虫草及蛹虫草对大鼠离体肝细胞能荷的影响

以ADP为底物观察到虫草及蠕虫草提取液能降低大鼠离体肝细胞反应体系内AMP的浓度,并能提高ATP浓度.在1.0～5.0mg生药/ml范围内,虫草液使细胞能荷值增加13.27％～37.35％;蠕虫草液使细胞能荷值增加24.08％～43.73％,P<0.05。     

Endogenous hydrogen sulfide regulates oxidant/antioxidant balance in rat with chronic obstructive pulmonary disease

AIM: To investigate the role of hydrogen sulfide (H₂S) in the protection against oxidative stress in rats with chronic obstructive pulmonary disease (COPD).METHODS: The rat model of COPD was established by cigarette smoking (CS) combined with lipopolysaccharide (LPS) instillation. Thirty-two healthy male Sprague-Dawley rats were randomly divided into 4 groups: control group, CS+LPS group, CS+LPS+NaHS (H₂S donor) group and CS+LPS+PPG (DL-propargylglycine, an inhibitor of cystathionine-γ-lyase) group. After 30 days, the lung functions of the rats were measured, the histological changes of lungs were observed under light microscope and the pathological scores were calculated. The H₂S level in plasma and the protein expression of cystathionine-γ-lyase (CSE) in the lung tissues were measured. The content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and catalase (CAT) were detected to reflect oxidative stress.RESULTS: Compared with control group, the peak expiratory flow (PEF) decreased by 24% and intra-pressure (IP) increased by 66% in CS+LPS group. The pathological scores of the lung tissues also increased. Compared with CS+LPS group, no change in the lung function was observed after given NaHS or PPG, but the pathological scores decreased in CS+ LPS+ NaHS group. Compared with control group, the content of H₂S in plasma was increased by 26% on day 16. Compared with CS+LPS group, the content of H₂S in plasma of CS+LPS+PPG group was decreased by 22% after 30 days. Compared with control group, the protein expression of CSE increased, and no statistical difference among CS+LPS group, CS+ LPS+ NaHS group and CS+LPS+PPG group was observed. Compared with control group, MDA content in the lung tissues was increased by 24% in CS+LPS group, the activity of SOD was increased by 47% and the activity of CAT was increased by 52%. Compared with CS+LPS group, the MDA content in CS+LPS+NaHS group was decreased by 21%, and no statistical difference in the activity of SOD and CAT was observed. The activity of SOD decreased by 33% after given PPG.CONCLUSION: H₂S plays a role as anti-oxidant in the rats with COPD. The CSE/ H₂S pathway may be involved in the development of COPD.     

冬虫夏草深层发酵的研究

通过时冬虫夏草菌（ＣｏｒｄｙｃｅｐｓＳｉｎｅｎｓｉｓ）的深层发酵条件如ｐＨ、培养温度、接种量及转速的探讨及对深层发酵培养基碳氮源和无机盐的筛选，确定了其深层培养所需条件及最佳碳氮源和无机盐。实验结果表明：冬虫夏草菌深层发酵的最适培养条件应为２４℃、２００ｒ／ｍｉｎ、ｐＨ５．０左右，接种量５～１５％，菌丝生长的最佳碳源为葡萄糖，最佳氮源为蛋白和氨基酸复合粉，最佳无机盐为磷酸二氢钾、硫酸镁和氯化钙。     

冬虫夏草生理活性成分的研究进展

冬虫夏草是极其名贵的药用真菌之一,主要含有虫草多糖、核苷类物质、甾醇类物质、氨基酸、活性肽、虫草酸等生理活性物质,对于人体健康具有重要的意义。对近年来冬虫夏草的生理活性成分、成分分析方法以及药理作用等方面的研究进行归纳与论述,为冬虫夏草在食品和药品上的进一步研究与开发提供参考。     

Effects of N-acetylcysteine on Clara cells and CC16 in rat COPD model

AIM: To investigate the effects of N-acetylcysteine (NAC) on the number of Clara cells and secretion of Clara cells secretory protein (CC16) in rat chrohic obstructive pulmonary disesae (COPD) model.METHODS: Thirty rats were divided into control, COPD and NAC groups (n=10). The change of Clara cell ultrastructure was detected through transmission electron microscope. The number of Clara cells and synthesis of CC16 were measured by immunohistochemistry. The CC16 levels in bronchoalveolar lavage fluid (BALF) and serum were tested by ELISA. The level of CC16 mRNA in lung was determined by RT-PCR.RESULTS: The percentage of Clara cells in terminal bronchioles in the COPD group was significantly decreased than that in the control (P<0.01), and the percentage in NAC group was significantly higher than that in COPD group (P<0.01). The levels of CC16 in the BALF and serum in COPD group were significantly lower than those in control group (P<0.01, respectively), and the levels of CC16 in NAC group were significantly higher than those in COPD group (P<0.05, respectively). The expression of CC16 mRNA in COPD group was weaker than that in control group and NAC group (P<0.01, respectively).CONCLUSION: The number of Clara cells and the secretion of CC16 decrease in a rat model of COPD. Antioxidant NAC can enhance the synthesis and secretion of CC16, which may be a mechanism for the suppression of airway inflammation.     

Diagnostic value of Th17 cells in symptom severity and prognosis of chronic obstructive pulmonary disease

AIM: To observed the correlation between Th17 cell level and the symptom severity and prognostic factors of chronic obstructive pulmonary disease (COPD), and to explore the clinical application value of Th17 cell level in assessing the prognosis of patients with COPD. METHODS: The patients with diagnosed COPD (n=110) in our hospital during May 2013 to December 2014, and 40 healthy subjects were enrolled in the study. According to the Global Initiative for Chronic Obstructive Lung Disease (GOLD), the COPD patients were divided into group A (low risk, less symptoms), group B (low risk, more symptoms), group C (high risk, less symptoms) and group D (high risk, more symptoms), which were given inhaled corticosteroid/long-acting β₂ agonist or corticosteroid/long-acting β₂ agonist+long-acting antimuscarinic agent treatment for 3 months. The proportion of Th17 cells, cytokines (IL-17 and IL-6), the COPD assessment test (CAT) score, age, body mass index, pulmonary function and the times of acute exacerbation of COPD in previous 1 year were observed before and after treatment. The correlation analysis between the level of Th17 cells and other clinical characteristics was performed. RESULTS: Th17 cell, IL-17 and IL-6 levels in COPD group were significantly higher than those in control group (P < 0.05). With the increase in the severity of COPD symptoms, Th17 cells, cytokines (IL-17 and IL-6) and CAT score in groups B and D were significantly higher than those in groups A and C (P < 0.05). The univariate analysis showed that the levels of Th17 cells in groups B and D before treatment were positively correlated with the CAT score (P < 0.05), which were negatively correlated with FEV₁ , FEV₁% Pred, FVC and FVC% Pred. The levels of Th17 cells were not correlated with the CAT score, FEV₁, FEV₁% Pred, FVC and FVC% Pred in groups A and C. The levels of Th17 cells after treatment were positively correlated with the CAT score, which were negatively correlated with FEV₁ , FEV₁% Pred, FVC and FVC% Pred (P < 0.05). CONCLUSION: The peripheral Th17 cell level has a good correlation with IL-17, IL-6, CAT score and pulmonary function in COPD patients, suggesting a potential value to predict the symptom severity and prognosis of COPD.     

冬虫夏草的分离及培养技术研究

实验通过对云南中甸采集的新鲜冬虫夏草全株进行菌种的分，纯化，并与云南丽江采集分离的冬虫夏草进行比较，该实验成功分离出冬虫夏草的优良菌种，并获得冬虫夏草的子座。     

发酵型冬虫夏草菌茶的制备工艺研究

以绿茶为主要原料,配伍麦麸组成固体培养基,接种冬虫夏草菌进行固体发酵培养,加工冬虫夏草菌茶。结果显示,冬虫夏草菌在茶叶为主料添加麦麸的培养基上长势良好,通过L9（34）正交试验,以菌茶粗多糖含量为指标,筛选出培养基最优配方为茶叶60 g、麦麸30 g、碳酸钙0.4 g、蔗糖1.2 g。在料水比为5∶3（w·v-1）培养基上,接种12%（mL·g-1）冬虫夏草液体菌种,24℃培养12 d,55℃烘干,粉碎包装。成品菌茶汤为酒红色,香气清淡,无絮状物沉淀,pH值7.28,菌茶粗多糖含量为7.566%,茶多酚含量为1.488%。     

不同产地冬虫夏草菌对寄主幼虫侵染力研究

比较不同产地冬虫夏草菌对寄主幼虫的侵染力,筛选高侵染力的菌株,提高寄主幼虫感染率,为人工培殖冬虫夏草奠定基础。试验结果表明,小金产冬虫夏草菌菌株感染率最高为72.22%,西藏比如产冬虫夏草菌菌株感染最低为45.56%;贡嘎蝠蛾感染率最高为75.56%,比如蝠蛾感染率最低为58.89%,剑川蝠蛾不能感染;除老君山虫草外,各地产冬虫夏草子实体无差别。不同产地冬虫夏草菌的侵染力有差异。     

比色法测定冬虫夏草和蛹虫草中多糖和甘露醇的含量

应用苯酚-硫酸法、3,5-二硝基水杨酸法和高碘酸钠氧化法建立虫草多糖和甘露醇的定量分析方法,并比较市售天然冬虫夏草和人工栽培蛹虫草在多糖和甘露醇含量上的差异。结果发现,用比色法测定三种天然虫草和六种蛹虫草中多糖和甘露醇的含量,其多糖平均加样回收率分别为:94.7%(RSD为2.29%),甘露醇的含量平均加样回收率为98.8%(RSD为1.79%)。实验表明,本研究方法可用于虫草多糖和甘露醇的定量分析。     

冬虫夏草菌丝体免疫特性测试

以发酵生产的冬虫夏草菌丝体为研究材料,通过测试其对荷瘤小鼠超氧化物歧化酶(SOD)和脂质过氧化物(LPO)水平的影响和观察小鼠组织细胞的病理变化,测试冬虫夏草的免疫功能。试验表明,冬虫夏草菌丝体能显著增加荷瘤小鼠脑组织和全血中SOD含量,从而提高机体免疫力,明显减少荷瘤小鼠S180癌细胞面积,降低癌细胞重量,促进癌细胞高度分化,致使癌细胞坏死,对肿瘤生长表现出较强的抑制力。     

Association of SOX5 polymorphisms in chronic obstructive pulmonary disease-related pulmonary hypertension

SOX5) gene single nucleotide polymorphisms (SNPs) among stable chronic obstructive pulmonary disease (COPD) patients,COPD with pulmonary hypertension (PH) patients and healthy controls,and to explore the association of the SOX5 SNPs in COPD-related PH.METHODS: From April 2013 to April 2015,250 patients with stable COPD were enrolled continuously in Ningxia People's Hospital according to COPD treatment guidelines (2013 edition).All the patients received echocardiography,and were divided into COPD with PH group[pulmonary artery systolic pressure (PASP)≥50 mmHg,n=103] and COPD without PH group (PASP<50 mmHg,n=147).The healthy persons (matched for age,sex,race and smoking index,n=127) were selected as control group at the same period.Genotyping of SOX5 gene rs10842262 and rs11046966 loci was performed using MassARRAY genotyping system (Sequenom).Genotype frequencies were calculated.RESULTS: Age,sex and smoking index showed no significantly difference between control group and COPD group,neither between COPD with PH group and COPD without PH group.Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci between control group and COPD group was of significant difference (P<0.05).Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci showed no significant difference between COPD with PH group and COPD without PH group.CONCLUSION: SOX5 gene rs10842262 and rs11046966 loci may play an important role in COPD,but not in COPD-related PH.     

环境因子对冬虫夏草子实体生长发育的影响

研究冬虫夏草子实体的生长发育和形成规律,为冬虫夏草人工培殖及资源保护提供理论依据。将老熟待僵化的幼虫,在不同温度、基质含水量和光照条件下培殖,观察子实体形成与伸展情况。研究表明,子实体分化与形成需要0℃左右低温刺激,变温有利于子实体的形成;子实体的生长还需要有充足的水分及高湿环境,在子实体生长期间,应保持基质含水量为60%~80%,相对湿度95%~100%为宜;微弱的光照都能诱导子实体产生,子实体有向光性生长的特性。环境因子对冬虫夏草子实体形成及伸展具有重要作用。     

虫草人工控制液体培养生态因子研究

通过人工控制液体培养虫草试验,研究了虫草人工控制液体栽培最佳碳、氮、无机盐、维生素种类、浓度及配比,以及最适pH和最适温度。结果表明:虫草液体培养的最适碳、氮组合为葡萄糖5.0 g/L、白糖3.0 g/L、蛋白胨2.0 g/L、牛肉膏1.0 g/L;最适无机盐和维生素是KH2PO41.0 g/L、MgSO4.7H2O0.5 g/L、CaCl2.2H2O0.5 g/L、VB10.1 g/L;最适pH5.5;最适温度是24℃。     

冬虫夏草及其菌丝体研究进展

本文从冬虫夏草及其菌丝体的药化、药理(含临床应用)、冬虫夏草的人工栽培及菌丝体的深层培养诸方面概述了近年来的研究工作,对今后的研究工作提出建议与设想。     

Function of dendritic cells in cord blood

AIM: To explore the function of dendritic cells in cord blood.METHODS: Dendritic cell precursor subsets pDC/pDC(CD11c+CD123- /CD11c-CD123+) in cord blood and adult peripheral blood were analyzed gated by CD34-Lin- HLA-DR+ cells and the levels of IL-12p40,IL-10,IFN-γ,IL-4 in the serums were tested by ELISA.RESULTS: The level of IL-12p40 in cord blood serum was higher than that in peripheral blood.pDC1/pDC2,IL-10,IFN-γ,IL-4 in cord blood were similar to that in peripheral blood.CONCLUSION: Function development of dendritic cells in cord blood may be consummate.     

Role of Th17/IL-17A in chronic inflammatory airway diseases

The idea has been popular for a long time that Th1/Th2 imbalance is the major cause of many diseases. However, the Th1/Th2 paradigm has encountered increasing challenge since the discovery of a novel subset of Th cells, Th17. Th17 cells secrete a series of cytokines (IL-17A~F, IL-21 and IL-22), which is quite different from those produced by Th1 and Th2 cells. It is now generally accepted that Th17/IL-17A plays a pivotal role in autoimmune and host defense. Although first discovered in autoimmune diseases, emerging studies begin to explore the way in which Th17/IL-17A acts in chronic inflammatory airway diseases, such as asthma and chronic obstructive pulmanary disease. In this review, we will summarize the differentiation and function of Th17, and introduce the progress in the correlation between Th17/IL-17A and chronic inflammatory airway diseases. Further elucidating the mechanism of Th17/IL-17A-related pathophysiological changes will contribute to prevention and treatment of chronic inflammatory airway diseases.