Objective To study the clinical significance of peripheral blood PD-1 (CD279) and its ligands(CD274 and CD273)between patients with anti-HBe positive chronic hepatitis B (CHB) or anti-HBs positive group of normal glutamic acid alanine aminotransferase (ALT).Methods A total of 70 subjects of normal ALT,including 23 patients with anti-HBe positive,22 anti-HBs positive and 25 normal controls,were enrolled.The expression level of CD279,CD274 and CD273 in peripheral lymphocyte were detected by flow cytometry.Compare the group differences by multivariate analysis of variance.Results The expression level of CD279 in control group,anti-HBe positive group and anti-HBs positive group was (33.3±5.4)％,(48.0±9.1) ％,(42.8±9.0) ％,respectively.The expression level of CD274 in control group,anti-HBe positive group and anti-HBs positive group was (59.4 ±4.4) ％,(71.3± 10.3) ％ and (62.9 ± 10.2) ％,respectively.The expression level of CD273 in control group,anti-HBe positive group and anti-HBs positive group was (5.3±2.2)％,(15.6±5.3)％ and (5.1±1.0)％,respectively.The expression level of CD279 and CD274 decreased gradually from anti-HBe positive group to anti-HBs positive group and then to normal control group.Three dependent variables in the overall difference was statistically significant (Pillai trace =0.988,F=3090.105,P=0.000),η partial square was 0.458.The difference for CD273 expression level of anti-HBs positive group compared with control group was not statistically significant (P =0.082) ; for other groups pairwise comparisons were statistically significant(P＜0.01).Conclusion Monitoring the alteration of PD-1 and its ligands of the HBeAg seroconversion CHB patients and the anti-HBs positive population can better understand the changes in course of HBV infection,thereby improving the prognosis of patients.with HBV infection.

ObjectiveTo summarize the one-year follow-up clinical result of patients undergoing Endoscopic vein harvest (EVH) technology to collect greater saphenous vein( GSV )in coronary artery bypass operation (CABG),and to assess the related factors influencing the outcome.Methods248 patients underwent off-pump coronary artery bypass grafting (OPCAG) from May 2009 to May 2010.Among these patients,136 patients with coronary artery disease received EVH technology to gain GSV,and 112 patients received conventional open vein harvesting (OVH).Then 71 EVH (group 1 ) patients and 64 OVH ( group 2 ) patients had one-year follow-up analysis.We compared and evaluated the data of two groups about operation information,lower limb wound complication,bridge blood vessels patency rate,and psychologic status.ResultsThe date of wound surface size,wound healing and appearance,wound infection rate,the second debridement rate and the overall lower limb wound complication rate show that EVH was better than OVH.After one-year follow-up,we contrasted the patency rate of bridge blood vessels between two groups.The patency rate of arterial bridge blood vessels was 96.8％ ( venous bridge blood vessels was 85.7％ )in group of EVH when we followed up 63 patients;while the patency rate of arterial bridge blood vessels was 94.9％ ( venous bridge blood vessels was 86.4％ ) in group of OVH that we followed up 59 patients.We compared the outcome between two groups,and found that there were no statistical significance in angina recurrence rate,patency rate of venous graft and patency rate of arterial graft.But the psychologic status was different,the EVH group was better than OVH group.ConclusionCompared to OVH,EVH technique has more advantages.Through taking a more secure method in the collection process of the bridging vein material,EVH group also maintained a satisfactory one-year graft patency rate.

Objective To explore the experience in the diagnosis and treatment of elastofibroma dorsi.Methods Clinical data of 19 cases of pathologically confirmed elastofibroma from October 2001 to October 2011 were reviewed.The clinical features,the specific radiological characteristics,the typical pathological alterations and the short-term and long-term effects were analyzed.Results All the lesions were located within the muscles in the subscapular region.There were 3 cases with bilateral lesions and 16 cases with unilateral lesions.Seven patients complained of local pain and feeling of foreign body when activated the upper limb,while 12 were asymptomatic.Except for the early 6 cases,accurate diagnosis was made in all the other 13 cases before the histological exams solely based on the physical examination and imaging findings.Marginal excision was done for all the cases:the diameters of the masses were between 3 centimeters to 10 centimeters.Fluid accumulation complications was found in 1 case and resolved by repeated paracentesis;errhysis complication,was found in 1 case and resolved by compression,applying hemostatics and blood transfusion.No recurrence was found during the follow-up period(6 months to 48 months).Conclusion Elastofibroma dorsi is usually found in the subscapular region in elderly women.The diagnosis can be made on the basis of unique imaging characteristics and physical examination before histological examine.Surgical marginal excision can achieve good short-term and long-term effects.

Objective To investigate the design and clinical result of the modified medial fasciocutaneous flap of lower leg in the different zonation. Methods The length of lower leg was divided into three zonations equally.According to the different zonation of the fasciocutaneous flap,the 45 patients were divided into three groups from Jan.2005 to Feb.2008.The defects of the patients were repaired with the flaps.The sizes of the flaps ranged from 5 cm×3 cm to 25 cm×10 cm.Results The flaps survived completely in 43 cases.The distal sides of flaps necrosed partially in 2 cases in the upper third of the lower leg.The necrosed part of the flap was repaired after the change dressings.The colour and texture of flaps were excellent,the appearance and function were satisfactory after a follow up of 4-20 months.Conclusion The modified medial fasciocutaneous flap of the lower leg is easy to design and dissect,blood supply is reliable without sacrifice of the major arteries.The flap can be used according to the different location of the fascia pedicel.It is the idea flap to repair the soft tissue defects of the front of knee joint,the leg,the ankle and the foot.

Objective:To explore the effects of Bufalin on the growth and proliferation of human glioma cells U251. Methods:Methyl thiazolyl tetrazolium(MTT)assay was used to detect the effect of Bufalin on the proliferation of human glioma cells U251. An in-verted microscope was used to observe the changes of cell number,morphology and activity. AnnexinV/ PI was used to measure the in-duction of cell apoptosis caused by Bufalin. Results:Bufalin at different concentrations(0. 001 - 10. 0μmol·L - 1 )inhibited the pro-liferation of U251 cells in a dose and time-dependent manner. Compared with that of the control group,the apoptosis rate of Bufalin group was increased significantly(P < 0. 01). Conclusion:Bufalin can inhibit the growth and proliferation of U251 cells in a dose and time-dependent manner,and induce the apoptosis of U251 cells.

Objective To investigate the neuroprotective effects of Fasudil in cerebral I/R injury in mice.Methods 51 C57BL/6J mice was divided into two groups,CMC treated group (n=26) and Fasudil treated group (n=25),randomly.The mice were treated with Fasudil (10 mg/kg) or CMC (0.5％ CMC 10 ml/kg) separately.Then the treated mice were subjected to 60 min of focal ischemia and 18 h reperfusion.The infarct volume of brain was analyzed by TTC staining with MCID image system.BBB permeability was assessed by Evans blue extravasation and albumin leakage which was detected by immuno-blotting assay.The activity of MMP9 was analyzed by zymography.Results The infarct volume in CMC group ((99.07±6.53) mm3) was larger than that in Fasudil group ((57.02±8.93) mm3),the difference was statistically significant (P＜0.01).The activity of MMP9 in the mice treated with Fasudil was lower than that in CMC group.Compared with the CMC group(albumin (2.95±0.77),Evans blue (5.15±0.24)),the albumin and Evans blue content in the Fasudil treated group (albumin (1.04±0.18),Evans blue (1.96±0.31))reduced significanctly(all P＜0.01).Conclusion Fasudil protects I/R damage by inhibiting the activity of MMP9 to maintain blood-brain barrier permeability.

Objective To observe the effects of Jianpi Liqi Huashi Prescription on hepatocyte injury,oxidative stress and nitrative stress in mice with non-alcoholic steatohepatitis(NASH);To explore its mechanism in the treatment of NASH.Methods Totally 32 male C57BL/6J mice were randomly divided into control group,model group and TCM low-and high-dosage groups,with 8 mice in each group.The control group was fed with ordinary diet,and the other groups were fed with high-fat diet,for consecutive 16 weeks.Starting from the 13th week,the control group and model group were given 0.4%CMC-Na solution by intragastric administration and the TCM groups were given corresponding doses of drugs by intragastric administration,respectively.Biochemical instrument was used to detect serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)contents,HE staining and oil red O staining were used to detect liver histopathology,triglyceride(TG)content and myeloperoxidase(MPO)activity in liver tissue were detected through test kit.immunofluorescence staining was used to detect positive expression of CD11b in liver tissue,the mRNA expressions of NOX1 and NCF1 were detected by RT-PCR,and the protein expressions of inducible nitric oxide synthase(iNOS)and 3-nitrotyrosine(3-NT)in liver tissue were detected by Western blot.Results Compared with the control group,the hepatocytes in the model group showed diffuse steatosis,hepatocyte swelling and inflammatory cell infiltration;a large number of fat droplets were formed by oil red O staining;serum ALT and AST contents significantly increased(P<0.05),the TG content and MPO activity in liver tissue significantly increased(P<0.05);the positive expression of CD11b in liver tissue increased;the mRNA expressions of NOX1 and NCF1 in liver tissue significantly increased(P<0.05);the expressions of iNOS and 3-NT protein in liver tissue significantly increased(P<0.05).Compared with the model group,the degree of liver steatosis,the level of inflammatory cell infiltration and the number of lipid droplets in TCM groups decreased significantly;serum ALT and AST contents significantly decreased(P<0.05);the TG contents and MPO activity in liver tissue significantly decreased(P<0.05);the positive expression of CD11b in liver tissue decreased;the mRNA expressions of NOX1 and NCF1 in liver tissue significantly decreased(P<0.05);the expression of 3-NT protein in liver tissue significantly decreased(P<0.05);the expression of iNOS protein in liver tissue of mice in TCM high-dosage group significantly decreased(P<0.05).Conclusion Jianpi Liqi Huashi Prescription may relieve liver cell damage,lipid deposition and inflammation in NASH mice by alleviating oxidative stress and nitrative stress in the liver,and play a role in the treatment of NASH.

BACKGROUND:Oxidative injury is considered to be one of the important factors of cerebral ischemia-reperfusion injury.Superoxide dismutase 2(SOD2)is a key mitochondrial antioxidant molecule,and fenofibrate can regulate the expression of SOD2 by activating peroxisome proliferator-activated receptor α. OBJECTIVE:To explore whether the mechanism of fenofibrate in the treatment of cerebral ischemia-reperfusion injury depends on the expression of SOD2. METHODS:The TALENs system was used to construct SOD2 transgenic mice.The transgenic mice were genotyped by PCR and DNA sequencing techniques.The expression of SOD2 protein in transgenic mice was detected by western blot assay.Wild-type and SOD2 transgenic mice were randomly divided into four groups:wild-type control group(n=6),wild-type fenofibrate group(n=6),SOD2 transgenic control group(n=5)and SOD2 transgenic fenofibrate group(n=5).A mouse model of middle cerebral artery occlusion was prepared using the suture-occlusion method.After 90 minutes of ischemia,the thread was removed to reperfuse cerebral blood flow for 30 minutes.A cerebral blood flow monitor was used to monitor local cerebral blood flow.Brain tissue slices were taken for 2,3,5-triphenyltetrazolium chloride staining to analyze the situation of cerebral infarction in each group. RESULTS AND CONCLUSION:After PCR and DNA sequencing analysis,nine SOD2+/+ transgenic mice were successfully constructed.After cerebral ischemia-reperfusion,the wild-type fenofibrate group showed partial recovery of cerebral blood flow and significantly reduced cerebral infarction volume compared with the wild-type control group(P<0.001).There was no significant difference in cerebral blood flow and cerebral infarction volume between the SOD2 transgenic fenofibrate group and the SOD2 transgenic control group.The SOD2 transgenic control was superior to the wild-type control group in terms of improving cerebral blood flow and cerebral infarction(P<0.001).There were also no significant differences in cerebral blood flow and cerebral infarction volume between the wild-type fenofibrate group and the SOD2 transgenic control group and between the wild-type fenofibrate group and the SOD2 transgenic fenofibrate group.To conclude,the expression of SOD2 is one of the mechanisms of fenofibrate in the treatment of cerebral ischemia-reperfusion injury.

ObjectiveTo explore the mechanism of modified Shuyuwan in treating vascular dementia （VaD） complicated with depression in mice. MethodThe VaD model was established by bilateral carotid artery stenosis （BCAS） in seven 3-month-old male C57/BL6 mice. The regional cerebral blood flow （rCBF） of mice was measured by laser speckle imaging before and after BCAS surgery. Then， the BCAS method was combined with chronic unpredictable mild stress （CUMS） to establish a mouse model of VaD complicated with depression. BCAS/CUMS mice were assigned into BCAS/CUMS， fluoxetine （0.01 g·kg^-1）， and high-， medium-， and low-dose （20， 10， 5 g·kg^-1， respectively） modified Shuyuwan groups. The shame group underwent sham operation without CUMS （n=10）. The tail suspension test and sucrose preference test were carried out to examine the depressive behaviors of mice. The distribution and expression of myelin-associated glycoprotein （MAG）， myelin basic protein （MBP）， neurofilament heavy polypeptide （NF200）， and anti-non-phosphorylated neurofilament epitope antibody （SMI32） in the corpus callosum （CC） were detected by the immunofluorescence assay. Western blot was employed to determine the protein levels of monocarboxylate transporter 1 （MCT1）， MBP， MAG， oligodendrocyte glycoprotein （MOG）， amyloid precursor protein （APP）， NF200， contactin-associated protein （Caspr）， and voltage-gated sodium channel （Nav1.6） in the corpus callosum. The level of lactic acid in the serum was measured by the lactic acid assay kit， and the ultrastructure of myelin was observed by ultraprojective electron microscope. ResultLaser speckle imaging showed that rCBF decreased immediately 10 min after BCAS surgery （P<0.01）， and the rCBF was still cerebral hypoperfusion and did not return to the preoperative level 2 weeks after surgery. Behavioral test results showed that compared with the sham group， the BCAS/CUMS group presented decreased percentage of sucrose preference （P<0.01） and prolonged immobile time in the tail suspension test （P<0.01）. Compared with the BCAS/CUMS group， fluoxetine and modified Shuyuwan increased the percentage of sucrose preference （P<0.01） and shortened the immobile time in the tail suspension test （P<0.01）. The level of lactic acid was the highest in the BCAS/CUMS mice （P<0.01）， and modified Shuyuwan lowered the lactic acid level （P<0.01）. Immunofluorescence results showed that compared with the sham group， the BCAS/CUMS group presented decreased fluorescence intensity of MAG， MBP and NF200 and increased fluorescence intensity of SMI32 in the corpus callosum， and such changes were reversed by modified Shuyuwan at different doses and fluoxetine. Western blot results showed that compared with the sham group， the BCAS/CUMS modeling down-regulated the protein levels of MCT1， MBP， MOG， MAG， NF20， and Caspr （P<0.05， P<0.01） and up-regulated the protein levels of APP and Nav1.6 in the corpus callosum， and the above trends were reversed by modified Shuyuwan （P<0.05， P<0.01）. Compared with the sham group， BCAS/CUMS modeling led to myelin ultrastructure damage and axon atrophy， which were alleviated by modified Shuyuwan. ConclusionModified Shuyuwan can ameliorate the transport disorder of lactic acid between myelin sheath and axon by upregulating the expressin of MCT1， promote the regeneration of myelin sheath in the corpus callosum， and improve the integrity of myelin sheath structure， thereby alleviating depression in VaD mice.

ObjectiveTo investigate the effect and mechanism of modified Shuyuwan （SYW） on hippocampal myelin sheath injury in vascular dementia （VD） model rats. MethodSixty male SD rats of SPF grade were randomly divided into sham operation group， model group， and high-， medium- and low-dose modified SYW groups， with 12 rats in each group. The VD model was induced by bilateral carotid artery ligation in rats of the groups except for those of the sham operation group. After modeling， rats were screened by the water maze test， followed by drug treatment by gavage. Specifically， rats in the modified SYW groups were treated with modified SYW at 10， 5， 2.5 g·kg^-1·d^-1， accordingly， and those in other groups were administered with the same amount of normal saline. After intragastric administration for 28 days， the spatial learning and memory abilities of rats were detected by the water maze test. The hippocampal neuron structure was observed by hematoxylin-eosin （HE） staining. The content of hippocampal tumor necrosis factor （TNF）-α， interleukin-6 （IL-6）， and glutamate （Glu） was observed by biochemical detection. The hippocampal expression of myelin basic protein （MBP）， astrocyte activation marker glial fibrillary acidic protein （GFAP）， and connexin 43 （Cx43） was detected by immunofluorescence detection. The myelin sheath structure in the hippocampus was observed by the electron microscope. The α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor （AMPAR） and Cx43 protein expression was detected by Western blot. ResultCompared with the sham operation group， the model group showed prolonged escape latency （P<0.01）， decreased times of crossing the original platform and percentage of target quadrant detention time （P<0.01）， disordered neuron structure in the hippocampal CA1 region， loose myelin sheath lamella with blurry edge， up-regulated expression levels of TNF-α， IL-6， and Glu in the hippocampal CA1 region， especially Glu （P<0.01）， reduced expression of AMPAR （P<0.01）， increased protein expression of p-AMPAR and Cx43 （P<0.01）， significantly dwindled protein expression of MBP in the myelin sheath， and enhanced fluorescence co-labeled by GFAP and Cx43. Compared with the model group， the modified SYW groups showed shortened escape latency （P<0.05）， increased times of crossing the original platform and percentage of target quadrant detention time （P<0.05）， closely arranged hippocampal neuron structure， denser myelin sheath lamella with clear edge， down-regulated expression levels of TNF-α， IL-6， and Glu in the hippocampal CA1 region， especially Glu （P<0.01）， up-regulated AMPAR （P<0.01）， reduced protein expression of p-AMPAR and Cx43， especially in the high-dose group （P<0.01）， significantly elevated protein expression of MBP in the myelin sheath， and weakened fluorescence co-labeled by GFAP and Cx43， especially in the high-dose group. ConclusionModified SYW can improve the learning and memory abilities of VD rats， and the mechanism may be related to the inhibition of Cx43 expression， reduction of the release of Glu， inhibition of AMPAR-mediated inflammatory response to reduce the production of astrocyte marker GFAP， and promotion of the expression of MBP protein to alleviate myelin injury.