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Small intestinal submucosa improves islet survival and function during in vitro culture

World Journal of Gastroenterology 2005年第46期11卷 7378-7383页

作者： xiao-hui tian wu-jun xue xiao-ming ding xin-lu pang yan teng pu-xun tian xin-shun feng Center of Renal Transplantation The First Hospital of Xi'an Jiaotong UniversityXi'an 710061 Shaanxi Province China

AIM： To evaluate the recovery and function of isolated rat pancreatic islets during in vitro culture with small intestinal submucosa （SIS）. METHODS： Pancreatic islets were isolated from Wistar rats by standard surgical procurement followed by intraductal collagenase distension, mechanical dissociation and Euroficoll purification. purified islets were cultured in plates coated with multilayer SIS （SIS-treated group） or without multilayer SIS （standard cultured group） for 7 and 14 d in standard islet culture media of RPMI 1640. After isolation and culture, islets from both experimental groups were stained with dithizone and counted. Recovery of islets was determined by the ratio of counts after the culture to the yield of islets immediately following islet isolation. Viability of islets after the culture was assessed by the glucose challenge best with low （2.7 mmol/L） and high glucose （16.7 mmol/L） solution supplemented with 50 mmol/L 3-isobutyl-1- methylxanthine （IBMX） solution. Apoptosis of islet cells after the culture was measured by relative quantification of histone-complexed DNA fragments using ELISA. RESULTS： After 7 or 14 d of in vitro tissue culture, the recovery of islets in SIS-treated group was significantly higher than that cultured in plates without SIS coating. The recovery of islets in SIS-treated group was about twice more than that of in the control group. In SIS treated group, there was no significant difference in the recovery of islets between short- and long-term periods of culture （95.8±1.0% vs 90.8±1.5%, P〉0.05）. When incubated with high glucose （16.7 mmol/L） solution, insulin secretion in SIS-treated group showed a higher increase than that in control group after 14 d of culture （20.7±1.1 mU/L vs 11.8±1.1 mU/L, P〈0.05）. When islets were placed in high glucose solution containing IBMX, stimulated insulin secretion was higher in SlS-treated group than in control group. Calculated stimulation index of SlS-treated group was about 23 times of

关键词： Islet culture Islet survival Islet function Small intestinal submucosa

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Allograft rejection-related gene expression in the endothelial cells of renal transplantation recipients after cytomegalovirus infection

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第11期10卷 820-828页

作者： yang LI Hang yan wu-jun xue pu-xun tian xiao-ming ding xiao-ming PAN xin-shun feng xiao-hui tian He-li XIANG jun HOU Department of Renal Transplantation Center of Nephropathy the First Affiliated Hospital School of Medicine Xi 'an Jiaotong University Xi 'an 710061 China The two authors contributed equally to this work

Objective： To explore the effects of cytomegalovirus （CMV） infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods： Endothelial cells （ECs） were cultured and stimulated by a variety of factors： A, normal control group; B, inactivated human cytomegalovirus （HCMV） infection group; C, HCMV infection group; D, HCMV supematant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 （ICAM-1） and major histocompability complex （MHC） class I and class II antigens was detected by flow cytometry （FCM） and immuno- histochemistry. Results： We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D （P〉0.05）. Although the expression levels of ICAM-1 were not significantly different between groups C and E （P〉0.05）, the ICAM-1 expression in these two groups was significantly higher than that in group A （P〈0.05）. ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E （P〉0.05）. Human leucocyte antigen （HLA）-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant dif- ferences of HLA-ABC and HLA-DR expression among groups A, B and D （P〉0.05）. However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported （P〈0.05）. Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C （P〈0.05）. Conclusion： CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.

关键词： Renal transplantation Cytomegalovirus (CMV) Rejection Intercellular adhesion molecule-1 (ICAM-1) Major histocompability complex (MHC)

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Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro

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Chinese Medical Journal 2005年第22期118卷 1857-1862页

作者： teng yan xue wu-jun ding xiao-ming feng xin-shun XIANG He-li JIANG Ya-zhuo tian pu-xun Department of Kidney Transplantation First Affiliated Hospital of Xi＇an Jiaotong University Xi＇an 710061 China

Background Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered ＇ nurse cells＇ in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro. Methods Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets＇ function when they were co-cultured for 21 days in vitro. Results In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95 % and they remained highly cytoactive for a long time in vitro （P〉0. 05 ）. Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment （P〈0.01 ）. Conclusions A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets＇ function when they were co-cultured in vitro. They could be a helper cell in islet transplanta

关键词： adult Sertoli cells trophic support allogeneic islet transplantation

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Polyglycolic Acid Fibrous Scaffold Improving Endothelial Cell Coating and Vascularization of Islet

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Chinese Medical Journal 2017年第7期130卷 832-839页

作者： yang Li Ping Fan xiao-ming ding xiao-hui tian xin-shun feng Hang yan xiao-ming Pan pu-xun tian Jin Zheng Chen-Guang ding wu-jun xue Department of Renal Transplantation Center of Nephrology The First Affiliated Hospital of Xi'an Jiaotong University School of Medicine Xi'an Shaanxi 710061 China Department of Rheumatism and Immunology The First Affiliated Hospital of Xi'an Jiaotong University School of Medicine Xi'an Shaanxi 710061 China

Background：Improving islet graft revascularization has become a crucial task for prolonging islet graft *** cells （ECs） are the basis of new microvessels in an isolated islet,and EC coating has been demonstrated to improve the vascularization and survival of an ***,the traditional method of EC coating of islets has low efficiency in *** study was conducted to evaluate the effect of a polyglycolic acid （PGA） scaffold on the efficiency of islet coating by ECs and the angiogenesis in the coated islet ***：A PGA fibrous scaffold was used for EC coating of islet culture and was evaluated for its efficiency of EC coating on islets and islet graft ***：In in vitro experiments,we found that apoptosis index of ECs-coating islet in PGA group （27% ± 8%） was significantly lower than that in control group （83% ± 20%,P 〈 0.05） after 7 days *** index was significantly greater in the PGA group than in the control group at day 7 after ECs-coating （2.07 ± 0.31 vs.1.80 ± 0.23,P 〈 0.05）.vascular endothelial growth factor （VEGF） level in the PGA group was significantly higher than the coating in the control group after 7 days culture （52.10 ± 13.50 ng/ml vs.16.30 ± 8.10 ng/ml,P 〈 0.05）.Because of a tight,circumvallated,adhesive and three-dimensional growth microenvironment,islet cultured in a PGA scaffold had higher coating efficiency showing stronger staining intensity of enzyme than those in the control group after 14 days of culture following *** in vivo study,PGA scaffold significantly prolonged the average survival time of EC-coated islet graft after transplantation compared with control group （15.30 ± 5.60 days vs.8.30 ± 2.45 days,P 〈 0.05）.The angiogenesis and area of survived grafts were more in the PGA group compared with the control group by measuring the mean microvessel density （8.60 ± 1.21/mm2 vs.5.20 ± 0.87/mm2,P 〈 0.05）.In addition,expression of VEGF and tyrosin-protein kinase receptor

关键词： Endothelial Cells Islet Polyglycolic Acid Fibrous Revascularization

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The clinical value of enzyme-multiplied immunoassay technique monitoring the plasma concentrations of cyclosporine A after renal transplantation

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Journal of Pharmaceutical Analysis 2011年第2期1卷 139-142页

作者： xiao-hui luo wu-jun xue pu-xun tian xiao-ming ding Hang yan He-Li Xiang yang Li Department of Renal Transplant Center of Nephropathy the First Affiliated Hospital Medical School of Xi'an Jiaotong University Xi'an 710061 China.

The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows：〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%）（P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%）（P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.

关键词： enzyme-multiplied immunoassay technique renal transplantation cyclosporin A blood concentration monitoring

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Application of next generation sequencing in the screening of monogenic diseases in China,2021:a consensus among Chinese newborn screening experts

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World Journal of Pediatrics 2022年第4期18卷 235-242页

作者： Fan Tong Jian Wang Rui xiao Bing-Bing wu Chao-Chun Zou ding-Wen wu Hua Wang hui Zou Lian-Shu Han Lin yang Lin Zou ming-yan Hei Ru-Lai yang tian-ming Yuan Wei Wen xin-Wen Huang xue-Fan Gu yan-Ling yang Yong-Lan Huang Yong-jun Zhang Yong-Guo Yu Zheng-feng Xu Wen-Hao Zhou Zheng-yan Zhao Children’s Hospital Zhejiang University School of MedicineNational Clinical Research Center for Child Health3333 Binsheng RoadBinjiang DistrictHangzhou 310052China Shanghai Children’s Medical Center Shanghai Jiaotong University School of MedicineShanghaiChina National Engineering Laboratory for Key Technology of Birth Defect Control and Prevention Screening and Diagnostic R and D CenterHangzhouChina Center for Molecular Medicine Children’s Hospital of Fudan University399 Wan Yuan RoadMin Xing DistrictShanghai 200000China National Health Commission Key Laboratory for Birth Defect Research and Prevention Maternal and Child Health Hospital of Hunan ProvinceChangshaChina Jinan Maternity and Child Care Hospital Affiliated To Shandong First Medical University JinanChina xinhua Hospital Shanghai Institute for Pediatric ResearchShanghai Jiao Tong University School of MedicineShanghaiChina Clinical Genetic Center Children’s Hospital of Fudan UniversityShanghaiChina Children’s Hospital Shanghai Jiaotong UniversityShanghaiChina Neonatal Center Beijing Children’s HospitalCapital Medical UniversityBeijingChina Affiliated Shenzhen Maternity and Child Healthcare Hospital Southern Medical UniversityShenzhenChina xinhua Hospital School of MedicineShanghai Jiaotong UniversityShanghaiChina Peking University First Hospital BeijingChina Guangzhou Women and Children’s Medical Center GuangzhouChina Center of Genetic Medicine Women’s Hospital of Nanjing Medical UniversityNanjing Maternity and Child Health Care HospitalNanjingChina

Newborn screening(NBS)refers to a maternal and newborn healthcare technology,in which special examinations of congenital and genetic diseases that could seriously impact the health of newborns,are implemented during the neonatal period to provide early diagnosis and treatment[1].With a history of more than 60 years,NBS has advanced greatly due to technological progress resulting in significant improvement in the number of diseases covered by NBS and in screening efficiency[2-7].

关键词： diseases diagnosis congenital

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Exploring an Integrative Therapy for Treating COVID-19:A Randomized Controlled Trial

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Chinese Journal of Integrative Medicine 2020年第9期26卷 648-655页

作者： WANG Jia-bo WANG Zhong-xia JING Jing ZHAO Peng DONG Jing-hui ZHOU Yong-feng yanG Guang NIU ming ZHAO Xu JIANG tian-jun Bl Jing-feng XU Zhe ZHANG Ping wu Dan BAI Zhao-fang GUO Yu-ming YU Si-miao SUN Yong-qiang ZHANG Zi-teng ZHAN xiao-yan LI Peng-yan ding Jin-biao ZHAO Peng-fei SONG xue-ai TANG Jian-yuan HE Dong-chu CHEN Zhu QIN En-qiang WANG Rui-lin xiao xiao-he China Military Institute of Chinese Medicine The Fifth Medical Center of Chinese PLA General HospitalBeijing(100039)China Integrative Medical Center The Fifth Medical Center of Chinese PLA General HospitalBeijing(100039)China Treatment and Research Center for Infectious Diseases The Fifth Medical Center of Chinese PLA General HospitalBeijing(100039)China National Clinical Research Center for Infectious Diseases Beijing(100039)China Department of Radiology The Fifth Medical Center of Chinese PLA General HospitalBeijing(100039)China Research Center for Clinical and Translational Medicine The Fifth Medical Center of Chinese PLA General HospitalBeijing(100039)China Sichuan Evidence-Based Medicine Center of Traditional Chinese Medicine Affliated Hospital Chengdu University of Traditional Chinese MedicineChengdu(610075)China College of Life Science Beijing University of Chinese MedicineBeijing(100029)China Department of Integrated Treatment General Hospital of Central Theater CommandWuhan(430070)China

Objectives:To develop a new Chinese medicine(CM)-based drug and to evaluate its safety and effect for suppressing acute respiratory distress syndrome(ARDS)in COVID-19 ***:A putative ARDS-suppressing drug Keguan-1 was first developed and then evaluated by a randomized,controlled two-arm *** two arms of the trial consist of a control therapy(alpha interferon inhalation,50(x g twice daily;and lopinavir/ritonavir,400 and 100 mg twice daily,respectively)and a testing therapy(control therapy plus Keguan-119.4 g twice daily)by random number table at 1:1 ratio with 24 cases each *** 2-week treatment,adverse events,time to fever resolution,ARDS development,and lung injury on newly diagnosed COVID-19 patients were ***:An analysis of the data from the first 30 participants showed that the control arm and the testing arm did not exhibit any significant differences in terms of adverse *** on this result,the study was expanded to include a total of 48 participants(24 cases each arm).The results show that compared with the control arm,the testing arm exhibited a significant improvement in time to fever resolution(P=0.035),and a significant reduction in the development of ARDS(P=0.048).Conclusions:Keguan-1-based integrative therapy was safe and superior to the standard therapy in suppressing the development of ARDS in COVID-19 patients.(Trial registration *** 04251871 at ***).

关键词： COVID-19 SARS-CoV-2 acute respiratory distress syndrome Chinese medicine

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重组腺病毒介导hCGPx转染致血管内皮细胞产生抗氧化损伤作用的研究

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南方医科大学学报 2006年第10期26卷 1417-1420页

作者：项和立薛武军侯军田普训滕琰潘晓鸣丁小明冯新顺西安交通大学医学院第一附属医院肾移植科陕西西安710061

目的研究重组腺病毒介导的人胞浆型谷胱甘肽过氧化物酶(hCGPx)转染对血管内皮细胞ECV304氧化损伤保护作用。方法将含hCGPxcDNA的质粒pGEM-T-hCGPx和重组腺病毒载体pACCMV-pLpA穿梭质粒进行基因重组,构建成pACCMV-hCGPx穿梭质粒后,与包... 详细信息

目的研究重组腺病毒介导的人胞浆型谷胱甘肽过氧化物酶(hCGPx)转染对血管内皮细胞ECV304氧化损伤保护作用。方法将含hCGPxcDNA的质粒pGEM-T-hCGPx和重组腺病毒载体pACCMV-pLpA穿梭质粒进行基因重组,构建成pACCMV-hCGPx穿梭质粒后,与包装质粒pJM17共转染293细胞,构建成重组腺病毒AdCMV-hCGPx。用AdCMV-hCGPx转染体外培养的ECV304细胞并分为转染24、48和72h组,以转染空载体的细胞为对照组,检测转染细胞的基因表达水平。各组ECV304细胞经H2O2氧化损伤处理后,分别对细胞的活力和凋亡进行检测分析。结果各转染组细胞转染基因表达率均显著高于对照组(P<0.01)。经H2O2氧化损伤处理后,AdCMV-hCGPx转染组细胞活力较对照组明显增强,凋亡受到抑制。结论重组腺病毒介导的hCGPx转染可保护ECV304细胞抵抗氧化损伤,具有明确的细胞保护作用,其具体保护机制可能与抗氧化和抑制细胞凋亡有关。

关键词：人胞浆型谷胱甘肽过氧化物酶基因转染血管内皮细胞氧化损伤

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重组hCGPx腺病毒对人肾小管上皮细胞(HK-2)缺氧再复氧损伤的保护作用

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细胞与分子免疫学杂志 2006年第4期22卷 472-474页

作者：项和立薛武军侯军田普训滕琰潘晓鸣丁小明西安交通大学医学院第一附属医院肾移植科陕西西安710061

目的:研究重组腺病毒介导的人胞质型谷胱甘肽过氧化物酶(hCGPx)转染对人肾小管上皮细胞缺氧再复氧损伤的保护作用。方法:将含hCGPXcDNA的质粒pGEM-T-hCG-Px和重组腺病毒穿梭质粒pACCMV-pLpA重组,构建pACC-MV-hCGPx穿梭质粒后,与包装质粒... 详细信息

目的:研究重组腺病毒介导的人胞质型谷胱甘肽过氧化物酶(hCGPx)转染对人肾小管上皮细胞缺氧再复氧损伤的保护作用。方法:将含hCGPXcDNA的质粒pGEM-T-hCG-Px和重组腺病毒穿梭质粒pACCMV-pLpA重组,构建pACC-MV-hCGPx穿梭质粒后,与包装质粒pJM17共转染293细胞,构建重组腺病毒AdCMV-hCGPx。以重组腺病毒载体AdCMV-hCGPx转染体外培养的人肾小管上皮细胞株HK-2,以转染空载体的HK-2细胞为对照组,检测转染细胞中CGPx的表达水平。将HK-2细胞经缺氧再复氧损伤处理后,分别检测细胞的存活率、凋亡率及死亡率。结果:各转染组细胞中CGPx的表达率显著高于对照组(P<0.01)。经缺氧再复氧损伤处理后,AdCMV-hCGPx转染组细胞的存活率较对照组明显增强,死亡细胞明显减少,细胞凋亡明显受到抑制。结论:重组腺病毒介导的hCGPx转染人肾小管上皮细胞可保护缺氧再复氧引起的损伤。

关键词：人胞质型谷胱甘肽过氧化物酶转染肾小管上皮细胞缺氧再复氧损伤

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肾移植后不同免疫抑制方案的效果及不良反应的临床分析

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中华器官移植杂志 2011年第4期32卷 201-204页

作者：田普训薛武军丁小明潘晓鸣燕航冯新顺侯军项和立田晓辉西安交通大学医学院第一附属医院肾脏病医院肾移植科西安交通大学器官移植研究所 710061

目的总结。肾移植后使用不同免疫抑制方案的效果和不良反应，以提高人／肾的长期存活率。方法对单中心3102例肾移植受者的临床资料进行回顾性分析，所采用的免疫抑制方案有环孢素A（CsA）＋硫唑嘌呤（Aza）＋泼尼松（Pred）、低剂量CsA... 详细信息

目的总结。肾移植后使用不同免疫抑制方案的效果和不良反应，以提高人／肾的长期存活率。方法对单中心3102例肾移植受者的临床资料进行回顾性分析，所采用的免疫抑制方案有环孢素A（CsA）＋硫唑嘌呤（Aza）＋泼尼松（Pred）、低剂量CsA＋吗替麦考酚酯（MMF）＋Pred、低剂量他克莫司（Tac）＋MMF＋Pred、低剂量CsA（或Tac）＋西罗莫司（SRL）＋Pred等方案，分析各方案的效果和不良反应。结果低剂量CsA＋MMF＋Pred方案的人／肾1、5、10年存活率均高于CsA＋AZa＋Pred方案，而高血压、震颤、高尿酸、肝。肾毒性、白细胞下降等的发生率显著低于CsA＋Aza＋Pred方案（P〈0．05），腹泻发生率显著高于CsA＋Aza＋Pred方案（P〈0．05）。低剂量Tac＋MMF＋Pred方案的高血糖发生率显著高于低剂量CsA＋MMF＋Pred方案（P〈0．05），多毛症发生率显著低于低剂量CsA＋MMF＋Pred方案（P〈0．05）；低剂量CsA（或Tac）＋SRL＋Pred方案的腹泻、高尿酸血症、肝肾毒性和多毛症等的发生率显著低于低剂量CsA（或Tac）＋MMF＋Pred方案（P〈0．05），但高血脂发生率显著高于后者（P〈0．05）。以低剂量Tac为基础的方案者高血糖发生率显著应用低剂量CsA者。结论低剂量CsA（或Tac）＋MMF＋Pred方案改善了肾移植受者和移植肾的存活，降低了不良反应发生率，尤以低剂量Tac＋MMF＋Pred方案为优；调整免疫抑制方案或剂量，改善饮食习惯，加强锻炼，优化降血压、降血脂、控制血糖的治疗措施对预防和控制不良反应尤为重要。

关键词：肾移植免疫抑制剂不良反应回顾性研究

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