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Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: A natural protocol and its use for patch clamp experiments

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World Journal of Gastroenterology 2005年第44期11卷 6968-6974页

作者： san-hua leng fu-er lu Institute of Integrative Traditional Chinese and Western Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM： To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study. METHODS： The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine ceils with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic β-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels （KATP）, voltage-dependent potassium channels （Kv）, and voltage-dependent calcium channels （KcA） in β-cells were identified by patch clamp technique. RESULTS： After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly （MTT value from 0.024 ±0.003 to　3.028±0.003, insulin secretion from 2.6±0.6 to 3.1±0.8 mIU/L）. Then MTT value and insulin secretion increased quickly from d 5 to d 10 （MTT value from 0.028 ±0.003 to 0.052±0.008, insulin secretion from 3.1±0.8 to 18.3±2.6 mIU/L）, then reached high plateau （MTT value 〉0.052±0.008, insulin secretion 〉18.3±2.6 mIU/L）. In contrast, for the isolated adult pancreatic islet cells, both insulin release and MTT value were stable in the first 4 d （MTT value from 0.029±0.01 to 0.031±0.011, insulin secretion from 13.9±3.1 to 14.3±3.3 mIU/L）, but afterwards they reduced gradually （MTT value 〈0.031 ±0.011, insulin secretion 〈8.2±1.5 mIU/L）, and the pancreatic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic β-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from KATP, Kv, and KCA. CONCluSION： Islet cells different

关键词： Pancreatic duct cells Pancreatic precursor cells Insulin-producing cells Patch clamp Experimental protocol ATP sensitive potassium channels Voltagedependent potassium channels Voltage-dependent calcium channels

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Therapeutic effects of berberine in impaired glucose tolerance rats and its influence on insulin secretion

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Acta Pharmacologica Sinica 2004年第4期25卷 98-104页

作者： san-hua leng, fu-er lu2, Li-jun XU Institute of Integrated Traditional Chinese & Western Medicine, Tongji Hospital, TongjiMedical College, huazhong University of Science & Technology, Wuhan 430030, China Institute of Integrated Traditional Chinese & Western Medicine Tongji Hospital Tongji Medical College Huazhong University of Science & Technology Wuhan 430030

AIM: To explore the anti-diabetic effects of berberine and its influence on insulin secretion. METHODS: Impaired glucose tolerance rats induced by iv injection of streptozotocin 30 mg/kgwere treated with berberine 187.5 and 562.5 mg/kg while fed with high fat laboratory chow. After rats were treated for 4 weeks, oral glucose tolerance was determined, and for 8 weeks, the fasting blood glucose, insulin, lipid series were determined. In insulin secretion experiments, berberine 93.75, 187.5, and 562.5 mg/kg was administered orally to BALB/c mice at a bolus. The murine serum was collected 2 h after the berberine administration for insulin determination. Insulin released from HIT-T15 cells and pancreatic islets incubated with berberine 1-100 μmol/L for 12 h was determined. RESULTS: The levels of fasting blood glucose (7.4±1.5 or 7.3±1.3 vs 9.3±1.3 mmol/L), triglycerides (0.61±0.22 or 0.63± 0.17 vs 1.8±0.7 mmol/L), total cholesterol (1.8±0.3 or 1.9±0.3 vs 2.2±0.2 mmol/L), free fatty acid (456±93 or 460±72 vs 550±113 μmol/L) and apolipoprotein B (0.37±0.02 or 0.42±0.05 vs 0.46±0.04 g/L) were reduced greatly in berberine-treated groups at doses of 187.5 and 562.5 mg·kg-1·d-1, respectively as compared with those in control group (Pereas high densitylipoprotein-cholesterol (1.5±0.3 or 1.4±0.3vs 1.1±0.1 g/L), apolipoprotein AI (0.80±0.08 or 0.87±0.08 vs 0.71±0.06 g/L) were significantly increased (Plucose tolerance was improved. In vitro experiment showed that berberine 1-10 μmol/L facilitated insulin secretion of HIT-T15 cells and murine pancreatic islets in a dose-dependent manner. Meanwhile murine serum insulin level (27.5±2.7 or 29±4 or 29±4 vs 24.3±2.8 pIU/L) was undoubtedly promoted and blood glucose (4.52±0.31 or 4.45±0.29 or 4.30±0.19 vs 4.87±0.21 mmol/L) was reduced after berberine administration at doses of 93.75, 187.5, and 562.5 mg/kg, respectively in the BALB/c mice. CONCluSION: Berberine possesses anti-diabetic effec

关键词： berberine diabetes mellitus glucose tolerance test insulin HIT-T15 cells pancreatic islets lipids

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Facilitating effects of berberine on rat pancreatic islets through modulating hepatic nuclear factor 4 alpha expression and glucokinase activity

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World Journal of Gastroenterology 2008年第39期14卷 6004-6011页

作者： Zhi-Quan Wang fu-er lu san-hua leng Xin-Sheng Fang Guang Chen Zeng-Si Wang Li-Ping Dong Zhong-Qing Yan Institute of Integrated Traditional Chinese and Weestern Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM： To observe the effect of berberine on insulin secretion in rat pancreatic islets and to explore its possible molecular mechanism. METHODS： Pdmary rat islets were isolated from male Sprague-Dawley rats by collagenase digestion and treated with different concentrations （1, 3, 10 and 30 μmol/L） of berberine or 1 μmol/L Glibenclamide （GB） for 24 h. Glucose-stimulated insulin secretion （GSIS） assay was conducted and insulin was determined by radioimmunoassay. 3-（4,5-Dimethylthiazol-2-yl）- 2,5-diphenyltetrazolium bromide （MTT） assay was performed to evaluate cytotoxicity. The mRNA level of hepatic nuclear factor 4 alpha （HAIF4α） was determined by reverse transcription polymerase chain reaction （RT-PCR）. Indirect immunofluorescence staining and Western blot analysis were employed to detect protein expression of HNF4α in the islets. Glucokinase （GK） activity was measured by spectrophotometric method. RESULTS： Berberine enhanced GSIS rather than basal insulin secretion dose-dependently in rat islets and showed no significant cytotoxicity on islet cells at the concentration of 10 μmol/L. Both mRNA and protein expressions of HNF4α were up-regulated by berberine in a dose-dependent manner, and GK activity was also increased accordingly. However, GB demonstrated no regulatory effects on HNF4α expression or GK activity. CONCluSION： Berberine can enhance GSIS in rat islets, and probably exerts the insulinotropic effect via a pathway involving HNF4α and GK, which is distinct from sulphonylureas （SUs）.

关键词： Berberine Sulphonylureas Hepatocyte nuclear factor 4 alpha Glucokinase Pancreatic islet

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人在回路深度学习垂体分割模型的建立

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放射学实践 2024年第3期39卷 330-334页

作者：冷渌清花蕊石峰陈明吴玉桥朱珠成都影和医学影像诊断中心成都610058 上海联影智能医疗科技有限公司上海200030 四川省遂宁市中心医院放射科四川629099

目的:采用深度学习方法,通过人在回路的方式进行迭代式标注-训练,建立垂体分割模型,实现垂体体积人工智能(AI)测量。方法:将1285例颅脑3D T 1WI图像按5~15岁、16~25岁、26~50岁、51~70岁年龄段分组,每个年龄组随机选择80例,分成4批次进... 详细信息

目的:采用深度学习方法,通过人在回路的方式进行迭代式标注-训练,建立垂体分割模型,实现垂体体积人工智能(AI)测量。方法:将1285例颅脑3D T 1WI图像按5~15岁、16~25岁、26~50岁、51~70岁年龄段分组,每个年龄组随机选择80例,分成4批次进行试验。初始每组选择3例图像进行人工预标注神经垂体和腺垂体,输入计算机进行学习,获取初始模型。应用模型对一批数据进行分割,获得分割后的神经垂体、腺垂体与垂体总体积数据,将分割结果进行人工校准,获得校准后相对应的体积数据作为金标准。用前一组校准后的分割图像进行计算机迭代式学习优化模型,再用优化后模型对新一组图像分割与校准,重复上述过程,最终将校准前后差异没有统计学意义的数据认定深度学习建模成功。数据采用配对t检验、Dice和Spearman相关性分析进行统计。结果:从第2批次开始,除5~15岁年龄段外,其它年龄段神经垂体体积在校准前后的差异没有统计学意义,腺垂体与垂体总体积的差异有统计学意义(P<0.05)。第4批次,各年龄段神经垂体、腺垂体与垂体总体积在校准前后的差异均无统计学意义(P=0.137~0.928),Dice值大于0.941,Spearman相关系数大于0.969。结论:通过迭代式训练进行深度学习建模可构建垂体分割模型,实现垂体体积AI自动测量。

关键词：垂体深度学习人在回路自动分割迭代式训练

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To explore the mechanism of Yigong san anti-gastric cancer and immune regulation

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World Journal of Gastrointestinal Oncology 2024年第5期16卷 1965-1994页

作者： Dou-Dou lu Ling Yuan Zhao-Zhao Wang Jian-Jun Zhao Yu-hua Du Na Ning Guo-Qing Chen Shi-Cong huang Yi Yang Zhe Zhang Yi Nan School of Clinical Medicine Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina College of Pharmacy Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina Traditional Chinese Medicine College Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina Department of Chinese Medical Gastrointestinal China-Japan Friendship HospitalBeijing 100029China Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina

BACKGROUND Yigong san(YGS)is a representative prescription for the treatment of digestive disorders,which has been used in clinic for more than 1000 ***,the mechanism of its anti-gastric cancer and regulate immunity are still remains *** To explore the mechanism of YGS anti-gastric cancer and immune *** Firstly,collect the active ingredients and targets of YGS,and the differentially expressed genes of gastric ***,constructed a protein-protein interaction network between the targets of drugs and diseases,and screened hub *** the clinical relevance,mutation and repair,tumor microenvironment and drug sensitivity of the hub gene were ***,molecular docking was used to verify the binding ability of YGS active ingredient and hub *** Firstly,obtained 55 common targets of gastric cancer and *** Kyoto Encyclopedia of Genes and Genomes screened the microtubule-associated protein kinase signaling axis as the key pathway and IL6,EGFR,MMP2,MMP9 and TGFB1 as the hub *** 5 hub genes were involved in gastric carcinogenesis,staging,typing and prognosis,and their mutations promote gastric cancer ***,molecular docking results confirmed that the components of YGS can effectively bind to therapeutic *** YGS has the effect of anti-gastric cancer and immune regulation.

关键词： Gastric cancer Yigong san Mechanism Immune

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Development of a nomogram for predicting liver transplantation prognosis in hepatocellular carcinoma

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World Journal of Gastroenterology 2024年第21期30卷 2763-2776页

作者： Li He Wan-Sheng Ji Hai-Long Jin Wen-Jing lu Yuan-Yuan Zhang hua-Guang Wang Yu-Yu Liu Shuang Qiu Meng Xu Zi-Peng Lei Qian Zheng Xiao-Li Yang Qing Zhang Department of Organ Transplantation The Third Medical Centre of Chinese PLA General HospitalBeijing 100039China School of Clinical Medicine Shandong Second Medical UniversityWeifang 261053Shandong ProvinceChina Clinical Research Center The Affiliated Hospital of Shandong Second Medical UniversityWeifang 261053Shandong ProvinceChina Department of Laboratory Medicine The Third Medical Centre of Chinese PLA General HospitalBeijing 100039China Physiatry Department Naval Aviation UniversityYantai 100071Shandong ProvinceChina

BACKGROUND At present,liver transplantation(LT)is one of the best treatments for hepatocellular carcinoma(HCC).Accurately predicting the survival status after LT can significantly improve the survival rate after LT,and ensure the best way to make rational use of liver *** To develop a model for predicting prognosis after LT in patients with *** Clinical data and follow-up information of 160 patients with HCC who underwent LT were collected and *** expression levels of alphafetoprotein(AFP),des-gamma-carboxy prothrombin,Golgi protein 73,cytokeratin-18 epitopes M30 and M65 were measured using a fully automated chemiluminescence *** best cutoff value of biomarkers was determined using the Youden *** regression analysis was used to identify the independent risk factors.A forest model was constructed using the random forest *** evaluated the accuracy of the nomogram using the area under the curve,using the calibration curve to assess consistency.A decision curve analysis(DCA)was used to evaluate the clinical utility of the *** The total tumor diameter(TTD),vascular invasion(VI),AFP,and cytokeratin-18 epitopes M30(CK18-M30)were identified as important risk factors for outcome after *** nomogram had a higher predictive accuracy than the Milan,University of California,san Francisco,and Hangzhou *** calibration curve analyses indicated a good *** survival and recurrence-free survival(RFS)of high-risk groups were significantly lower than those of low-and middle-risk groups(Per clinical *** The study developed a predictive nomogram based on TTD,VI,AFP,and CK18-M30 that could accurately predict overall survival and RFS after *** can screen for patients with better postoperative prognosis,and improve longterm survival for LT patients.

关键词： Hepatocellular carcinoma Liver transplantation Liver cancer Nomogram Prognosis

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Aquaporin-4 deficiency attenuates acute lesions but aggravates delayed lesions and microgliosis after cryoinjury to mouse brain

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Neuroscience Bulletin 2012年第1期28卷 61-68页

作者： Wen-Zhen Shi Chun-Zhen Zhao Bing Zhao Xiao-Liang Zheng san-hua Fang Yun-Bi lu Wei-Ping Zhang Zhong Chen er-Qing Wei Department of Pharmacology and Institute of Neurobiology School of Medicine Zhejiang University Hangzhou 310058China

Objective To determine whether aquaporin-4 （AQP4） regulates acute lesions, delayed lesions, and the associated microglial activation after cryoinjury to the brain. Methods Brain cryoinjury was applied to AQP4 knockout （KO） and wild-type mice. At 24 h and on days 7 and 14 after cryoinjury, lesion volume, neuronal loss, and densities of microglia and astrocytes were determined, and their changes were compared between AQP4 KO and wild-type mice. Results Lesion volume and neuronal loss in AQP4 KO mice were milder at 24 h following cryoinjury, but worsened on days 7 and 14, compared to those in wild-type mice. Besides, microglial density increased more, and astrocyte proliferation and glial scar formation were attenuated on days 7 and 14 in AQP4 KO mice. Conclusion AQP4 deficiency ameliorates acute lesions, but worsens delayed lesions, perhaps due to the microgliosis in the late phase.

关键词： aquaporin-4 gene deficiency cryoinjury microglia astrocyte

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Aggravated inflammation and increased expression of cysteinyl leuko-triene receptors in the brain after focal cerebral ischemia in AQP4-deficient mice

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Neuroscience Bulletin 2012年第6期28卷 680-692页

作者： Wen-Zhen Shi Chun-Zhen Zhao Bing Zhao Qiao-Juan Shi Li-Hui Zhang Yan-Fang Wang san-hua Fang Yun-Bi lu Wei-Ping Zhang er-Qing Wei Institute of Neuroscience and Department of Pharmacology School of MedicineZhejiang University Hangzhou Key Laboratory of Neurobiology and Department of Pharmacology School of Basic MedicineHangzhou Normal University

Objective Aquaporin-4 （AQP4）, the main water channel protein in the brain, plays a critical role in water homeostasis and brain edema. Here, we investigated its role in the inflammatory responses after focal cerebral ischemia. Methods In AQP4-knockout （KO） and wild-type mice, focal cerebral ischemia was induced by 30 rain of middle cerebral arterial occlusion （MCAO）. Ischemic neuronal injury and cellular inflammatory responses, as well as the expression and localization of cysteinyl leukotriene CysLT2 and CysLT~ receptors, were determined at 24 and 72 h after MCAO. Results AQP4-KO mice showed more neuronal loss, more severe microglial activation and neutrophil infiltration, but less astrocyte proliferation in the brain after MCAO than wild-type mice. In addition, the protein levels of both CysLT1 and CysLT2 receptors were up-regulated in the ischemic brain, and the up-regulation was more pronounced in AQP4-KO mice. The CysLT1 and CysLT2 receptors were primarily localized in neurons, microglia and neutrophils; those localized in microglia and neutrophils were enhanced in AQP4-KO mice. Conclusion AQP4 may play an inhibitory role in postischemic inflammation.

关键词： aquaporin 4 gene deficiency inflammation cysteinyl leukotriene receptor microglia astrocyte focal cerebral ischemia

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Mutagenic and Estrogenic Effects of Organic Compounds in Water Treated by Different Processes: A Pilot Study

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Biomedical and Environmental Sciences 2015年第8期28卷 571-581页

作者： lu Yi LYU Xue Min XIAO san hua YANG Xiao Ming WANG Ya Zhou TANG Fei Institute of Environmental Medicine MOE Key Laboratory of Environmental and Health School of Public HealthTongji Medical College of Huazhong Science and Technology University

Objective In this study, a pilot-scale investigation was conducted to examine and compare the biotoxicity of the organic compounds in effluents from five treatment processes (P1-P5) where each process was combination of preoxidation (O3), coagulation, sedimentation, sand filtration, ozonation, granular activated carbon, biological activated carbon and chlorination (NaClO). Methods Organic compounds were extracted by XAD-2 resins and eluted with acetone and dichlormethane (DCM). The eluents were evaporated and redissolved with DMSO or DCM. The mutagenicity and estrogenicity of the extracts were assayed with the Ames test and yeast estrogen screen (YES assay), respectively. The organic compounds were detected by GC-MS. Results The results indicated that the mutation ratio (MR) of organic compounds in source water was higher than that for treated water. GC-MS showed that more than 48 organic compounds were identified in all samples and that treated water had significantly fewer types and concentrations of organic compounds than source water. Conclusion To different extents, all water treatment processes could reduce both the mutagenicity and estrogenicity, relative to source water. P2, P3, and P5 reduced mutagenicity more effectively, while P1 reduced estrogenicity, most effectively. Water treatment processes in this pilot plant had weak abilities to remove Di-n-butyl phthalate or 1, 2-Benzene dicarboxylic acid.

关键词： Water treatment processes Organic compounds Yeast estrogen screen(YES assay) Ames test GC-MS

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Caffeic acid ameliorates early and delayed brain injuries after focal cerebral ischemia in rats

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Acta Pharmacologica Sinica 2006年第9期27卷 1103-1110页

作者： Yu ZHOU san-hua FANG Yi-lu YE Li-sheng CHU Wei-ping ZHANG Meng-ling WANG er-qing WEI~2 Department of Pharmacology,School of Medicine,Zhejiang University,Hangzhou 310031,China Department of Pharmacology School of Medicine Zhejiang University Hangzhou China

Aim:To investigate the effects of caffeic acid on early and delayed injuries afterfocal cerebral ischemia in rats,and the possible relation to ***:Transient focal cerebral ischernia was induced by middlecerebral artery occlusion in Sprague-Dawley *** acid(10 and 50 mg/kg)was ip injected for 5 d after *** brain injuries were observed,and thelevels of cysteinyl leukotrienes and leukotriene B4in the brain tissue ***:Caffeic acid(50 mg/kg)ameliorated neurological dysfunctionand neuron loss,and decreased infarct volume 24 h after ischemia;it attenuatedbrain atrophy,infarct volume,and particularly astrocyte proliferation 14 d *** addition,it reduced the production of leukotrienes(5-lipoxygenasemetabolites)in the ischemic hemispheres 3 h and 7 d after ***:Caffeic acid has protective effect on both early and delayed injuries after focalcerebral ischemia in rats;and this effect may partly relate to 5-lipoxygenaseinhibition.

关键词： caffeic acid focal cerebral ischemia neuron astrocyte 5-lipoxygenase leukotriene

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