输注供者抗原特异性调节性T淋巴细胞延长小鼠胰岛移植物的存活时间

目的 研究体外扩增获得的供者抗原特异性调节性T淋巴细胞(Treg细胞)对小鼠同种胰岛移植物存活时间的影响.方法 建立从Balb/c小鼠到C57小鼠的胰岛移植模型,通过观察移植后小鼠血糖情况来判断移植胰岛的存活时间.受鼠制成糖尿病模型后分为3组.对照组:不给予任何处理,只观察血糖变化;单纯胰岛移植组:移植胰岛,但不输注Treg细胞;实验组:术前1 d静脉给予1×106个供者特异性Treg细胞,然后行胰岛移植.结果胰岛移植后,对照组小鼠血糖持续高于16.7 mmol/L;单纯胰岛移植组小鼠血糖于术后1～2 d全部降至正常,到7～11 d时陆续开始升高,并维持在术前水平,存活时间为(8.50±1.6)d;实验组小鼠血糖于术后2 d内降至正常,之后维持在较低水平,至第21天开始有小鼠血糖升高超过16.7 mmol/L,存活时间为(26.2±3.9)d,明显长于单纯胰岛移植组(P＜0.05).结论 输注供者抗原特异性Treg细胞可以明显延长移植胰岛的存活时间,在胰岛移植耐受中有积极的作用.

Abstract：

Objective To investigate the effects of donor-specific regulatory T cells (Treg) transfusion on islet allograft survival. Methods Allogeneic fresh islets from Balb/c mice were transplanted to streptozotocin-induced diabetic C57 mice. The survival of islet allografts was observed. The experiment was divided into 3 groups: control group, nothing had been done to the recipients; simple islet transplantation group, the recipients received the islet transplantation only; experimental group, the recipients were given 1 ×106 Treg, then received islet transplantation. Results Blood glucose (BG) was above 16. 7 mmol/L after islet transplantation in control group; In simple islet transplantation group,BG level returned to normal level 1 to 2 days after transplantation, and hyperglycemia appeared 7 to 11 days after transplantation and maintained as the same as that before transplantation; In experimental group, BG level returned to normal level 2 days after transplantation and maintained at a low level,and at the 21st day after transplantation BG level was over 16. 7mmol/L in some recipients. Islet allograft survival in experimental group was significantly prolonged as compared with simple islet transplantation group. Conclusion Donor-specific Treg transfusion could prolong the islet allograft survival,and maybe have positive effect on tolerance induction of islet transplantation.     

环孢素A转换为他克莫司对慢性移植肾肾病患者的治疗效果

目的 探讨由环孢素A(CsA)转换为他克莫司(Tac)为主的免疫抑制方案对慢性移植肾肾病(CAN)患者的治疗效果.方法 选择接受同种肾移植后发生CAN的患者153例,患者肾移植后均采用CsA、吗替麦考酚酯(MMF)及泼尼松(Pred)的免疫抑制方案.根据是否以Tac替换CsA将患者分为两组.(1)CsA组:45例,进入研究后患者维持原免疫抑制方案.(2)Tac组:108例,进入研究后将CsA转换为Tac,停用CsA后立即开始服用Tac,MMF和Pred的用法同CsA组.对所有患者随访12个月,观察人/移植肾存活率、急性排斥反应发生率、移植肾功能、24 h尿蛋白定量、移植肾穿刺病理学活检及免疫抑制剂的不良反应等指标.结果 随访12个月时,CsA组和Tac组患者存活率均为100%,移植肾存活率分别为86.6%和93.5%(P＜0.05);急性排斥反应发生率分别为4.4%(2/45)和3.7%(4/108)(P＞0.05),6例发生急性排斥反应的患者均经甲泼尼龙冲击治疗3 d后逆转.Tac组患者移植肾功能明显改善,并且出现重度蛋白尿、重度肾间质纤维化和肾小管萎缩的患者比例较CsA组显著减少(P＜0.05).Tac组有13.8%(15例)的患者出现轻度血糖增高,发生率显著高于CsA组的4.4%(2例)(P＜0.05);Tac组有22.2%(24例)的患者发生高血压,发生率显著低于CsA组的55.6%(25例)(P＜0.05);17例因使用CsA而出现牙龈增生和多毛症者,经转换治疗后,症状均明显好转.结论 由CsA转换为Tac为主的免疫抑制方案能够显著改善CAN患者的移植肾功能,延缓CAN的发展,转换过程中未发生严重Tac不良反应并且改善了使用CsA时出现的不良反应.

Abstract：

Objective To investigate the effect of conversion from cyclosporine A (CsA) to tacrolimus (Tac) on chronic allograft nephropathy (CAN). Methods 153 CAN patients undergoing kidney transplantation received CsA, mycophenolate mofetil (MMF) and prednisone (CsA-MMF-Pred) regimen after kidney transplantation, and divided into 2 groups according to whether CsA were maintained in the immunosuppressive regimen: CsA + MMF + Pred group (CsA group, n = 45); Tac + MMF + Pred group (Tac group, n = 108). The patients were followed up with patient/kidney survival rate, acute rejection incidence, renal function, 24-h proteinuria and adverse events of immunosuppressive drugs for 12 months. Results Compared with CsA group, the transplanted kidney survival rate was significantly higher in Tac group (93. 5 % vs 86.6 %, P＜0. 05). Acute rejection (AR) was diagnosed in 4. 4 % (2/45) of recipients in CsA group and 3. 7 % (4/108) in Tac group (P＞0. 05) respectively. Acute rejection (2 cases in CsA group and 4 in Tac group) was reversed by 500 mg of methylprednisolone for consecutive 3 days, and the patients in Tac group showed a significantly lower degree of interstitial fibrosis and tubular atrophy (IF/TA) (P＜0. 05).Renal allograft functions and 24-h proteinuria during a follow-up period of 12 months were significantly improved in Tac group (P ＜ 0. 05). Incidence of mild hyperglycemia in Tac Group (13.8 %, 15/108) was significantly higher than in CsA group (4.4 %, 2/45), and that of hypertension in Tac group (22. 2 %, 24/108) was significantly lower than in CsA group (55.6 %,25/45). CsA-related side effects (such as hirsutism and gingival hypertrophy) in 17 patients were greatly improved after conversion from CsA to Tac treatment. Conclusion The conversion from CsA to Tac on the patients with CAN can improve renal allograft function, retard the progression of renal allograft dysfunction, reduce the incidence of CsA-related side effects and not generate serious adverse effects of Tac.     

NOD小鼠未成熟树突状细胞体外诱导淋巴细胞生成并扩增调节性T细胞的研究

目的 探索非肥胖糖尿病(non-obese diabetic,NOD)小鼠未成熟树突状细胞(immature dendritic cell,imDC)诱导并扩增调节性T细胞(regulatory T cells,Treg)的优化方案.方法 分离、培养NOD小鼠骨髓来源的imDC和淋巴细胞,并进行混合淋巴细胞培养,分别加...     

中医治疗颈椎综合征临床疗效分析

目的观察中药内服与中药内服加局部外敷治疗颈椎综合征的疗效。方法随机将94例颈椎综合征分为观察组63例和对照组31例,观察组采用中药内服加局部外敷治疗,对照组单用中药内服治疗。两组均10d为1个疗程,连续治疗2个疗程,比较两组疗效。结果两组治疗都有良好的疗效,但观察组治愈率和总有效率均高于对照组,两组比较差异有统计学意义（P〈0.01）。结论中药内服加局部外敷治疗颈椎综合征,疗效优于单用内服中药治疗。     

小鼠胰岛的分离与纯化

目的 探讨稳定、高效的小鼠胰岛细胞的分离纯化方法.方法 采用胆总管内灌注不同浓度胶原酶(分别为0.5、1.0、1.5 g/L)消化胰腺的方法分离BALB/C小鼠胰岛,Ficoll-400不连续密度梯度离心法纯化胰岛.双硫腙(DTZ)对胰岛进行特异性染色计算胰岛产量及纯度,葡萄糖刺激释放试验体外测胰岛功能.结果 不同浓度的胶原酶在不同时间内消化胰腺后收获的胰岛数量有较大的差异,其中采用0.5 g/L胶原酶V、38 ℃水浴消化20 min组收获量最大为(230±20)个胰岛细胞团,纯度约为90%.DTZ染色后胰岛呈腥红色,形态完整.葡萄糖刺激释放实验示高糖刺激后胰岛素释放量为低糖刺激后的2.3倍.结论 胶原酶浓度、消化时间和温度是影响小鼠胰岛分离结果的重要因素,当胶原酶浓度为0.5 g/L,消化时间20 min时可获得数量较多,纯度较好的胰岛细胞.

Abstract：

Objective To investigate the stable and efficient method of isolation and purification from mice pancreas. Methods BALB/C mouse islets were isolated by different concentrations of collagenase digestion (0. 5, 1. 0, 1. 5 g/L respectively) and purificated by Ficoll density gradient centrifugation.The number, purity and vitality of the islets were analyzed. The production and purity of the islets were checked by Dithizone immunofluorescence staining. The glucose-induced insulin secretion was detected by enzyme linked immunosorbent assay (ELISA) for islet function in vitro. Results Different number of islets was obtained by mice pancreas digestion with different concentrations of collagenase and for different digestion durations.After the mouse pancreata were digested in 38 C and with 0. 5 g/L collagenase V for 20rmin, maximum number of islets was obtained, and the purity of the final preparation was ＞ 95%. After culture in vitro, insulin release of islets under high glucose stimulation was 2. 3 times of that under low glucose stimulation. Conclusion Concentration of collagenase, temperature, and digestion duration were important factors of islet isolation and purification from mice pancreas. More production and higher purity of islets were obtained under the concentration of 0. 5 g/L collagenase Ⅴ for 20 min.     

三氧化二砷对裸鼠人乳腺癌移植瘤的作用及机制

目的探讨三氧化二砷（As2O3）对乳腺癌移植瘤的作用及其机制。方法建立BALB／C—nu／nu裸鼠ER阴性MDA—MB-435s人乳腺癌移植瘤模型,分别以不同浓度的As2O3、顺铂和生理盐水（NS）腹腔内注射,观察移植瘤的瘤重及抑瘤率。并用免疫组化检测乳腺癌石蜡标本中PTEN和Caspase-7的表达。结果As2O3高剂量组、As2O3低剂量组、顺铂组均能明显抑制裸鼠皮下肿瘤的生长,抑瘤率分别为48．68％、32．80％、66．67％。用药后移植瘤组织中PTEN和Caspase-7蛋白阳性细胞数显著增多（P〈0．05）。结论As2O3能明显抑制乳腺癌移植瘤的生长,其作用机制可能是通过上调PTEN和Caspase-7的表达,促进细胞的凋亡来抑制人乳腺癌细胞的生长。     

肾移植术后微小病毒B19复发感染诊疗分析

目的 总结肾移植术后受者感染微小病毒B19(human parvovirus B19,HPV-B19)且多次复发的诊断方法、临床表现及防治措施。方法 2020年6月至2022年5月于华中科技大学同济医学院附属同济医院器官移植中心行肾移植术后感染HPV-B19的37例受者中有8例复发,本研究对这8例受者的临床表现、诊疗措施、实验室检查及预后进行回顾性分析和总结。结果 8例受者经过多疗程静脉注射丙种球蛋白（intravenous immunoglobulin G,IVIG）、转换免疫抑制剂及降低免疫抑制强度等综合治疗后,贫血症状改善,血红蛋白（hemoglobin,Hb）水平稳定,并在复发时使用同样治疗措施有效。8例受者最后1次复发后继续随访1年,8例受者再无复发且Hb水平稳定[（136.8±12.0）g/L],整个感染期间肾功能稳定。结论 肾移植术后受者因免疫力低下,易感染HPV-B19且容易复发。当受者感染HPV-B19后,应定期监测Hb及网织红细胞水平,尤其是治疗好转的受者一旦再次出现Hb进行性下降应立即引起重视,进一步通过聚合酶链反应（polymerase chain reacti...     

胰腺切除后自体胰岛移植治疗慢性胰腺炎

慢性胰腺炎的处理一直是外科难题之一,八十年代后国外开始尝试一种新的治疗方法,即用胰腺全切或次全切来控制患者的疼痛,同时从切下的胰腺中分离出胰岛并行自体移植以防止糖尿病的发生,其疗效在部分患者已得到肯定。本文介绍了这一方法的技术及效果,指出若能改进胰岛分离方法,提高胰岛收获量甚至移植同种或异种胰岛,有可能使这一疗法更为完善,效果更好。     

胆管腔内超声联合胆汁肿瘤标志物对胆管狭窄病因的诊断价值探讨

目的探讨胆管腔内超声（IDUS）联合胆汁肿瘤标志物测定对胆管狭窄良恶性的鉴别诊断价值。方法57例胆管狭窄患者（良性狭窄8例,恶性狭窄49例）行胆管腔内超声检查,同时行血清及胆汁肿瘤标志物[CA19-9、癌胚抗原（CEA）]测定,以手术病理结果为金标准,统计分析腹部超声、CT、磁共振胰胆管成像术（MRCP）、IDUS以及IDUS联合胆汁肿瘤标记物鉴别诊断胆管狭窄良恶性的敏感度、特异度、阳性预测值、阴性预测值及准确度。结果IDUS和IDUS联合胆汁肿瘤标记物（胆管良恶性鉴别诊断的分界值CA19-9值和CEA值分别为107kU／L和66．71斗∥L）鉴别诊断胆管狭窄良恶性的特异度分别为63．6％（7／11）和77．8％（7／9）（P〉0．05）,阳性预测值分别为91．8％（45／49）和95．9％（47／49）（P〉0．05）,准确度分别为91．2％（52／57）和94．7％（54／57）（P〉0．05）,均显著高于腹部超声、CT和MRCP,差异均有统计学意义（P〈0．05）。IDUS联合胆汁CEA（远端胆管狭窄良恶性鉴别诊断的分界值为71μg/L）鉴别诊断远端胆管狭窄良恶性的准确度为97．9％（46／47）,明显高于IDUS的87．2％（41／47）,差异有统计学意义（P〈0．05）。结论IDUS联合胆汁肿瘤标记物测定对胆管良恶性狭窄性质的鉴别有较高的价值,联合胆汁CEA测定能够在IDUS基础上进一步提高远端胆管恶性狭窄诊断的准确度。     

去分化脂肪肉瘤预后风险模型的建立与分析

目的 基于美国国家癌症研究所监测、流行病学和最终结果(SEER)数据库，构建预后列线图用于预测去分化脂肪肉瘤(DDLPS)3、5和8 a的总生存(OS)率。方法 从SEER数据库中提取DDLPS患者的临床信息，随机分为建模组和验证组。通过LASSO回归和多因素Cox回归筛选变量，将筛选出的变量纳入预测模型并构建列线图。使用一致性指数(C指数)、时间依赖曲线下面积(时间依赖AUC)和校准曲线评估模型的区分度和校准度。通过决策曲线分析(DCA)、净重新分类指数(NRI)、综合判别改善指数(IDI)进一步比较列线图和AJCC分期之间的净收益和预测准确性。根据列线图计算总得分，对患者进行风险分层，通过Kaplan-Meier曲线和log-rank检验比较分层间OS率差异。结果 共纳入1 172例DDLPS患者，筛选出年龄、原发位置、T分期、N分期、M分期、FNCLCC分级、手术和放疗8个预后变量。时间依赖AUC(>0.7)和C指数(建模组为0.741,验证组为0.764)表明列线图有良好的区分度。校准曲线表明预测生存概率和实际生存率具有良好的一致性。IDI和NRI表明，列线图的准确性优于...