Directed differentiation of Balb/C mouse embryonic stem cells into pancreatic islet-like cell clusters in vitro: observation by atomic force microscope]

OBJECTIVE: To observe the morphological changes of Balb/C mouse embryonic stem cells following directed differentiation into pancreatic islet-like cell clusters (PICC) in vitro using atomic force microscope (AFM). METHODS: Balb/C mouse embryonic stem cells were first cultured into embryonic bodies (EBs) and allowed to differentiate spontaneously for 4 days. The cells were then transferred to gelatin-coated dishes for the EBs to attach and spread on the tissue culture plates, in the course of which a series of cell growth factors such as basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1) and nicotinamide were added into the culture medium at specific time points to induce directed differentiation of the stem cells into PICC. Immunocytochemistry was employed to detect the cells positive for insulin and glucagon, which were observed with AFM. RESULTS: The embryonic stem cells developed into cell clusters of different sizes, in which the cells were tightly arranged. Islet B cells were numerous in the center of clusters and darkly stained, but fewer in the peripherals with lighter stains. Islet A cells expressing glucagon were relatively fewer in the cell clusters, found mainly in the peripherals. Scanning of the insulin-positive clusters by AFM revealed large quantity of tissue fibers resembling nerve fibers that formed a reticular structure in disorderly arrangement. Numerous round granules were observed in the cytoplasm of almost identical sizes ranging from 0.5 to 1.0 mum in diameter. CONCLUSION: The cell clusters obtained by directed differentiation are mature in both morphology and function with also well organized structures.     

三维数字减影血管造影在颅内动脉瘤诊疗中的应用价值

目的：探讨3D—DSA在颅内动脉瘤诊断和治疗中的应用价值。方法：对蛛网膜下腔出血患者常规行DSA检查，发现病变或可疑病变后行3D—DSA检查。结果：26例颅内动脉瘤中检出动脉瘤31枚，其中后交通9枚，C1段3枚，C2段5枚，C3段3枚，前交通6枚，大脑中3枚。椎动脉动2枚。3例2D—DSA未见病变3D—DSA确诊，5例因血管重叠3D—DSA排除了动脉瘤。6枚GDC栓塞，5枚手术夹闭，2枚颈内动脉可脱性球囊封阻术。结论：3D—DSA极大提高了脑血管造影检查的准确性和可靠性，为动脉瘤的介入治疗和手术夹闭提供了可靠保障，在临床诊疗中具有很高的应用价值。     

Construction of Eukaryotic Expression Plasmid of Human PRX3 and Its Expression in HEK-293FT Cells

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could he expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.     

膀胱出口梗阻后逼尿肌收缩蛋白表达和膀胱重量的改变

目的探讨膀胱出口梗阻(BOO)后逼尿肌收缩蛋白表达和膀胱重量的改变及意义。方法BOO组病人16例,筛选条件为入院诊断良性前列腺增生症(BPH)并经尿动力学压力-流率检查证实为高压低流型;对照组5例,为外伤等情况入院并排除有下尿路梗阻病史者。BOO组所有病例均行耻骨上经膀胱前列腺摘除术,术前B超检查测定膀胱重量和前列腺内外径比值,术中切取膀胱上壁组织2cm×1cm×1cm大小,标本行RT-PCR反应,检测膀胱逼尿肌中肌动蛋白和肌球蛋白mRNA的表达,并比较其与膀胱重量间的线性相关性。结果BOO组与对照组膀胱重量分别为(92.15±34.89)g和(56.08±20.35)g,(P<0.05);前列腺内外径比值分别为(0.57±0.16)和(0.18±0.06),(P<0.01);与对照组相比,BOO组肌动蛋白和肌球蛋白mRNA的表达量均有显著增加,分别为(40.32±59.67)×106和(6.59±5.62)×106,(P值均<0.01);且两者表达量与膀胱重量之间均有明显线性正相关性(P<0.05)。结论逼尿肌中肌动蛋白和肌球蛋白的表达与膀胱逼尿肌的功能状态密切相关。     

载脂蛋白AⅠ-CⅢ-AⅣ基因簇Xmn Ⅰ、Msp Ⅰ位点多态性与胆固醇结石病关系的研究

目的探讨载脂蛋白（Apo）AⅠ—CⅢ-AⅣ基因簇多态性与胆囊胆固醇结石病的关系，揭示胆固醇结石病的遗传易感性。方法应用多聚酶链反应技术（PCR）对胆固醇结石病患者及正常对照者ApoAⅠ—CⅢ-AⅣ基因簇的限制性片段长度多态性进行研究，检测ApoAⅠ-CⅢ-AⅣ基因位点XmnAⅠ及MspAⅠ。结果胆石组和对照组均以X1、M1等位基因为主，多为X1X1、M1M1纯合子基因型。胆石组各基因型和各等位基因频率与对照组比较，差异无统计学意义（P〉0．05）。胆石组女性患者正常型基因X1X1纯合子频率和正常等位基因X1的频率低于对照组女性，突变型基因X1X2杂合子、X2X2纯合子以及突变型等位基因X2的频率均高于对照组女性，差异有统计学意义（P〈0．05）。胆石组男性患者正常型基因M1M1纯合子频率低于对照组男性，而突变型基因M1M2杂合子频率高于对照组男性，差异有统计学意义（P〈0．05）。结论ApoAⅠ—CⅢ-AⅣ基因簇XmnⅠ位点的多态性可能与女性胆囊胆固醇结石病有关，而MspⅠ位点的多态性可能与男性胆囊胆固醇结石病有关。     

单独肝尾状叶血管瘤切除术

肝尾状叶由于解剖位置特殊,位置深,难以显露,手术难度大,是肝脏外科领域手术操作的难点与研究热点.随着肝血流控制技术的发展、肝实质离断技术的提高,肝尾状叶肿瘤切除率明显提高[1].2006年4月至2008年10月,我科完成单独肝尾状叶血管瘤切除术9例,现将手术技巧与疗效报道如下.     

复方丹参滴丸对不稳定型心绞痛患者血浆内皮素和血小板α-膜颗粒蛋白的影响

目的观察复方丹参滴丸对不稳定型心绞痛患者的临床疗效及血浆内皮素（ET）和血小板α-膜颗粒蛋白（GMP-140）水平的影响。方法168例不稳定型心绞痛患者分为常规治疗组（79例）和复方丹参滴丸组（89例）。所有入选患者均接受硝酸酯类、美托洛尔（或钙拮抗剂）及阿司匹林治疗，复方丹参滴丸组在上述治疗基础上加用复方丹参滴丸10粒，每日3次，连用8周。观察统计两组治疗前1周和治疗结束后1周心绞痛发作次数及自拟的疼痛强度及持续时间记分值，检测治疗前后患者血浆ET和GMP-140水平的变化。结果复方丹参滴丸组和常规治疗组比较，治疗后疼痛发作频率、疼痛强度及持续时间记分均有明显减少，而复方丹参滴丸组的降低程度比常规治疗组明显（P〈0．05）。两组可改善不稳定型心绞痛患者心电图ST段的偏移，而复方丹参滴丸组的疗效比常规治疗组显著（P〈0．05）。两组治疗后血浆ET和GMP-140水平均明显减低，而复方丹参滴丸组的疗效比常规治疗组显著（P〈0．01）。结论复方丹参滴丸治疗不稳定型心绞痛患者疗效明显高于常规治疗，可能与改善血管内皮功能、抑制血小板聚集密切相关。     

The effect of the septal area in acupuncture analgesia]

This review summarized some articles on the effect of the septal area in acupuncture analgesia. The data showed that the pain threshold of animal was increased when septal area was stimulated by electro-acupuncture, and that electrical stimulation of septal area had a marked inhibitory effect on the pain discharges of cells in parafascicular nucleus of thalamus, lateral habenular nucleus, periaqueductal gray and dorsal raphe nucleus. The septal area play an important role in acupuncture analgesia. The majority of the cholinergic neurons in septal area are located in nucleus of the vertical limb of the diagonal band (VDB); gamma-aminobutyric acid of septal area is mainly found in the diagonal band nucleus(td); Dopamine is present in high levels in td and lateral septal nucleus(S1) of septal area; The S1 contain high densities enkephalin-containing neuronal cell bodies and terminals; In addition, substance P and norepinephrine are also high levels in the septal area. These substance above-mentioned have a relations with acupuncture analgesia of septal area. A large number of serotonin-containing neurons are found in the raphe nuclei. The serotonin play an important role in acupuncture analgesia. The serotonin-containing neurons in dorsal raphe nucleus project to S1. The fiber connections of the raphe nuclei with the td are reciprocation. The periaqueductal gray is a important structure on pain modulation. It projects to septal area and receives the fibers from S1. A number of adrenergic neurons are located within the locus coeruleus. The locus coeruleus participate pain modulation and acupuncture analgesia. The neuro-anatomy study demonstrated that locus coeruleus projects to septal area.(ABSTRACT TRUNCATED AT 250 WORDS)     

青霉烷砜对氨苄青霉素的增效作用

观察加用青霉烷砜后氨苄青霉素对痢疾杆菌的体外杀菌效果。结果表明,加用青霉烷砜后对69株痢疾杆菌的杀菌效果,从单纯氨苄青霉素的杀菌率59.4％(41/69)提高到98.6％(68/69),有显著增效作用(P<0.005)。各株痢疾杆菌对国产和墨西哥产氨苄青霉素的敏感性无差异。证明青霉烷砜对氨苄青霉素有增效作用,对原耐氨苄青霉素痢疾杆菌的杀菌率达96.4％(27/28)。对耐药痢疾杆菌治疗药物的选择提供了有益的参考。     

微波快速过氧化物酶染色法与灌注法用于微循环形态学研究之比较

以健康Ｗｉｓｔａｒ大白鼠为材料，对微波快速内源性过氧化物酶染色与灌注法显示微血管的方法进行了对比研究。结果，过氧化物酶组织化学法具有不用灌注，操作简单，所用时间短，对血管无扩张、破裂等人为改变的特点，保持了微血管真实的自然形态和管径大小，可以定量或半定量地判定组织器官活体时的血液循环状况。可以用于人及动物的大脑、脊髓、皮肤、耳及食管等组织内微血管形态学研究和定量分析。并对过氧化物酶显示微血管的原理和微波辐射促进染色的原理及特点进行了分析、讨论。