ABSTRACT
ObjectiveTo explore the possible mechanism of the Yiqi Jiedu formula （YQ） in treating ischemic stroke （IS） from the perspective of the microbial-gut-brain axis （MGBA）. MethodRats were randomly divided into five groups， with six in each group， including sham surgery group， model group， and low， medium， and high dose YQ groups （1， 5， and 25 mg·kg-1）. Except for the sham surgery group， all other groups were established with a middle cerebral artery occlusion （MCAO） model using the thread occlusion method. The success of modeling was determined through neurobehavioral scoring， and the protective effect of YQ on IS was evaluated. Then， the changes in gut microbiota before and after MCAO modeling and YQ administration were compared using 16S rDNA sequencing technology， and the possible biological pathways related to the effect of this formula were analyzed. The expression of inflammatory factors such as interleukin-6 （IL-6）， interleukin-17A （IL-17A）， and interleukin-10 （IL-10） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression of tight junction proteins ZO-1 and Occludin in brain and intestinal tissue， and hematoxylin-eosin staining （HE） was used to observe pathological changes in the cerebral cortex and colon， so as to validate the possible mechanism of action. ResultYQ significantly improved the neurobehavioral score of MCAO rats （P<0.01） and played a good regulatory role in intestinal microbial disorders caused by enriched pathogens and opportunistic pathogens during the acute phase. Among them， significantly changed microorganisms include Morgentia， Escherichia Shigella， Adlercreutzia， and Androbacter. Bioinformatics analysis found that these bacteria may be related to the regulation of inflammation in the brain. Compared with the blank group， the detection of inflammatory factors in the serum of IS model rats showed an increase in inflammatory factors IL-6 and IL-17A （P<0.01） and a decrease in the content of anti-inflammatory factor IL-10 （P<0.01）. Compared with the model group， the content of inflammatory factors IL-6 and IL-17A in the serum of the treatment group decreased （P<0.05）， and that of anti-inflammatory factor IL-10 increased （P<0.01）. The expression results of barrier proteins ZO-1 and Occludin in brain and intestinal tissue showed that the expression levels of both decreased in IS model rats （P<0.05）， while the expression levels of both increased in the treatment group （P<0.05）. ConclusionAcute cerebral ischemia can lead to an imbalance of intestinal microbiota and damage to the intestinal barrier， and it can increase intestinal permeability. YQ can regulate intestinal microbiota imbalance caused by ischemia， inhibit systemic inflammatory response， and improve the disruption of the gut-blood brain barrier， preventing secondary cascade damage to brain tissue caused by inflammation. The MGBA may be an important mechanism against the IS.
ABSTRACT
Objective To study the pharmacological function of the aconitine in treating adjuvant arthritis(AA).Methods Fifty rats were randomly divided into the control group, AA model group, methotrexate group(0.5 mg/kg), and aconitine groups of different dosages (25, 100μg/kg). Except the control group, each group was injection of intradermal Freund's complete adjuvant (0.1 ml) into right hindpaw of rats to establish adjuvant arthritis model. On the 10th day after model onset, the aconitine were administered by gavage with 25, 100μg/kg once daily, and the methotrexate group was administered with 0.5 mg/kg methotrexate twice per week, and all groups were treated for 19 days. After the last administration, the foot swelling was measured by toe volume meter, arthritis index was calculated by 5-grade scoring method, spleen and thymus index were calculated, and the pathological changes of right ankle joint were observed by HE staining.Results After the model establishment, compared with the model group, the degree of swelling of the ankle at 20 days (668.7 ± 144.5μl, 566.9 ± 179.3μl vs. 912.1 ± 200.5μl), 24 days (833.1 ± 144.0μl, 803.9 ± 213.4μlvs.1069.5 ± 164.6μl) and 28 days (736.4 ± 115.0μl, 835.7 ± 170.1μlvs. 1107.2 ± 215.8μl) in the aconitine groups significantly decreased (P<0.05 orP<0.01). After the model establishment, compared with the model group, arthritic index scores at 18 days (3.1 ± 0.7, 3.2 ± 0.4vs. 3.8 ± 0.6), 24 days (3.1 ± 0.5, 3.4 ± 0.5vs.3.9 ± 0.3), 28 days (2.7 ± 0.6, 3.2 ± 0.9 vs. 4.0 ± 0.0) in the aconitine groups significantly decreased (P<0.05). Compared with the model group, the spleen index (3.5 ± 0.4, 3.3 ± 0.4, 4.0 ± 0.6vs.4.9 ± 0.5) respectively in the low and high dose group of aconitine and methotrexate group (P<0.01).Conclusion Aconitine has a certain degree therapeutic effect on AA rats.
ABSTRACT
Objective To investigate the effects of artesunate combined with arsenic trioxide (ATO) on the proliferation and apoptosis of NB4 cells.Methods The NB4 cells were treated with different concentrations of artesunate and arsenic trioxide respectively for 48 h.The cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method.The cells were divided into 4 groups:control group,artesunate group,arsenic trioxide group,and the combination of artesunate and arsenic trioxide group.The cell cycle and apoptosis were detected by flow cytometry (FCM).The protein expression levels of Bcl-2 and Bax were detected by Western blot.Results The MTT results showed that compared with the control group,the proliferation inhibition rates of 0.25,0.50,1.00,2.00,4.00 μmol/L artesunate group (19.26％ ± 3.59％,36.53％ ± 2.67％,61.32％ ± 2.50％,70.30％ ± 3.15％,86.92 ± 0.02％) significantly increased (P＜0.05);the proliferation inhibition rates of 1,2,4,8,16 μmol/L arsenic trioxide group (12.69％ ± 2.43％,64.26％ ± 2.02％,85.10％ ± 2.67％,92.06％ ± 2.21％,93.67％ ± 3.36％) significantly increased (P＜0.05);and the proliferation inhibition rate (40.17％ ± 5.49％ vs.32.23％ ± 3.52％) of combination of artesunate and arsenic trioxide group significantly higher than the arsenic trioxide group (P＜0.05).Compared with the arsenic trioxide group,the percentage of G0/G1 phase cells (74.20％ ± 1.43％ vs.66.14％ ± 1.78％),the apoptosis rate (58.00％ ± 2.41％ vs.34.57％ ± 1.22％),and the expression level of Bax protein (1.35 ± 0.09 vs.1.13 ± 0.09) in the combination of artesunate and arsenic trioxide group significantly increased (P＜0.05),the expression level of Bcl-2 protein (0.45 ± 0.09 vs.1.03 ± 0.10) in the combination of artesunate and arsenic trioxide group significantly decreased (P＜0.05).Conclusions Artesunate can significantly enhance the proliferation inhibition and apoptosis induced by arsenic trioxide on NB4 cells.The possible mechanism of proliferation inhibition and apoptosis of NB4 cells by artesunate combined with arsenic trioxide may be related to reduce the expression of anti-apoptotic protein Bcl-2 and increase the expression of apoptotic protein Bax.
ABSTRACT
Objective To explore the correlation of traditional Chinese medical syndrome elements with plasma connective tissue growth factor ( CTGF) and platelet-derived growth factor ( PDGF) in early liver cirrhosis induced by type B hepatitis. Methods The distribution of traditional Chinese medical syndrome elements in early liver cirrhosis induced by type B hepatitis was analyzed, plasma contents of CTGF and PDGF were detected by enzyme-linked immunosorbent assay ( ELISA) , and the correlation of syndrome elements with CTGF and PDGF was discriminated. Results ( 1) The distribution of traditional Chinese medical syndrome elements in early liver cirrhosis induced by type B hepatitis showed as follows: the syndrome elements involved the viscera of liver and spleen, and the pathogenesis was characterized as dampness, heat, qi stagnation, and yin deficiency. ( 2) CTGF was closely related with spleen, gallbladder and dampness, with OR value being 1.598, 1.567, 2.797, respectively. PDGF was closely related with heat, with OR value being 1.134. Conclusion Early liver cirrhosis induced by type B hepatitis mainly affects the viscera of liver and spleen, the pathogenesis is characterized by dampness, heat, qi stagnation, and yin deficiency. The patients with higher CTGF are apt to show the pathological changes of spleen, gallbladder, dampness, and have the syndrome el-ements of spleen, gallbladder, dampness. The patients with higher PDGF are apt to show the pathological changes of heat, and have the syndrome element of heat.
ABSTRACT
Objective: To investigate the incidence of coutrast media iodixanol-induced delayed adverse reaction with the risk factors in general clinical practice. Methods: A total of 20,185 patients with contrast iodixanol were recruited from 95 medical centers in China. The risk factors for adverse drug reaction as hypertension, asthma, previous contrast reaction were assessed;the administrative processes as route, injection manner, lfow rate of injection, prior heating of iodixanol were monitored and the demographic information was documented. The immediate adverse reaction within 1 hour of media administration and the delayed adverse reaction from 1 hour to 7 days after administration were recorded. The risk factors for iodixanol-induced delayed adverse reaction were studied by singlevariate and multivariate logistic regression analysis. Results: The overall iodixanol-induced adverse reaction rate was 1.52%, of which the immediate reaction was 0.58%and delayed reaction was 0.97%. The major delayed reaction was mild and it mostly happened in skin (0.68%) including rash, pruritus and urticaria. Multivariate logistic regression analysis revealed that male gender (OR=0.71, P=0.036), age (OR=0.82, P=0.001), route of administration (OR=0.21, P<0.001), prior heating of iodixanol (OR=1.44, P=0.036), lfow rate of injection (OR=1.28, P=0.001) and previous contrast reaction (OR=16.04, P<0.001) were the independent risk factors for delayed adverse reactions.
ABSTRACT
Quality assurance system for postgraduate education including the implementation system,monitoring and information gathering and feedback system,resource guarantee system was established according to the characteristics of postgraduate education and combined with theory of total quality management.The implementation scheme for total quality management-oriented postgraduate education was formulated from the aspects of training plan,objective requirements,etc.
ABSTRACT
Objective To explore the underlying mechanism of potential effect of sildenafil on porcine pulmonary artery smooth muscle cells (SMCs) proliferation induced by serotonin. Methods Porcine pulmonary artery SMCs at 3-5 passages were randomly divided into 4 groups: control group (CON), 5-HT group (HT), sildenafil group (SIL) and sildenfil-5HT combined group (S-HT). Pulmonary artery SMCs at exponential growth phase were serum starved with 0.2% FBS for 72 h, followed by sterile PBS, serotonin (10 μmol/L) and sildenafil (1 μmol/L) incubation in CON group, HT group and SIL group, respectively. In combined group (S-HT): pulmonary artery SMCs were serum starved and then exposed to sildenafil for 20 min, followed by serotonin incubation for indicated time. The phosphorylation of extracellular signal regulated kinase (ERK1/ERK2) was measured by Western blot at indicated time points. Flow active cell sorting (FACS) was used to evaluate the ratio of S phase cells in the cell cycle after 24 h of treatment. MTT color metric method was used to analyze SMCs proliferative index after 72 h of treatment. Results Compared with CON group, the phosphorylation of ERK1/ERK2, the percentage of cells in S phase, and the cell proliferation index increased remarkably after incubation with 5-HT (P<0.05). Preincubation with sildenafil followed by serotonin enhanced the phosphorylation of ERK1/ERK2 (p-ERK1/ERK2) and further elevated the percentage of cells in S phase and the cell proliferation index compared with that of HT group. While the total ERK1/ERK2 (t-ERK1/ERK2) was not statistically different among these groups. Conclusions Sildenafil potentiates the proliferative effect of serotonin on pulmonary arterial SMCs via upregulating phosphorylation of ERK1/ERK2.
ABSTRACT
Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain, touch and temperature senses. To comprehensively identify proteins of plasma membrane (PM) from small amount of dorsal root ganglion (DRG) neurons, a proteomics strategy that utilizes aqueous polymer two-phase partition in combination with differential velocity centrifugation was adopted to enrich the PM, followed by SDS-PAGE, CapLC-MS/MS and bioinformatics analysis. Western blot analysis showed that the concentration of PM in purified plasma membrane(PPM) was 2.3 times higher than that in crude plasma membrane(CPM), 15 times higher than that in whole tissue lysate (WTL). By searching against the rat IPI protein sequence database, a total of 729 non-redundant proteins were identified from the PM preparation, of which 547 had a gene ontology (GO) annotation indicating a cellular component, and 159 (21.8%) were unambiguously identified as PM proteins. A data set of plasma membrane proteins of DRG as well as a tool to study PM proteins were provided in a small amounts of sample.
ABSTRACT
Objective: To assess the immunological effects by orally administering chicken type Ⅱ collagen(CCⅡ) on collagen-induced arthritis(CIA)mice. To assess the effect on producing IL-1 of peritoneal macrophage in adjuvant arthritis rats by orally administering CCⅡ. Methods: Arthritis were induced in Kunming mice by immunization with chicken type Ⅱ collagen with Freund's complete adjuvant, followed by an interperitoneal injection of CCⅡ 3 weeks later.Chicken type Ⅱ collagen was orally administered from 5 days prior to the induction of arthritis to 14 days after inducing arthritis model. The animals were examined visually twice weekly for polyarthritic signs of swollen and erythemic limbs. Quantitation of antibody level of CIA mice was measured by ELISA method. Subpopulations of T lymphocytes in mice were evaluated by flow cytometry method. IL-1 assay was evaluated by ELISA method. Results: Joint swelling was significantly reduced at a dose of 5 μg.kg-1 and 50 μg.kg-1 of CCⅡ, but not at 250 μg.kg-1. The level of anti-collagen antibodies was also reduced at a dose of 5 μg.kg-1 and 50μg.kg-1 (OD value from CIA model control 0.242±0.073 to CCⅡ 5 μg.kg-1 0.123±0.029 and CCⅡ 50 μg.kg-1 0.110±0.075 respectively). Subpopulations of T-lymphocytes were changed by orally administering of CCⅡ, and the ratio of L3T4+/Lyt-2+ was lowered (the ratio from 1.71 of CIA model control to 1.21, 1.51 of administered CⅡ 5 μg.kg-1, 50μg.kg-1 respectively.) after administering CCⅡ. IL-1 level can be reduced (the value from adjuvant arthritis model control 62.8±0.9 to 43.4±1.3, 49.7±0 ng.L-1 administered CⅡ 5 μg.kg-1, 50μg.kg-1 respectively). Conclusion: Arthritis sign in CIA animal model can be suppressed by oral CCⅡ. The effects may be involved by influencing the mechanisms both humoral and cellular immunity. The effects occurred at lower doses of CCⅡ. These results demonstrated the biologic relevance of by-stander suppression associated with oral tolerance, and the potential use of this approach to treat human inflammatory joint diseases.
ABSTRACT
Objective:To study the antimicrobial susceptibility of periodontopathic bacteria associated with failing implants to commonly used antibiotics in periodontal therapy and dental practice.Methods:Antimicrobial susceptibility of bateria isolates was determined by the method of microbroth dilution and the activities of 8 kinds of antibiotics against international laboratory strains and clinical isolates of Provotella intermedia (Pro.I), Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), Fusobacterium nucleatum (Fn) were examined.Results:Ofloxacin, clindamycin, tinidazole and metronidazole showed stronger antibacterial activity aginst Pro.I,Pg,Aa and Fn with MIC_ 90 ≤0.5 ?g/ml and MBC_ 90 ≤0.5 ?g/ml.Conclusion:Commonly-used antibiotics are potential against bacteria isolated around failing implants.
ABSTRACT
Heme was extracted from fresh pig blood and made into powder in black coffee color. The product contained about 1.2%-1.8% iron and was added into food to raise its iron content.Three experiments were performed on young mice, piglets and anemic children. After feeding this product, their hemoglobin was all raised significantly (P