目的：探讨血清α2-巨球蛋白（α2-MG ）在评估2型糖尿病肾病中的应用价值。方法空腹采集血样，其中确诊的2型糖尿病肾病者46例，2型糖尿病无肾病者73例，同期健康对照者66例，用免疫比浊检测血清α2-MG、尿微量清蛋白水平，用高效液相色谱法检测糖化血红蛋白（HbA1c）水平，并进行组间比较，分析糖尿病肾病患者血清α2-MG 与尿微量清蛋白和 HbA1c 的相关性。结果血清α2-MG、尿微量清蛋白在糖尿病肾病者分别为（2．25±0．53）g／L 和（199．8±100．9）mg／L，明显高于健康对照者和糖尿病无肾病者，差异有统计学意义（P ＜0．05）；糖尿病肾病者中血清α2-MG 水平与尿微量清蛋白呈显著正相关（r ＝0．514，P ＜0．05）。结论检测血清α2-MG 水平对诊断、预测糖尿病肾脏损伤具有重要的应用价值。

Objective To investigate whether pancreatic progenitors and islets differentiated from human embryonic stem cells(hESCs) could correct hyperglycemia in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice.Methods We obtained pancreatic progenitors and islets derived from hES cells line YT1 according to the optimized four-stage differentiation protocol.Stage 1:definitive endoderm formation; Stage 2:pan creatic specialization; Stage 3:amplification of pancreatic progenitors,Stage 4:maturation of pancreatic islets.To observe the morphological changes of each stage and immunofluorescent expression of pancreatic and duodenal homeobox gene(PDX-1),glucagon,insulin,C-peptide,glucose transporter-2(Glut-2).The differentiated cells from stage 3 and stage 4 were then transplanted into one of the epididymal fat pads(EFP) of NOD/SCID mice.The survival and function of the graft were measured by immunohistochemistry and blood glucose monitor.Results The stage 4-differentiated pancreatic islets expressed mature β cell-specific markers such as glucagon,insulin and Glut 2,and even PDX-1 and C-peptide-double-positive.The stage 4-pancreatic islets had nearly 17.1％ insulin-positive cells as assayed by flow cytometry analysis.Differentiated pancreatic islets released insulin/C-peptide in response to glucose stimulation.After implantation into EFP of NOD/SCID mice,hES cell-derived human pancreatic progenitors and islets corrected hyperglycemia for at least 12 weeks.Conclusions Pancreatic progenitors and islets differentiated from hESCs can correct hyperglycemia in NOD/SCID mice.

BACKGROUND:Human pancreatic islet transplantation is a prospective curative treatment for diabetes. However, the lack of donor pancreases greatly limits this approach. One approach to overcome the limited supply of donor pancreases is to generate functional islets from human embryonic stem cells (hESCs), a cell line with unlimited proliferative capacity, through rapid directed differentiation. This study investigated whether pancreatic insulin-producing cells (IPCs) differentiated from hESCs could correct hyperglycemia in severe combined immunodeficient (SCID)/non-obese diabetic (NOD) mice, an animal model of diabetes.METHODS:We generated pancreatic IPCs from two hESC lines, YT1 and YT2, using an optimized four-stage differentiation protocol in a chemically defined culture system. Then, about 5-7 × 10(6) differentiated cells were transplanted into the epididymal fat pad of SCID/NOD mice (n = 20). The control group were transplanted with undifferentiated hESCs (n = 6). Graft survival and function were assessed using immunohistochemistry, and measuring serum human C-peptide and blood glucose levels.RESULTS:The pancreatic IPCs were generated by the four-stage differentiation protocol using hESCs. About 17.1% of differentiated cells expressed insulin, as determined by flow cytometry. These cells secreted insulin/C-peptide following glucose stimulation, similarly to adult human islets. Most of these IPCs co-expressed mature β cell-specific markers, including human C-peptide, GLUT2, PDX1, insulin, and glucagon. After implantation into the epididymal fat pad of SCID/NOD mice, the hESC-derived pancreatic IPCs corrected hyperglycemia for ≥ 8 weeks. None of the animals transplanted with pancreatic IPCs developed tumors during the time. The mean survival of recipients was increased by implanted IPCs as compared to implanted undifferentiated hESCs (P<0.0001).CONCLUSIONS:The results of this study confirmed that human terminally differentiated pancreatic IPCs derived from hESCs can correct hyperglycemia in SCID/NOD mice for ≥8 weeks.

Objective To investigate whether pancreatic progenitors differentiated from human embryonic stem(hES) cells could correct hyperglycemia in non-obese diabetic(NOD)/severe combined immunodeficient(SCID) mice or not.Methods Pancreatic islets derived from hES cells line YT1 according to the optimized four-stage differentiation protocol in a chemical-defined culture system were observed.In the first stage,activin A and wortmannin were utilized to induce definitive endoderm formation.In the second stage,the differentiated endoderm cells were treated with all-trans retinoic acid(RA),NOGGIN and basic fibroblast growth factor(bFGF) to induce pancreatic specialization.In the third stage,pancreatic progenitors were induced by epidermal growth factor(EGF)-regulated expansion.Finally,in the fourth stage,a cocktail of factors was utilized to induce mature insulin-producing cells.The differentiated human pancreatic islet cells of(3-5)×106 were then transplanted into one of the epididymal fat pads(EFP) of NOD/SCID mice.Graft survival and function were measured by blood glucose levels.Results The differentiated hES cells obtained by the four-stage approach comprised nearly 17.1% insulin-positive cells and 3.8% C-peptide positive as assayed by flow cytometry analysis,which released insulin/C-peptide in response to glucose stimuli in a manner comparable to that of adult human islets.Most of these insulin-producing cells co-expressed mature β cell-specific markers such as PDX1,insulin,C-peptide and glucagon.After implantation into EFP of NOD/SCID mice,hES cell-derived pancreatic islets corrected hyperglycemia for an least eight weeks.Conclusion Human terminal differentiation pancreatic islet cells can correct hyperglycemia in NOD/SCID mice.

Human cancer-associated UniGene sets (NCBI GeneBank) provide a platform for identifying differentially-expressed genes in human cancers. The present study identified and characterized a set of human cancer-associated genes using the Digital Differential Display (DDD) and functional analysis tools. A total of 1,904 genes were differentially expressed in 15 cancer types, including genes that had been previously shown to be specific in certain human cancers. A total of 274 genes were uniquely expressed in certain cancer types, including 37 genes that were highly expressed in the human testes and epididymis. These genes mainly functioned as ribosomal proteins, enzymes, receptors, secretory proteins and cell adhesion molecules. The most common domains that were encoded by the cancer-associated genes were those of cytochrome P450 CYP2D6, serpin and apolipoprotein A-I. A further gene ontology (GO) enrichment analysis revealed seven major functional clusters, which corresponded to the enriched pathways involved in cancer. The present study provides a source of cancer-associated genes and their functions. The results provide new insights into cancer biology and the involvement of highly-expressed epididymal genes in cancer biomarkers.

目的探讨体外诱导人胚胎干细胞(hESs)定向分化为胰岛细胞的可行性。方法体外分三阶段诱导hESCs定向分化为胰岛细胞。第一阶段:予以活化素-A、渥曼青霉素诱导分化形成定型内胚层;第二阶段:采用维甲酸(RA)、NOGGIN、碱性成纤维细胞生长因子(bFGF)诱导胰腺细胞定向分化;以EGF扩增胰腺祖细胞;第三阶段:采用尼克酰胺、唾液素4、bFGF及骨形成蛋白(BMP4)促进胰岛细胞成熟;观察诱导各阶段细胞形态变化,免疫荧光法鉴定胰十二指肠同源异型盒基因(PDX-1)、胰高糖素、胰岛素、C肽、葡萄糖转运子2(glut-2)表达。结果诱导14 d时hESs出现胰高糖素荧光表达;20 d时细胞出现PDX-1和C肽共表达;22 d形成的成熟胰岛细胞出现glut-2和胰岛素的阳性表达;胰岛素阳性细胞占17.1%,C肽阳性细胞占3.8%。结论体外可简单高效的诱导hESs定向分化为成熟胰岛细胞。

Objective To discuss the methods of separation,purify and culture of neonatal mice cardiomyocytes(CM) for inducing mesenchymal stem cells(MSCs) to CM by coculture,and found the way to identify the cells.Methods CM from ventricular tissues of neonatal mice were separated with enzyme digestion,purified by differential and adherent,and treated with 5-bromodeoxyuridine(5-Brdu).The expressions of CM mRNA and protein were detected with reverse transcription-polymerase chain reaction(RT-PCR) and immunofluorescence staining,and the ultras-tructural characteristics of CM were observed with transmission electron microscope after culture for 7 days.Results CM with well morphology and spontaneous beating obtained by the use of 0.08% trypsin and 0.04% collagenase type Ⅱ,98 % cells are alive,and then CM were arranged in loose and jumping synchronously by adherent culture for 3 days.The percentage of positive cell(CM) were 99.07%,marked with anti-cardiac Troponin T(anti-cTnT) antibody by immuofluorescence staining.The expression of myocardial myosin heavy chain α/β(MHC α/β),cTnT gene in CM were detected apparently by RT-PCR with MSCs as negative control detetednegative control.The Ultra-structure of few CM can be observed,used 3%-4% Agar to pre-embed and then got the extra-thins lice.Conclusions Our studies found a way to obtain and identify neonatal mouse CM with nomal tructure and function,and provide a foundation for further inducing MSCs by cocultrue with CM,and identifying the cells after differentiation.

Cancer is a complex disease caused by multiple factors including genetic mutations, and environmental factors. Cancer-associated proteins are potential biomarkers or targets for diagnostic and therapeutic interventions in cancer. The Universal Protein Resourse (UniPort) is a well-annotated comprehensive resourse for protein sequence records. In the present study, we performed data mining of Uniprot proteins as a proteomic resource. we generated a catalog of 1653 cancer-associated proteins including 344 secretory proteins and 300 cell surface proteins. Integrated bioinformatic analysis including ontological classification, functional enrichment and pathway construction were performed. These proteins could serve as a reference for further studies to discover cancer targets, and the enriched bioinformatic analysis provides new insights into cancer proteomics research.

Pancreatic stem cells were isolated and cultured from aborted human fetal pancreases of gestational age 14-20 weeks. They were seeded at a density of 1 × 10(4) in serum-free media for differentiation into neuron-like cells, expressing β-tubulin III and glial fibrillary acidic protein. These neuron-like cells displayed a synapse-like morphology and appeared to form a neuronal network. Pancreatic stem cells were also seeded at a density of 1 × 10(5) for differentiation into islet-like cells, expressing insulin and glucagon, with an islet-like morphology. These cells had glucose-stimulated secretion of human insulin and C-peptide. Results suggest that pancreatic stem cells can be differentiated into neuron-like and islet-like cells.

UKPDS及DCCT等一系列大规模的糖尿病强化治疗研究证实严格的血糖控制可以明显减少微血管及大血管并发症,而要完全控制糖尿病只有通过胰腺移植或胰岛移植.但胰岛移植需要足够的供体,目前胰腺供体严重匮乏,且受配型的限制,成功配型的机率过低,另外移植后长期的免疫抑制治疗给病人带来诸如排斥、感染、肿瘤等一系列问题,因此有必要寻找一种新的胰岛细胞来源,近年对胚胎干细胞(ES)及成体干细胞(AES)的研究为糖尿病的细胞替代治疗带来了新的思路.

Objective To study and to optimize culture conditions of islet-like cells induced in vitro from fetus bone marrow(BM) mesenchymal stem cells(MSCs).Methods BM was obtained from miscarried human fetus.The MSCs between three to eight passages were used to differentiate into islet-like clusters-through three stages of culture supplemented with 2mercaptoethanol,epidermal growth factor(EGF),basic fibroblast growth factor(bFGF),B_(27) and nicotinamide.Results The 2nd stage cells expressed nestin and/or panceatic and duod-enal homeobox 1(PDX-1),and the 3rd stage cells formed islet-like clusters expressing insulin and glucagon together with positive dithizone staining.Specific insulin secretion could be detected(81.3±23.6μu /ml) from differentiated MSCs which have the capacity to respond to different glucose concentrations.Conclusion Fetal bone marrow MSCs can be differentiated into pancreatic islet-like clusters,and 20mmol/L nicotinamide could be the optimal concentration in culture.

OBJECTIVE To confirm previous effort to identify type 2 diabetes susceptibility genes in a Northern Chinese population by conducting a new genome scan with both an increased number of type 2 diabetes families and a new set of microsatellite markers within the previously localized regions. METHODS A genome scan method was applied. After multiplexed PCR, electrophoreses, genescan and genotyping analysis, we obtained size information for all loci, and then a further study was done by both parametric and non-parametric linkage analysis to investigate the P values and Z values of these loci. RESULTS We surveyed 34 microsatellite markers which distributed within 5 regions along chromosome 1, and a total of 12,000 genotypes were screened. Evidence of linkage with diabetes was identified for 8 of the 34 loci. All P values of the 8 loci were lower than 0.05, and the highest Z value was 2.17. A very interesting finding is that all 5 markers at the p- terminal 1p36.3-1p36.23 region, spanning a long range of 16.9 cM, were identified to have a low P value of less than 0.05, which suggests that this region may contain multiple susceptibility genes. Regions 4 and 5 also confirmed the previous findings, and we narrowed these two regions to a 2.7 cM and 2.5 cM regions, respectively. CONCLUSIONS We further confirmed the results gained in the previous genome-wide scan using an increased number of NIDDM families and a new set of microsatellite markers lying within the initially localized regions. The fact that all 5 loci at the p- terminal region displayed a low P value of less than 0.05 suggests that more than 1 susceptibility gene may reside in this region.

目的　对86例老年糖尿病人机会感染进行研究，探讨其发病机制、常见机会致病菌及临床特点。　方法　从住院的老年糖尿病人中筛选机会感染者86例，分析其致病菌、感染部位、血糖、血浆蛋白、血纤维蛋白原、血红蛋白及尿蛋白等。　结果　主要危险因素是高血纤维蛋白原、低白蛋白血症和高血糖等。常见的感染部位是泌尿系统、呼吸系统、双足、躯体皮肤及败血症；常见的机会致病菌有葡萄球菌、大肠埃希菌、肺炎克雷白菌及链球菌。　结论　老年糖尿病人易发生机会感染，致病菌呈多样性，在抗感染治疗中应积极改善器官功能和支持治疗。

本文旨在讨论老年糖尿病人发生复数菌感染的常见部位,致病菌及危险因素,为老年糖尿病人感染的诊断和治疗提供参考.

目的探讨老年糖尿病患者康复过程中严重皮肤感染发病机制、临床表现及常见致病菌.方法回顾性分析38例皮肤损伤原因、感染好发部位及常见致病菌,分析入院时生化指标.结果皮肤损伤多因护理不当和保护知识缺乏,感染多发生在足部,其次是背部及臀部;病原菌多为条件致病菌,以金黄色葡萄球菌最多见;其中高血糖、高纤维蛋白原、低白蛋白血症及贫血为独立危险因素.结论老年糖尿病患者康复治疗过程中易发生皮肤感染,对糖尿病患者教育及护理是预防严重皮肤感染的关键.

糖尿病患者多有较严重的动脉硬化,因此脑卒中的发生率增高.本文报告28例糖尿病性昏迷合并急性脑血管病(脑卒中)的情况,探讨早期诊断的可能性. 1 资料与方法

调查Ⅱ型糖尿病家系22个,其一级系属116人,己诊为Ⅱ型糖尿病者54人,余行75g葡萄糖耐量及胰岛素释放试验,结果新发现糖尿病4人,糖耐量异常者16人.将新诊糖尿病和己诊糖尿病例为DM组(n=16),新诊糖耐量异常者为IGT组(n=16),糖耐量正常者列为NGT组(n=31).经统计分析发现DM与IGT存在明显高胰岛素血症,胰岛素敏感性(ISI)明显低于正常组.

我们于1997～1998年间对山东沿海地区408例2型糖尿病患者进行血管并发症患病率调查,并探讨其危险因素,现报告如下.

To analyse the diabetic nephropathy(DN) and its related irsk factors from diabetic vascular complication study in Shandong sea board.171 of them were found to have DN and 237 were non DN.The results showed that the older age,duration,glucose,blood pressure level,fibrinogen,triglyceride(TG) and cholesterol of DN group were significantly higher than non DN group.DN group was significantly higher in the ocerrence of diabetic retinopathy,coronary heart disease than that non DN group.Using multiple regression analysis showed that the significant related factors were duration,high fibrinogen,high TG and high systolic presure for DN.The results suggest that close control and correction of these abnormalities were very important to prevent diabetic vascular complications.