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.

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. [

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

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.

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

AIM:To establish a simple and accurate HPLC method for quality evaluation and determination of 8 active constituents, i.e. Danshensu, Protocatechualdehyde,rosmarinic acid,Salvianolic acid B, Dihydrotanshinone, Cryptotanshinone, Tanshinone Ⅰand Tanshinone ⅡA in Radix Salviae Miltiorrhizae, at the same time, in order to study the correlations of the contents of the 8 constituents. METHODS: Detected at 286 nm, analytes were separated on a ODS C_ 18 column(150 mm?4.6 mm, 10 ?m) eluted gradually with methanol and 1% tetrafuran in 1% formic acid water solution(3∶97)on the increasement of methanol from 3% to 70% in the whole analysis. After determination, SPSS 13.0 software was used to calculate the correlation between the 8 constituents. RESULTS: Under the above conditions, HPLC method was constructed to quantify 8 constituents,and their correlation coefficients were calculated. CONCLUSION: The result shows that the contents of the 8 constituents in Salvia miltiorrhiza Bge. vary greatly from different origins; and there are significant correlations between the contents of hydrophilic constituents such as Salvianolic acid B,Danshensu,rosmarinic acid, so do hydrophobic constituents like Cryptotanshinone,Tanshinone Ⅰ,Tanshinone ⅡA,but there is no correlation between the hydrophilic constituents and hydrophobic constituents.

ObjectiveTo examine the relationship between HIF-1αt and VEGF expression and the clinicopathological characteristics in hypopharyngeal squamous cell carcinoma. Methods The expression status of HIF-1α and VEGF were examined by immunohistochemistric method (IHC) in 62 tumor tissue and 19 paracancerous normal tissue.The relationships between the expression of HIF-1α and VEGF and clinicopathological characteristic were analyzed. Results HIF-1αt and VEGF expression were higher in hypopharyngeal carcinoma tissues than those in paracancerous normal tissues (66. 1％:26. 3％ ;x2 =18. 02,P ＜0. 05 ;67. 7％ : 31.6％ ; x2 =19.22, P ＜ 0. 05 ). The expression intensity of HIF-1 α was related to T stage, N stage and TNM stage ( x2 =4. 23,5.83,9.94,all P ＜0. 05). The expression intensity of VEGF was related to metastasis, T stage, N stage and TNM stage (x2 =5.62,7. 38,15.75,4. 29 ,all P ＜0. 05 ). There was minus relationship between overall survival and expression level of HIF-1 α and/or VEGF (x2 =29. 25, P＜0.01; x2 =24.88, P＜ 0.01 ).On multivariate analysis,HIF-1α expression and T stage were independent prognostic factors for overall survival (x2 =4.80,5.74, all P＜0. 05).ConclusionsHIF-1α and VEGF may be considered as a parameter in evaluation of progression, metastasis and prognosis of hypopharyngeal carcinoma and also may be a direction of molecular target therapy.

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.