Objective: To observe the effect of curcumin derivatives L6H4 on rats with abnormalities of glucose and lipid metabolism, and preliminarily study the mechanism of it.Methods: Twenty-four male SD rats were randomly allocated into three groups: normal control group (NC group,n=8), high fatty group (HF group, n=8), and fatty treatment group (FT group,n=8). To fed the rats in HF group and FT group with high fat and surgar diet for 12 weeks, from the 5th week with drug treatment for 8 weeks, at the end of 12th week, the fol-lowing indexes were measured respectively in each group: serum biochemical indexes, liver and skeletal muscle triglycerides, and related indicators of sugar and lipid matabolism. Observing the liver pathological changes at the same time.Results:①Compared HF group with NC group: weight gain (P<0.05), liver weight, fasting blood glucose (FBG), fasting insulin (FINS), insulin resistance index (HOME-IR), serum triglycerides, liver triglycer-ide, skeletal muscle triglycerides, serum ALT, FFA, high-sensitivity c-reactive protein (hs-CRP) and interleukin 6 increased (P<0.01), adiponectin and leptin decreased (P<0.01,P<0.05, respectively).②Compared the FT group with HF group: weight and ALT loss (P<0.01), liver weight and liver index had no obvious difference, FBG de-creased (P<0.01), FINS (P>0.05), the HOME-IR decreased (P<0.01), serum triglycerides, liver triglycerides and&nbsp;skeletal muscle triglyceride were reduced (P<0.01,P<0.01, andP<0.05, respectively), FFA and hs-CRP reduced (P<0.01), fibroblast growth factor 21 (FGF-21) increased (P<0.01), adiponectin, interleukin 6 increased with P<0.01 andP<0.05, respectively.③Inlfammatory injury in hepatic tissue of HF groups was slight, while fatty de-generation in hepatic tissue and skeletal muscle were obvious, the condition was signiifcantly reduced in the FT group.Conclusion: Curcumin derivatives L6H4 has the effect of controlling weight, protecting the liver function and improving the condition of insulin resistance on high fat and sugar-fed rats.

临床实习对于检验医学专业学生的教育来说是一个极其重要的阶段，实习的过程是将学校学到检验医学理论知识和临床工作实践相结合、由书面的知识向实践工作能力转化的过程，为今后从事临床检验工作打下重要的基础。作为附属医院检验科，每年都接受很多来自检验医学专业的实习生，免疫检验室是目前检验科开展项目多，岗位设置也比较多的实验室。随着自动化的提高，更多地应用先进检验仪器设备，并不断更新，但并不意味着手工操作被淘汰，很多项目还是使用手工操作来检测。本文将免疫实验室带教过程中一些，体会进行总结和分析。

目的:制备M13K07辅助噬菌体多克隆抗体.方法:将M13K07辅助噬菌体进行大量扩增,以此作为免疫原于1、3、5周进行兔免疫,收集0周和免疫后2、4、6、8周免疫血清,用间接ELISA法对免免疫血清的效价进行检测,进一步用免疫斑点实验检测免多克隆抗体与M13K07辅助噬菌体的特异性结合能力.结果:M13K07辅助噬菌体免疫兔可产生高效价的多克隆抗体,效价达1∶3 200,并能特异性识别M13K07辅助噬菌体.结论:成功制备了高效价的M13K07辅助噬菌体多克隆抗体,为噬菌体展示技术研究提供了检测抗体.

在讨论传统临床生化检验课程教学存在问题的基础上,提出了以立体化的教材资源、网络化的精品资源共享课教学平台为支撑的教学资源和数字化战略为整体的解决方案。开展立体化教学的新设想,能适应临床生化检验快速发展的教学要求。其中,转变教育教学观念、不断更新教学内容和全面改革教学方法是开展立体化教学的基本素质要求。

[ ] Objective To investigate the effects of gossypol on diabetic nephropathy and the possible mechanism. Methods Thirty male Sprague-Dawley rats were divided randomly into three groups: normal control group, type 2 diabetic group and gossypol-treated group. After being fed with high-fat feeding for 4 weeks, the latter two groups were injected with low dosage streptozotocin(30mg/Kg)intraperitoneal y to induce type 2 diabetes. Diabetic rats in gossypol-treated group were given gossypol at the dosage of 15mg/kg.d for 4 weeks by gavage, afterwards once per week for 8 week. The rats were sacrificed after fasting for 12h and 24-h urine samples were col ected. The concentrations of blood glucose, blood urea nitrogen(BUN)and 24-h urine protein were measured. The morphological changes of the kidney were studied by light microscopy and transmission elec-tron microscopy. The mRNA expression of SGK1, Col-Ⅳ kidney tissue was assayed by semi-quantity RT-PCR. Results Com-pared with normal control group, the level of glucose, BUN and the 24-h urine protein in diabetic group were significantly in-creased (P<0.01). The volume of glomerulus was enlarged, the deposition of PAS positive substance in the glomerular intersti-tium increased under light microscopy, and the glomerular basal membrane was thickened significantly under electron mi-croscopy in diabetic group, but they were ameliorated in gossypol-treated group. The mRNA expression of SGK (P<0.05)and Col-Ⅳ(P<0.01)was increased significantly compared with normal control group. After the treatment of gossypol, the mRNA ex-pression of Col- Ⅳ was decreased compared with type 2 diabetic group (P<0.05). Conclusion Gossypol may delay the progress of diabetic nephropathy by down-regulating mRNA expression level of Col-Ⅳin kidney of type 2 diabetic rats.

<正>自动生化分析仪是以紫外分光光度法为主要分析技术,将取样、加试剂、混匀、保温反应、吸光度检测、结果计算、可靠性判断、数据显示和打印、数据传输、清洗等步骤结合起来实现自动化的仪器,主要用于测定肝功能、肾功能、血糖、血脂、血清酶活性以及特定蛋白等常规生化指标。1957年由Technicon公司根据Skeggs医生提出的方案,设计了第一台商品化的连续流动式自动生化分析仪,是20世纪50~80年代市场占有率最高的仪器,由于通道少、分析速度慢、试剂浪费大,逐渐被淘汰。20世纪70年代初期设计出离心式自动生化分析仪,随后的20年中这类分析仪在中国非常流行,因一个离心盘一般只分析一个项目,反应盘中的凹槽无自动清洗功能,分析速度慢,故其应用逐渐

Objective:Determine the detection limit of hydrothorax and ascites total protein determining with biuret,as well as hydrothorax and ascites albumin determining with bromocresol green method,and perform linearity evaluation of the range of detection.Methods: Prepare blank sample,select samples that are close to the detection limit and those that have high pathology value.Perform total protein determining with biuret,and albumin determining with bromocresol green method on Hitachi 7600 Automatic Biochemic Analyzer,and analyze the lower limit of detection and biologic limit of detection.Apply EP-6P and polynomial to evaluate linearity.Results: By determining hydrothorax and ascites total protein with biuret,the lower limit of detection is determined to be 0.47 g/L,and the biologic limit of detection is 1.33 g/L;The linearity from 2.5 g/L to 49.7 g/L is accepted by clinic.By determining hydrothorax and ascites albumin with bromocresol green method,the lower limit of detection is determined to be 0.44 g/L,and the biologic limit of detection is 0.98 g/L.The linearity from 1.2 g/L to 35.2 g/L is accepted by clinic.Conclusion: The method of determining hydrothorax and ascites total protein with biuret and the method of determining hydrothorax and ascites albumin with bromocresol green method are acceptable clinically.

Objective:To study the value of CK-MB in diagnosing neonatal asphyxia with brain damage.Methods: 69 patients(22 neonatal asphyxia,27 neonatal asphyxia with brain damage,20 normal) were evaluated with total CK,CK-MB,calculate CK-MB ratio using automatic biochemical analyser and agarose sugar electrophoresis method.Results:(1)The results of CK-MB% measured by automatic biochemical analyser were higher than the agarose sugar electrophoresis in the control group and the test group(P0.01).Compared between two groups,results were indifferent(P0.05).(2) Compared the brain damage group with neonatal asphyxia group,results were indifferent(P0.05).Conclusion: The results of CK-MB% measured by automatic biochemical analyser were significantly higher than the agarose sugar electrophoresis.The results of CK-MB% may not be regarded as a biochemical indicator in diagnosing neonatal asphyxia with brain damage.

Objective To compare the enzymic method and Jaffe kinetic creatinine assay for the determination of creatinine clearance(CrCl) and discuss whether using foreign CrCl reference interval is appropriate.Methods The levels of serum creatinine,urine creatinine and CrCl were determined in 266 patients by sarcosine oxidase enzymic method and Jaffe kinetic creatinine assay.The results were divided into 4 groups by the CrCl level of Jaffe kinetic creatinine assay.Their CrCl levels were <25,26-50,51-80 and >81 mL/min in Group 1-4 respectively.The two methods′ results of serum creatinine,urine creatinine and CrCl and their difference were analyzed statistically.Results The creatinine concentrations in serum and urine determined by the enzymic method were significantly lower than those determined by Jaffe kinetic creatinine assay(P<0.001).However,the CrCl(except Group 1) determined by the enzymic method was much higher than that by Jaffe kinetic creatinine assay(P<0.001).The relative deviation of serum creatinine between the two methods was small and was close to the relative deviation of urine creatinine in Group 1,so that CrCl values by both methods had no significant difference.The relative deviation of serum creatinine gradually increased in Group 2-4,however,the relative deviation of urine creatinine almost unchanged.The deviation of CrCl between the two methods would be larger and larger(P<0.001).Conclusions When CrCl <25 mL/min,CrCl by the two methods has no difference.On the contrary,when CrCl >26 mL/min,the difference of CrCl appears and will be growing.The deviation of CrCl in normal group is large,so it is unappropriate to use foreign CrCl reference interval.

Objective To express in prokaryotic system and to analyze the antigenic specificity of EB virus(EBV) latent membrane protein 2(LMP2) multi-epitopes gene rich of T cell and B cell epitopes.Methods Using on-line prediction service, T cell epitopes and B cell epitopes of EB virus latent membrane protein 2 were predicted. The genes rich of CTL and th cell epitopes were selected as the candidate gene sequences, while B cell epitopes around them were taken into account. The finial selected multi-epitope gene was synthesized after being optimized according to prokaryotic codon bias and inserted into prokaryotic expression vector pET32a( + ) to get the recombinant plasmid: pET32a( + )/EBV-LMP2 multi-epitopes. After transformed into E. coli BL21 (DE3) and induced by IPTG, the target multi-epitopes gene can be expressed as Trx-His fusion protein. The expression products can be identified by SDS-PAGE and Western blot. Moreover, rabbit serum antibody to EBV membrane protein and nasopharyngeal carcinoma(NPC) patient serum were used respectively to detect the antigenic specificity of the multi-epitopes. Meanwhile, 6-8 weeks female BALB/c mice were immunized with EBV-LMP2 multi-epitope at 2 week intervals, three times in all, Trx-His protein and PBS were set as the control groups. At the second week after the last immunization, the mice were sacrificed. LDH and indirect ELISA were taken to detect the specific spleen CTL activityand specific IgG in serum, which reflected the immunogenicity of the EBV-LMP2 multi-epitope. Results Two amino acid sequences which locate at the LMP2 (aa195 -232 ) and LMP2 (aa419-436 ) were selected and connected in series to be the target gene. The recombinant plasmid containing EBV-LMP2 multi-epitope gene successfully constructed and the target protein was expressed in E. coli BL21 ( DE3 ). The relative molecular mass(Mr) of The expression products is about 27 × 103 , which matches up to the expected Mr. The antigenic specificity of the multi-epitopes protein was identified by Western blot and the multi-epitopes protein also can be detected by rabbit serum antibody to EBV membrane protein and NPC patient serum respectively. In the result of the animal experiment, EBV-LMP2 multi-epitope was able to induce the specific CTL activity in BALB/c mice. With the increasing of the effector: target ( E: T) 1: 5,1: 10, 1: 25, the CTL activity was also increased wih( 12.52% + 2.59% ), (21.80% + 1.08% ), (23.68% + 3.74% ) respectively; EBV-LMP2 multi-epitope was able to induce LMP2-specific antibody response(A490 =0.258 +0.040) as compared with Trx-His protein(A490 =0.095 +0.011) and PBS(A490 =0.068 +0.014,P＜0.05＝. Conclusion The EBV-LMP2 multi-epitopes gene was designed successfully and expressed precisely in prokaryotic expression system with good antigenicity and immunogenicity.

目的研究酚磺乙胺对两种不同酶法检测血清肌酐的干扰。方法严格按照CLSI推荐的EP7-A干扰实验程序,设计配对差异试验和患者标本偏差分析考察对肌酐酶偶联肌氨酸氧化酶法和肌酐亚胺水解酶偶联谷氨酸脱氢酶法检测血清肌酐的影响。结果酚磺乙胺浓度在0.003～0.3g/L范围内,肌酐酶偶联肌氨酸氧化酶法测定肌酐对照组标本和药物加入标本组肌酐检测结果差异有显著性(<0.01),其干扰值在血清肌酐高、中、低浓度水平均大于―1.96,病人标本偏差分析结果显示用药患者肌酐结果与非用药组差异有显著性(<0.01)。而以肌酐亚胺水解酶偶联谷氨酸脱氢酶法检测,两组标本之间结果差异无显著性。结论酚磺乙胺对肌酐酶偶联肌氨酸氧化酶法法检测肌酐有负干扰,对肌酐亚胺水解酶偶联谷氨酸脱氢酶法检测无干扰。

AIM:To study the effect and mechanism of extract of ginkgo biloba(EGB) on liver glucocorticoid receptor(GR) expression in type 2 diabetic rats.METHODS:Thirty male Sprague-Dawley rats were divided randomly into three groups:normal control group(n=10),type 2 diabetic group(n=10) and ginkgo biloba treated group(n=10).After fed with high-fat feeding for 4 weeks,the later two groups were injected with streptozotocin at a dose of 30 mg/kg intraperitoneally to induce type 2 diabetic rat model.The EGB treated group was gavaged with EGB at the dose of 50 mg·kg-1·d-1 for 12 weeks.At the end of experiment,the rats were sacrificed,the blood glucose,serum lipid and blood insulin were measured.The morphology of liver tissue was observed under light microscopy with HE staining.GR mRNA expression in liver was measured by RT-PCR.The level of GR protein expression in liver tissue was detected by immunohistochemistry.RESULTS:EGB reduced the levels of blood glucose,blood lipids,blood insulin in diabetic rats.EGB also relieved fatty degeneration and necrosis of the hepatic cells,ameliorated infiltration of inflammatory cells in the liver;and decreased GR expression at both mRNA and protein levels in diabetic liver.CONCLUSION:EGB may inhibit GR expression in liver of type 2 diabetic rats,which results in decreasing the level of blood glucose,blood lipid,blood insulin and relieving the liver damage in type diabetic rats.

AIM:To study the effect of gossypol on the cognitive function of type 2 diabetic rats,and to explore its mechanism. METHODS:Thirty male Sprague-Dawley rats were divided into three groups randomly:normal group,type 2 diabetic group and gossypol treated group. After fed with high-fat diet for 4 weeks,the later two groups were injected with streptozotocin intraperitoneally to establish type 2 diabetic rat model. The animals in gossypol treated group were given gossypol at dosage of 15 mg/kg once per day for 4 weeks by gavage. Since 5th week,the times of gavages were changed into once per week at the same dosage and lasted to 12th week. Learning and memory abilities of rats were assayed with Morris water maze test. The concentration of blood glucose was measured by biochemical method. The levels of serum corticosterone and insulin were detected by enzyme-linked immunosorbent assay and radioimmunoassay,respectively. The protein expressions of 11β-HSD1 and GR in cerebral cortex and hippocampus were determined by Western blotting. The morphological changes of cerebral cortex and hippocampus were observed under light microscope and transmission electronic microscope,respectively. RESULTS:Compared to normal group,the karyopyknosis,dilation of golgiosome and mitochondria swelling of neuron from cerebral cortex and hippocampus were prominent in diabetic group. The concentrations of blood glucose,serum corticosterone and insulin increased significantly (P0.01). Protein expression of GR decreased (P0.05),11β-HSD1 protein tended to increase. Platform searching score was lower (P0.01) and escape latency was longer (P0.01) in diabetic group. After treated with gossypol,the concentrations of blood glucose,serum corticosterone and insulin declined (P0.01). The protein expression of 11β-HSD1 was decreased (P0.05) and GR was increased (P0.05). Escape latency was shorter (P0.01) and platform searching score was increased (P0.01). CONCLUSION:Gossypol may improve the cognitive function of type 2 diabetic rats. Decreasing the level of 11β-HSD1 and increasing GR protein in the brain may be involved in the mechanism.

Objective:To study the reliability of the method of detecting the glycosylated hemoglobin A1c in whole blood with latex particle intensified immunity transmission turbidity.Methods:Used the method on latex particle intensified immunity transmission turbidity to detect the glyeosylated hemoglobin A1C and also evaluated the precision,comparative test and the result of different operation.Results:The inter-CV and the intra-CV for GHb were 0.9% and 3.63%.The average recovery rates were 105.3% and 103.7% respectively.And it had great relativity with latex particle agglutination inhibition.The different operation had little interference.Conclusion:Immunity transmission turbidity with latex particle intensified has a good precision and accuracy.It is a rapid method which can act as a clinical routine method for measuring GHb.

目的:为实现不同检测系统间血清酶[丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、各氨酰肽酶(GGT)、淀粉酶(AMY)、肌酸激酶(CK)、乳酸脱氢酶(LDH)]测定结果具可比性.方法:参照NCCLS批准的<EP-9A指南文件>,以1/4 CLIA′88为系统间偏差临床是否可接受的标准,以由cfas量值传递的新鲜混合血清为校准待评系统的临时校准品,分别在三个不同生化检测系统上以患者新鲜血清对七项常见血清酶作校准前、校准后的方法学比对试验,并对结果进行直线回归分析和偏倚评估.结果:棱准前系统2的ALT、CK、LDH、AMY的SE%>1/4 CUA′88,临床不能接受,ALP、GGT部分可接受:系统3的ALT、GGT临床不能接受.对两待评系统用临时校准品校准后再做比对评估,两待评系统所有项目与目标系统的相对偏差都小于允许范围(1/4CLIA′88),临床全部可接受.结论:经量值传递后的患者定值血清可用来校正不同检测系统的酶检测,从而使相互的测定结果具可比性.

Objective:To analyze the concentration of non-high density lipoprotein cholesterol(non-HDL-C) and atherogenic index of plasma(AIP) from Wen Zhou City of the normal population.Methods:The 30448 people with health examination from 2002 to 2006 were analyzed,the blood samples from the population were detected TG,TCH,HDL-C,LDL-C,and calculated the AIP,non-HDL-C.Results:In these people,the concentration of AIP,non-HDL-C from the men was higher than that of the women,and gradually stepped up following the age;AIP was positive correlation with the TG,non-HDL-C,LDL-C and non-HDL-C was positive correlation with the TG,TC,LDL-C,AIP,The non-HDL-C,AIP were negative correlation with the HDL-C.Conclusion:About the population of Wen Zhou City,the concentration of AIP,non-HDL-C from the men was higher than that of the women,and by degrees heightened following the age,so we must think highly of the population.

Objective To analyze the levels of non-high density lipoprotein cholesterol(non-HDL-C)and atherogenic index of plasma(AIP)in people with normal concentrations of triglyceride(TG)and total cholesterol(TCH)so as to provide basis for the prevention and control of lipid abnormality.Methods 14,855 people with normal concentrations of triglyceride(TG)and total cholesterol(TCH)were selected from total 45470 people who took physical examination in our hospital in 2001,in which the level of high density lipoprotein cholesterol(HDL-C)was detected.And then non-HDL-C,LDL-C,AIP were calculated and compared among the groups divided by gender and age.Result The level of non-HDL-C and AIP were significantly high in men than in women,and increased with age.AIP was positively correlated with TG,non-HDL-C and LDL-C;non-HLD-C was positively correlated with TG,TC,LDL-C and AIP.The both of AIP,non-HDL-C and HDL-C showed negative correlation.Conclusion TG and TC are risk factors of atherosclerosis in normal population.The risk is higher in men than in women,and increases with age.

Objective To investigate the significance of determination for binding ratio of L-PHA(leukoagglutinating phytohemagglutinin) with serum alkaline phosphatase in the diagnosis of primary hepatic cancer.Methods The binding ratios of L-PHA with serum alkaline phosphatase in 50 healthy individuals,103 cases of primary hepatic cancer,22 cases of hepatocirrhosis,36 cases of chronic hepatitis,18 cases of acute hepatitis and 13 cases of digestive tract tumor were determined by lectin affinity chromatography.Results The binding ratio of L-PHA with serum alkaline phosphatase in primary hepatic cancer(16.01% 5.42%) were significantly increased as compared with the healthy control(7.50% 2.53%)(P 0.01).The binding ratios were positively correlated with the tumor size and AFP level.Conclusion The binding ratio of L-PHA with serum alkaline phosphatase is a potential parameter for the laboratory diagnosis of primary hepatic cancer.

目前医院检验科已普遍采用自动生化分析仪,手工检测已经成为历史,生化检验中重点工作是对所检验指标做出确的评价。因此,临床生化检验实验课内容也要做出相应的调整,笔者对实验课内容作了修改和补充,分别设计了每次实验课的具体内容和课程目标,从而改变了过去只简单地对手工操作技能的反复训练的实验模式,以适合现代临床实验室要求的工作。

目前医院检验科生化检验中手工检测已经成为历史,自动生化分析仪完成了绝大部分的工作,工作人员的重点工作是对所检验指标的方法学性能做出很好的评价,并在检测过程中采取质量控制手段。医学检验专业的临床生化检验实验课内容也理应做出相应的调整,为此我们对实验课内容作了较大的修改和补充,以适合现代临床实验室工作的要求。