the tibial nerve and the common peroneal nerve prospectively using different motion probing gradients (MPGs).Methods A total of 21 healthy volunteers underwent DW-MRN at the knee (unilateral imaging) on a 3.0 T magnetic resonance system with unidirectional MPGs.The protocol included anteriorposterior unidirectional,right-left unidirectional,three-directional and six-directional MPGs.The apparent SNR and CNR of tibial nerve and common peroneal nerve were calculated.Three-dimensional MIP images of the nerves were evaluated blindly by two radiologists using a four-point grading scale on basis of entirety depiction and the signal intensity.Significance was determined by using Friedman and paired Wilcoxon tests.Results The SNR of tibial nerves on DW-MRN with anterior-posterior,right-left,three directional and six directional MPGs were 4.17 (2.70-5.65),4.35 (0.47-4.69),3.46 (2.27-4.62) and 3.30 (2.06-4.43),respectively.CNR were 0.61 (0.46-0.70),0.63 (0.36-0.73),0.55 (0.39-0.64) and 0.53(0.35-0.63),respectively.The scores of tibial nerve image quality were 4.0 (2.0-4.0),4.0 (3.0-4.0),2.5 (2.0-3.5),2.0 (1.0-2.5),respectively.Interobserver agreement was good and the Kappa value was 0.69 (P ＜ 0.05).The SNR of the common peroneal nerves on DW-MRN with anteriorposterior,right-left,three directional and six directional MPGs were 3.05 (2.30-4.20),3.05 (2.26-4.34),2.72 (1.84-13.80) and 2.68 (1.87-3.67),respectively.CNR were 0.51 (0.39-0.62),0.51 (0.39-0.63),0.46(0.30-0.86) and 0.46(0.30-0.57),respectively.The scores of the common peroneal nerve image quality were 3.5 (2.0-4.0),4.0 (2.0-4.0),2.0 (1.0-3.0) and 2.0 (1.0-2.5),respectively.Interobserver agreement was good and the Kappa value was 0.70(P ＜0.05).For SNR,CNR and nerve image quality of the tibial nerves and the common peroneal nerves,there were significant differences among different MPGs (x2 =215.01,215.01,60.49 and 182.82,182.82,60.22,respectively,P ＜ 0.05).SNR,CNR and nerve image quality of the tibial nerves and the common peroneal nerves on DW-MRN with unidirectional MPGs were better than those with three-directional and sixdirectional MPGs (Z =-6.90-4.03,P ＜ 0.05).The SNR and CNR of the tibial nerves on DW-MRN with right-left direction were better than those with anterior-posterior unidirectional MPGs (Z =-3.53,-3.41,P ＜ 0.05),but there was no significant difference of image quality between right-left and anteriorposterior directional MPGs (Z =-0.58,P ＞ 0.05).DW-MRN of the tibial nerves with three-directional MPGs was better than that with six-directional MPGs (Z =-3.00,-2.80,-3.92,P ＜ 0.05).There were no significant differences between right-left and anterior-posterior unidirectional MPGs,or between three-directional and six-directional MPGs of common peroneal nerves (Z =-1.94--0.31,P ＞ 0.05).Conclusions DW-MRN has capability to provide three-dimensional visualization of the tibial and common peroneal nerves,and the image quality with unidirectional MPGs is better than that with three or six directional MPGs.

[ ABSTRACT] AIM:To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 ( Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation.METHODS:C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation.The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology.The expression levels of IL-17, RORγt and IFN-γin the spleen were measured by real-time PCR.Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γin peripheral blood were measured by flow cytometry.RESULTS:1α, 25-dihydroxyvitamin D3 signifi-cantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ.In the pe-ripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION:The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejec-tion in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.

Objective To investigate the relationship between the serum chemokine IP-10 and RANTES levels and Interferon therapeutic early response in patients with chronic hepatitis B(CHB).Methods 50 patients with chronic hepatitis B were chosen into interferon therapy.After 12 weeks,they were devided into three groups:complete response A、partial response B、non response C group.HBV-DNA was detected by PCR;The serum chemokines( IP-10 and RANTES) were measured by Luminex Liquichip technology.Results The base HBV DNA and RANTES levels of three groups weren't significantly different (P ＜0.05) ;The base ALT and IP-10 levels of A group were significantly higher than that in B and C group( P ＜ 0.05).The IP-10、RANTES contents of A group in therapeutic 4th week were significantly lower than that before interferon therapy(P ＜ 0.05 );There were no significant differences in B、C group (P ＞0.05) ;The levels changes of IP-10、RANTES、HBV DNA and ALT in therapeutic 12th week were significantly different between the three groups ( P ＜ 0.05 ),The level of ALT in 50 patients has positive correlation with IP-10 level (P ＜ 0.05) ;The level of HBV DNA in 50 patients had positive correlation with RANTES level( P ＜ 0.05 ) ;The base level of IP-10 had positive correlation with the change of HBV-DNA contents in therapeutic 12th week( P ＜0.05 ) ;The change of ALT level in reponse patients in therapeutic 12th week had positive correlation with the change of IP-10 、RANTES levels( P ＜ 0.05 ).Conclusion The decrease of IP-10,RANTES level in CHB patients received 12weeks interferon-α therapy could lead to reduce liver inflammation;The base IP-10 level probably was relevant to the early response in CHB patients received interferon-α therapy.