A型肉毒毒素多表位串联体的设计、表达、纯化及鉴定

目的:设计、表达两个A型肉毒毒素多表位串联体.方法:从文献报道的A型肉毒毒素(botulinum neurotoxin type A,BoNT/A)的表位及生物信息学方法预测得到的B细胞表位中遴选表位,并加入适当的辅助性元件,设计多表位串联体A、B.对其基因进行优化后,经重叠PCR方法合成串联体A、B的全长基因.分别插入原核表达载体pQE-30,再转化E.coli M15[pREP4]感受态细胞中进行诱导表达,以金属螯合亲和层析法纯化重组蛋白,并进行鉴定.结果与结论:成功设计并构建了两个多表位串联体A、B,并在E.coli中以包涵体形式获得高效表达.Ni-NTA法纯化后分别获得纯度大于92.2%、99%的重组串联体A、B蛋白,并经透析复性法复性.Western印迹和间接ELISA显示纯化、复性后的重组蛋白与抗天然BoNT/A马血清有特异性结合反应.     

中央型小细胞肺癌的CT诊断及鉴别诊断

目的:探讨中央型小细胞肺癌(Small Cell Lung Cancer,SCLC)的CT表现特点,并找出与非小细胞肺癌(NSCLC)、肺结核、结节病、纵隔淋巴瘤的鉴别诊断要点。方法:回顾性分析经病理证实的57例中央型SCLC、60例NSCLC,5例肺结核,2例结节病,3例纵隔淋巴病的胸部平片及CT资料,全部病例均行胸部平片和CT平扫、增强检查。结果:57例中央型SCLC,CT主要表现为沿支气管长轴生长的肺内肿块,肺门与纵隔广泛淋巴结肿大,而肺不张相对少见。NSCLC60例主要表现为肺门肿块,支气管阻塞性改变相对多见。5例肺结核表现为肺内结节和肺门、纵隔淋巴结肿大,伴有同侧和/或对侧肺内结核病灶。2例结节病,表现为双侧肺门对称性淋巴结肿大。3例纵隔淋巴病表现为中前纵隔的弥漫性软组织肿块。结论:沿支气管长轴生长的肺内肿块、肺门及纵隔淋巴结肿大。支气管阻塞改变少见,随访1～2个月,肿块体积增大1倍以上,是中央型SCLC的CT诊断要点,据此特征诊断正确率达80％。结合临床与其他检查方法可与NSCLC、肺结核、结节病、纵隔淋巴瘤鉴别。     

氟对体外培养人牙乳头间充质细胞合成I型胶原的影响

目的:明确氟对人牙乳头细胞合成 I型胶原的影响。 方法:对人牙乳头细胞进行体外培养 ,分别给予0 .0、0 .2× 10 - 6、1.0× 10 - 6、5 .0× 10 - 6、2 5 .0× 10 - 6 g/ L 氟处理 ,采用免疫细胞化学方法染色 ,图象分析氟对细胞合成 I型胶原的影响。 结果:I型胶原在对照组和 4种氟浓度处理组细胞表达均为阳性。图象分析结果表明 ,4种氟浓度对体外培养的人牙乳头细胞合成 I型胶原量与对照组相比差异无统计学意义 (P>0 .0 5 )。结论:过量氟对牙本质基质 型胶原的影响可能不是通过影响成牙本质细胞内合成实现的     

糖尿病脑内胰岛移植临床初步研究(附三例报告)

本文报告胰岛细胞脑内移植治疗I型糖尿病共3例,分别经过3个月、4个月、6个月的临床观察,其临床症状明显改善或消失,空腹血糖由移植前平均12.38mmol/L下降至7.77mmol/L;3例患者普通胰岛素用量平均每日62.3~u,移植后第21～30天均完全停用胰岛素,其中1例已持续撤离胰岛素治疗达5个月。观察结果提示:胰岛移植物在患者的脑内成活,并具有良好的内分泌功能。     

改良《颈性眩晕症状与功能评估量表》在椎动脉型颈椎病中医临床中的应用及评价

目的探讨改良《颈性眩晕症状与功能评估量表》在椎动脉型颈椎病(CSA)的中医临床中的应用可行性。方法151例CSA患者随机分为针灸推拿治疗组和单针灸对照组,治疗前后采用改良《颈性眩晕症状与功能评估量表》进行量化评估,并对该量表在本次中医临床中的效度及信度进行初步检验。结果治疗组治疗后症状与功能评分比治疗前有显著提高(p<0.01),治疗组改善指数明显高于对照组(p<0.05)。经检验,该量表也有可以接受的准则效度(r在0.8-0.9之间,p<0.01)和再测信度(r在0.6-0.9之间,p<0.01)。结论针灸推拿结合治疗CSA具有确切的临床疗效,能明显改善患者症状、颈椎功能和生活质量。改良《颈性眩晕症状与功能评估量表》在CSA中医临床中具有可信性、有效性。     

二甲双胍和罗格列酮对肥胖型PCOS治疗作用的对照性研究

目的观察罗格列酮和二甲双胍联合氯米芬治疗肥胖型多囊卵巢综合征患者的内分泌改善及生殖功能的临床疗效。方法50例肥胖型PCOS患者分别给予罗格列酮4mg/d和二甲双胍1500mg/d联合氯米芬100mg/d,治疗3个月,比较治疗前后体重指数、内分泌参数、腰臀比和Hom a IR的变化。结果用罗格列酮治疗后排卵率为88%,周期排卵率为64.29%,优势卵泡平均个数为1.8±0.8个,妊娠率为56%,而用二甲双胍治疗后分别为72%、54.84%、1.1±0.6个、48%。两者治疗后能使LH、LH/FSH、T的血清浓度明显下降,SHBG的浓度明显上升,Hom a IR明显改善。结论罗格列酮和二甲双胍联合氯米芬治疗肥胖型多囊卵巢综合征疗效可靠。二甲双胍有降低体重作用、价格便宜,适用于肥胖型P-COS伴胰岛素抵抗不严重者;罗格列酮在胰岛素增敏作用优于二甲双胍,适用于胰岛素抵抗较严重的PCOS患者。     

Incretin mimetics as a novel therapeutic option for hepatic steatosis.

BACKGROUND: Fat accumulation in the liver or non-alcoholic fatty liver disease (NAFLD) is regarded as a key pathogenic factor and component of the metabolic syndrome. It was reported that administration of the incretin mimetic exenatide reversed hepatic steatosis in an obese mouse model. We had the opportunity to study the effect of additional exenatide administration on liver fat content in a patient with type 2 diabetes. CASE REPORT: A 59-year-old male with poorly controlled type 2 diabetes was treated with exenatide in addition to metformin monotherapy. Following 44 weeks of exenatide therapy, mean the liver fat measured by liver spectroscopy declined from 15.8% to 4.3%. This dramatic decrease in liver fat was accompanied by significant beneficial changes in several cardiovascular disease risk factors and improvement of all liver enzymes, in particular alanine aminotransferase, the most important marker of liver steatosis. CONCLUSION: This case report suggests that the incretin mimetic exenatide decreases hepatic fat accumulation and may play a role in the future treatment of NAFLD, and the associated insulin resistance and cardiovascular risk factors in an ever-growing high-risk population.     

Increased response of renal perfusion to the antioxidant vitamin C in type 2 diabetes.

BACKGROUND: Reactive oxygen species play a major role in the development of endothelial dysfunction. It is as yet unspecified whether increased oxidative stress contributes to endothelial dysfunction of the renal vasculature in patients with type 2 diabetes. METHODS: Renal haemodynamics were studied in 20 patients with type 2 diabetes and arterial hypertension (age 62 +/- 5 years) and 20 non-diabetic hypertensive patients at baseline and following infusions of the nitric oxide synthase inhibitor, N(G)-monomethyl-L-arginine (L-NMMA; 4.25 mg/kg); the substrate of nitric oxide synthase, L-arginine (100 mg/kg); and the antioxidant, vitamin C (3 g, co-infused with L-arginine 100 mg/kg). RESULTS: The response of renal plasma flow (RPF) to L-NMMA (-54 +/- 62 and -45 +/- 42 ml/min/1.73 m(2); P = NS) and L-arginine (+46 +/- 36 and +49 +/- 25 ml/min/1.73 m(2); P = NS) was not different between diabetic and non-diabetic patients. In contrast, vitamin C induced a more pronounced increase in RPF in diabetic than in non-diabetic patients when co-infused with L-arginine (+71+/-47 and +43+/-33 ml/min/1.73 m(2); P<0.05). CONCLUSIONS: The difference in the response of renal perfusion to an antioxidant suggests increased formation of reactive oxygen species and thereby reduced nitric oxide bioavailability in the renal vasculature of patients with type 2 diabetes.     

Monoclonal Antibody Specific for TIRC7 Induces Donor-specific Anergy and Prevents Rejection of Cardiac Allografts in Mice

T cell immune response c-DNA (TIRC7) is up-regulated during the early stages of T-cell activation in response to alloantigens. In this study, we analyzed the effects of newly developed monoclonal antibodies (mAb) against TIRC7 in acute cardiac allograft rejection. Fully vascularized heterotopic allogeneic heart transplantation was performed in mice across a full-mismatch barrier (C57Bl/10 into CBA). Recipients received seven injections (day 0-7) of a novel anti-TIRC7 mAb or remained untreated. Graft survival, histology and ex vivo lymphocyte functions were tested. Targeting of TIRC7 with an anti-TIRC7 mAb diminishes lymphocyte infiltration into grafts resulting in delay of morphological graft damage and prolongation of allograft survival. The lymphocytes from anti-TIRC7 mAb-treated animals exhibit hypo-responsiveness without evidence of lymphocyte depletion against the donor allo-antigens. Proliferation and expression of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) were down-regulated while interleukin-4 (IL-4) and IL-10 expression were spared. Moreover, anti-TIRC7 mAb enhanced up-regulation of CTLA-4 expression but suppressed up-regulation of CD25 on stimulated lymphocytes in vitro and in vivo. Ligation of TIRC7 has important effects on the regulation of co-stimulatory signaling pathways associated with suppressing of T-cell activation. Targeting of TIRC7 may therefore provide a novel therapeutic approach for modulating T cell immune responses during organ transplantation.     

2型糖尿病患者的智能障碍

目的：调查没有临床神经损害体征的2型糖尿病患者早期的高级神经功能改变，方法：分别对52例2型糖尿病患者和32名健康对照者进行了韦氏智力量表的相似、数字广度、数字符号、木块图等4项分测验测定，采用二合一(相似、木块图)的方法计算简式智商，同时进行事件相关电位检查，结果：糖尿病患者组的相似、木块图分测验成绩和简式智商明显较健康对照组低下，ERP检查发现病人组N2、P3潜伏期较对照组明显延长，结论：2型糖尿病可导致高级神经功能损害。并且这种损害可能要早于周围神经损害。