鼠伤寒沙门菌spvC基因缺陷变异株的制备

目的: 为进一步研究鼠伤寒沙门菌毒力基因spvC的功能,制备鼠伤寒沙门菌spvC基因缺陷变异株。方法: 根据鼠伤寒沙门菌spvC基因序列,设计PCR特异性引物,制备spvC基因缺陷性同源性核苷酸片段,导入自杀质粒pCVD442后再导入鼠伤寒沙门菌野生株,进行同源重组,用PCR观察重组现象,将完全重组的菌株作为spvC基因的缺陷变异株,并通过核苷酸序列分析加以确定。结果： PCR及序列分析证实,缺陷变异株的spvC基因缺失711个碱基。结论： 成功构建鼠伤寒沙门菌spvC基因缺陷变异株,为进一步研究其在鼠伤寒沙门菌中的功能奠定了基础。     

伤寒沙门菌fljA样基因缺陷变异株的制备

目的：为探讨z66鞭毛抗原阳性伤寒沙门菌的fljA样基因的功能,制备沙门菌fljA样基因缺陷变异株。方法：根据伤寒沙门菌fljA样基因序列,设计PCR特异性引物并在5末端加接酶BglII位点,扩增fljA样基因上、下游片段,用BglII消化后,定向连接成fljA样基因的缺损性同源性核苷酸片段,先克隆至pGEM—T质粒,再经BnmHI酶切后导入自杀质粒pGMB151,电转化入伤寒沙门菌(GIFU 10007),进行同源重组。用PCR观察重组现象,将完全重组的菌株作为fljA样基因的缺陷变异株,并通过相应的核苷酸序列分析加以确定。结果：PCR及序列分析证实,缺陷变异株的fljA样基因缺失231个碱基。结论：成功构建伤寒沙门菌flja样基因缺陷变异株,为进一步研究其在伤寒沙门菌鞭毛抗原表达的调控作用奠定了基础。     

伤寒沙门菌rpoE基因缺陷变异株的制备及其生存能力

目的为深入研究rpoE基因在伤寒沙门菌侵袭致病中的作用,制备rpoE基因缺陷变异株;观察rpoE基因缺陷变异株在应激条件下的生存能力。方法根据伤寒沙门菌rpoE基因序列,设计PCR特异性引物,制备rpoE基因缺陷性同源性核苷酸片段导入自杀质粒PGMB151后再导入伤寒沙门菌野生株,进行同源重组,用PCR观察重组现象;绘制生长曲线,对比rpoE基因缺陷变异株与野生株在应激条件下的生存情况。结果PCR及序列分析证实,缺陷变异株的rpoE基因缺失288个碱基;生长曲线提示rpoE基因缺陷变异株在氧、酸、高渗应激条件下生存能力明显低于野生株,差异有统计学意义（P〈0.05）。结论成功构建了伤寒沙门菌rpoE基因缺陷株,并发现在氧、酸、高渗应激条件下其生存能力明显降低,为进一步研究其在伤寒沙门菌中的功能奠定了基础。     

伤寒沙门菌OmpR缺陷变异株制备与分泌蛋白初析

目的观察伤寒沙门菌调节蛋白OmpR的调节作用，制备伤寒沙门菌OmpR缺陷变异株，并比较分析野生株和OmpR缺陷变异株在不同渗透压环境培养下的分泌蛋白。方法用自杀质粒pGMB151介导同源性重组方法制备伤寒沙门菌OmpR缺陷变异株，用PCR方法观察重组现象，变异株经测序和外膜蛋白的SDS—PAG电泳鉴定，用SDS—PAG电泳结合Western blot比较分析野生株和OmpR缺陷变异株在高、低渗透压环境培养下的SipC等分泌蛋白。结果成功制备伤寒沙门菌OmpR缺陷变异株，并发现变异株中，包括SipC、鞭毛素蛋白在内的多种蛋白的表达分泌有明显差异。结论伤寒沙门菌OmpR在不同的渗透压下分别影响SipC、鞭毛素蛋白等多种蛋白的表达分泌。     

伤寒沙门菌细胞壁缺陷突变株16SrRNA基因的检测与分析

目的 探讨伤寒沙门菌细胞壁缺陷突变株的菌种相关性或亲缘性.方法 分离提取伤寒沙门菌细胞壁缺陷突变株及其亲代细菌型的染色体DNA,采用16SrRNA基因引物进行PCR扩增,对扩增的16SrDNA产物进行琼脂糖凝胶电泳和碱基序列测定.结果 经PCR扩增后伤寒沙门菌细胞壁缺陷突变株及其亲代细菌型均获得了大小约230 bp的特异性片段,其产物经琼脂糖凝胶电泳可见在230 bp处出现与其亲代细菌型一致的DNA条带,碱基序列测定显示具有同其亲代细菌型一致的核苷酸序列.结论 伤寒沙门菌的细胞壁缺陷突变株虽然丧失了常规细菌学可检测的绝大多数表型特征,但检测16SrRNA基因能快速、灵敏地对其进行菌种相关性或亲缘性的分析与鉴定,并且也有利于对各种不能自发返祖的细菌稳定L型进行菌种相关性或亲缘性的分析与鉴定.     

PmrA对伤寒沙门菌高渗应激后期基因表达调节初探

目的:探讨伤寒沙门菌调节因子PmrA在高渗应激后期对基因表达调节的影响.方法:应用自杀质粒介导的同源重组方法制备伤寒沙门菌pmrA基因缺陷变异株;利用伤寒沙门菌全基因组芯片分析技术,比较伤寒沙门菌野生株和pmrA基因缺陷变异株在高渗应激后期的基因表达谱差异,并选择部分差异表达基因进行RT-RCR验证.结果:经PCR及序列分析证实,伤寒沙门菌pmrA基因缺陷变异株制备成功;基因表达谱比较分析结果表明伤寒沙门菌pmrA基因缺陷变异株在高渗应激后期有81个基因表达上调,有22 个基因表达下调.结论:PmrA在伤寒沙门菌高渗应激后期的基因表达调节中发挥重要作用.     

伤寒沙门菌fljB∶∶lacZ变异株的制备

目的:为研究伤寒沙门菌z66鞭毛抗原编码基因fljB的表达调节机制,建立该fljB基因的β-半乳糖苷酶基因(lacZ)插入变异株。方法:采用自杀质粒介导的同源重组方法进行制备。设计加接KpnⅠ和SalⅠ的特异性引物,用PCR扩增获得fljB基因的上、下游同源性DNA片段,并与用KpnⅠ和SalⅠ酶切pSV-β-半乳糖苷酶载体(pSV-β-galactosidase vector)的片段定向连接,将连接片段克隆至自杀质粒pGMB151,再通过电击法将重组质粒转入伤寒沙门菌,在含X-Gal的蔗糖平板上筛选获得同源重组的变异株。结果:经筛选获得阳性克隆,经PCR及序列分析证实,fljB基因中的763 bp被3550 bp的lacZ片段替代。结论:成功构建伤寒沙门菌flj∶∶lacZ突变株,为进一步研究fljB基因表达调节机制奠定了基础。     

OsmY缺失对伤寒沙门菌在高渗应激早期基因表达的影响

目的:研究伤寒沙门菌OsmY在高渗应激早期对其他基因表达的调节。方法:通过同源重组的方法利用自杀质粒制备伤寒沙门菌osmY基因缺陷变异株;采用伤寒沙门菌全基因组芯片比较野生株和osmY基因缺陷变异株在高渗应激早期的基因表达差异,并对其中部分表达差异基因进行实时定量PCR验证。结果:成功制备伤寒沙门菌osmY基因缺陷变异株;基因芯片结果显示,在高渗应激早期,与野生株相比,伤寒沙门菌osmY基因缺陷变异株有128个基因表达下调,有27个基因表达上调。实时定量PCR与芯片结果一致。结论:OsmY可作为一调节因子在伤寒沙门菌高渗应激早期对基因表达起重要调节作用。     

Hfq对伤寒沙门菌高渗应激早期基因表达的调节

目的：探查伤寒沙门菌hfq基因在高渗应激早期对其他基因表达的影响。方法：用自杀质粒介导的同源重组方法制备伤寒沙门菌hfq基因缺陷变异株;利用伤寒沙门菌全基因组芯片技术,比较伤寒沙门菌野生株和咆向基因缺陷变异株在高渗应激早期的基因表达谱差异,并选择部分表达差异基因进行实时定量PCR验证。结果：伤寒沙门菌hfq基因缺陷变异株制备成功;基因表达谱比较分析结果表明伤寒沙门菌hfq基因缺陷变异株在高渗应激早期有62个基因表达上调,有32个基因表达下调。实时定量PCR结果与芯片分析一致。结论：Hfq作为一个调节因子在伤寒沙门菌高渗应激早期对基因表达起重要调节作用。     

mig-14对伤寒沙门菌在高渗应激早期基因表达的调节

目的:探查伤寒沙门菌mig-14在高渗应激下对若干基因表达的调节作用.方法:用自杀质粒介导的同源重组方法制备伤寒沙门菌mig-14基因缺陷变异株,用基因芯片分析高渗应激早期野生株和mig-14缺陷株基因表达差异,并对部分结果采用实时荧光定量PCR进行验证.结果:成功制备伤寒沙门菌mig-14缺陷株,高渗应激早期mig-14变异株与野生株相比有77个基因表达下调和72个基因表达上调. 结论:mig-14可能是一种调节基因,主要参与调节细菌的物质和能量代谢.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.