Immune effects of different forms of vaccines of meningococcus serogroup B outer membrane protein 0315 / 中国免疫学杂志

Objective:To evaluate preliminarily immunocompetence and immunoprotection of a NMB0315 nucleic acid vaccine,a recombinant protein vaccine and a nucleic acid vaccine plus a recombinant protein vaccine against Neisseria meningitidis serogroup B in mice, and to provide reliable experimental basis for further exploration of the effective immunization methods and pathways of NMB0315 vaccine. Methods:The NMB0315 nucleic acid vaccine [ pcDNA3. 1 (+)/NMB0315 ] and recombinant protein vaccine(pET-30a/NMB0315)were prepared. Female BALB/c mice were inoculated with a NMB0315 DNA vaccine followed by boosting with recombinant protein NMB0315 through intramuscular and intraperitoneal immunization respectively. Next, humoral immunologic response and cellullar immunologic response were detected in female BALB/c mice by ELISA. The survival rate of BALB/c mice was used to evaluate immunoprotection of the vaccines in mice. Results:Specific IgG,IgG1,IgG2a,and sIgA,induced by the NMB0315 DNA vaccine(pNMB0315-CpG),protein NMB0315 vaccine(rNMB0315-FA),NMB0315 DNA vaccine prime-protein boost at week 8, were detected by indirect ELISA,the A450 values were up to(0. 505±0. 042,0. 513±0. 022,0. 342±0. 017,0. 250±0. 015),(0. 823± 0. 061,0. 807±0. 045,0. 596±0. 027,0. 450±0. 028)and(0. 694±0. 053,0. 711±0. 032,0. 455±0. 021,0. 386±0. 024)respectively, which was significantly higher than the PBS control(P<0. 05). The antibody level of protein vaccine was significantly higher than the nucleic acid vaccine group and combined immunization group ( P<0. 05 ) . The stimulation index and IFN-γ level of combined immunization group were significantly higher than the protein vaccine group and nucleic acid vaccine group(P<0. 05). The bactericidal titer of nucleic acid vaccine group, protein vaccine group and combined immunization group reached 1 :64, 1 :128 and 1 :128 respectively,and the protection rates were 70%,95% and 80% respectively. The IgG2a/IgG1 ratios of the nucleic acid vaccine group, the recombinant protein vaccine group and the combined immunization vaccine group were all less than 1 at week 2, 4, 6, 8. Conclusion:The humoral immunity effects( including mucosal immune) induced by the NMB0315 vaccines form high to low were as follows:the recombinant protein vaccine group, the combined immunization vaccine group, the nucleic acid vaccine;and the cellular immune effects from high to low were as follows:the combined immunization vaccine group, the nucleic acid vaccine group, the recombinant protein vaccine group;The protection effects induced by the NMB0315 vaccines in BALB/c mice within 72 hours from high to low were as follows:the recombinant protein vaccine group, the combined immunization vaccine group, the nucleic acid vaccine group.

Immune effects of different forms of vaccines of meningococcus serogroup B outer membrane protein 0315 / 中国免疫学杂志

Objective:To evaluate preliminarily immunocompetence and immunoprotection of a NMB0315 nucleic acid vaccine,a recombinant protein vaccine and a nucleic acid vaccine plus a recombinant protein vaccine against Neisseria meningitidis serogroup B in mice, and to provide reliable experimental basis for further exploration of the effective immunization methods and pathways of NMB0315 vaccine. Methods:The NMB0315 nucleic acid vaccine [ pcDNA3. 1 (+)/NMB0315 ] and recombinant protein vaccine(pET-30a/NMB0315)were prepared. Female BALB/c mice were inoculated with a NMB0315 DNA vaccine followed by boosting with recombinant protein NMB0315 through intramuscular and intraperitoneal immunization respectively. Next, humoral immunologic response and cellullar immunologic response were detected in female BALB/c mice by ELISA. The survival rate of BALB/c mice was used to evaluate immunoprotection of the vaccines in mice. Results:Specific IgG,IgG1,IgG2a,and sIgA,induced by the NMB0315 DNA vaccine(pNMB0315-CpG),protein NMB0315 vaccine(rNMB0315-FA),NMB0315 DNA vaccine prime-protein boost at week 8, were detected by indirect ELISA,the A450 values were up to(0. 505±0. 042,0. 513±0. 022,0. 342±0. 017,0. 250±0. 015),(0. 823± 0. 061,0. 807±0. 045,0. 596±0. 027,0. 450±0. 028)and(0. 694±0. 053,0. 711±0. 032,0. 455±0. 021,0. 386±0. 024)respectively, which was significantly higher than the PBS control(P<0. 05). The antibody level of protein vaccine was significantly higher than the nucleic acid vaccine group and combined immunization group ( P<0. 05 ) . The stimulation index and IFN-γ level of combined immunization group were significantly higher than the protein vaccine group and nucleic acid vaccine group(P<0. 05). The bactericidal titer of nucleic acid vaccine group, protein vaccine group and combined immunization group reached 1 :64, 1 :128 and 1 :128 respectively,and the protection rates were 70%,95% and 80% respectively. The IgG2a/IgG1 ratios of the nucleic acid vaccine group, the recombinant protein vaccine group and the combined immunization vaccine group were all less than 1 at week 2, 4, 6, 8. Conclusion:The humoral immunity effects( including mucosal immune) induced by the NMB0315 vaccines form high to low were as follows:the recombinant protein vaccine group, the combined immunization vaccine group, the nucleic acid vaccine;and the cellular immune effects from high to low were as follows:the combined immunization vaccine group, the nucleic acid vaccine group, the recombinant protein vaccine group;The protection effects induced by the NMB0315 vaccines in BALB/c mice within 72 hours from high to low were as follows:the recombinant protein vaccine group, the combined immunization vaccine group, the nucleic acid vaccine group.

Effect of VEGF overexpression on the Bcl-2 of TAMs/MCF-7 cell co-culture system / 中国现代普通外科进展

Objective:To explore the effect of VEGF overexpression on the Bcl-2 of TAMs /MCF-7 cell co-culture system.Methods:Application of PMA and IL-4 cells induced THP-1 cell differentiation into TAMs in vitro;TAMs and MCF-7 cell were co-cultured in non-contact Transwell system.MCF-7 cells' proliferation status after co-culture were detected by MTT method;Effect of VEGF over-expression in co-cultured system on Bcl-2 level s in the two cell lines by Western blot assay.Results:PMA and IL-4 induced THP-1 cell become TAMs in vitro.After co-cultured with TAMs 24 h,48 h,MCF-7 cell's proliferation activity increased by 16.16 ％ and 33.99％ vs the control group respectively.TAMs,MCF-7 cells were added VEGF and the supematant of co-culture system respectively,then Bcl-2 levels in both cells were significantly higher,the difference was statistically significant (P＜0.05).Conclusions Tumors secrete VEGF and other chemokines to recruit and activate TAMs.Proliferation,apoptosis and progression of tumor was affected by VEGF/Bcl-2 paracrine loop in tumor microenvironment.

Key technologies of clinical study of traditional Chinese medicine on pediatric functional abdominal pain / 药物评价研究

Functional abdominal pain is one of the most common problems in functional gastrointestinal disorders,and it's also one of the diseases benefit most from traditional Chinese medicine (TCM) treatment.This paper illustrates some key considerations on the study design of traditional Chinese medicine intended for the treatment of FAP based on the latest treatment and evaluation guidelines,technical guidance,professional authority works as well as the latest clinical studies,and combined with experience of clinical trial design.It hopes to offer helps for research designers in this genera.

Aberrant methylation of hMLH1 gene promoter in papillary thyroid cancer and its clinical significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the aberrant promoter methylation of hMLH1 gene promoter and its clinical significance in papillary thyroid cancer (PTC).</p><p><b>METHODS</b>methylation of hMLH1 gene promoter in the cancer tissue and matched tumor-adjacent normal tissue of 152 PTC patients were detected by real-time methylation specific PCR (qMSP). The relationship between the methylation of hMLH1 gene promoter and clinicopathological features was analyzed.</p><p><b>RESULTS</b>The methylation rate of hMLH1 gene promoter in cancer tissues was 37.5% (57/152), of which 33 cases were totally methylated and 24 cases were partially methylated. The methylation rate of adjacent normal tissues was 5.3% (8/152)(all were partially methylated). The methylation rate of PTC tissues was significantly higher than that in the tumor-adjacent normal tissue (P < 0.01). The promoter methylation of hMLH1 gene in PTC was significantly correlated with age, size and number of the primary lesion, local invasion, T stage and lymph node metastasis (P < 0.05) , but not correlated with gender and clinical stage (P > 0.05).</p><p><b>CONCLUSION</b>Promoter methylation of hMLH1 gene is a common molecular event in PTC tissue, and it is significantly correlated with the progression of PTC.</p>

A reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human blood type B→O conversion / 中国实验血液学杂志

This study was aimed to prepare a reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human B to O blood group conversion. Based on the construction of recombinant E. Coli (DE3) which can express α-galactosidase, the inducing time and inducer concentration were optimized for high expression of α-galactosidase. Then, the expression products in supernatant were purified by cation and anion exchange column chromatography. The purified α-galactosidase was used to treat B group red blood cells in phosphate buffer (pH 6.8) for 2 hours to prepare O group red blood cells. The results showed that the optimal inducing conditions for α-galactosidase expression were IPTG 0.1 mmol/L, 37°C and 2 hours. The specific enzyme activity of purified protein increased from 0.42 U/mg to 2.1 U/mg as compared with pre-purification. And, the conditions of B to O blood group conversion were 26°C, pH 6.8 (neutral pH condition) and 2 hours. Moreover, 225 µg of the enzyme could converse 1 ml B red blood cells to O completely. It is concluded that the technology of expression and purification of recombinant α-galactosidase has been established, and the purified protein can converse B red blood cells to O completely, which means that an effective enzyme conversing B red blood cells to O has been obtained.

Bcl-2 inhibitor ABT-737 enhances the cisplatin-induced apoptosis in breast cancer T47D cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of ABT-737 combined with cisplatin on apoptosis of breast cancer cell line T47D cells.</p><p><b>METHODS</b>T47D cells cultured in vitro was used for this experiment. Cell proliferation was measured by MTT assay. The expression of apoptosis-related protein was determined by Western blot. Morphological changes of apoptotic cells were observed by fluorescence microscopy. The apoptosis rate was examined by flow cytometry.</p><p><b>RESULTS</b>The MTT assay showed that ABT-737 significantly decreased the IC(50) of cisplatin in T47D cells [(26.00 ± 1.41) µmol/L of single cisplatin vs. (13.00 ± 1.11) µmol/L of combination (ABT-737 + cisplatin)]. As a single agent, ABT-737 did not inhibit the proliferation of T47D cells, but enhanced the inhibitory effect of cisplatin in a dose-dependent manner. The detection of the cleavage of PARP showed that ABT-737 lowered the doses of cisplatin to induce apoptosis and shortened the induction time of apoptosis in T47D cells. Compared with the single use of cisplatin, the combination of ABT-737 and cisplatin accelerated the cleavage of PARP and caspase3, but did not alter the expression levels of Bcl-2, Bcl-X(L), and Bax. Both flow cytometry and fluorescence microscopy showed that ABT-737 combined with cisplatin significantly increased the apoptosis induction in T47D cells (2.3% ± 0.1 % in the control, 30.0% ± 0.8% in the cisplatin alone, and 49.0% ± 0.5% in the cisplatin + ABT-737 groups, P < 0.05).</p><p><b>CONCLUSION</b>The Bcl-2 inhibitor ABT-737 can significantly enhance cisplatin-induced apoptosis in human breast cancer T47D cells in vitro.</p>

Progress in research on preparation of antibodies against rhesus D antigen / 军事医学科学院院刊

The RhD antigen is expressed only in human red blood cells (RBC).Its immunogenicity and clinical application are only next to ABO blood group system, and is widely used both for blood typing and prevention of hemolytic disease of the newborn. The traditional anti-Rh(D) is derived by fractionation of plasma from individuals who have been sensitized by pregnancy or transfusion, or have been deliberately immunized to produce anti-Rh(D). Because of the limited source of plasma, researchers began the study of monoclonal and recombinant antibody. Monoclonal and recombinant anti-Rh(D) antibodies may provide alternatives to the current plasma derived polyclonal IgG anti-Rh(D), but up to now,none of them have yet proved effective in humans for prevention of RhD immunity and hemolytic disease of the newborn.

Analysis of Pediatric Antibiotics in Our Hospital during the Period of 2002～2005 / 中国药房

OBJECTIVE:To evaluate status quo of the application of antibiotics for children and its trend.METHODS:The data about the amount and money of antibiotics used from2002to2005in out patient pharmacy of our hospital are collected and analyzed.RESULTS:Cephalothin,penicillin and azithromycin were widely used in our hospital and the using situation was reasonable at large,but some problems still existed.CONCLUSION:Rational using of antibiotics need efforts of all the departments in the hospital.