Objective To construct five types of recombinant modified vaccinia virus Ankara (MVA) carrying genes encoding antigen 85A (Ag85A), early secretory antigenic target (ESAT-6) or IL-2 and to investigate the immunogenicity of these recombinant MVA in mice. Methods The genes encoding Ag85A and ESAT-6 were amplified by PCR from Mycobacterium tuberculosis H37Rv genomic DNA. The am-plified DNA fragments were sub-cloned into vector p18. The recombinant plasmids were introduced respec-tively into MVA by homologous recombination. The five types of recombinant MVA clone were selected in the MPA selection medium and their expressions of TB antigens were confirmed by Western blot. The recombi-nant MVA were used to immunize BALB/c mice. Ag85A-ESAT-6 fusion protein induced proliferation of spleno-lymphecytes from immunized mice was detected by MTT test. The concentration of IFN-γ in superna-tant of spleno-lymphocytes cultures was also analyzed by ELISA. The delayed type hypersensitivity (DTH) response to tuberculosis antigens was measured after injection of tuberculin purified protein derivative (PPD) in hind footpads of immunized mice. Results The five types of recombinant MVA were successfully con-structed by confirming their expression of TB antigen with Western blot. After co-cultured with the fusion protein Ag85A-ESAT-6 in vitro, the spleno-lymphocytes from recombinant MVA immunized mice showed sig-nificant proliferation activity (P<0.01) and increased IFN-γ secretion (P<0.05) while the same cells showed no response when co-cultured with saline. The spleno-lymphocytes from recombinant MVA immu-nized mice also demenstrated significant proliferation activity (P<0.01) and increased IFN-γ secretion (P<0.01) under Ag85A-ESAT-6 inducement, compared with the spleno-lymphocytes from mice immunized with wild type MVA or saline. The significant DTH response to PPD was detected in recombinant MVA-immunized mice (P<0.05). Conclusion The five types of recombinant MVA carrying genes of TB anti-gens were constructed. The constructed recombinant MVA could induce specific cellular immunity against tu-berculosis antigen in mice.

OBJECTIVETo study the effect of Yishen Daluo Decoction (YDD) on the expression of protein lipoprotein (PLP), oligodendrocyte transcription factor 1 (Olig 1), and oligodendrocyte transcription factor 2 (Olig2) in mice with experimental autoimmune encephalomyelitis (EAE).

METHODSTotally 40 mice were randomly divided into 4 groups, i.e., the normal group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 mice in each group. Each mouse in the model, CM, and WM groups was subcutaneously injected with 200 microL antigen emulsion (containing 150 micro g PLP139 -151 and 400 micro g H37RA) in two parts at the upper abdomen on the first day. 100 microLBordetella pertussis juice (containing 0. 6 x 10(6) Bordetella pertussis) was injected by caudal vein on the first and the third day. On the 7th day after modeling, each mouse in the normal group and the model group was intragastrically given normal saline (0. 1 mL/10 g). YDD (0. 2 g crude drug/10 g) was intragastrically given to mice in the CM group, and prednisone (0. 039 mg/10 g) was intragastrically given to mice in the WM group. All mice were intervened for 54 days. Changes of PLP, Olig1, and Olig2 in the brain tissue of EAE mice were detected by Western blot. Results The levels of PLP and Olig2 in the brain tissue of the model group were less than those of the normal group (P <0.05). Compared with the model group, the levels of PLP, Olig1, and Olig2 in the brain tissue increased in the CM group (P <0.05); the levels of PLP and Olig2 in the brain tissue increased in the WM group (P <0.05). Compared with the WM group, the level of Olig1 in the brain tissue increased in the CM group (P <0.05).

CONCLUSIONYDD could enhance remyelination by elevating the levels of Olig1 and Olig2 in the brain tissue of EAE mice.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Brain ; Drugs, Chinese Herbal ; pharmacology ; Encephalomyelitis, Autoimmune, Experimental ; metabolism ; Gene Expression ; Mice ; Nerve Tissue Proteins ; metabolism ; Oligodendrocyte Transcription Factor 2 ; Transcription Factors

OBJECTIVETo study the effect of Qinggan Huoxue recipe (QGHXR) in treating alcoholic liver disease (ALD).

METHODSBy adopting the multi-centered, randomized and controlled method, the patients were divided in to the QGHXR group (60 patients) treated orally by QGHXR, the XCHG group (30 patients) treated orally by Xiaochaihu granule and the control group (30 patients) treated orally by conventional therapy such as glucurolactone, vitamin C. The changes in symptoms, signs, liver function, blood lipid, liver fibrosis markers, cytokines, lipid superoxidation parameters and B-untrasonographic figure after treatment of the two groups were observed.

RESULTSThe total therapeutic efficacy of QGHXR, improvements in anorexia, nausea, vomiting and jaundice as well as effect in reducing blood levels of alanine transaminase (ALT), aspartate aminotransferase (AST) and triglyceride (TG) were superior in the QGHXR group to those in the other two groups (P < 0.01), and the effect in decreasing gamma-glutamyl transferase (GGT) and very low-density lipoprotein (VLDL) in the QGHXR group was more significant than that in the control group (P < 0.01). QGHXR also showed effects in lowering levels of liver fibrosis markers and cytokines, alleviating the anti-lipid superoxidation damage in liver, and could markedly improve the degree of fatty liver.

CONCLUSIONQGHXR shows obvious therapeutic effect in treating ALD, the mechanism could possibly be related with its effects in antagonizing lipid superoxidation, stabilizing hepatic cell membrane, adjusting the lipid metabolic disturbance of liver, regulating immune function, anti-liver fibrosis and promoting the intrahepatic metabolism of alcohol.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver, Alcoholic ; drug therapy ; Female ; Humans ; Liver Cirrhosis, Alcoholic ; drug therapy ; Liver Diseases, Alcoholic ; drug therapy ; Male ; Middle Aged ; Phytotherapy

The text introduces an electromyogram-guided treatment instrument with simple operation and lower cost, and it is easy to find the lesion muscle. Its clinical tests have shown a satisfying result.
Electromyography ; instrumentation ; Equipment Design ; Musculoskeletal Manipulations ; instrumentation

In order to study how to activate T cells and their immunological characteristics, the anti-CD3 and anti-CD28 McAbs were used to stimulate PBMNC, then their related immunological changes, such as lymphocyte transformation function, the percentage of CD8(+)CD25(+) cells and TGF-beta expression were deleted by lymphocyte transformation assay, flow cytometry and RT-PCR respectively. The results showed that in costimulation with anti-CD28 antibody stimulation, the activity of anti-CD3 antibody was significantly enhanced, the ratio of CD8(+)CD25(+) cells of T cells was obviously increased, while TGF-beta expression was down-regulated. It was concluded that the anti-CD28 antibody costimulation could provide stimulatory signal II, which make T cells more active, while the expression of TGF-beta significantly down-regulated.
Antibodies, Monoclonal ; pharmacology ; CD28 Antigens ; immunology ; CD3 Complex ; immunology ; Down-Regulation ; Humans ; Leukocytes, Mononuclear ; immunology ; metabolism ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; immunology ; Transforming Growth Factor beta ; biosynthesis

AIMTo evaluate the effects of administration of hyperbaric oxygenation(HBO) when initiated at different time after acute transient ischemia. Apoptosis in the ischemic penumbra was further investigated to search for the possible mechanism.

METHODSThe male SD rats were randomly assigned to the following groups: control, HBO therapy initiated 3 h after ischemia, HBO therapy initiated 6 h after ischemia, HBO therapy initiated 12 h after ischemia. All animals were subjected to 90 min intraluminal middle cerebral artery occlusion (MCAO) with the regional cerebral blood flow monitored in vivo by laser Doppler flowmetry. HBO treatment was performed in a pressure chamber with 100% O2, 3 arm for 1 h. Neurological deficits and infarct volumes were assessed at 24 hours after ischemia. The immunohistochemical changes of apoptosis in the penumbra were evaluated by detecting the expression of cleaved Caspase-3, cleaved Caspase-9, Bcl-2, Bax and TUNEL staining.

RESULTSHBO therapy initiated at 3 and 6 hours after ischemia significantly improved the neurological function and reduced infarct volume. Meanwhile, it increased the expression of Bcl-2 protein and decreased the expression of activated Caspase-3, activated Caspase-9 and TUNEL-positive cells. However, HBO therapy administrated 12 hours after ischemia aggravated the neurological deficits and enlarged infarct volume, while it showed no significant reduction of apoptotic change compared with control.

CONCLUSIONThere is a therapeutic window for the use of HBO in acute transient cerebral ischemia in rats. HBO-treatment is highly effective in reducing infarct volume when initiated up to 6h after the onset of ischemia. Inhibition of apoptotic cell death in the penumbra appears to be the underlying protective effect of early therapy.

Animals ; Apoptosis ; Brain Ischemia ; metabolism ; pathology ; therapy ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cerebral Infarction ; metabolism ; pathology ; therapy ; Hyperbaric Oxygenation ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective:Through determination of urinary arsenic metabolites in high water arsenic exposed areas of Jilin and Shanxi provinces, to explore the mode and possible influencing factors of arsenic metabolism in different populations.Methods:From October 2018 to August 2019, a cluster sampling was carried out in villages (arsenic in drinking water ≥0.05 mg/L) of some townships (towns) in Lyuliang City, Shanxi Province and Baicheng City, Jilin Province for epidemiological investigation and general health examination. The residents over 35 years old drinking water from local centralized water supply and small well water sources were selected as arsenic exposure group, and people (nearby low-arsenic water source areas) with the same diet and living habits and similar economic conditions were selected as control group. Urine samples were collected. Liquid chromatography-atomic fluorescence spectrometry(LC-AFS) technology was used to separate and detect 4 species of arsenic compounds, including trivalent inorganic arsenic (iAs Ⅲ), pentavalent inorganic arsenic (iAs Ⅴ), methylated arsine (MMA), and dimethylated arsine (DMA). Total arsenic (tAs), inorganic arsenic percentage (iAs%), MMA percentage (MMA%), DMA percentage (DMA%), primary methylation index (PMI) and the secondary methylation index (SMI) were calculated. The influencing factors of arsenic metabolism were analyzed by multiple linear regression. Results:A total of 1 415 villagers were investigated, including 1 256 in arsenic exposure group and 159 in control group. Compared with the control group, there were no significant differences in age, gender ratio and occupation distribution between arsenic exposure group and control group ( P > 0.05), but there were significant differences in smoking, drinking, body mass index (BMI) and education level distribution ( P < 0.05). The median of urinary tAs, iAs%, MMA%, DMA%, PMI and SMI in control group and arsenic exposure group were 12.86 μg/L, 15.03, 5.23, 76.35, 84.97, 93.68 and 69.68 μg/L, 10.24, 8.37, 79.31, 89.76, 90.65, respectively, the levels of urinary tAs, DMA% and PMI in arsenic exposed group were higher than those in control group, while iAs% and SMI were lower than those in control group, the differences were statistically significant ( U=- 13.87, - 4.30, - 6.64, - 6.64, - 1.99, P < 0.05). After analysis of the factors influencing urinary arsenic metabolism in the population, we found that age and BMI had an impact on iAs% ( β=- 0.08, - 0.08, P < 0.05); gender, drinking, BMI and education level were influencing factors of MMA% ( β =- 0.11, - 0.09, - 0.07, 0.08, P < 0.05); DMA% was mainly affected by age, gender, BMI and education level ( β = 0.06, 0.09, 0.10, - 0.09, P < 0.05); PMI was mainly affected by age and BMI ( β = 0.08, 0.08, P < 0.05); while SMI was affected by gender, drinking, BMI and education level ( β=0.09, 0.08, 0.08, - 0.09, P < 0.05). Conclusions:The urinary arsenic metabolism models of different arsenic exposed groups are different. Age, gender, smoking, drinking, BMI and education level may be influencing factors of different arsenic metabolism models.

OBJECTIVETo investigate the effect of hyperbaric oxygen(HBO)therapy on mitochondrial free radicals after transient focal cerebral ischemia in rats.

METHODSThe male SD rats were randomly assigned into two groups, control and HBO groups. All animals were subjected to 90 min intra-luminal middle cerebral artery occlusion (MCAO) with the regional cerebral blood flow monitored in vivo by laser Doppler flowmetry. HBO treatment was performed in a pressure chamber with 100% O(2)(3 ATM 1 h) 3 h after ischemia. Twenty-four hours after ischemia, mitochondria in the ischemic core and penumbra were isolated and the contents of H(2)O(2), O(2)(*-), MDA, SOD, GSH-PX and GSH in mitochondria were measured respectively.

RESULTAfter cerebral ischemia-reperfusion, contents of mitochondrial H(2)O(2), O(2)(*-), MDA increased, while the SOD, GSH-PX and GSH in the mitochondria decreased significantly both in the ischemic core and the ischemic penumbra, compared with those in the normal controls(P<0.05). In the ischemic penumbra, HBO therapy increased significantly the content of O(2)(*-)(P<0.05), enhanced the activity of SOD, and decreased the level of MDA (P<0.05). However, HBO therapy did not change the level of MDA, though it also increased the content of O(2)(*-) and the activity of SOD in the ischemic core. HBO therapy had no significant effect on the contents of H(2)O(2), GSH-PX and GSH in the ischemic mitochondria.

CONCLUSIONHBO therapy initiated early after acute transient cerebral ischemia in rats can increase the mitochondrial free radicals level, but also increase the activity of the anti-radical enzymes. HBO treatment inhibits the lipid peroxidation damage of mitochondria in the ischemic penumbra, but not in the ischemic core, which indicates that the mitochondrial function plays a role in the reaction of the free radical in the ischemic area after HBO therapy.

Animals ; Free Radicals ; metabolism ; Glutathione Peroxidase ; metabolism ; Hyperbaric Oxygenation ; methods ; Infarction, Middle Cerebral Artery ; metabolism ; therapy ; Ischemic Attack, Transient ; metabolism ; therapy ; Male ; Mitochondria ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

AIMTo investigate whether the selective AT1 receptor antagonist irbesartan exerts neuroprotective effect on focal cerebral ischemia in normotensive rats.

METHODSCerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 90 min followed by reperfusion, with the monitoring of laser Doppler flowmetry. To avoid the interaction with peripheral AT1 receptors, irbesartan was infused intracerebroventricularly (ICV) at a dose which effectively inhibited brain- but not vascular AT1 receptors. Neurological status was evaluated daily after MCAO. Rats were killed and brain samples were collected for the measurement of infarct size and immunohistochemical evaluation of apoptosis by deoxynucleotidyltransferase-mediated biotinylated UTP nick end labeling (TUNEL) and expression of activated Caspase-3 and the cleavage fragment of poly (ADP-ribose) polymerase (PARP).

RESULTSTreatment with irbesartan improved significantly the neurobehavioral functions after cerebral ischemia. The infarct size was reduced about 42% on day 7 after MCAO (P < 0.05). Meanwhile,irbesartan treatment significantly decreased the number of TUNEL-positive cells in the penumbra. The expression of activated Caspase-3 and the downstream cleavage fragment of poly (ADP-ribose) polymerase in the penumbra were also inhibited by irbesartan therapy on day 3 after transient cerebral ischemia.

CONCLUSIONAngiotensin AT1 receptor antagonist exhibits neuroprotection against transient cerebral ischemia in the brain. The neuroprotective effects in ischemic tissue may be associated with its inhibition of apoptotic cell death in the penumbra.

Angiotensin Receptor Antagonists ; pharmacology ; Animals ; Biphenyl Compounds ; pharmacology ; Cerebral Infarction ; pathology ; Ischemic Attack, Transient ; drug therapy ; pathology ; Lateral Ventricles ; Male ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Wistar ; Tetrazoles ; pharmacology

Adult ; Aged ; Antigens, CD ; biosynthesis ; genetics ; B7-H1 Antigen ; CD28 Antigens ; biosynthesis ; genetics ; Female ; Hepatitis B, Chronic ; immunology ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Middle Aged