Regulatory effect of Ganoderma lucidum polysaccharides on H2O2-induced apoptosis and mitochondrial dysfunction in SH-SY5Y cells / 中国组织工程研究

BACKGROUND:Current studies have confirmed that Ganoderma lucidum polysaccharides can promote nerve regeneration in neurodegeneration-related diseases.The occurrence of neurodegenerative diseases is closely related to mitochondrial dysfunction,but the role of Ganoderma lucidum polysaccharides on the regulation of apoptosis and mitochondrial function in neurodegenerative diseases is not yet clarified. OBJECTIVE:To explore the regulatory effects and mechanisms of Ganoderma lucidum polysaccharides on apoptosis and mitochondrial dysfunction in H2O2-induced SH-SY5Y cells. METHODS:SH-SY5Y cells were divided into three groups:control group,H2O2 group,and Ganoderma lucidum polysaccharides group.Cells in the control group were normally cultured.Cells in the H2O2 group were treated with 300 μmol/L H2O2 for 24 hours.In the Ganoderma lucidum polysaccharides group,the intervention with 300 μg/L Ganoderma lucidum polysaccharides was conducted first for 1-2 hours,followed by the addition of 300 μmol/L H2O2 for 24 hours.The mitochondrial membrane potential was detected by JC-1 kit.Apoptosis was detected by TUNEL staining kit.The activities of malondialdehyde and superoxide dismutase were detected by malondialdehyde test kit and superoxide dismutase test kit,respectively.The apoptosis and expression of mitochondrial dynamics-related proteins were detected by immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the mitochondrial membrane potential and superoxide dismutase activity were significantly reduced,as well as apoptotic rate and malondialdehyde levels were significantly increased in the H2O2 group(P<0.05).After treatment with Ganoderma lucidum polysaccharides,the membrane potential and superoxide dismutase activities were significantly increased,and apoptotic rate and malondialdehyde levels were significantly reduced compared with the H2O2 group(P<0.05).(2)The expression levels of pro-apoptotic proteins Bax and Caspase-3 were significantly increased,but the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the H2O2 group compared with the control group(P<0.05).Compared with the H2O2 group,the levels of Bax and Caspase-3 were significantly decreased,but the expression of anti-apoptotic protein Bcl-2 was significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(3)Compared with the control group,the expression of mitochondrial splitting proteins Fis1 and p-Drp1 was significantly increased,but the expression of mitochondrial fusion proteins OPA1,Mfn1,and Mfn2 was decreased in the H2O2 group(P<0.05).Compared with the H2O2 group,Fis1 and p-Drp1 expression was significantly reduced,but the expression levels of OPA1,Mfn1,and Mfn2 were significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(4)The above results confirm that Ganoderma lucidum polysaccharides can attenuate H2O2-induced oxidative stress damage and apoptosis in SH-SY5Y cells by ameliorating mitochondrial dysfunction.

The role and mechanism of TLRs/MyD88/NF-κB signaling pathway in multiple sclerosis / 中国组织工程研究

BACKGROUND:Multiple sclerosis is a chronic inflammatory demyelinating disease of the central nervous system mediated by T cells.The Toll-like receptors(TLRs)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa-B(NF-κB)signaling pathway plays an important role in the development of the disease.Exploring the specific mechanism of the signaling pathway is essential for further treatment of the disease and improving the prognosis of patients. OBJECTIVE:To review the TLRs/MyD88/NF-κB signaling pathway and its role in multiple sclerosis/experimental autoimmune encephalomyelitis models,which provides new ideas and strategies for the treatment of multiple sclerosis by inhibiting the TLRs/MyD88/NF-κB signaling pathway. METHODS:The literature related to the topic from January 2002 to December 2022 was searched in CNKI,WanFang and PubMed databases.A total of 61 articles were finally included for analysis. RESULTS AND CONCLUSION:The TLRs/MyD88/NF-κB signaling pathway is an important pathway that triggers a pro-inflammatory immune response.The TLRs/MyD88/NF-κB signaling pathway plays an important role in the development of multiple sclerosis by regulating the antigen presentation of dendritic cells,destroying the integrity of the blood-brain barrier,and promoting the activation of T cells,B cells and microglia.By targeting TLRs,MyD88 and NF-κB molecules,inhibiting the activation or signal transduction of TLRs,MyD88 and NF-κB,and reducing the secretion of pro-inflammatory factors,multiple sclerosis can be treated.Animal studies have shown that active ingredients of traditional Chinese medicines,such as flavonoids and glycosides,and traditional Chinese medicine compound formulas,such as Buyang Huanwu Tang,can also treat experimental autoimmune encephalomyelitis by regulating the TLRs/MyD88/NF-κB signaling pathway,which points to the direction of searching for medicines targeting the TLRs/MyD88/NF-κB signaling pathway for the treatment of multiple sclerosis.

Astragaloside inhibits astrocyte activation and inflammatory response induced by inflammation / 中国组织工程研究

BACKGROUND:Astrocytes play an important role in the pathology of central nervous system diseases.The phenotypic and functional changes in astrocytes suggest that it may be an effective therapeutic target for central nervous system diseases.Our previous studies have confirmed that astragaloside can inhibit the lipopolysaccharide-induced astrocyte inflammatory response.Whether astragaloside can regulate the phenotype and function of astrocytes through Notch-1 and its downstream signaling pathway remains unclear. OBJECTIVE:To explore the effect of astragaloside on astrocyte activation and inflammatory response induced by inflammation and its possible mechanism. METHODS:Cerebral cortex astrocytes derived from neonatal C57BL/6 mouse were cultured in vitro.CCK-8 assay was used to determine the optimum concentration of astragaloside and Notch active inhibitor DAPT.The astrocytes were divided into five groups:PBS group,lipopolysaccharide group,lipopolysaccharide + astragaloside group,lipopolysaccharide + DAPT group and lipopolysaccharide + DAPT + astragaloside group.The secretion level of inflammatory factors was detected by ELISA,and the level of nitric oxide was detected by Griess method.The astrocytes and splenic mononuclear cells were co-cultured in Transwell chamber to observe the migration of CD4T cells.The expression of astrocyte activation marker GFAP,A1 marker C3 and A2 marker S100A10 as well as Notch 1 and Jag-1 was detected by immunofluorescence staining.The expressions of CFB,C3,S100A10,PTX3,Notch-1,Jag-1,and Hes were detected by western blot assay. RESULTS AND CONCLUSION:(1)According to the results of CCK8 assay,the final concentration of astragaloside was selected as 25 μmol/L and the final concentration of DAPT was 50 μmol/L for follow-up experiments.(2)Compared with PBS group,interleukin-6,interleukin-12 and nitric oxide secretion levels in the lipopolysaccharide group were significantly increased(P<0.05,P<0.05,P<0.01).Compared with the lipopolysaccharide group,interleukin-6(all P<0.05),interleukin-12(P>0.05,P<0.05,P<0.05)and nitric oxide(P<0.05,P<0.01,P<0.01)secretion significantly reduced in the lipopolysaccharide + astragaloside group,lipopolysaccharide +DAPT group,lipopolysaccharide + DAPT + astragaloside group.(3)Compared with the PBS group,the expression of GFAP that is the marker of activated astrocytes and the migration of CD4 T cells were significantly increased in the lipopolysaccharide group(P<0.01).Compared with the lipopolysaccharide group,astrocyte activation was significantly inhibited and CD4 T cell migration was significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group(P<0.05,P<0.05,P<0.01).(4)Compared with the PBS group,the expressions of A1 markers C3 and CFB in the lipopolysaccharide group were increased(P<0.01,P<0.05),and the expressions of A2 markers S100A10 and PTX3 were decreased(P<0.01,P<0.05).Compared with the lipopolysaccharide group,C3(all P<0.01)and CFB(both P<0.05)were significantly reduced and S100A10(all P<0.01)and PTX3(P<0.05,P<0.05 and P>0.05)were increased in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(5)Compared with the PBS group,the expressions of Jag-1,Notch-1 and Hes in the lipopolysaccharide group were significantly increased(all P<0.01).Compared with the lipopolysaccharide group,the expressions of Jag-1(all P<0.01),Notch-1(all P<0.01)and Hes(P<0.05,P<0.01 and P<0.01)were significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(6)The results indicate that astragaloside can promote the transformation of astrocytes from A1 to A2 by regulating Notch-1 signaling pathway,reduce the secretion of inflammatory factors and the migration of CD4 T cells,and thus inhibit astrocyte activation and inflammatory response.

Immunomodulatory effect of astragaloside IV on T cells of experimental autoimmune encephalomyelitis mice / 中国组织工程研究

BACKGROUND:In the initial stage of multiple sclerosis,central immune cells activate and release a large number of inflammatory factors,causing white matter demyelination and even involving gray matter neurons.The equilibrium of differentiation between different subsets of CD4+ T cells plays an important role in the progression of experimental autoimmune encephalomyelitis.The previous results of the research group showed that the active ingredient astragalus glycoprotein in astragalus can regulate the immune response in experimental autoimmune encephalomyelitis mice,and whether it has a regulatory effect on the differentiation of T cell subsets has not been determined. OBJECTIVE:To explore the therapeutic effects and immune regulatory mechanisms of astragaloside IV on experimental autoimmune encephalomyelitis mice. METHODS:Female C57BL/6 mice were divided into the normal control group,experimental autoimmune encephalomyelitis disease model group,and astragaloside IV treatment group(n=8 per group).Myelin oligodendrocyte glycoprotein peptides 35-55 were used for experimental autoimmune encephalomyelitis model induction in the last two groups.On day 10 to 28 after immunization,the astragaloside IV treatment group was treated with 40 mg/kg per day astragaloside IV intragastrically.Body weight and clinical scores of mice in each group were recorded from the immunization day to the 28th day.On the 28th day after immunization,the mouse spinal cord was taken and made into frozen sections for hematoxylin-eosin staining and Lux fast blue staining to observe pathological changes in the spinal cord.Percentage of splenic T cell subsets was detected using flow cytometry.Western blot assay was used to determine the protein expression of interferon-γ,interleukin-17 and interleukin-6 in the spinal cord.Levels of interferon-γ,interleukin-17,interleukin-6 and interleukin-4 in supernatants of cultured splenocytes were determined by ELISA. RESULTS AND CONCLUSION:(1)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could reduce the degree of weight loss in experimental autoimmune encephalomyelitis mice(P<0.05),ameliorate clinical symptoms(P<0.05),inhibit the infiltration of inflammatory cells and alleviate myelin loss(P<0.01,P<0.05).(2)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could inhibit the proportion of CD4+T cell subsets expressing interferon-γ(P<0.001)and interleukin-17(P<0.001),but increase percentages of CD4+ interleukin-10+(P<0.001)and CD4+ transforming growth factor-β+(P<0.01)T cell subsets.(3)Astragaloside IV could inhibit the expression of interferon-γ(P<0.05,P<0.01),interleukin-17(P<0.05,P<0.05),and interleukin-6(P<0.05,P<0.05)in the spinal cord and spleen,and up-regulate the expression of interleukin-4(P<0.01)in spleen.(4)These findings confirm that astragaloside IV alleviates clinical symptoms in experimental autoimmune encephalomyelitis mice,which may be related to regulating the splenic T cell subsets,therefore,inhibiting the infiltration of inflammatory cells into the center and reducing the demyelination.

Knock-down of ROCK2 gene improves cognitive function and reduces neuronal apoptosis in AD mice by promoting mitochondrial fusion and inhibiting its division / 细胞与分子免疫学杂志

Objective To explore the effect of knocking down Rho-associated coiled-coil kinase (ROCK2) gene on the cognitive function of amyloid precursor protein/presenilin-1 (APP/PS1) double transgenic mice and its mechanism. Methods APP/PS1 double transgenic mice were randomly divided into AD model group (AD group), ROCK2 gene knock-down group (shROCK2 group), ROCK2 gene knock-down control group (shNCgroup), and wild-type C57BL/6 mice of the same age served as the wild-type control (WT group). Morris water maze and Y maze were employed to test the cognitive function of mice. Neuron morphology was detected by Nissl staining. Immunofluorescence histochemical staining was used to detect the expression of phosphorylated dynamin-related protein 1 (p-Drp1) and mitochondrial fusion 1 (Mfn1). Western blot analysis was used to detect the expression ROCK2, cleaved-caspase-3 (c-caspase-3), B-cell lymphoma 2 (Bcl2), Bcl2-related protein X (BAX), p-Drp1, mitochondrial fission 1 (Fis1), optic atrophy 1 (OPA1), Mfn1 and Mfn2. Results Compared with AD group mice, the expression of ROCK2 in shROCK2 group mice was significantly reduced; the cognitive function was significantly improved with the number of neurons in the hippocampal CA3 and DG areas increasing, and nissl bodies were deeply stained; the expression of c-caspase-3 and BAX was decreased, while the expression of Bcl2 was increased; the expression of mitochondrial division related proteins p-Drp1 and Fis1 were decreased, while the expression of mitochondrial fusion-related proteins OPA1, Mfn1 and Mfn2 were increased. Conclusion Knock-down of ROCK2 gene can significantly improve the cognitive function and inhibit the apoptosis of nerve cells of APP/PS1 mice. The mechanism may be related to promoting mitochondrial fusion and inhibiting its division.

Efficacy of the Mini-Mental State Examination versus the Montreal Cognitive Assessment in screening cognitive impairment in patients with lacunar cerebral infarction / 中国基层医药

Objective:To investigate the efficacy of the Mini-Mental State Scale (MMSE) versus the Montreal Cognitive Assessment Scale (MoCA) in screening cognitive impairment in patients with a lacunar cerebral infarction. Methods:138 eligible patients who received treatment in the Affiliated Hospital of Shanxi Datong University from January 2018 to October 2019 were recruited for this study. They received cognitive function evaluation by the MMSE and MoCA. These patients were grouped according to the median number of age or the median number of years of education. The sensitivity and consistency of the MMSE versus MoCA in screening cognitive impairment in patients with a lacunar cerebral infarction were analyzed using the χ2 test. The total cognitive scores of the MMSE and MoCA, and the scores of each cognitive domain such as memory, execution, visual space, attention, language, and orientation, were compared between groups using multiple linear regression analysis. Results:The sensitivity of MoCA in screening for cognitive impairment in low-age, high-age, low-year-education, and high-year-education groups and the whole population of patients with a lacunar cerebral infarction was 76.5%, 75.7%, 74.2%, 77.8%, 76.1%, respectively, which were significantly higher than those of MMSE (44.1%, 65.7%, 60.6%, 50.0%, 55.1%, χ2 = 12.17, 13.13, 9.33, 15.75, 23.86, all P < 0.01). The Kappa coefficients of low-age, high-age, low-year-education and high-year-education groups were 0.336, 0.391, 0.358, 0.389, and 0.373, respectively, all of which were less than 0.4 (all P < 0.01). These findings suggest that the consistency of the two scales in screening cognitive impairment is poor. The cognitive impairment detection rate by the MMSE was significantly higher in the high-age group than in the low-age group (65.7% vs. 44.1%, χ2 = 6.50, P < 0.05). The total cognitive scores of MMSE and MoCA and the scores of memory, execution, visual space, attention, language, and orientation in patients with a lacunar cerebral infarction were significantly lower in the high-age group or low-year-education group than in the low-age group ( tMMSE = 3.61, 2.49, 3.12, 4.26, 1.70, 3.69, 2.24, all P < 0.01; tMoCA = 3.83, 1.75, 3.28, 3.80, 2.21, 4.08, 2.52, all P < 0.05) or high-year-education group ( tMMSE = -2.87, -2.32, -0.85, -2.54, -0.73, -2.57, -2.96, all P < 0.01; tMoCA = -2.95, -1.12, -3.39, -1.54, -1.52, -3.09, -3.02, all P < 0.05). Conclusion:Combined application of MMSE and MoCA has a high clinical value in screening cognitive impairment in patients with a lacunar cerebral infarction. High-age patients with a lacunar cerebral infarction who receive low-year education have memory, execution, visual space, attention, language, and orientation impairments.

Correlation between cognitive function and living ability of older adult patients living in a mining community / 中国基层医药

Objective:To investigate the correlation between cognitive function and living ability of older adult patients living in a mining community.Methods:A total of 180 older adult patients living in a mining community who received treatment during July-October 2019 were included in this study. They were randomly divided into the low-age group (< 68 years old, n = 94) and the high-age group (≥ 68 years old, n = 86). Cognitive function and living ability were evaluated using the Mini-Mental State Examination (MMSE), The Montreal Cognitive Assessment (MoCA), and the Activity of Daily Living Scale (ADL). The relationship between cognitive function and living ability was investigated using hierarchical analysis and Pearson correlation analysis. Results:The proportions of older adult patients with abnormal cognitive function identified by the MMSE and MoCA were 39.4% and 66.0%, respectively in the low-age group, and they were 32.6% and 61.6%, respectively in the high-age group. The MoCA had a greater performance in identifying abnormal cognitive function in each group than the MMSE ( χ2 = 26.69, 10.18, both P < 0.001). There were no significant differences in proportions of older adult patients with abnormal cognitive function identified by the MMSE and MoCA between low-age and high-age groups ( χ2 = 0.90, 0.36, both P > 0.05). The proportion of older adult patients with abnormal living ability was not significantly different between low-age and high-age groups (4.3% vs. 10.5%, χ2 = 2.58, P > 0.05). Compared with patients negative for MMSE items, living ability and instrumental activity of daily living increased by 7.0% and 9.4% in low-age patients positive for MMSE items (both P < 0.05). Compared with patients negative for MoCA items, living ability increased by 3.5% in low-age patients positive for MoCA items ( P < 0.05). Correlation analysis revealed that total scores of MMSE and MoCA were significantly negatively correlated with ADL score ( r = -0.26, -0.27, both P < 0.001) and instrumental activity of daily living score ( r = -0.27, -0.27, P < 0.001). Conclusion:Cognitive function and living ability are correlated in older adult patients living in a mining community. We should pay attention to the screening results of cognitive disorder in older adult patients and improve their living ability by improving their cognitive function.

Characteristics and related risk factors of mild cognitive impairment in middle-aged and elderly adults in a coal mine community / 中国医师杂志

Objective:To investigate the characteristic of mild cognitive impairment (MCI) in the adults aged 48 years and over in a coal mine community, and to analyze its associated risk factors.Methods:From July to October 2019, a questionnaire survey for basic information was conducted among 180 middle-aged and elderly adults who met the inclusion criteria in the Datong coal mine community. The cognitive function was evaluated by Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). The effects of gender, age, years of education, sleep, living alone, physical exercise, social activities, smoking and drinking status, body mass index and chronic diseases on cognitive level were analyzed by single factor stratification and multiple linear regression.Results:There was no significant difference in the positive rate of MCI screened by MMSE and MoCA in the age groups of 48-<64, 64-<72 and 72-90 (original and corrected P>0.05); The positive rate of MCI in MoCA screening (64.4%, 66.7%, 60.9%) was significantly higher than that in MMSE (35.6%, 45.6%, 28.1%) (all P<0.05); MMSE was positively correlated with MoCA score ( r=0.762, P<0.001). With the increase of age, the scores of memory, execution and visual space detected by MoCA decreased significantly (all P<0.05), while the scores of attention, language and orientation did not change significantly (all P>0.05). Univariate stratification showed that the significant influencing factors of MMSE or MoCA scores were gender, age, years of education and sleep status (all P<0.05). Multiple linear regression analysis showed that gender ( βMMSE=-0.192; βMoCA=-0.140), years of education ( βMMSE=0.209; βMoCA=0.328) and sleep status( βMMSE=-0.162; βMoCA=-0.136) were risk factors affecting MMSE and MoCA scores ( P<0.05). Conclusions:More middle-aged and elderly adults with MCI might be observed in a coal mine community, and the main characteristics of MCI are impaired memory, executive function and visual space. To prevent and reduce the occurrence of dementia, early interventions of MCI should be carried out among the adults with female, old age, low years of education and poor sleep quality.

Comparisons of serum procalcitonin and fibrinogen degradation product levels between type 2 diabetes mellitus patients with Escherichia coli bloodstream and urinary tract infections / 中国基层医药

Objective:To compare serum procalcitonin and fibrinogen degradation product levels between type 2 diabetes mellitus patients with Escherichia coli bloodstream and urinary tract infections. Methods:The clinical data of 82 type 2 diabetes mellitus patients with Escherichia coli infections who received treatment between December 2014 and December 2019 in the First Affiliated Hospital of Datong University (The Fifth People's Hospital of Datong) were retrospectively analyzed. These patients were assigned to bloodstream infection ( n = 40) and urinary tract infection ( n = 42) according to the way of Escherichia coli infection. Serum procalcitonin and fibrinogen degradation product levels, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, C-reactive protein, white blood cell count, D-Dimer level, antithrombin III activity, and electrolytes were determined and compared between the two groups. Correlation between procalcitonin and other variables was analyzed. Multiple linear regression analysis was performed with procalcitonin level as a dependent variable and other relevant indexes as independent variables. Results:Body temperature, white blood cell count, neutrophil count, monocyte count, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, procalcitonin level, C-reactive protein level, fibrinogen degradation product level, and D-Dimer level in the bloodstream injection group were (39.49 ± 0.64) ℃, (14.92 ± 11.78) × 10 9/L, (13.39 ± 11.60) × 10 9/L, (0.72 ± 0.36) ×10 9/L, (14.86 ± 10.52), (199.15 ± 160.69), (22.81 ± 17.86) μg/L, (133.44 ± 63.63) mg/L, (49.71 ± 41.44) mg/L, (16.56 ± 12.20) mg/L, respectively, which were significantly higher than those in the urinary tract infection group [(37.12 ± 1.20) ℃, (9.04 ± 3.95) × 10 9/L, (6.25 ± 4.02) × 10 9/L, (0.42 ± 0.29) × 10 9/L, (3.67 ± 3.34), (120.01 ± 44.08), (4.46 ± 8.69) μg/L, (39.22 ± 22.16) mg/L, (3.81 ± 3.41) mg/L, (0.84 ± 0.75) mg/L), t = 7.356, 2.578, 3.162, 2.958, 5.538, 2.591, 2.810, 4.825, 2.902, 2.375, all P < 0.05]. Platelet count, lymphocyte count, blood sodium level and antithrombin Ⅲ activity in the bloodstream infection group were (167.50 ± 104.93) × 10 9/L, (1.06 ± 0.58) × 10 9/L, (130.89 ± 6.50) mmol/L, (57.88 ± 16.28)% , which were significantly lower than those in the urinary tract infection group [(239.40 ± 82.52)× 10 9/L, (2.14 ± 0.71) × 10 9/L, (138.46 ± 5.96) mmol/L, (90.11 ± 8.90)%, t = -2.853, -6.313, -4.046, -7.350, all P < 0.05]. Correlation analysis revealed that serum procalcitonin level was positively correlated with body temperature ( r = 0.387), white blood cell count ( r = 0.355), neutrophil count ( r = 0.368), C-reactive protein ( r = 0.605), fibrinogen degradation product level ( r = 0.616), D-Dimer level ( r = 0.486) (all P < 0.05), and it was negatively correlated with sodium level ( r = -0.319) and antithrombin Ⅲ activity ( r = -0.465) (both P < 0.05). Multiple linear regression analysis results revealed that fibrinogen degradation product level and body temperature were greatly correlated with procalcitonin level. Conclusion:Inflammatory indicators procalcitonin level, body temperature, white blood cell count, neutrophil count, C-reactive protein, fibrinogen degradation product level and D-Dimer level were remarkably higher in type 2 diabetes mellitus patients with Escherichia coli bloodstream infection than those in type 2 diabetes mellitus patients with Escherichia coli urinary tract infection. Procalcitonin level was greatly correlated with body temperature and fibrinogen degradation product level.

Relationship between visceral fat area and brachial-ankle pulse wave velocity in patients with diabetic nephropathy / 中国基层医药

Objective:To investigate the clinical characteristics of different stages of type 2 diabetic nephropathy(DN), and to explore the possible factors affecting visceral fat area (VFA).Methods:From September 2018 to March 2019, 464 patients with type 2 diabetes who were hospitalized in the First Affiliated Hospital of Datong University were selected.Among them, 315 patients with urinary albumin/creatinine ratio(UACR)<30 mg/g were selected as normal proteinuria group, 72 patients with UACR 30-299 mg/g were selected as microalbuminuria group, 45 patients with UACR>300 mg/g were selected as massive proteinuria group, and 32 patients with serum creatinine higher than the reference value were selected as renal failure group.The serum creatinine of the first three groups was in the normal range.The clinical data of these patients such as blood pressure, body mass index(BMI), VFA, subcutaneous fat area(SFA), brachial-ankle pulse wave velocity(BAPWV), blood lipid, renal function and blood sugar were collected and compared among the four groups.Using VFA as strain and other indicators as independent variables, multivariate linear regression analysis was carried out.Results:There were statistically significant differences among the four groups in age, height, weight, BMI, head circumference, neck circumference, waist circumference, hip circumference and waist-hip ratio ( F=15.580, 4.679, 6.186, 3.553, 3.153, 2.689, 5.170, 3.114, 3.535, all P<0.05). The VFA of the normal proteinuria group, microalbuminuria group, massive proteinuria group and renal failure group were (102.25±37.09)cm 2, (104.12±40.93)cm 2, (119.63±48.82)cm 2, (110.54±41.58)cm 2, respectively, and the BAPWV were (1 546.97±330.18)cm/s, (1 595.52 ±381.27)cm/s, (1 459.63±285.61)cm/s, (1 703.89±318.64)cm/s, the differences were statistically significant among the four groups( F=3.344, 4.020, all P<0.05). There were statistically significant differences in alanine aminotransferase, creatinine, uric acid, total cholesterol, red blood cell, hemoglobin, ratio of neutrophils to lymphocytes (NLR) and ratio of platelets to lymphocytes (PLR) among the four groups ( F=3.405, 15.535, 6.552, 2.803, 6.158, 15.580, 3.764, 3.262, all P<0.05). With VFA as strain, multivariate linear regression analysis showed that waist circumference, BMI, TG and BAPWV were risk factors for VFA. Conclusion:DN is associated with multiple obesity-related indicators and inflammatory indicators such as NLR, PLR; VFA is associated with BAPWV.

Clinical effect of anisodamine combined with gabapentin on painful diabetic neuropathy / 中国基层医药

Objective To assess the clinical effect of anisodamine combined with gabapentin on painful diabetic neuropathy(PDN).Methods From June 2013 to February 2017,120 patients with PDN in the Fifth People's Hospital of Datong were assigned into anisodamine group,gabapentin group and combined group according to the digital table,with 40 cases in each group,the patients were treated with anisodamine,gabapentin and combination therapy respectively.The abnormal foot plantar pressure,MCV and SCV of median nerve and common peroneus nerve,VAS and total effective rate of three groups were compared.Results After treatment,the indicators of the combined group were improved more significantly than those of the anisodamine group and gabapentin group in abnormal foot plantar pressure[(6.10 ± 1.66) vs.(10.80 ± 2.64) vs.(6.37 ± 1.44),F =14.602,P ＜ 0.01],total effective rate (92.5％ vs.62.2％ vs.75.0％,x2 =10.155,P=0.006),MCV[(50.33 ±5.54)m/s vs.(41.12 ±4.47)m/s vs.(42.32 ±4.40)m/s,F=9.404,P =0.001],and SCV of median nerve[(48.51 ±6.19)m/s vs.(41.81 ±5.72)m/s vs.(41.76 ± 7.17) m/s,F =3.728,P =0.037],MCV of common peroneus nerve [(40.60 ± 5.69) m/s vs.(32.04 ± 4.47) m/s vs.(33.52 ± 7.76) m/s,F =5.614,P =0.009] and SCV of common peroneus nerve [(42.72 ± 4.97) m/s vs.(36.21 ± 6.16) m/s vs.(35.45 ± 5.54) m/s,F =4.265,P =0.025].After treatment,the VAS scores of the three groups decreased,which were (4.49 ± 1.61) points,(5.82 ± 1.58) points,(6.37 ± 1.44) points,respectively,the difference was statistically significant (F =14.602,P =0.000),and the decrease was more significant in the combined group.Conclusion Compared with the anisodamine group and gabapentin group,the combined treatment is more effective on PDN.

Clinical effect of anisodamine combined with gabapentin on painful diabetic neuropathy / 中国基层医药

Objective@#To assess the clinical effect of anisodamine combined with gabapentin on painful diabetic neuropathy(PDN).@*Methods@#From June 2013 to February 2017, 120 patients with PDN in the Fifth People's Hospital of Datong were assigned into anisodamine group, gabapentin group and combined group according to the digital table, with 40 cases in each group, the patients were treated with anisodamine, gabapentin and combination therapy respectively.The abnormal foot plantar pressure, MCV and SCV of median nerve and common peroneus nerve, VAS and total effective rate of three groups were compared.@*Results@#After treatment, the indicators of the combined group were improved more significantly than those of the anisodamine group and gabapentin group in abnormal foot plantar pressure[(6.10±1.66) vs.(10.80±2.64) vs.(6.37±1.44), F=14.602, P<0.01], total effective rate (92.5% vs.62.2% vs.75.0%, χ2=10.155, P=0.006), MCV[(50.33±5.54)m/s vs.(41.12±4.47)m/s vs.(42.32±4.40)m/s, F=9.404, P=0.001], and SCV of median nerve[(48.51±6.19)m/s vs.(41.81±5.72)m/s vs.(41.76±7.17)m/s, F=3.728, P=0.037], MCV of common peroneus nerve[(40.60±5.69)m/s vs.(32.04±4.47)m/s vs.(33.52±7.76)m/s, F=5.614, P=0.009]and SCV of common peroneus nerve[(42.72±4.97)m/s vs.(36.21±6.16)m/s vs.(35.45±5.54)m/s, F=4.265, P=0.025]. After treatment, the VAS scores of the three groups decreased, which were (4.49±1.61)points, (5.82±1.58)points, (6.37±1.44)points, respectively, the difference was statistically significant(F=14.602, P=0.000), and the decrease was more significant in the combined group.@*Conclusion@#Compared with the anisodamine group and gabapentin group, the combined treatment is more effective on PDN.

Clinical observation of venenum bufonis combined with antibiotics in the treatment of bacterial liver abscess / 中国基层医药

Objective To evaluate the clinical effect of venenum bufonis combined with antibiotics in the treatment of bacterial liver abscess .Methods 60 patients with bacterial liver abscess were enrolled ,and all patients took percutaneous transhepatic puncture guided by ultrasound .They were randomly divided into two groups according to the digital table ,30 cases in each group .The control group received antibiotics treatment ,and the treatment group received venenum bufonis plus antibiotics intravenous drip ,once daily ,one course of treatment lasted seven days .The body temperature , blood routine , procalcitonin and other project changes of the two groups were detected .Results Compared with the control group,the effective rate of the treatment group was higher (96.7％ vs.80.0％,χ2 =4.043,P=0.044).In the treatment group,the time of body temperature returning to normal [(7.00 ±1.67)d vs. (9.00 ±1.41)d],leukocyte recovery time [(7.83 ±2.32) d vs.(9.82 ±1.94) d],procalcitonin recovery time [(7.00 ±1.67)d vs.(9.00 ±1.41)d],symptom disappearance time [(5.17 ±1.72)d vs.(7.50 ±1.87)d],disap-pearance time of abscess[(12.00 ±3.41)d vs.(16.00 ±2.37)d]were shorter than those in the control group(t=-2.601,-2.890,-2.236,-2.248,-2.362,P=0.026,0.016,0.049,0.044,0.041).Conclusion Venenum bufonis combined with antibiotics can significantly increase the curative rate and accelerate infection control , there-fore,it is worthy of popularizing in clinical practice .

Effects of astragaloside Ⅳ on inflammatory responses in astrocytes induced by lipopolysaccharide in vitro and its mechanism / 中华微生物学和免疫学杂志

Objective To investigate whether astragaloside ( AST) Ⅳ could inhibit lipopolysac-charide (LPS)-induced activation of astrocytes and the possible mechanism. Methods Effects of different concentrations of AST Ⅳ on astrocyte viability were determined by MTT to select the optimum concentration for the following experiments. Primary astrocytes were induced by LPS to construct the inflammatory model. Astrocytes were divided into three groups: PBS, LPS and LPS+ASTⅣ(LPS+ASTⅣ) groups. The release of nitric oxide (NO) was detected by Griess method. Expression of glial fibrillary acidic protein ( GFAP), an astrocyte marker, and TLR4 were analyzed by immunohistochemistry and Western blot. Cytokines of IL-6, TNF-α, IL-4 and IL-10 in the supernatants of cell culture for 24 h collected after stimulation were meas-ured by ELISA. Results ASTⅣsignificantly inhibited the LPS-induced activation of astrocytes and NO re-lease (P<0. 01), suppressed the expression of TLR4 (P<0. 05) and reduced the secretion of IL-6 (P<0. 01) and TNF-α (P<0. 01), but increased the secretion of IL-4 and IL-10 (P<0. 05 and P<0. 01). Con-clusion AST Ⅳ could significantly inhibit the LPS-induced activation of astrocytes and suppress inflamma-tory responses, and the possible mechanism might be related to reduced secretion of inflammatory factors af-ter blocking the expression of TLR4.

Immunoregulatory effect of BuyangHuanwu Decoction on experimental autoimmune encephalomyelitis mice / 中国免疫学杂志

Objective:To explore the anti-inflammatory therapeutic effect and possible immunoregulatory mechanism of Buyang Huanwu Decoction (BYHWD) on the development of experimental autoimmune encephalomyelitis (EAE). Methods:Female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides ( MOG35-55 ) ,and randomly divided into saline group,BYHWD group,with 13 mice in each group. At the 3th day,25 ml/kg of saline was orally given to each mouse of saline group,50 g/kg ig of crude BYHWD was orally given to each mouse in BYHWD group for 25 days. Clinical score and body mass were recorded every day. Inflammatory cell infiltrations of spinal cord were observed by HE staining Myelin staining observes the demyelination situa-tion. And the expression of ROCKⅡ in spleen was detected by immunofluorescence staining. The subtypes of CD4+ T cells were analyzed by flow cytometry. Western blot was used to detect the expression of TLR4,Myd88,NF-κB,COX-2,ROCKⅡ in spinal cord and ROCKⅡ in brain. Results: The neurologicalscore significantly decreased in EAE mouse of BYHWD group compared with the saline group (P<0. 001) . BYHWD inhibited the inflammatory cell infiltration and demyelination in the nervous centralis(P<0. 05). The treatment of BYHWD effectively reduced the increased the proportion of CD25+(P<0. 05),IL-10+(P<0. 05),TGF-β+(P<0. 01), IFN-γ+( P<0. 05 ) CD4+T cells , and inhibited the expression of peripheral and central ROCKⅡ( P<0. 05 ) ;BYHWD reduced the expression of TLR4,MyD88,NF-κB,COX-2 in spinal cord (P<0. 05). Conclusion:BYHWD can exert anti-inflammatory and immune regulation effect by the inhibition of ROCKⅡ/TLR4/ NF-κB signaling pathway and regulation of the proportion of peripheral T cell sub-sets.

Immunoregulatory effect of Buyang-Huanwu decoction on monocyte-mac-rophages in mice with experimental autoimmune encephalomyelitis / 中国病理生理杂志

AIM:To explore the therapeutic effect of Buyang-Huanwu decoction (BYHWD) on experimental au-toimmune encephalomyelitis ( EAE) and its immunoregulatory effect on monocyte-macrophages .METHODS: Chronic EAE was induced by myelin oligodendrocyte glycoprotein peptide fragment 35-55 ( MOG35-55 ) in the female C57BL/6 mice, which were randomly divided into saline group and BYHWD group .On day 3 after immunization , the mice in BYHWD group were orally administrated with BYHWD , while normal saline was given to the control mice .The clinical score and body mass were recorded every other day .At day 17 after immunization , the mice were sacrificed and spinal cords were obtained for HE staining and myelin staining .The M1 and M2 macrophage phenotypes of splenic cells were detected by flow cytometry and immunofluorescence staining .The protein expression of iNOS , TNF-α, arginase and IL-10 in the spinal cord macro-phages was determined by Western blotting .RESULTS:BYHWD delayed the onset of EAE , reduced the clinical scores of EAE, inhibited the inflammatory cell infiltration and demyelination in the spinal cord , and promoted the conversion of M 1 macrophages into M2 phenotype in the spinal cord and spleen .CONCLUSION:BYHWD intervention attenuates the be-havioral and pathological changes in the EAE mice , and its mechanism may be related to the macrophage conversion .

Immunoregulative effect of Fasudil on encephalomyelitic T cells in experimental autoimmune encephalomyelitis mice / 中南大学学报(医学版)

OBJECTIVE@#To explore the therapeutic effect of Fasudil-modified splenic mononuclear cells (MNCs) in experimental autoimmune encephalomyelitis (EAE) and the possible mechanisms.
@*METHODS@#C57BL/6 female mice were immunized with myelin oligodendrocyte glycoprotein peptide 35-55 to establish active immunity EAE model. Splenic MNCs were isolated on the 9th day after immunization and treated with or without Fasudil for 72 h in vitro. These cells were collected for analysis of the variance of T cell subtypes, the level of cytokines and the activity of Rho kinase (ROCK). MNCs (5×107 cells) were resuspended in 500 µL of phosphate buffer solution (PBS) and transferred into EAE model (intraperitoneal injection), which was divided into a PBS-MNCs group and a Fasudil-MNCs group. Changes of body weight and clinical symptom scores were observed.
@*RESULTS@#Splenic encephalitogenic MNCs from EAE mice on the 9th day after immunization could establish passive transfer EAE model. But Fasudil-treated MNCs did not trigger EAE development. Compared with the PBS-MNCs group, the loss of body weight was less in the Fasudil-MNCs group. The in vitro experiment indicated that Fasudil could suppress the activity of ROCK on MNCs (P<0.01), decrease the percentage of CD4+ T cells with the expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) (IFN-γ: P<0.01; IL-17: P<0.05), while increase the secretion of CD4+ T cells with the expression of transforming growth factor-β (TGF-β) and IL-10 (all P<0.001) . Furthermore, Fasudil could inhibit the release of IL-17 (P<0.001) and enhance the level of IL-10 (P<0.05).
@*CONCLUSION@#Fasudil-modified cell therapy affects the occurrence and development of EAE by inhibiting the inflammatory reaction of helper T cell 1 (Th1) and Th17 while enhancing the immunoregulative effect of Th2.

Clinical observation of diabetic foot ulcer treated with Shuxuetong plus external application of deproteinised ;calf blood serum injection / 中国基层医药

Objective To evaluate the clinical effect of Shuxuetong plus external application of deproteinised calf blood serum injection on diabetic ulcer foot.Methods 60 patients with diabetic foot ulcer were enrolled:30 cases (treatment group)were randomly assigned to receive deproteinised calf blood serum injection(external use)plus Shuxuetong(iv gtt),and the other 30 cases (control group)received routine western medicine therapy.One course of treatment lasted one month.In addition,both two groups received insulin to control blood sugar and antibiotics to control infection,with foot ulcer treated with debridement,drainage,and removal of necrotic tissue etc.Results Com-pared with the control group,the effective rate was higher in the treatment group(86.7% vs.60.0%,χ2 =5.455,P =0.020).The occurrence time of granulation tissue[(6.60 ±1.14)d vs.(10.80 ±2.17)d,t =-3.834,P =0.008] and wound healing time[(23.62 ±6.14)d vs.(35.65 ±4.45)d,t =-4.405,P =0.002]were shorter in the treat-ment group.The wound area in the two groups reduced after treatment,and the wound area in the treatment group reduced significantly[(3.48 ±1.16)cm2 vs.(8.60 ±2.51)cm2 ,t =-4.143,P =0.007].Conclusion As com-pared with the routine therapy with western medicine,the combination therapy of deproteinised calf blood serum injec-tion plus Shuxuetong has better efficacy for diabetic ulcer foot.

The immunoregulatory effect of Rho kinase inhibitor Fasudil on macrophages in a mouse model of ;experimental autoimmune encephalomyelitis / 中华微生物学和免疫学杂志

Objective To investigate the immunoregulatory effect of Fasudil-modified macrophages on cell transferred experimental autoimmune encephalomyelitis ( EAE) in a mouse model.Methods Fe-male C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE.The encephalomyelitic mononuclear cells ( MNCs) were isolated from spleen of mice with EAE on day 9 after immunization and treated with or without Fasudil for 72 h in vitro.Several assays including the flow cytometry analysis, Griess reaction and ELISA were performed to analyze the M1 and M2 phenotypes of macrophages, the production of NO and the levels of cytokines, respectively.The cultured MNCs (5×107 cells) were resuspended in 500μl of PBS and transferred into na?ve C57BL/6 recipients via intraperitoneal injection.Two groups including the PBS-MNCs group and the Fasudil-MNCs group were set up.The body weights and clinical scores of the mice in each group were recorded in every other days after the induction of EAE in the recipients.Results The Fasudil treated MNCs affected the induction of EAE in adoptive cell transferred mice.The expression of CD16/32, iNOS and IL-12 on F4/80-macrophages were decreased, while the expression of CD206, CD23 and IL-10 on F4/80-macrophages were increased upon the treatment of Fasudil, indicating that Fasudil im-proved the differentiation of macrophages from M1 to M2 phenotypes.Moreover, Fasudil inhibited the pro-duction of NO and enhanced the expression of Arginase-1.Conclusion Fasudil ameliorated the clinical se-verity of EAE in mice by promoting the transformation of macrophages from M1 to M2 phenotype.