Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

Background: Nummular headache is a new category of primary headache disorder characterized by consistent location, size, and shape of painful areas. The pathogenesis is uncertain. Bifocal painful areas are rare manifestations but may expand the clinical diversity of nummular headache. Methods: The clinical characteristics of 5 bifocal nummular headache patients were reported and those of 11 patients in previous studies were reviewed. Bifocal nummular headache was classifi ed into two types. Type I was defi ned as a simultaneous activation of two painful areas while type II was defi ned as two painful areas occurring in different times. Results: All 16 patients were female, with mean age of onset and initial presentation of 54.7 years and 58.2 years, respectively. There were seven type I and nine type II patients. The parietal area, especially the tuber parietale, was the leading site of involvement in both types of patients. The shape and size of painful areas were also similar between these two groups. There was an equal frequency of ipsilateral and contralateral painful areas. The pain intensity was similar in both types of patients but was milder in new painful areas than in previous painful areas in type II patients. Conclusions: Bifocal nummular headache suggests a central role of nummular headache but does not debunk the peripheral theory of nummular headache. The accumulated fi ndings in bifocal NH patients do not support a generalization of pain occurrence or a reproduction of local process of epicranial neuralgia at multiple sites in nummular headache.

Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

AIM: In order to explore the pathogenesis of ulcerative colitis (UC), an experimental colitis in mouse was induced by the hapten dinitrochlorobenzene (DNCB), and the activity of leukocyte migration inhibitory factor (LMIF) was measured at the same time. METHODS: 67 BALB/c mice were randomly divided into control (60% ethanol) and DNCB groups. After they were sensitized by smearing 3.3% DNCB on the abdominal skin, they were challenged with DNCB at concentration of 0.1%, 0.2% and 0.4% respectively by instillation once a day. The weight, stool viscosity and hematochezia were observed and accumulated as disease active index (DAI) score. The pathological changes in colon tissue were judged macropathologically and by means of microscope. LMIF activity was determined by the absorbance (A) of migrated leukocytes. RESULTS: Compared to control group, the increases in DAI accumulate score, pathologic score, and LMIF activity in DNCB groups were observed. CONCLUSION: Mouse colitis was induced by DNCB, which was accompanied by an increase in LMIF activity. [

Ascorbic Acid ; therapeutic use ; Bandages ; Calcium Gluconate ; therapeutic use ; Humans ; Levofloxacin ; therapeutic use ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Sarcoidosis ; drug therapy ; Skin Ulcer ; drug therapy

Objective To summarize the experience of intraoperative care for the patients undergoing microsurgery via retrosigmoid keyhole approach, so as to improve the treatment effect and minimize complications. Method The clinical records on the intraoperative care of 100 patients undergoing microsurgery via retrosigmoid keyhole approach were retrospectively analyzed. Result Using the keyhole technique and small incision, the cerebellopontine angle lesions were exposed clearly. Moreover, the proper intraoperational cooperation shortened the duration of operation, reduced bleeding and minimized postoperative complications such as cerebrospinal fluid otorrhea, intracranial infection, intracranial hemorrhage, and subcutaneous hydrops. Conclusion Proper operational postures and intraoperative cooperation can make full use of the limited space of the microsurgery via retrosigmoid keyhole approach and ensure the success of the operation.

This study was aimed to establish the TLC identification method of Radix Mirab ilis himalaic a. The β-sitosterol and daucosterol were used as the reference substances. The single-factor test was used. A variety of factors which affected TLC were systematically investigated to filter out the best TLC conditions for identification of different batches of medicines. The results showed that the best TLC conditions were as follows: silica gel G plates, extraction solvent (methanol), reagent (5% sulfuric acid in ethanol), extraction method (ultrasonic extraction with methanol), ex-tracted time (30 min), the agent (petroleum ether-ethyl acetate-acetone (5:2:1)) and sample volume (6 μL). It was concluded that the method, which had high separation degree, was reproducible and simple. It can be used as the quality control of Radix Mirab ilis himalaic a.

Objective To analyze the stress distribution of the weakened root dentine restored with titanium post-core crown and fiber post-core crown by finite element method.Methods The sample was scanned by Inveon Micro-CT technology from incised margin to root apex at 21 μm interval to obtain transverse data of DICOM format.The three-dimensional finite element model was created by rebuilding the morphology of maxillary central incisor with Inveon Research Workplace as well as Mimics,Geomagic and Catia software.The model was meshed and loaded to analyze the stress distribution on the weakened dentine of root by finite element method.Results The maximum stress,shearing stress and equivalent stress of the root restored with fiber post-core crown were 60%less than the stresses of the root restored with titanium post-core crown in the experiment.The stress on various parts of the root restored with titanium post-core crown was higher than that of the root restored with fiber post-core crown.Furthermore,the stress concentration occurred mainly on the labial interface of the upper 1/3 of root. Especially when the root was restored with titanium post-core crown,the stress value reached 37 .73 MPa,which was 1.9 times that of fiber post-core crown.Conclusion The finite element analysis indicates that the fiber post-core crown is better than titanium post-core crown in avoiding stress concentration and root fracture in the restoration of weakened root of maxillary central incisors.

AIM:To explore the role of sphingosine 1-phosphate (S1P) in the dysfunction of vascular endo-thelial cells exposed to high glucose.METHODS: In human aortic endothelial cells cultured under high-glucose ( 22 mmol/L glucose) medium, nitric oxide ( NO) level, polymorphonuclear neutrophil-endothelial cell adhesion rate, protein level of intercellular adhesion molecule-1 ( ICAM-1) , migration of endothelial cells and Akt/endothelial nitric oxide syn-thase ( eNOS) pathway activation were observed after S1P, sphingosine kinase-1 inhibitor and/or Akt inhibitor treatments. RESULTS:S1P decreased NO level, increased polymorphonuclear neutrophil adhesive rate, enhanced ICAM-1 protein level, and inhibited migration of endothelial cells and activation of Akt/eNOS pathway in endothelial cells cultured under high-glucose condition.Sphingosine kinase-1 inhibitor, which reduced S1P content, significantly improved the above endo-thelial cell function indexes and restored the activation of Akt/eNOS pathway.CONCLUSION: S1P promoted high glu-cose-induced dysfunction of endothelial cells probably by inhibiting the activation of Akt/eNOS signal pathway.Targeting S1P is expected to become one of potential treatment strategies to reduce endothelial cell dysfunction.

Objective To investigate the biological effects of exosomes secreted by KYSE410 cells on migration and invasion of KYSE410,KYSE510,YES2 cells and the possible mechanisms underlying the phenotype change.Methods The exosomes were isolated from the conditional supernatant of esophageal cancer cell line KYSE410 by ultracentrifugation.The morphology of exosomes was observed by transmission electron microscopy (TEM).Western blotting was used to detect the protein markers of exosomes.The uptaken of fluorescence-labeled KYSE410 exosomes by KYSE410,KYSE510 and YES2 was also recorded under confocal microscopy.Migration and invasion ability of the three esophageal carcinoma cell lines and the effects of exosomes from KYSE410 on migration and invasion of KYSE410,KYSE510 and YES2 cells were analyzed by Transwell chamber,respectively.The alteration of Wnt/β-catenin and PI3K/Akt pathway-related proteins were detected by Western blotting.Results The membrane structure of KYSE410 derived exosomes could be observed with its diameter ranged between 30-100nm.The invasion and migration ability of three esophageal cancer cells are KYSE410＞ KYSE510＞ YES2.KYSE410 exosomes promoted the migration and invasion of KYSE410,KYSE510 and YES2 cells.Conclusions Concentrated exosomes derived from the highly migratory and invasive esophageal cancer cell line KYSE410 promoted the migration and invasion potentials of itself and esophageal cancer cell lines KYSE510 and YES2,which possibly exerted the effects by activating Wnt/β-catenin and PI3K/Akt signaling pathways.