Objective To prepare the recombinant epitopes of HSV-gB and HSV-gD protein and provides a new antigen protein for the development of herpes simplex virus(HSV)vaccine.Methods The epitopes of HSV-gB and HSV-gD protein were analyzed by epitope prediction software.A novel gene named X which encoded 9 predicted epitopes of HSV-Gb and HSV-gD protein was designed and synthesized using chemical method.X gene was cloned into vector PET-28a(+),expressed in Escherichia cob' BL21(DE3),and analyzed by Western blot.Results X gene was successfully designed and expressed in Escherichia coli BL21(DE3).Western blot analysis showed that recombinant X protein,which was with His marker,can be detected by anti-His antibody.Conclusion In this study we establish a newmethod to express recombinant epitope protein,which may be a new protein for developing vaccine against HSV infection.

Objective To study the effect of Sarcandra Glabra on the expression of signal transduction molecules of TGF-β1/Smads signaling pathway in miniature pig of radiation-induced lung injury.Methods 75 miniature pigs were divided into control group,radiation group and radiation plus medication group randomly.At 1 week before exposure of right lung with 15 Gy γ-rays,the miniature pigs in radiation plus medication group were given Sarcandra glabra,while those in the other groups received an equal amount of saline.Right lung were taken at weeks 2,4,8,12 and 24 after irradiation,the pathological changes in the lung tissue were observed by HE staining,and the expression of mRNA and protein of TGF-β1,Smad2,Smad3,and Smad7 were detected by real-time PCR and western blotting,respectively.Results Sarcandra glabra reduced the inflammation and fibrosis of the lung tissue in miniature pig after irradiation.Compared with control group,the expressions of TGF-β1 and Smad3 were significantly increased at 2 weeks after irradiation(P ＜ 0.05),Smad2 and Smad7 were increased at 8 and 12 weeks after irradiation(P ＜ 0.05),respectively,in the radiation group.Compared with the radiation group,the expressions of TGF-β1 and Smad2 were significantly decreased(P ＜ 0.05) from the fourth and eighth week,respectively,Smad3 had no obvious change while Smad7 was significantly increased from the second week in the radiation plus medication group (P ＜ 0.05).Conclusions Sarcandra Glabra plays protective effect on radiation-induced lung injury in miniature pig by regulating TGF-β1,Smad2 and Smad7 expressions in the TGF-β1/Smads signaling pathway.

Objective To investigate the mechanism of hypedipidemia on pancreas of rats by comparative proteomic analysis.Methods Ten male SD rats were randomly divided into two groups,the experimental group Was fed with high lipid forage and the control group Was fed with normal food.Pancreatic samples from the two groups were harvested six weeks later.Differential protein analysis Was performed using differential in-gel electrophoresis(DIGE),and characterizing the protein biomarkers using tandem mass spectrometry.Western blot Was used to confirm the expression of significantly changed proteins.Results Compared to the normal pancreas tissue,a total of 3 protein spot-features were found to be significantly increased and 11 significantly decreased in pancreatic samples with hyperlipidemia.Significantly increased proteins in hyperlipidemia pancreatic samples were arginaseⅡ,ribonuclease inhibitor and glyeine amidinotransferase,which increased by 2.19,1.82 and 1.12 fold,respectively.Significantly decreased proteins in hyperlipidemia group were tyrosyl-tRNA synthetase,alpha-amylase,triacylglycerol lipase,DJ-1protein,Cu,Zn superoxide dismutase,which dicreased by 2.48,2.37,1.85,1.73 and 1.65 folds,respectively.Western blot analysis revealed increased arginase Ⅱ levels and decreased alpha-amylase in pancreatic samples with hyperlipidemia.Conclusions Pancreas wag possibly injured by hyperlipidemia via increase of arginase Ⅱ.Decreased amylase and lipase may be the protection mechanism of pancreas.

This text briefly introduces tutorial system's history evolution and development and expounds its necessity and realistic meaning,emphasizing that practising such a guidance program is necessary for developing students' innovative ability and individuality,and for strengthening the construction of the teachign staff,which is the call of students and the responsibility of education,and the inevitable choice under credit system.It focuses on how to set up effective and practical tutorial system of the undergraduate course from the construction of tutors'troops,the feasible measures and system,and tutor stimulative mechanism.

Objective To summarize the clinical characteristics of primary pigmented nodular adrenocortical disease (PPNAD).Methods Eight patients with PPNAD from 2001 to 2009 in PUMC Hospital were reviewed,their clinical data were collected.Results PPNAD often occurred in adolescents.62.5% of patients with PPNAD were complicated with Carney complex(CNC).In addition to general features of Cushing's syndrome,amenorrhea and growth retardation in stature were very frequent in clinical manifestations of PPNAD.Plasma ACTH was undetectable,circadian rhythm of plasma cortisol was disappeared,glucocorticoid excretion was increased paradoxically during the dexamethasone suppression test in 50% patients with PPNAD.Adrenal imaging from 75% patients revealed normal-sized adrenal glands or suspectable micronodular changes.Adrenal pathologic analysis revealed numerous brown cortical nodules containing lipofuscin pigmentation.Unilateral adrenalectomy may relieve symptoms of Cushing's syndrome,but plasma ACTH and circadian rhythm of plasma cortisol were difficult to recovere.Hypercorticoidism might re-occure after unilateral adrenalectomy.Conclusion PPNAD should be bewared in ACTH independent Cushing's syndrome patients without apparent adrenal mass,and CNC should be screened and followed up.

Objective To explore the influence and mechanism of cytokines and protein expression of alveolar epithelial type (ACE) Ⅱ cells in Bama minipigs' right-thorax with a single 15 Gy dose irradiation.Methods All minipigs received either right thoracic irradiation or sham-irradiation under anesthesia.At 4,8,12 and 24 week post-irradiation,5 minipigs respective and random from irradiarion groups and control group were sacrificed to remove the lungs.The protein expression of surfactant associated protein (SP)-A,transforming growth factor (TGF)-β1,Vimentin and E-cadherin were detected by Western blot.The protein expression of α-smooth muscle actin (SMA) was detected by immunohistochemistry.The co-localization of SP-A and α-SMA was visualized by double immunofluorescence staining.Results At 4,8,12 and 24 week post-irradiation,a significant increase in the protein expression of α-SMA,TGF-β1 and Vimentin were observed in irradiated lung compared to sham-irradiated controls(α-SMA:t =2.46-3.26,P ＜0.05;TGF-β1:t =2.96-3.52,P ＜0.05;Vimentin:t =3.24-5.05,P ＜ 0.05).By contrast,the protein expression of SP-A and E-cadherin in irradiation group was lower than it in control group (SP-A:t =3.62-4.65,P ＜ 0.05;E-cadherin:t =2.53-4.15,P ＜ 0.05).Moreover,at 8 week after irradiation,under confocal laser scanning microscope,the co-localization of SP-A and α-SMA was observed in irradiated alveolar epithelium cells,and it was not observed in sham-irradiated controls.Conclusions These data demonstrate that E-cadherin,SP-A and TGF-β1 may act as sensitive predictors of radiation-induced lung injury(RILI).Irradiation may lead to ACE Ⅱ cells achieving a mesenchymal phenotype,namely,epithelial to mesenchymal cells transition occurs,and ACE Ⅱ cells play the important part in the development of RILI by epithelial-mesenchymal transition.

Objective To investigate the clinical efficacy of scalp acupuncture plus electroacupuncture in treating migraine without aura.Methods Three hundred and twenty-six patients with migraine without aura were randomly allocated to a treatment group of 157 cases and a control group of 169 cases. The treatment group received scalp acupuncture plus electroacupuncture and the control group, medication alone. The Pain Rating Index (PRI) score and the TCM syndrome rating scale score were scored in the two groups of patients before and after treatment. The numbers of taking medicine and the numbers of headache attacks were compared between the two groups of patients during treatment. An analysis was made of factors influencing the therapeutic effect on migraine. Results There were statistically significant differences in the PRI score and the TCM syndrome rating scale score at the end of treatment and at one, two and three months of follow-up compared with before treatment in the two groups (P<0.05). There were statistically significant differences in the PRI score and the TCM syndrome rating scale score at the end of treatment and at one and two months after it between the treatment and control groups (P<0.05). There was a statistically significant difference in the weekly number of taking medicine between the treatment and control groups during treatment (P<0.05). There was a statistically significant difference in the number of headache attacks between the treatment and control groups at two, three and four weeks of treatment (P<0.05). In the treatment group, the therapeutic effect at one, two and three months of follow-up was related to both TCM syndrome type and the VAS score; the therapeutic effect at two months of follow-up was related to the sex; the therapeutic effect at three months of follow-up was related to the educational level.Conclusion Scalp acupuncture plus electroacupuncture is an effective way to treat migraine without aura.

Objective To evaluate the effect of early enteral nutrition (EN) on the pancreatic exocrine secretion in dogs with acute necrotizing panereatitis (ANP). Methods ANP model was induced by injection of mixtured solution of 5% sodium tanrecholate and trypsin into the pancreatic duct. Thirty dogs were randomly divided into total parenteral nutrition (TPN) group (n=5), duodenal PEPTI-2000VARIENT (DP) group (n=5), duodenal Nutrison MuhiFibre (DN) group (n=5), jejunal PEPTI-2000VARIENT (JP) group (n=5), jejunal Nutrison MuhiFibre (JN) group (n=5). The dogs were treated by either TPN or EN 24 hours after ANP model induction and the nutrition support lasted for 5 days. Serum amylase, LDH, lipase, secretin (SEC), cholecystokinin (CCK) and gastrin were measured at 1, 2, 3, 4, 5 d. Pancreatic juice was collected for 3 hours after TPN or EN started, and the amount of pancreatic juice and levels of proteinase, amylase, lipase, HCO3-, K+, Cl-, Na+ were determined. Dogs in each group were sacrificed at day 7. Histological and ultra-structure changes of the pancreatic tissues were evaluated pathologically. Results The levels of serum amylase, LDH, lipase, CCK, amount of pancreatic secretion and K+, Cl+, Na+ were not significantly different among these groups. The levels of plasma SEC and gastrin, HCO3-, proteinase, amylase, lipase in the duodenal nutrition groups were significantly higher than those in TPN group (P<0.05). The above mentioned parameters in the jejunal nutrition group were significantly lower than those in the duodenal group (P<0.05) and higher than those in the TPN group without significant difference. Among the 2 jejunal nutrition groups, the levels of plasma gastrin, HCO3- in pancreatic juice, proteinase, amylase, lipase in the JP group were significantly higher than those in the JN group (P<0.05). The above mentioned parameters in the DP group were significantly lower than those in the DN group (P<0.05). The amount of pancreatic secretion, HCO3-,K+, Cl+, Na+ were not significantly different among these groups. The pathological changes were similar among these groups, and the extent of pathological changes was relatively better in the JP group. The amount and density of intracytoplasm zymogen granules of pancreatic acinar cell were not significantly lower than those in the TPN group. Conclusions The delivery of nutrients to the proximal jejunum with elemental low-fat diets did not increase the pancreatic exacrine activity.

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Objective To study the intra-and extra-hepatic collateral pathways in various types of Budd-Chiari syndrome (BCS) using liver acceleration volume acquisition (LAVA) enhanced MRI.Methods The clinical data and imaging findings of 240 patients with BCS were collected and analyzed.The types of BCS confirmed by DSA.Intra-and extra-hepatic collateral pathways were studied using LAVA enhanced MRI with a 3.0T scanner.Correlations of the intra-/extra-hepatic collateral pathways with the types of BCS were analyzed using the Chi-square test.Then,the degrees of correlation were calculated by the Cramet correction coefficient of contingency.Results Among the 240 patients,DSA confirmed 60 patients to have hepatic vein occlusion,39 patients to have inferior vena cava occlusion and 141 patients to have both hepatic vein and inferior vena cava occlusion.MRI demonstrated dilated accessory hepatic veins in 157 patients,intra-hepatic communicating branches in 69 patients,inferior phrenic veins in 43 patients,superficial epigastric veins in 135 patients,umbilical veins in 94 patients and hemiazygos/azygos veins in 195 patients.Accessory hepatic veins and hemiazygos/azygos veins as collateral pathways were associated with the types of BCS (x2 =30.239,P ＜ 0.05;x2 =51.295,P ＜ 0.05,respectively).The degrees of correction were 0.355 and 0.462,respectively.Accessory hepatic veins as collateral pathways were most common in the mix type,accounting for 79.4％.Hemiazygos/azygos veins were most common in the inferior vena cava occlusion type and the mix type,accounting for 92.3％ and 91.5 ％,respectively.Conclusions Accessory hepatic veins and hemiazygos/azygos veins as collateral pathways were associated with the types of BCS,while the intra-hepatic communicating branches,inferior phrenic veins,superficial epigastric veins and umbilical veins were not correlated with the types of BCS.LAVA may help to diagnose and determine the best choice of treatment for the various types of BCS.