Flammulina velutipes polysaccharide-iron (Ⅲ) complex used to treat iron deficiency anemia after being absorbed via GLUT2 and SGLT1 transporters

Iron deficiency anemia(IDA)is a common nutritional problem, but traditional iron supplements cause many adverse reactions. Thus, the development of a novel iron supplement might be significant for the treatment of IDA. This study aimed to study the transport mechanism of Flammulina velutipes polysaccharide-iron complex(FVP1-Fe(Ⅲ))in Caco-2 cells and the therapeutic effect on IDA rats, as well as the influence on gut microbiota in vivo. These results showed that in vitro, the uptake of FVP1-Fe(Ⅲ)was mediated by sodium-dependent glucose transporter-1(SGLT1)and facilitated glucose transporter-2(GLUT2)and GLUT2 played a dominant function. The multidrug resistance-associated protein-2(MRP-2)was involved in the efflux of FVP1-Fe(Ⅲ)across the Caco-2 cells. In vivo, FVP1-Fe(Ⅲ)had a better restorative effect on blood parameters and iron status indicators in rats with IDA as compared with FeSO_4 and exerted this effect by downregulating the expression of hepcidin. FVP1-Fe(Ⅲ)could also regulate gut microbiota dysbiosis in iron deficiency rats by returning the relative abundance of gut microbiota to the normal level. Besides, as a dietary factor, vitamin C(vit C)could enhance the therapeutic effect of FVP1-Fe(Ⅲ). These present findings showed that FVP1-Fe(Ⅲ)could be exploited as a novel iron supplement to treat IDA.

Establishment and evaluation of a mouse model of bronchial asthma with Yin deficiency syndrome

Objective: To establish and evaluate a mouse model of bronchial asthma with Yin deficiency syndrome. Methods: The mouse model of bronchial asthma with Yin deficiency syndrome was established by the treatment with injecting ovalbumin(OVA) two times to sensitize, inhaling OVA 14 times to stimulate, and using thyroxin through lavage during late stimulation. This model was evaluated through body weight, asthmatic behaviors, respiratory function, autonomous activity, lung pathology, and pulmonary fluid clearance. Results: OVA combined with thyroxin was an appropriate method to induce the mouse model with increased food and water intake, autonomous activity, asthmatic behaviors score, and respiratory rate, decreased body weight, tidal volume, and wet/dry ratio of lung, and changed with pathology of lung tissue. The changes of the above mentioned parameters indicated that the model was the bronchial asthma with Yin deficiency syndrome. Conclusion: The OVA combined with thyroxin is a good pattern to establish a mouse model of bronchial asthma with Yin deficiency syndrome successfully, which can highly simulate the clinical symptoms of this disease.

Novel translational rat models of dopamine transporter deficiency

Dopamine （DA） is one of the brain＇s fundamental neurotransmitters. Despite the fact that the dopaminergic synapses constitute less than 1% of all brain synapses, DA is implicated in a number of critical physiological functions and in the pathogenesis of important psychiatric diseases such as schizophrenia, attention-deficit/hyperactivity disorder （ADHD）, Parkinson＇s disease （PD） and others.

Rat Models of Fatty Liver due to Liver Depression and Spleen Deficiency

[Objectives] To establish animal models of fatty liver due to liver depression and spleen deficiency that were suitable for activity discovery of traditional Chinese medicine( TCM) compounds,efficacy evaluation,new drug research and development and correspondence between prescription and syndrome of TCM. [Methods] A syndrome score scale that was suitable for evaluating the rat models was established according to the evolvement rules of etiology and pathogenesis of TCM and the modern clinical pathological mechanism. At the same time,bifendate pills and Sanqi Zhigan pills were selected as drug counterevidence for the models. Rats in model groups were given different proportions of high-fat and low-protein fodder and different concentrations of alcohol every day,and intraperitoneally injected with porcine serum twice a week. Drug groups were given with bifendate pills and Sanqi Zhigan pills( 8. 1 mg/kg,and 2. 7 g/kg) respectively for 14 consecutive days.During the experiment,general status,weight,daily fodder intake and daily water intake of the animals were observed,and TCM syndromes were scored. After the experiment,the levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),γ-glutamine transpeptidase( γ-GT),total cholesterol( TC),triglycerides( TG),low density lipoprotein-cholesterol( LDL-C),and high density lipoprotein-cholesterol( HDL-C) in serum were detected,and the pathological changes in liver tissue were observed. [Results] Compared with the control group,body weight,daily fodder and water intake of rats increased slowly in the model group,and the levels of ALT,AST,γ-GT,TC,TG and LDL-C increased significantly,while the level of HDL-C dropped. Pathological examination showed that steatosis and fat granule were observed in hepatocytes of rats in the model group. Behavioral observation found that main symptoms and minor symptoms of rats in the model group conformed to the syndrome manifestation of liver depression and spleen deficiency,which suggested that the three model groups were established successfully. Among them,three rats died in model group 1,and one rat died in model group 2. The manifestation of all the above lesions can be alleviated by drug counterevidence. [Conclusions]The animal model of fatty liver due to liver depression and spleen deficiency shows an obviously lower mortality and shorter duration and can be used for research of correlation between prescription and syndrome and its mechanism.

Rat model for study of the essence of spleen deficiency and dampness in Chinese medicine

目的:通过构建脾虚水湿证型的动物模型探讨中医“脾虚”和“水湿”的本质。方法:以高脂低蛋白因素和游泳过度疲劳因素诱导脾虚水湿动物模型,选择临床常见的25项血清学指标进行检测,采用t检验、秩检验和回归分析对数据进行分析。结果:模型组大鼠的总蛋白、白蛋白、胃动素、干扰素-γ、白细胞介素-2、免疫球蛋白A、免疫球蛋白G、补体3水平均低于对照组(P=0.029,P=0.032,P<0.001,P<0.001,P<0.001,P<0.001,P<0.001,P<0.001)。模型组的血清肌酐、总胆固醇、醛固酮、抗利尿激素、胃泌素、生长抑素、IL-4、心房钠尿肽和血管活性肠肽水平均高于对照组(P<0.001,P=0.015,P<0.001,P<0.001,P<0.001,P<0.001,P<0.001,P<0.001,P<0.001)。两组在天冬氨酸转氨酶,丙氨酸转氨酶、球蛋白、白蛋白/球蛋白、血尿素氮、高密度脂蛋白,低密度脂蛋白水平无统计学差异。回归分析结果显示,模型组动物证候具有二维性,且此二维性在各生物学指标上难以明显分开。结论:脾虚常伴随水湿,细胞因子、免疫球蛋白、及胃肠激素是影响脾虚水湿的重要因素。

Electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure

Objective: To investigate the electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure.Methods: Guinea pigs model of iron deficiency anemia complicated with chronic heart failure in 10 guinea pigs of the experimental group was made by feeding a low iron diet,pure water and subcutaneous injection of isoproterenol. The control group consisting of 11 guinea pigs was given normal food, normal water and injected with normal saline. The left ventricular outflow tract model specimen was also prepared. The standard microelectrode technique was used to observe electrophysiological changes of autonomic cells in the outflow tract of left ventricular heart failure complicated with iron deficiency anemia in guinea pig model. The indicators of observation were maximal diastolic potential, action potential amplitude, 0 phase maximal depolarization velocity, 4 phase automatic depolarization velocity, repolarization 50% and 90%, and spontaneous discharge frequency.Results: Compared with the control group, 4 phase automatic depolarization velocity,spontaneous discharge frequency and 0 phase maximal depolarization velocity decreased significantly(P < 0.01) and action potential amplitude reduced(P < 0.01) in model group. Moreover, repolarization 50% and 90% increased(P < 0.01).Conclusions: There are electrophysiological abnormalities of the left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with heart failure.

Effects of Different Processed Products of Radix Codonopsis on Intestinal Flora of Rats with Spleen Deficiency

[Objectives]This study aimed to observe the effects of different processed products of Radix Codonopsis on intestinal flora in rats with spleen deficiency.[Methods]Rat models with spleen deficiency were established by bitter-cold purgation method with Radix et Rhizoma Rhei.Normal group,model group,Radix Codonopsis group,fried Radix Codonopsis group,rice-fried Radix Codonopsis group,honey-fried Radix Codonopsis group and bran-fired Radix Codonopsis group were designed.After subjecting to corresponding treatments,the changes in the quantity of intestinal microorganisms of the rats were detected.[Results]There was no significant change in the quantity of intestinal microorganisms of the rats in the normal group and model group.The rats in the rice-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased,and the quantity of Escherichia coli and Staphylococci reduced.The rats in the Radix Codonopsis group,fried Radix Codonopsis groups,honey-fried Radix Codonopsis group and bran-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased(the increases were smaller than those in the rice-fried Radix Codonopsis group),and the abundance of E.coli and Staphylococci reduced,close to normal levels.[Conclusions]Different processed products of Radix Codonopsis have obvious regulation effect on intestinal flora of rats with spleen deficiency,and the regulation effect of rice-fried Radix Codonopsis on rats with spleen deficiency is better than that of Radix Codonopsis,fried Radix Codonopsis,honey-fried Radix Codonopsis and bran-fried Radix Codonopsis.

Guipi decoction effects on brain somatostatin levels and receptor mRNA expression in rats with spleen deficiency

BACKGROUND： Somatostatin is abundant in the hypothalamus, cerebral cortex, limbic system, and mesencephalon. Somatostatin mRNA expression in the brain of rats with spleen deficiency is noticeably reduced, as well as attenuation of cognitive function. OBJECTIVE： To observe the interventional effect of Guipi decoction on somatostatin level and somatostatin receptor 1 （SSTRl） mRNA expression in different encephalic regions of rats with spleen deficiency, and to compare the interventional effects of Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet. DESIGN： A randomized controlled observation. SETTING： Basic Medical College, Beijing University of Traditional Chinese Medicine. MATERIALS： Fifty adult Wistar male rats, of clean grade, weighing （160 ± 10） g, were provided by Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. The protocol was performed in accordance with ethical guidelines for the use and care of animals. Somatostatin 1 polyclonal anti-rabbit antibody and SSTRl in situ hybridization kit were provided by Department of Neuroanatomy, Shanghai Second Military Medical University of Chinese PLA. The drug for developing rat models of spleen deficiency was composed of Dahuang, Houpu and Zhishi, and prepared at 2：1：1. Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet recipes were made according to previous studies. METHODS： This study was performed at the Basic Medical College, Beijing University of Traditional Chinese Medicine from March 2002 to March 2005. The rats were randomly divided into 5 groups, with 10 rats in each group： normal, model, Guipi decoction, Chaihu Shugan powd.er, and Tianwang Buxin pellet groups. Rat models of the latter 4 groups were developed by methods of purgation with bitter and cold nature drugs, improper diet, and overstrain. The rats received 7.5 g/kg of the drugs each morning and were fasted every other day, but were allowed free access to water at all times. The rats were forced to swim in 25 ℃ water until fatigued. Rats in the normal group were intragastrically administered the same amount of normal saline. Rats in the Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet groups were intragastrically administered 7.5 g/kg Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet, respectively, every afternoon. All rats were treated for 6 weeks. MAIN OUTCOME MEASURES： Somatostatin protein and SSTRI mRNA expression in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were determined by immunohistochemistry and in situ hybridization, respectively. RESULTS： Fifty rats were included in the final analysis. In the model group, expression of somatostatin protein and SSTRl mRNA in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were significantly less than in the normal group （P 〈 0.01）. Above-mentioned indices were identical in the Chaihu Shugan powder and model groups. However, expression of somatostatin protein and SSTRl mRNA were significantly higher in the Guipi decoction group compared to model group （P 〈 0.01）. In the Tianwang Buxin pellet group, SSTRl mRNA expression in rat ventral nucleus of hypothalamus and somatostatin level in rat hippocampal CAl region and cortex of prefrontal lobe, as well as ventral nucleus of hypothalamus, were significantly higher compared to model group （P 〈 0.01 ）. CONCLUSION： Somatostatin level and SSTRl mRNA expression in rats with spleen deficiency were lower than in normal rats. Guipi decoction and Tianwang Buxin pellet up-regulated somatostatin level and SSTRl mRNA expression.

Cost effectiveness of routine duodenal biopsies in iron deficiency anemia

AIM To investigate the cost effectiveness of routine small bowel biopsies(SBBs) in patients with iron deficiency anemia(IDA) independent of their celiac disease(CD) serology test results.METHODS We used a state transition Markov model. Two strategies were compared: routine SBBs during esophagogastroduodenoscopy(EGD) in all patients with IDA regardless their celiac serology status(strategy A) vs SBBs only in IDA patients with positive serology(strategy B). The main outcomes were quality adjusted life years(QALY),average cost and the incremental cost effectiveness ratio(ICER). One way sensitivity analysis was performed on all variables and two way sensitivity analysis on selected variables were done. In order to validate the results,a Monte Carlo simulation of 100 sample trials with 10,and an acceptability curve were performed.RESULTS Strategy A of routine SBBs yielded 19.888 QALYs with a cost of $218.10 compared to 19.887 QALYs and $234.17 in strategy B. In terms of cost-effectiveness,strategy A was the dominant strategy,as long as the cost of SBBs stayed less than $67. In addition,the ICER of strategy A was preferable,providing the cost of biopsy stays under $77. Monte Carlo simulation demonstrated that strategy A yielded the same QALY but with lower costs than strategy B. CONCLUSION Our model suggests that EGD with routine SBBs is a cost-effective approach with improved QALYs in patients with IDA when the prevalence of CD is 5% or greater. SBBs should be a routine screening tool for CD among patients with IDA,regardless of their celiac antibody status.

Electrophysiological changes of Papillary Muscles in Guinea Pigs with iron deficiency anemia and heart failure

Objective:To investigate the changes of left ventricular papillary muscle action potentials in guinea pigs with iron deficiency anemia and heart failure.Methods:A total of 20 cases of iron deficiency anemia with heart failure were treated with experimental group and 10 normal guinea pigs as control group. Blood samples were collected to determine hemoglobin content, red blood cell number and whole blood iron index, and the changes of cardiac function and hemodynamics were detected by 6240 biological signal collection system to determine whether the model was successful or not, Intracellular microelectrode technique was used to determine the action potentials of the papillary muscles in the model group and the control group. the potential amplitudes (APA), overshoot values (APA), maximum depolarization rate (Vmax), 20 % of repolarization, 50 % and 90 % of repolarization (APD20, APD50 and APD90 ) and the average velocity of repolarization were measured. Compare statistical difference between the model group and the control group.Results: 14 cases of model group survived completely, compared with control group, APD50 and APD90 prolonged, and the average velocity decreased.Conclusions: the action potential repolarization duration in the guinea pig papillary muscle of iron deficiency anemia with heart failure is prolonged, and the average repolarization velocity is slow.

中老年冠心病心绞痛患者中医虚证诊断模型的探索与构建

目的 构建中老年冠心病心绞痛中医虚证的诊断预测模型。方法 选取2021年1月至2022年12月南京中医药大学泰州附属医院住院的160例冠心病心绞痛中老年(50~85岁)患者的临床资料,统计年龄、性别、体重指数、吸烟、饮酒、血常规、血脂、肝肾功能、冠状动脉血管病变程度,以及高血压病、糖尿病、胃肠道疾病、慢性阻塞性肺疾病、脑血管疾病等合并症作为自变量,每位患者均给出虚实辨证,其结果作为因变量。应用单因素logistic回归筛选变量,多因素logistic回归分析建立模型,可视化模型。使用受试者操作特征曲线、校准曲线及决策曲线分析评估模型。结果 160例患者实证83例,虚证77例。多因素回归结果显示:年龄>67~85岁[OR=2.776,95%CI(1.219,6.321),P=0.015]、糖尿病[OR=2.230,95%CI(0.960,5.178),P=0.062]、Genisi评分4.50~7.50[OR=10.126,95%CI(2.574,39.843),P=0.001]、Genisi评分8.00~15.75 [OR=13.510,95%CI(3.441,53.040),P<0.001]、Genisi评分16.00~80.00[OR=53.032,95%CI(11.769,230.027),P<0.001]是冠心病心绞痛虚证的预测因子。模型方程为:Logit(虚证概率)=-4.231+1.021×年龄(>67~85岁)+0.802×患糖尿病+2.315×Gensini等级(4.50~7.50)+2.603×Gensini等级(8.00~15.75)+3.952×Gensini等级(16.00~80.00)。该模型受试者曲线下面积值为0.852(95%CI:0.795~0.910)。校准度曲线提示准确度较好,决策曲线分析表现在该预测变量在6%~85%广大风险范围内展现出较好的正向收益率。结论 本研究构建了一个中老年冠心病心绞痛虚证诊断预测模型,有较好的临床预测能力。

不同剂量腺嘌呤建立脾肾阳虚型慢性肾衰竭大鼠模型的实验研究

目的比较不同剂量腺嘌呤灌胃对模型大鼠一般状态、生化指标及肾脏病理组织的影响,确定腺嘌呤灌胃制备脾肾阳虚型慢性肾衰竭(Chronic kidney disease,CRF)大鼠模型的最佳造模剂量。方法将24只SD雄性大鼠随机分为正常组与模型组,模型组大鼠分为M1组、M2组和M3组,每组6只。正常组给予等量生理盐水灌胃,M1组以腺嘌呤150 mg·kg^(-1)·d^(-1)灌胃、M2组以腺嘌呤200 mg·kg^(-1)·d^(-1)灌胃,M3组以腺嘌呤300 mg·kg^(-1)·d^(-1)灌胃。连续干预3周,造模前及造模后每周检测肌酐(Serum creatinine,Scr)、尿素氮(Blood urea nitrogen,BUN)、24 h尿蛋白定量(24-hour urinary protein quantification,24 h-UTP)水平,3周后取肾脏病理组织采用苏木-伊红(HE)和马松(Masson)染色法对大鼠肾脏组织病理学进行模型评估。结果与正常组大鼠比较,模型组大鼠均出现不同程度的精神萎靡不振,活动度降低,弓背蜷缩,毛发稀疏无光泽,耳缘及脚趾苍白发凉,小便频数,大便稀溏等现象。与正常组大鼠比较,各模型组大鼠体重均有不同程度的减轻(P<0.05),造模第2周,与M1组大鼠体重比较,M2组、M3组大鼠差异均有统计学意义(P<0.05)。与正常组大鼠比较,各模型组大鼠24 h尿量明显增多(P<0.05),Scr、BUN及24 h-UTP水平均升高(P<0.05),且M3组大鼠各指标升高最明显。各模型组大鼠Scr、BUN及24 h-UTP比较差异均有统计学意义(P<0.05)。肾脏病理染色结果显示各模型组大鼠的肾脏组织出现不同程度的改变,镜下见肾小球形态基本完整,部分肾小球结构破坏;部分近肾小管空泡变性严重,上皮萎缩,部分肾小管管腔扩张,管腔内见红染无结构的蛋白管型样物及较多红细胞;随着腺嘌呤灌注剂量增加,肾小球内可见大量黄绿色结晶沉淀物,存在肾间质炎性细胞浸润和纤维组织增生。结论腺嘌呤灌胃可成功建立脾肾阳虚型慢性肾衰竭动物模型,其中200 mg·kg^(-1)·d^(-1)灌胃组更接近临床脾肾阳虚型慢性肾衰竭发展进程,且死亡率较低,符合脾肾阳虚型慢性肾衰竭生理病理学特点。

脾虚证实验动物模型构建及评价方法的研究述评

中医症候实验动物模型的构建在中医药作用机理研究方面起到重要支撑作用,近年来脾虚证动物模型的构建和评价文献报道越来越多,但是构建方法标准不一,评价指标客观化不足,本文将从动物选取、模型建立、宏观表征、行为学实验、脾虚的客观指标等方面总结脾虚动物模型的构建和评价方法,以期为脾虚证实验动物模型的构建提供理论指导,为脾虚动物模型平台的选取提供文献参考。

脾虚证动物模型研究述评

中医界运用现代科学方法进行相关研究已有多年历史,在病证动物模型上颇有收获。本文对脾虚证动物模型发展历程进行回顾,对动物选择、造模方法、评价指标、消化系统病证结合模型进行总结和评论。模型动物多选用以大鼠为代表的小型啮齿类。目前,多基于病因病机造模,而较少使用化学药物与物理因素造模,复合因素造模法最常见。评价指标则分为中医证型指标、与“脾”功能和特性有关量化指标,以及方剂反证。功能性腹泻/消化不良为代表的脾虚证病证结合模型是目前研究的重心。脾虚证动物模型研究成绩斐然的同时仍存在许多不足,如缺乏脾虚证模型行业标准和共识,评价指标缺乏系统性和整体性,脾虚理论的现代阐释缺乏统一,学界亟需解决上述问题以促进发展。

基于多因素造模法改良脾胃气虚型糖尿病胃轻瘫大鼠模型制备方法研究

目的:采用多因素造模法联合链脲佐菌素(STZ)配合高糖高脂饲料建立脾胃气虚型糖尿病胃轻瘫(DGP)大鼠模型,并对该模型进行评价。方法:20只SPF级Wistar大鼠依据随机数字表随机分为空白组8只和造模组12只,以多因素造模法复制脾胃气虚型大鼠模型,制定脾胃气虚型模型评价量表,将符合条件大鼠纳入实验;采用小剂量多次注射STZ联合高糖高脂饲料复制DGP大鼠模型,于实验6周末取材。观察大鼠随机血糖、体质量、胃排空率、小肠推进率;Elisa法检测血清中血清淀粉样蛋白(SAA)、D-木糖、乳酸脱氢酶(LDH)的含量;光镜下观察脾脏病理结构改变。结果:与空白组比较,造模组大鼠一般情况(精神状态、粪便、饮食量)较差、体质量降低,差异有统计学意义(P<0.05);造模组随机血糖显著升高,差异有统计学意义(P<0.01);与空白组比较,造模组胃排空率、小肠推进率、SAA、D-木糖、LDH含量降低,差异有统计学意义(P<0.05,P<0.01);光镜下显示造模组脾脏被膜增厚,红髓和白髓分界限不清,淋巴细胞减少。脾胃气虚型DGP证型评价:造模组大鼠有神疲倦怠、体质量降低、蜷缩、活动度差、眯眼、毛色暗淡、舌色偏暗等脾胃气虚证表现。结论:多因素造模法建立的大鼠模型符合脾胃气虚型DGP大鼠标准。

水液缺乏型干眼模型小鼠睑板腺的变化

背景:近年来,越来越多的研究关注于睑板腺中腺泡细胞的增殖异常分化,认为这一过程与干眼症的发生和发展密切相关。睑板腺管腔阻塞和腺泡萎缩等结构和功能异常,可引起或加重干眼。因此,研究干眼中睑板腺变化对于深入理解干眼症的发病机制、寻找治疗和预防干眼症的新靶点具有重要的意义。目的:探究水液缺乏型干眼模型小鼠中睑板腺的变化。方法:选取6-8周雌性C57/B6小鼠32只,随机将其分成2组,每组16只。实验组小鼠采用同时切除眶外和眶内的泪腺的方法建造水液缺乏型干眼小鼠模型,对照组不做手术处理。正常饲养2周后于裂隙灯下观察两组小鼠角膜的改变,同时检测泪液分泌量。麻醉后脱颈处死两组小鼠后取睑板腺,观察睑板腺大体形态的变化,制成冰冻切片,苏木精-伊红染色观察睑板腺的结构,油红染色评价睑板腺的功能,免疫荧光染色和RT-qPCR观察两组小鼠睑板腺中祖细胞角蛋白14、细胞核增殖相关抗原(Ki67)以及异常分化的小脯氨酸丰富蛋白1B的表达情况。结果与结论:①造模2周后,实验组小鼠角膜可见片状缺损,角膜缘新生血管生成且侵入角膜中央;②实验组小鼠泪液分泌量较对照组明显减少(P<0.05);③显微镜照相显示,实验组睑板腺导管中断萎缩,排列紊乱;④苏木精-伊红结果显示,实验组小鼠睑板腺中的脂质空泡较对照组明显增多;⑤油红染色中可见实验组小鼠脂质沉积;⑥免疫荧光和RT-qPCR结果显示,实验组小鼠睑板腺中祖细胞角蛋白14、Ki67、小脯氨酸丰富蛋白1B的表达较对照组明显增多(P<0.05);⑦提示水液缺乏型干眼可导致睑板腺的代偿性肥大、增殖增加、脂质代谢异常,同时可发生睑板腺异常分化。

阿尔茨海默病“肾虚酿毒伤髓”病机演变模型构建及其应用

目的 根据肾-骨髓-血细胞-脑髓相互影响的中西医理论,建立一种阿尔茨海默病(Alzheimer′s disease, AD)“肾虚酿毒伤髓”模型,筛查AD有效中药,并初步探讨该模型的临床应用。方法 由于AD患者B淋巴细胞可以促进神经细胞产生Aβ毒素。将肾虚型AD患者外周血提取B淋巴细胞永生化,建立淋巴母细胞系(lymphoblastoid cell lines, LCL),并与人神经上皮瘤细胞(SKNMC)共培养,产生Aβ毒素,制成阿尔茨海默病“肾虚酿毒伤髓”模型。通过免疫荧光法检测Aβ_(1-42)的表达量,筛查有抑制Aβ_(1-42)生成的中药。并初步探讨该模型的临床应用。结果 与正常对照组相比,肾虚型AD模型中神经细胞Aβ_(1-42)的生成明显增多,具有统计学意义(P<0.01)。与肾虚模型组相比,白术、人参、黄芪、生地黄、梓醇减少了神经细胞Aβ_(1-42)的生成,差异具有统计学意义(P<0.01)。结论 白术、人参、黄芪、生地黄、梓醇可以减少AD中Aβ_(1-42)的生成。该模型除了筛查有效中药外,可以作为AD患者“肾虚酿毒”严重程度的生物标记物,及评价中药干预的疗效指标。

脾虚湿盛证动物模型构建方法探析

近年来,脾虚湿盛证动物模型的构建已成为中医证候研究的热点。脾虚湿盛证病位在脾,病性为虚与湿,因此模型的构建较为复杂,目前尚无公认的动物模型。现从脾虚湿盛证模型构建的思路方法及模型评价指标2个维度为核心对当前研究进行探析与总结,研究表明脾虚湿盛证动物模型常选用饮食失宜、久居湿地及劳倦过度等因素从病因角度进行模型构建,造模方法分为单因素、双因素与混合多因素;脾虚湿盛证模型评价指标常以症状及表证进行宏观与证候评价、多通过疲劳指标、胃肠功能指标及水液代谢指标进行客观评价;并从脾虚湿盛模型证候命名、模型证候评价标准及构建模型的方法剖析目前模型构建存在的问题、以此讨论并分析脾虚湿盛证动物模型未来的发展方向,以期为脾虚湿盛证动物模型的研究提供参考,为研究者提供便利。

基于解痉多肽表达化生病证结合的小鼠模型探讨胃癌前病变脾胃虚实证候的本质

目的为了构建病证结合的小鼠模型及进行胃癌前病变(gastric precancerous lesions,GPL)脾胃虚实证候的本质研究。方法采用他莫昔芬(tamoxifen,TMXFR)诱导构建解痉多肽表达化生(spasmolytic peptide expression metagenesis,SPEM)小鼠模型,并在SPEM模型基础上分别模拟构建脾胃湿热证、脾气虚证模型,结合TMXFR诱导的SPEM模型具有自恢复的特性,以TMXFR组及生理盐水处理组作为对照组,监测小鼠一般情况,HE染色检测SPEM模型的构建及恢复情况,ELISA检测小鼠血清胃泌素水平,PCR检测各组小鼠胃黏膜中miR-7a-5p含量,免疫组化检测各组小鼠胃黏膜中白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-13(interleukin-13,IL-13)表达。结果病证结合的脾胃湿热证、脾气虚证组较TMXFR组、SPEM组织恢复明显延迟;脾胃湿热证较对照组及TMXFR组的胃泌素表达异常升高(P<0.05),SPEM模型组小鼠胃黏膜中miR-7a-5p较对照组水平降低(P<0.05),病证结合的脾胃虚实组miR-7a-5p表达较TMXFR有下降趋势;病证结合的脾胃虚实组IL-1β表达较TMXFR组、对照组升高(P<0.05),脾胃湿热证、脾气虚证组中IL-13表达有升高趋。结论此项研究结果侧面证实了在癌前病灶已形成的前提下,虽无幽门螺杆菌感染,在外感湿热之邪、进食肥甘厚腻、饥饱失常、过劳伤脾等诱因下,机体可突破自身代偿修复能力,延缓SPEM恢复甚至使GPL进一步进展。这可能跟miR-7a-5p介导炎症因子IL-1β使得Th1/Th2细胞失衡相关,但具体机制尚待进一步研究。

基于LASSO回归的心脏神经官能症刚虚证(肝肾阴亏、肝阳上亢证)关联因素筛选及诊断模型构建

目的:利用LASSO回归联合Nomogram构建心脏神经官能症刚虚证(肝肾阴亏、肝阳上亢证)的诊断模型。方法:采用单中心前瞻性研究,收集141例心脏神经官能症患者的临床资料,纳入分析变量包括年龄、民族、婚姻、教育程度、脑力/体力工作、体质量指数(BMI)、中医症状、中医五态人格量表各因子得分、汉密尔顿焦虑量表-14项、汉密尔顿抑郁量表-24项、症状自测评量表各因子均分。通过LASSO回归筛选与刚虚证诊断显著相关的影响因素,纳入二元多因素Logistic回归分析构建诊断模型,并对模型预测区分度及校准度评价,利用10重交叉验证进行内部验证,最后对模型进行Nomogram可视化,并根据ROC曲线确定诊断阈值。结果:共纳入刚虚证患者70例,非刚虚证患者71例。LASSO回归筛选出与刚虚证诊断相关性最显著的5个变量为女性、年龄、疲乏无力、善嗳气、五态人格中少阳积分。模型AUC为0.85,H-L检验为2.94(P=0.9824),提示模型区分度及校准度较好,10重交叉内部验证结果提示AUC为0.82。结论:LASSO回归联合Nomogram构建的诊断模型可协助诊断心脏神经官能症刚虚证,但研究结果的准确性尚待大样本临床研究进一步验证。