西藏地区农牧民生活习俗与包囊虫病的关系调查

目的　探讨西藏地区农、牧民生活习俗与包囊虫病的关系。方法　自制问卷对 7个县 2 40个家庭 163 1名成员与包囊虫感染有关的生活习俗进行调查分析。结果　饭前便后洗手的人仅有 16 1%,餐具一般都用天然水洗 ,大部家庭用天然绵羊绒擦洗茶具。大多数人都喝过河溪水 ,池塘水 ,人人都吃半生不熟的肉 ,半数以上的吃生肉和生血肠 ,绝大多数人喝未经消毒的鲜牛、羊奶。 10 0 %家庭养狗 ,所有人员与狗、羊、牛、马接触密切。包囊虫病人占调查人数 0 43 %( 7/ 163 1) ,有典型症状者占 0 67%( 11/ 163 1)。结论　当地居民感染包囊虫与不良生活习俗有关     

类风湿性关节炎患者氧化还原物水平的测定

OBJECTIVE: To examine the oxidation or reduction products in patients with rheumatism arthritis (RA), and investigate the relationship between oxidation or reduction products and occurrence and development of RA. METHODS: The serum levels of total ascorbic acid (TAA), dehydroascorbic acid (DHAA)/TAA, vitamin E, advanced oxidation protein products (AOPP) and malondialdehyde (MDA) were detected by high-performance liquid chromatography with electrochemical detection in 83 RA patients and 30 healthy adults. Correlation analysis of AOPP, MDA and hs-CRP was performed. RESULTS: Compared with normal control group, significantly higher serum MDA, DHAA/TAA, and AOPP levels were detected in RA patients (P<0.05), but vitamin E showed no significant difference (P<0.05). Linear regression analysis showed that MDA (P<0.01) was positively but AOPP (P>0.05) negatively correlated to hs-CRP. CONCLUSIONS: Oxidation or reduction products in serum of RA patients increases significantly, which may be an important mechanism for the occurrence and development of RA. Serum AOPP and MDA levels can reflect the oxidation status in RA patients.     

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.     

凋亡素原核表达载体的构建和表达

目的 构建凋亡素原核表达系统，以制备抗原物质凋亡素融合蛋白。方法 通过PCR方法，以pcDNA-VP3质粒为模板，扩增出凋亡素VP3基因。将其克隆到原核表达载体pET-DsbA的多克隆位点，构建成凋亡素的高效原核表达栽体pET-DsbA-VP3，将该质粒转化到大肠杆菌E．coliBL21(DE3)plysS中，以异丙基硫代-β-D-半乳糖苷(isopropylthio-β-D-galactoside，IPTG)对其进行诱导表达，聚丙烯酰胺凝胶电泳分析目的蛋白。结果 转化有凋亡素原核表达载体pET-DsbA-VP3的大肠杆菌E．coliBL21(DE3)plysS经IPTG诱导后，聚丙烯酰胺凝胶电泳出现38300的目的蛋白条带。结论 凋亡素原核表达栽体pET-DsbA-VP3能高效表达出凋亡素融合蛋白。     

Epidermal growth factor receptor inhibition modulates the nuclear localization and cytotoxicity of the Auger electron emitting radiopharmaceutical 111In-DTPA human epidermal growth factor.

(111)In-DTPA-human epidermal growth factor ((111)In-DTPA-hEGF [DTPA is diethylenetriaminepentaacetic acid]) is an Auger electron-emitting radiopharmaceutical that targets EGF receptor (EGFR)-positive cancer. The purpose of this study was to determine the effect of EGFR inhibition by gefitinib on the internalization, nuclear translocation, and cytotoxicity of (111)In-DTPA-hEGF in EGFR-overexpressing MDA-MB-468 human breast cancer cells. METHODS: Western blot analysis was used to determine the optimum concentration of gefitinib to abolish EGFR activation. Internalization and nuclear translocation of fluorescein isothiocyanate-labeled hEGF were evaluated by confocal microscopy in MDA-MB-468 cells (1.3 x 10(6) EGFRs/cell) in the presence or absence of 1 microM gefitinib. The proportion of radioactivity partitioning into the cytoplasm and nucleus of MDA-MB-468 cells after incubation with (111)In-DTPA-hEGF for 24 h at 37 degrees C in the presence or absence of 1 microM gefitinib was measured by cell fractionation. DNA double-strand breaks caused by (111)In were quantified using the gamma-H2AX assay, and radiation-absorbed doses were estimated. Clonogenic survival assays were used to measure the cytotoxicity of (111)In-DTPA-hEGF alone or in combination with gefitinib. RESULTS: Gefitinib (1 microM) completely abolished EGFR phosphorylation in MDA-MB-468 cells. Internalization and nuclear translocation of fluorescein isothiocyanate-labeled EGF were not diminished in gefitinib-treated cells compared with controls. The proportion of internalized (111)In that localized in the nucleus was statistically significantly greater when (111)In-DTPA-hEGF was combined with gefitinib compared with (111)In-DTPA-hEGF alone (mean +/- SD: 26.0% +/- 5.5% vs. 14.6% +/- 4.0%, respectively; P < 0.05). Induction of gamma-H2AX foci was greater in MDA-MB-468 cells that were treated with (111)In-DTPA-hEGF (250 ng/mL, 1.5 MBq/mL) plus gefitinib (1 microM ) compared with those treated with (111)In-DTPA-hEGF alone (mean +/- SD: 35 +/- 4 vs. 24 +/- 5 foci per nucleus, respectively). In clonogenic assays, a significant reduction in the surviving fraction was observed when (111)In-DTPA-hEGF (5 ng/mL, 6 MBq/microg) was combined with gefitinib (1 microM ) compared with (111)In-DTPA-hEGF alone (42.9% +/- 5.7% vs. 22.9% +/- 3.6%, respectively; P < 0.01). CONCLUSION: The efficacy of (111)In-DTPA-hEGF depends on internalization and nuclear uptake of the radionuclide. Nuclear uptake, DNA damage, and cytotoxicity are enhanced when (111)In-DTPA-hEGF is combined with gefitinib. These results suggest a potential therapeutic role for peptide receptor radionuclide therapy in combination with tyrosine kinase inhibitors.     

尺神经腕背支营养血管逆行皮瓣的临床应用

目的探讨应用尺神经腕背支营养血管逆行皮瓣修复小指皮肤缺损的临床效果及手术操作要点。方法以尺神经腕背支营养血管远端为蒂,选择小指尺侧分支为皮瓣轴心血管,将皮瓣向小指远端转移,修复小指掌背侧皮肤缺损8例。结果8例全部成活,其中1例因创面止血不彻底,皮瓣受压导致远端部分坏死,经换药后痊愈。结论尺神经腕背支走向较恒定,本组未发现变异。沿皮神经干有纵行的皮神经旁血管网及皮神经干内血管网,此皮瓣血供可靠的,皮瓣切取容易,对供区影响小,是修复小指皮肤缺损的理想方法。     

X线引导下经皮通用脊柱系统内固定治疗胸腰椎骨折

目的探讨X线引导下经皮通用脊柱系统(USS)内固定治疗胸腰椎骨折的可行性.方法 2002年10月～2004年5月对无须减压胸腰椎骨折8例采用后路经皮USS椎弓根螺钉植入,在椎旁肌深层置入固定棒行伤椎复位固定.结果手术时间120～240 min,平均140 min.术中出血量20～90 ml,平均40 ml.后凸Cobb's角术前(21.5±4.3)°、术后(2.7±1.5)°(t=17.541,P=0.001),椎体塌陷术前18.5%±4.1%、术后2.4%±1.0%(t=16.504,P=0.001).术后3周佩戴腰围支具下床,复查Frankel分级,C级恢复至D级2例,D级恢复至E级1例,保持D级1例,保持E级4例.结论经皮USS内固定治疗无须减压的胸腰椎骨折创伤小,恢复快,但技术难度高,X线暴露时间长.     

纳洛酮对颅脑损伤后细胞凋亡的Caspase-3介导的信号传递机制的实验研究

目的观察纳洛酮对严重颅脑损伤后神经细胞Caspase-3表达的影响,探讨Caspase-3介导的信号传递机制.方法选用家兔77只,随机分成正常组、对照组、大剂量组和小剂量组,分别予非颅脑损伤、颅脑损伤、颅脑损伤后大剂量纳洛酮腹腔注射及颅脑损伤后小剂量纳洛酮腹腔注射处理,15天后均断头处死,于致伤处和对侧处取材,采用SP法制作切片,观察比较各组神经细胞Caspase-3表达阳性细胞数量.结果致伤处或对侧处,正常组、大剂量组、小剂量组和对照组神经细胞Caspase-3表达阳性细胞数量两两之间比较均有显著差异,值从小到大均为:正常组＜大剂量组＜小剂量组＜对照组.结论Caspase-3介导的信号传递机制可能参与纳洛酮抗颅脑损伤后神经细胞的凋亡过程.     

湿热环境下密闭性功能敷料护理伤口对β—内啡肽的影响

OBJECTIVE: To study the changes of plasma beta-endorphin (beta-EP) in pigs with traumatic injury after occlusive wound dressing supplemented with antiphlogistic and analgesic agents in hot and humid environments (HHE). METHODS: Traumatic models were established in 10 pigs, 5 of which received antiphlogistic- and analgesic-supplemented occlusive dressings of the wounds (experiment group, EG), while the rest pigs were assigned to control group (CG) to receive routine wound management. The pigs in both groups were then exposed to artificial HHE and at different time points during the exposure, the plasma beta-EP level, respiratory frequency and heart rates were measured respectively. RESULTS: The plasma beta-EP concentration of EG was significantly lower than that of CG (P <0.01) after the injury, but in both groups, the levels before the injury were similar to those measured at hour 8 during HHE exposure and at hour 24 following the injury. The variation range of the respiratory frequency and heart rates during HHE exposure were significantly smaller in EG than CG (P <0.01). CONCLUSION: This supplemented occlusive wound dressing can help restrain the peak of plasma beta-EP level and the variation range of respiratory frequency and heart rates of pigs exposed to HHE.