Objective To study the type variation of microglial activation in spinal dorsal horn of rats after sciatic nerve injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into the control and experimental groups, 24 rats in each group.The experimental group underwent ligation of sciatic nerve trunk to generate nerve injury in the rats.The pain behavior in the rats was measured at the 1th, 7th and 14th postoperative days, and the changes of microglial activation in the rat lumbar spinal cord dorsal horn was detected by immunofluorescence staining.qRT-PCR assay was used to validate the activation trends of M1 and M2 types of microglia cells.Results No significant changes were found in the microglial cells in the spinal cord dorsal horn of rats in the sham-operation group during 14 days after operation.In the sciatic nerve ligation group at 1 day after operation, no significant change was observed in the number of microglial cells, but the expression of marker of M1 microglia was significantly increased.At 7 and 14 days after operation, the number of microglial cells and the expression of M1 microglia marker in the spinal cord dorsal horn were increased significantly.Conclusions Microglia activation in the spinal dorsal horn starts at the first day after sciatic nerve injury, and lasts at least for two weeks after the operation.M1 microglia activation dominates during this period.

Objective To explore the role of spinal cord p300 in neuropathic pain induced by chronic constriction nerve injury (CCI) in rats.Methods Thirty two Sprague-Dawley (SD) rats were randomly divided into sham and CCI groups,14 days after surgery,immuno-fluorescence staining and Western blot were used to detect distribution and expression of p300 protein.After rats were successfully implanted with an intrathecal catheter and accepted CCI surgery,another 24 rats were randomly divided into three groups (n =8):dimethyl sulfoxide (DMSO) group,p300 acetyltransferase inhibitor C646 group,and control C37 group.Each rat were administered through the intrathecal catheter from day 7 to 14 and mechanical withdraw threshold were tested.Results (1) The p300 positive cells were detected mainly in neurons,and p300 protein in spinal cord of CCI group were significantly higher than sham group (P ＜0.05).(2) C646 alleviated significantly neuropathic pain in rats,without significant changes in pain threshold after injection of C37 and DMSO.Conclusions The p300 protein in spinal cord was involved in the development of neuropathic pain in rats,the mechanism may be referred to its acetyltransferase activity.

Objective To investigate the clinical application of scalp nerve block combined with target-controlled infusion in neurosurgical anesthesia.Methods 40 adult patients undergoing frontotemporal craniotomies were randomly divided into the ropivacaine scalp nerve block group (group R) and control group (group C).The patients in group R received scalp nerve block with 0.5％ ropivacaine before induction while those in group C didnt.We used propofol and remifentanil in target-controlled infusion and atracurium in constant infusion to maintain anesthesia.The heart rate(HR),mean arterial pressure (MAP),bispectral index (BIS) of different time,usage of propofol and remifentanil,extubation time,visual analogue scale,and complication were recorded.Results Both groups had stable hemodynamics.The usage of remifentanil in group R was less than that of group C (t =11.10,P ＜ 0.01).The difference of extubation time,usage of propofol,and incidence of complications were not statistically significant (P ＞ 0.05).The difference of visual analog scale (VAS) (2 hour and 6 hour after operation) was statistically significant (t =5.02,4.60,P ＜0.O1).Conclusions Scalp nerve block combined with target-controlled infusion is simple with less usage of remifentanil and better analgesic effect.

Intellectual property protection is an indispensable management measure in the process of knowledge innovation in the National Traditional Chinese Medicine (TCM) Clinical Research Bases. Longhua Hospital, a National TCM Clinical Research Base , has actively participated in studies and construction of TCM intellectual property policy , laws and regulations , and endeavored to enhance international research cooperation and broaden the international repercussions of TCM . Longhua Hospital made great efforts to strengthen management on intel-lectual property management team and measures, advocacy training, property rights formation and transactions. Intellectual property protection played an important role in the construction of the National TCM Clinical Re-search Base in Longhua Hospital .

Objective To explore the risk factors for early interhepatic recurrence and metastasis of hepatocellular carcinoma (HCC)on CT imaging before treatment.Methods 1 1 5 patients suffered from HCC from July 2003 to January 2009 were retrospec-tively enrolled for reviewing their clinical characteristics and CT signs.The status of metastasis and/or recurrence was followed reg-ularly.Signs on pre-treatment enhanced CT images were measured and analyzed.Analysis of variance and independent sampler t test were applied for Univariate survival analysis.Then multivariate analysis was carried out by the Logistic regression,Lon rank meth-od,and p-value < 0.05 was defined to be statistically significant.Results The early interhepatic recurrence and metastasis rate of the study group was 58.26%.With univariate analysis,tumor size,location,extent,capsule,satellite nodule,vascular invasion, AVM and necrosis were the risk factors for early recurrence and metastasis of HCC on pre-treatment enhanced CT imaging (P<0.05).Multi-variable Logistic regression analysis showed that tumor size,satellite nodule,vascular invasion capsule and were independently sig-nificant CT signs for early interhepatic recurrence and metastasis of HCC (P =0.031,0.005、0.037、0.048).Conclusion Pre-treat-ment enhanced CT imaging with HCC was closely related to early interhepatic recurrence and metastasis of the tumor.A tumor of larger size,with satellite nodules,without complete capsule and vascular invasion on CT may predict a tendency to early interhepatic recurrence and metastasis of HCC.

Objective To study the protective effect of codon optimized TPI DNA vaccine against Schistosoma japonicum infection.Methods Sixty female BALB/c mice were randomly divided into 5 groups.The mice were injected through musculus quadriceps fexoris with 100 ?g pcDNA 3.1 control(Group A), pcDNA3.1-TPI(Group B), pcDNA 3.1-TPI-mHSP70(Group C), pcDNA3.1-TPI.opt(Group D), and pcDNA3.1-TPI.opt-mHSP70(Group E) respectively.All mice were immunized for three times with an interval of two weeks.The mice were challenged with(40?1) cercariae of S.japonicum per mouse by abdominal skin penetration 4 weeks after the last immunization, and sacrificed at 42 days post-challenge, the number of worms or hepatic eggs was counted.Blood was taken for the detection of IgG, IgG1, and IgG2a 2 days before immunization and before challenge, respectively.Spleen cells of 2 mice from each group were cultured and stimulated with ConA and rSjCTPI peptide, and the supernatant was collected for detection of IL-2, IL-4, IL-5, IFN-?, and TNF by flow cytometry.Results ELISA showed that the mice in groups B, C, D, and E produced specific IgG and IgG1, IgG2a antibody isotypes, and the ratio of IgG2a/IgG1 was 1.73, 2.06, 2.44, and 3.09, respectively.The levels of IL-2, IFN-? and TNF in groups D and E were higher than that of groups B and C.The worm reduction rate and hepatic egg reduction rate in groups D(36.03%, 41.7%) and E(39.03%, 46.85%) were higher than those of groups B(26.28%, 28.35%) and C(28.38%, 31.39%)(P

Objective To evaluate the value of MRI performance for the differential diagnosis of atypical solitary metastatic malignant melanoma from spinal hemangioma.Methods Thirteen patients of atypical solitary metastatic malignant melanoma and 40 patients of spinal solitary hemangioma were retrospectively analyzed.Conventional MR imaging (T1WI,T2WI,and fat suppressed T2WI) and enhanced imaging were performed at 1.5T MRI.The signal intensities (SIs) of spinal lesions were qualitatively evaluated on conventional imaging and were described as hypointense,isointense,or hyperintense.The spinal lesions were qualitatively categorized into minimal enhancement,iso-enhancement,slightly hyper-enhancement,or strong enhancement on contrast-enhanced imaging.The lesions' maximum diameter was also measured and the mean value was obtained.Results The qualitative assessment of SIs on T1WI showed that 76.92％ (10/13),15.38％ (2/13) and 7.69 ％ (1/13) of atypical solitary metastatic malignant melanoma were hypointensity,isointensity and hyperintensity respectively.The qualitative evaluation of SIs on T2WI were found that 61.54％ (8/13) of atypical solitary metastatic malignant melanoma with hypointense,30.77％ (4/13) with isointensity and 7.69％ (1/13) with hyperintensity,respectively.About 92.31％ (12/13) of atypical solitary metastatic malignant melanoma displayed strong enhancement on contrast-enhanced imaging.There were significant differences in SIs on T1WI,T2WI and contrast-enhanced imaging between atypical solitary metastatic malignant melanoma and hemangioma (all P＜0.05).The maximum diameter of atypical solitary metastatic malignant melanoma was significantly higher than that of spinal hemangioma (P＜0.001).Conclusion MR imaging would be practicable for differentiation between atypical solitary metastatic malignant melanoma and hemangioma in spine.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

Objective To enhance the protective immunity effects of nucleic acid vaccines against Schistosoma japonicum infection by electroporation(EP)in vivo in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml.pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs were mixed by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1/EP control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP in vivo for three times at week 0,3,6;in Group C(pcDNA3.1/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccine/EP group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6;in Group E(cocktail DNA vaccine/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-? by flow cytometre.Results The worm reduction rates in Group C,D and E were 18.09%,45.00% and 57.09%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P