Objective To make a comparative study of the immunogenicity of multi-epitope DNA vaccine and BCG of Mycobacterium tuberculosis and therapeutic effects of the vaccines combined with chemotherapy in a mouse model infected with multi-drug resistant (MDR)Mycobacterium tuberculosis.Methods The BALB/c mice were randomly divided into Group A,Group B and Group C,which received subcutaneous immunization with PBS,BCG and multi-epitope DNA vaccine,respectively,once at weeks 1 ,3 and 5 .Specific IgG antibody,IL2 and IFN-γin mice serum were determined with ELISA after the final vaccination.At the same time,the splenic lymphocytes of mice were separated and the lymphocytes proliferation was determined by MTT method.To study the therapeutic effects of multi-epitope DNA vaccine,the mice were infected by intravenous injection in the tail vein with Mycobacterium tuberculosis clinical isolates that were resistant to isoniazid (INH)and rifampin (RFP).Four weeks later,the model mice were divided into Groups D,E and F.The mice in Group D and control group received saline injection. The mice in Group E were treated with RFP and INH.The mice in Group F were treated with multi-epitope DNA vaccine combined with INH and RFP for 10 weeks.The lungs,livers and spleens of mice were removed and weighed;the colony number of Mycobacterium tuberculosis in the lungs and spleens was counted.Results The antibody IgG,IL2 and IFN-γwere obviously higher in multi-epitope DNA vaccine group than in BCG group (P<0.05).Multi-epitope DNA vaccine combined with chemotherapy could obviously reduce the colony number of Mycobacterium tuberculosis in the lungs and spleens as well as indexes of the lungs,livers and spleens (P<0.05). Conclusion Mycobacterium tuberculosis Hsp70,Ag85A,and ESAT-6 multi-epitope DNA vaccine could induce strong specific immune response in mice that produced a high level of specific IgG antibody,IL2 and IFN-γ,specific lymphocyte proliferation,and significantly improve the efficacy of anti-tuberculosis drug resistant tuberculosis in mice.

Acupuncture Therapy ; Adipose Tissue ; injuries ; Adolescent ; Adult ; Combined Modality Therapy ; Female ; Humans ; Male ; Massage ; Young Adult

OBJECTIVE:To establish a method for simultaneous determination of gallic acid,protocatechuic acid,catechuic acid,corilagin and brevifolincarboxylic acid in Geranium carolinianum. METHODS:HPLC was performed on the column of Won-daSil C18 WR with mobile phase of acetonitrile-0.1% Phosphoric acid(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 280 nm,the column temperature was 40 ℃,and the volume injection was 20 μl. RESULTS:The linear range was 0.02-20.02 for gallic acid(r=0.999 9),0.01-20.10 for protocatechuic acid(r=0.999 7),0.02-19.78 for catechuic acid(r=0.999 6), 0.02-20.02 for corilagin(r=0.999 9)and 0.02-20.10 for brevifolincarboxylic acid(r=0.999 5);RSDs of precision,stability and re-producibility tests were lower than <3.0%;recoveries were 95.1%-100.6%(RSD=2.20%,n=6),95.8%-100.6%(RSD=1.74%,n=6),95.1%-101.9%(RSD=2.71%,n=6),97.7%-103.1%(RSD=2.04%,n=6)and 95.3%-99.0%(RSD=1.46%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the contents determination of gallic ac-id,protocatechuic acid,catechuic acid,corilagin and brevifolincarboxylic acid in G. carolinianum.

Objective To evaluate the role of nuclear factor kappa B (NF-κB) in sevofluraneinduced release of inflammatory factors in mouse microglias.Methods Microglial cells obtained from newborn C57BL/6 mice (aged 2-3 days) were seeded in 24-well plates (density 1 × 105 cells/ml, 1 ml/well) , a total of 80 wells.The cells were randomly divided into 4 groups using a random number table (n =20 each) : control group (group C);sevoflurane group (group S);NF-κB selective inhibitor pyrrolidine dithiocarbamate (PDTC) group (group P);PDTC + sevoflurane group (group P +S).In S and P+S groups, 4.1％ sevoflurane was inhaled for 6 h.In P+S group, 10 μmol/L PDTC was added at 1 h before sevoflurane inhalation.At 6 h of incubation with sevoflurane, NF-κB activity and expression and levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined.Results Compared with group C, the NF-κB activity and levels of IL-6 and TNF-α were significantly increased in group S, and no significant change in the above parameters was found in the other two groups.Compared with group S, the NF-κB activity and levels of IL-6 and TNF-α were significantly decreased in group P+S.There was no significant difference in NF-κB expression between the four groups.Conclusion NF-κB mediates sevoflurane-induced release of inflammatory factors in mouse microglias.

BACKGROUND: Studies have shown that the combined use of sufentanil and neural stem cell (NSC) transplantation can increase the number of newborn nerve fibers.OBJECTIVE: To investigate the effect of sufentanil on the hind limb function of rats with spinal cord injury after neural stem cell transplantation.METHODS: (1) Eighty adult female Sprague-Dawley rats were used to build spine cord injury model according to the modified Allen's method and divided into model group, sufentanil group, NSCs transplantation group and sufentanil combined with NSCs transplantation group (combined group). Extra 20 adult female Sprague-Dawley rats were not conducted any treatment as normal control group. (2) After 6 days of modeling, the model rats were subjected to subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of saline in the model group; subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of sufentanil (150 μg/kg) in the sufentanil group; subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of saline in the NSCs transplantation group; and subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of sufentanil (150 μg/kg) in the combined group. (3) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels were detected by RT-PCR, and the cell apoptosis changes around the spine cord injury area were determined with TUNEL staining method. (4) The motor functions of rats were tested by Basso, Beattie and Bresnahan score and inclined plane test after 1, 3 days and 1, 2, 3 and 4 weeks of modeling. (5) After 4 weeks of modeling, the histopatholgical changes in the area of spine cord injury were observed by hematoxylin-eosin staining method. The survival changes of NSCs labeled by CM-Dil were determined by fluorescence microscope. The regenerations and distributions of spinal nerve fibers were observed by fluorescein gold retrograde tagging.RESULTS AND CONCLUSION: (1) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels as well as the cell apoptotic rate in the combined group were significantly lower than those in the model, sufentanil and NSCs transplantation groups (P < 0.05). (2) After 2 weeks of modeling, the combined treatment significantly improved the hind limb motor functions of rats compared with the sufentanil and NSCs transplantation groups (P < 0.05), and the recovery of motor function was better in the sufentanil and NSCs transplantation groups than in the model group (P < 0.05). (3) After 4 weeks of modeling, the results of hematoxylin-eosin staining manifested that the spinal cord tissues lost and the magnified syringomyelias occurred in the model group. The syringomyelias in the sufentanil and NSCs transplantation groups were significantly smaller than that in the model group, and the syringomyelias almost disappeared in the combined group. (4) The number of positive cells was the most in the combined group, more in the NSCs transplantation group, but there were no positive cells labeled by CM-Dil in the sufentanil and negative control groups. (5) The number of positive neural fibers in the combined group was the highest followed by the sufentanil and NSCs transplantation groups, and the number of positive neural fibers in negative control group was the lowest. To conclude, sufentanil can improve the recovery of hind limb motor function by reducing the AQP4 and MMP9 expression levels in the injury area, promoting the survival of transplanted NSCs, and decreasing the local NSCs apoptosis after spinal cord injury.

Objective To investigate the relationship between interleukin 1β( IL?1β) , interleukin?33 ( IL?33) , neutrophil?to?lymphocyte ratio ( NLR ) and atrial fibrillation. Methods Eighty?two patients with nonvalvular atrial fibrillation treated in the department of cardiology in the Fourth People′s Hospital of Jinan from October 2015 to October 2016 were enrolled in the study,including 43 patients with persistent atrial fibrillation and 39 patients with paroxysmal atrial fibrillation. 50 healthy subjects were selected as the control group. Enzyme linked immunosorbent assay (ELISA) was used to determine the concentration of IL?1βand IL?33,and the left atrial diameter ( LAD) was measured by echocardiography. Results ( 1) The concentrations of IL?1β,NLR and LAD in the paroxysmal atrial fibrillation group were (24. 44±4. 89) ng/L,(2. 51±1. 22) %,(36. 16± 6. 12) mm,the concentrations of IL?1β,NLR and LAD in the persistent atrial fibrillation group were (26. 95±5. 86) ng/L,(5. 7±1. 8) %,(39. 36±4. 78) mm and the values in the control group were (19. 53±4. 51) ng/L,(1. 82 ± 0. 41 ) %, ( 33. 31 ± 2. 89 ) mm, respectively. The differences among the three groups were statistically significant ( F=16. 74,11. 82,14. 85,P<0. 01) . The indexes of paroxysmal atrial fibrillation group and persistent atrial fibrillation group were higher than those of the control group ( P<0. 01 ) . ( 2 ) In the persistent atrial fibrillation group, NLR and LAD were ( 5. 7 ± 1. 8 )% , and ( 39. 36 ± 4. 78 ) mm, higher than those of the paroxysmal atrial fibrillation group ( (2. 51±1. 22)%,(36. 16±6. 12) mm),the differences were statistically significant ( P<0. 01) . However,the level of IL?1βin the persistent atrial fibrillation group was not significantly different from that in the paroxysmal atrial fibrillation group ( (26. 95±5. 86) ng/L vs. (24. 44±4. 89) ng/L,P>0. 05). (3) The concentrations of IL?33 in the paroxysmal atrial fibrillation group,atrial fibrillation group, control group were ( 48. 31 ± 4. 72 ) ng/L, ( 50. 03 ± 2. 18 ) ng/L, ( 56. 87 ± 5. 12 ) ng/L, respectively. The difference among the three groups has no statistical significance ( F=2. 52, P>0. 05 ) . ( 4 ) NLR level was positively correlated with LAD ( r=0. 32,P=0. 002) . There was no significant correlation among IL?1β,IL?33 and LAD ( r=0. 16, P=0. 11, r=0. 02, P=0. 37 ) . Conclusion The levels of IL?1β, NLR and LAD in peripheral blood of patients with atrial fibrillation were significantly higher than those in patients with sinus rhythm,and there was a positive correlation between NLR and LAD.

Objective: To study the correlations between galectin-3, soluble ST2 (sST2) levels and chronic heart failure (CHF) classiifcation, traditional HF indicator and short-term death in relevant patients. Methods: This research included 2 groups: CHF group, containing 142 relevant patients treated in our hospital from 2014-02 to 2015-10 and Control group, containing 85 normal subjects from physical examination at the same period of time. Based on NYHA criterion, the patients were classiifed in NYHA grade II, III and IV respectively. Blood levels of N-terminal brain natriuretic peptide (NT-ProBNP), high-sensitivity C reactive protein (hs-CRP) and ultrasonic morphology were examined upon admission; protein expressions of galectin-3 and sST2 were assessed by ELISA. Results: The patients with NYHA grade III and IV had increased levels of galectin-3 and soluble sST2; galectin-3, sST2 were positively related to NT-ProBNP, hs-CRP and LVEDD, while negatively related to LVEF. Logistic regression analysis indicated that galectin-3 and sST2 were related to short-term death in CHF patients,P<0.05. Area under ROC curve of galectin-3 and sST2 for diagnosing CHF were 0.738 and 0.771,P<0.01. Conclusion: Galectin-3 and sST2 levels were related to traditional HF indicator and could be used for CHF diagnosis in relevant patients.

Objective To explore the pharmacological action of Bazheng mixture(BM) on infections disease of urinary system and effects of BM on immune function in mice.Methods After E.coli was injected into bladder of mice, the kidneys were examined for the anti-infection action of BM on ascending infection;and the effects of BM on immune function of mice were tested.Results BM ig remarkably decreased the percentage of kedney's infections area caused by ascending infection of E.coli [ED_(50) was(11.01?1.63)g/kg and the 95% CL was (9.50-12.76)(g/kg]);BM was also found effective in increasing the rate of carbon particle clearance and the index of carbon particle clearance,but BM had no significant effects on body fluid immunity and cell immunity in test for mice.Conclusion The mechanism of BM in treating infections disease of urinary system are mainly associated with the increasing rate of carbon particle clearance,and with bacteria clearance in urinary system.

Objective To determine whether the single nucleotide polymorphisms (SNPs) rs231775 and rs3087243 of the cytotoxic T lymphocyte antigen 4 (CTLA-4) gene are associated with susceptibility to Graves' disease (GD) in Chinese Han population.Methods Patients were enrolled from outpatient department and wards.Blood samples from each subject were collected to extract DNA,and the genotypes were determined by TagMan-MGB probe.Results The frequencies of allele G (OR =1.244,95％ CI 1.124-1.377,P＜0.01) and genotype GG (55.3 ％ vs 49.1％,OR =1.279,95 ％ CI 1.126-1.454,P＜0.01) of rs231775 in GD group were higher than those in control group.The frequencies of allele G (OR =1.303,95％ CI 1.166-1.457,P＜0.01) and genotype GG (76.8％ vs 71.8％,OR=1.302,95％ CI 1.143-1.484,P＜0.01) of rs3087243 in GD group were also higher.Conclusion GG genotypes in rs231775 and rs3087243 of CTLA-4 gene are related to the high risk of GD.

Objective:To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-pSIREN-RetroQ.Methods:Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-pSIREN-RetroQ interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results:We successfully design and built three TNFAIP8-shRNA-pSIREN-RetroQ interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.