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Objective To evaluate the clinical effect of Shuxuetong plus external application of deproteinised calf blood serum injection on diabetic ulcer foot.Methods 60 patients with diabetic foot ulcer were enrolled:30 cases (treatment group)were randomly assigned to receive deproteinised calf blood serum injection(external use)plus Shuxuetong(iv gtt),and the other 30 cases (control group)received routine western medicine therapy.One course of treatment lasted one month.In addition,both two groups received insulin to control blood sugar and antibiotics to control infection,with foot ulcer treated with debridement,drainage,and removal of necrotic tissue etc.Results Com-pared with the control group,the effective rate was higher in the treatment group(86.7% vs.60.0%,χ2 =5.455,P =0.020).The occurrence time of granulation tissue[(6.60 ±1.14)d vs.(10.80 ±2.17)d,t =-3.834,P =0.008] and wound healing time[(23.62 ±6.14)d vs.(35.65 ±4.45)d,t =-4.405,P =0.002]were shorter in the treat-ment group.The wound area in the two groups reduced after treatment,and the wound area in the treatment group reduced significantly[(3.48 ±1.16)cm2 vs.(8.60 ±2.51)cm2 ,t =-4.143,P =0.007].Conclusion As com-pared with the routine therapy with western medicine,the combination therapy of deproteinised calf blood serum injec-tion plus Shuxuetong has better efficacy for diabetic ulcer foot.

BACKGROUND: The animals commonly used to induce experimental allergic encephalomyelitis (EAE) in oversea laboratory are rodentia animals such as Lewis rats. But in China we are short of Lewis rats. The un-susceptive animal Wistar rats are inexpensive and plentiful. The adding of pertussis toxin may induce EAE successfully in EAE un-susceptive Wistar rats.OBJECTIVE: To investigate the influence of pertussis toxin injected atdifferent sites in inducing EAE model in un-susceptive Wistar rats.DESIGN: A randomized control animal experiment.SETTING: Institute of Brain Science, Shanxi Datong University.MATERIALS: The study was performed in the Institute of Brain Science of Shanxi Datong University from March to October in 2003. Fifty-eight und adjuvant (CFA) group (n=10).METHODS: Besides routine immunization, each rat in the foot dorsum EAE group and intraperitoneal EAE group was administrated with 005 mL pertussis toxin (containing 5.0×1010 thalli), which were given intraperitoneally and subcutaneously on one hind foot respectively, and the antigen in the CFA group was replaced by CFA.cidence rate and tine of atta ck: In the foot dorsum EAE group, the incidence of EAE was 87.5% (21/24), and the time of attack was at (10.25 ±1.67) days after immunization, which were significantly different from those in the intraperitoneal EAE group [35.7% (9/24), (14.8±l.79) days, P sum EAE group, the change of body mass was (-16.00±7.30) g and the symptomscpre was 3.4±0.7, and those in the intraperitoneal EAE group Therewere no or little infiltration of inflammatory cells in the encephalon and spinal cord of CFA rats. In the EAE rats, there were inflammatory cells infiltrated in the boundary of white matter and gray matter of lumbar intumescence, spinal pia mater, spinal parenehyma, and the boundary of cerebral cortex and medulla, even deep medulla, meninges and around lateral ventricle. There were also mild inflammations in the cerebellum,brainstem and optic chiasma, which were concordant with the observed asynchronism, tic, etc. Hematoxylin and eosin (HE) staining displayed that the infiltrated mononuclear cells assembled in perivascular spaces, which were identified by morphological criteria as lymphocyte and macrophages.Forming typical muff-like changes, the inflammation was less severe in intraperitoneal EAE group than in subcutaneous foot dorsum EAE group.CONCLUSION: The EAE model induced in Wistar rats by Pertussis toxin administered subcutaneously on foot dorsum has the representative course of diseases, pathology change and clinical manifestation and the incidence of diseases is high and the cost is low. So it is a more ideal EAE model inducing method.

Objective:To explore the anti-inflammatory therapeutic effect and possible immunoregulatory mechanism of Buyang Huanwu Decoction (BYHWD) on the development of experimental autoimmune encephalomyelitis (EAE). Methods:Female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides ( MOG35-55 ) ,and randomly divided into saline group,BYHWD group,with 13 mice in each group. At the 3th day,25 ml/kg of saline was orally given to each mouse of saline group,50 g/kg ig of crude BYHWD was orally given to each mouse in BYHWD group for 25 days. Clinical score and body mass were recorded every day. Inflammatory cell infiltrations of spinal cord were observed by HE staining Myelin staining observes the demyelination situa-tion. And the expression of ROCKⅡ in spleen was detected by immunofluorescence staining. The subtypes of CD4+ T cells were analyzed by flow cytometry. Western blot was used to detect the expression of TLR4,Myd88,NF-κB,COX-2,ROCKⅡ in spinal cord and ROCKⅡ in brain. Results: The neurologicalscore significantly decreased in EAE mouse of BYHWD group compared with the saline group (P<0. 001) . BYHWD inhibited the inflammatory cell infiltration and demyelination in the nervous centralis(P<0. 05). The treatment of BYHWD effectively reduced the increased the proportion of CD25+(P<0. 05),IL-10+(P<0. 05),TGF-β+(P<0. 01), IFN-γ+( P<0. 05 ) CD4+T cells , and inhibited the expression of peripheral and central ROCKⅡ( P<0. 05 ) ;BYHWD reduced the expression of TLR4,MyD88,NF-κB,COX-2 in spinal cord (P<0. 05). Conclusion:BYHWD can exert anti-inflammatory and immune regulation effect by the inhibition of ROCKⅡ/TLR4/ NF-κB signaling pathway and regulation of the proportion of peripheral T cell sub-sets.

AIM:To evaluate the effect of lipoic acid ( LA) on LPS-induced Parkinson disease ( PD) model of mice.METHODS:Female C57BL/6 mice of 10-month-old were randomly divided into saline control group , PD group and LA group.The PD mouse model was induced by intranasal instillation of LPS .Assays of tyrosine hydroxylase , microglia and nuclear factor kappa B ( NF-κB) were performed by the methods of immunohistochemistry and Western blotting .RE-SULTS:Intranasal LPS instillation exhibited basic characteristics of PD model .However, LA administration significantly improved motor dysfunction , protected dopaminergic neurons from damage , and inhibited NF-κB activation in inflammatory microglia in the substantia nigra area of the brain .CONCLUSION:LA may exert a profound neuroprotective effect by an-ti-neuroinflammatory reaction to arrest the progression of PD .

Objective To investigate the immunoregulatory effect of Fasudil-modified macrophages on cell transferred experimental autoimmune encephalomyelitis ( EAE) in a mouse model.Methods Fe-male C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE.The encephalomyelitic mononuclear cells ( MNCs) were isolated from spleen of mice with EAE on day 9 after immunization and treated with or without Fasudil for 72 h in vitro.Several assays including the flow cytometry analysis, Griess reaction and ELISA were performed to analyze the M1 and M2 phenotypes of macrophages, the production of NO and the levels of cytokines, respectively.The cultured MNCs (5×107 cells) were resuspended in 500μl of PBS and transferred into na?ve C57BL/6 recipients via intraperitoneal injection.Two groups including the PBS-MNCs group and the Fasudil-MNCs group were set up.The body weights and clinical scores of the mice in each group were recorded in every other days after the induction of EAE in the recipients.Results The Fasudil treated MNCs affected the induction of EAE in adoptive cell transferred mice.The expression of CD16/32, iNOS and IL-12 on F4/80-macrophages were decreased, while the expression of CD206, CD23 and IL-10 on F4/80-macrophages were increased upon the treatment of Fasudil, indicating that Fasudil im-proved the differentiation of macrophages from M1 to M2 phenotypes.Moreover, Fasudil inhibited the pro-duction of NO and enhanced the expression of Arginase-1.Conclusion Fasudil ameliorated the clinical se-verity of EAE in mice by promoting the transformation of macrophages from M1 to M2 phenotype.

Objective To evaluate the clinical effect of venenum bufonis combined with antibiotics in the treatment of bacterial liver abscess .Methods 60 patients with bacterial liver abscess were enrolled ,and all patients took percutaneous transhepatic puncture guided by ultrasound .They were randomly divided into two groups according to the digital table ,30 cases in each group .The control group received antibiotics treatment ,and the treatment group received venenum bufonis plus antibiotics intravenous drip ,once daily ,one course of treatment lasted seven days .The body temperature , blood routine , procalcitonin and other project changes of the two groups were detected .Results Compared with the control group,the effective rate of the treatment group was higher (96.7％ vs.80.0％,χ2 =4.043,P=0.044).In the treatment group,the time of body temperature returning to normal [(7.00 ±1.67)d vs. (9.00 ±1.41)d],leukocyte recovery time [(7.83 ±2.32) d vs.(9.82 ±1.94) d],procalcitonin recovery time [(7.00 ±1.67)d vs.(9.00 ±1.41)d],symptom disappearance time [(5.17 ±1.72)d vs.(7.50 ±1.87)d],disap-pearance time of abscess[(12.00 ±3.41)d vs.(16.00 ±2.37)d]were shorter than those in the control group(t=-2.601,-2.890,-2.236,-2.248,-2.362,P=0.026,0.016,0.049,0.044,0.041).Conclusion Venenum bufonis combined with antibiotics can significantly increase the curative rate and accelerate infection control , there-fore,it is worthy of popularizing in clinical practice .

AIM:To explore the therapeutic effect of Buyang-Huanwu decoction (BYHWD) on experimental au-toimmune encephalomyelitis ( EAE) and its immunoregulatory effect on monocyte-macrophages .METHODS: Chronic EAE was induced by myelin oligodendrocyte glycoprotein peptide fragment 35-55 ( MOG35-55 ) in the female C57BL/6 mice, which were randomly divided into saline group and BYHWD group .On day 3 after immunization , the mice in BYHWD group were orally administrated with BYHWD , while normal saline was given to the control mice .The clinical score and body mass were recorded every other day .At day 17 after immunization , the mice were sacrificed and spinal cords were obtained for HE staining and myelin staining .The M1 and M2 macrophage phenotypes of splenic cells were detected by flow cytometry and immunofluorescence staining .The protein expression of iNOS , TNF-α, arginase and IL-10 in the spinal cord macro-phages was determined by Western blotting .RESULTS:BYHWD delayed the onset of EAE , reduced the clinical scores of EAE, inhibited the inflammatory cell infiltration and demyelination in the spinal cord , and promoted the conversion of M 1 macrophages into M2 phenotype in the spinal cord and spleen .CONCLUSION:BYHWD intervention attenuates the be-havioral and pathological changes in the EAE mice , and its mechanism may be related to the macrophage conversion .

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor WAR 5 on the experimental autoimmune encephalomyelitis (EAE) and its possible mechanism.METHODS: Female C57BL/6 mice were randomly divided into EAE group and WAR5 group.EAE model was induced by the application of MOG 35-55 peptide.WAR5 was in-jected intraperitoneally every other day from post-immunization (PI) day 3 to PI day 27.The clinical score and body weight were recorded every other day .On PI day 28, the animals were sacrificed and spinal cords were obtained for HE and mye-lin staining .The splenocytes were isolated and the expression of CD 16/32 and CD206 were analyzed by flow cytometry . The protein extracts from the brains and spinal cords were collected for the measurement of inducible nitric oxide synthase ( iNOS) by Western blotting .RESULTS:The administration of WAR 5 delayed the onset of EAE and attenuated the clini-cal symptoms .The results of the pathological examination revealed that WAR 5 inhibited the infiltration of inflammatory cells and improved myelination in spinal cords , accompanied with the poralization of M 1 macrophages to M2 phenotype in the spleen.WAR5 inhibited the expression of iNOS in the central nervous system , especially in the spinal cords .CON-CLUSION:The therapeutic effect of WAR5 on EAE may be related to the shift of M1 macrophages to M2 phenotype and inhibition of inflammation in the central nerve system .

Objective To investigate whether astragaloside ( AST) Ⅳ could inhibit lipopolysac-charide (LPS)-induced activation of astrocytes and the possible mechanism. Methods Effects of different concentrations of AST Ⅳ on astrocyte viability were determined by MTT to select the optimum concentration for the following experiments. Primary astrocytes were induced by LPS to construct the inflammatory model. Astrocytes were divided into three groups: PBS, LPS and LPS+ASTⅣ(LPS+ASTⅣ) groups. The release of nitric oxide (NO) was detected by Griess method. Expression of glial fibrillary acidic protein ( GFAP), an astrocyte marker, and TLR4 were analyzed by immunohistochemistry and Western blot. Cytokines of IL-6, TNF-α, IL-4 and IL-10 in the supernatants of cell culture for 24 h collected after stimulation were meas-ured by ELISA. Results ASTⅣsignificantly inhibited the LPS-induced activation of astrocytes and NO re-lease (P<0. 01), suppressed the expression of TLR4 (P<0. 05) and reduced the secretion of IL-6 (P<0. 01) and TNF-α (P<0. 01), but increased the secretion of IL-4 and IL-10 (P<0. 05 and P<0. 01). Con-clusion AST Ⅳ could significantly inhibit the LPS-induced activation of astrocytes and suppress inflamma-tory responses, and the possible mechanism might be related to reduced secretion of inflammatory factors af-ter blocking the expression of TLR4.