体外模拟二氧化碳人工气腹对宫颈癌细胞生长的影响

目的 观察二氧化碳（CO2）人工气腹对宫颈癌细胞生长的影响，探讨其与乏氧的关系。方法 2005-10—2006-03华中科技大学同济医学院附属协和医院通过裸鼠右下腹腔注射宫颈癌细胞，注射宫颈癌细胞株（Hela）数量：4×10^9／L：将注射后40只裸鼠随机分成4组，每组10只：阴性对照组（常氧组）：仅给予右下腹腔注射宫颈癌细胞；CO2气腹模拟环境按CO2灌注压分3组：CO2灌注压4mmHg（1mmHg=0．133kPa）组，CO2灌注压10mmHg组，CO2灌注压16mmHg组．4周后观察裸鼠体重改变、切除瘤灶称瘤重、瘤体标本用免疫组化测定增殖细胞核抗原（PCNA）、Western-blot方法检测瘤体的缺氧诱导因子-1α（HIF-1α）表达．结果 与阴性对照组比较，CO2气腹各组裸鼠体重下降明显，而瘤体重量增加相对显著，并随着CO2灌注压的增高这种趋势更为显著（P〈0．05）；CO2气腹各组裸鼠PCNA表达显著高于阴性对照组（P〈0．01）；CO2气腹各组裸鼠HIF-1α蛋白的表达与阴性对照组比较明显增加，差异有统计学意义（P〈0．01）．结论 与阴性对照组比较，CO2人工气腹环境可促进宫颈癌细胞生长，其机制可能与HIF-1α激活了PCNA表达，促进了细胞增殖有关。     

Oncogenic aberrations in the p53 pathway are associated with a high S phase fraction and poor patient survival in B-cell Non-Hodgkin's lymphoma

The implications of aberrations in the p53 pathway for induction of apoptosis and regulation of S phase entry, and for patient survival, were investigated in 83 B-cell Non-Hodgkin's lymphomas. Eight cases had missense mutations in exons 5, 7, 8 and 9 as revealed by constant denaturant gel electrophoresis and sequencing. Fifteen cases had lost 1 TP53 allele as revealed by fluorescent in situ hybridization and comparative genomic hybridization. Ten cases expressed high levels of p53 as assessed by immunoblotting and immunohistochemistry. S phase fractions were higher, apoptotic fractions were the same and survival times were shorter in all aberration groups compared with the cases with no TP53/p53 aberrations. Since many tumors had more than one TP53/p53 aberration, the tumors were divided into groups with the following characteristics: no TP53/p53 aberrations; loss of one TP53 allele only (9 cases), TP53 point mutation (8 cases), high-level p53 expression and no TP53 mutation (3 cases). Tumors from the 3 latter groups had higher median S phase fractions (5%, 7.6%, and 5%, respectively, p<0.02) than the cases without any aberrations (1.1%), and survival time for these patients was much shorter (relative risks of 5.9, 8.9, and 6.6, respectively, p<0.003). Apoptotic fractions were similar in all these groups (p=0.09). Multivariate analysis showed that the presence of TP53/p53 aberrations is a strong and independent prognostic parameter in B-cell Non-Hodgkin's lymphoma.     

纵向观测二分类数据的广义线性模型分析

目的：利用广义线性模型对纵向观测二分类数据进行分析,充分考虑纵向观测间的相关性,给出一般分析方法。方法：采用Zeger和提出的广义估计方程,拟合logistic广义线性模型,讨论3种协方差矩阵结构。结果：同时获得回归参数、相关参数的估计,完成了较为实用的运行程序,并进行了实例分析。结论：医学研究和临床试验中经常接触到纵向观测数据,对这类数据需采用特殊的方法进行分析处理,以解决重复观测间的相关性问题。     

99Tcm直接法标记血管抑素

目的: 探索用99Tcm直接标记血管抑素(angiostatin, AS)的方法,并研究标记产物的体外稳定性及其生物活性. 方法:制备的AS经鉴定后,分别用2-巯基乙醇(2-ME)和氯化亚锡(SnCl2)还原法进行标记,并用正交设计筛选最佳合成条件;对标记产物用纸层析法及Sephadex G-50层析柱分离测标记率;产物中分别加入牛血清白蛋白(BSA)、生理盐水及不同摩尔比的半胱氨酸(Cys)以观察其体外稳定性;用人脐静脉内皮细胞系ECV304观察其生物活性. 结果:2-ME法最佳标记条件为AS 100 μg,PB(0.5 mol*L-1, pH 7.3) 1 mL, 2-ME 100 μg, 亚甲基二膦酸盐(MDP)(用1 mL生理盐水溶解) 10 μL,加入99TcmO4- 185 MBq,标记率可达(97.0±1.5)%; SnCl2法为AS 100 μg,硼酸缓冲液(0.1 mol*L-1, pH 9.0) 1 mL, 20 g*L-1 SnCl2 (1 mol*L-1盐酸作为溶剂) 20 μL加入MDP药盒,用去氧水稀释至1 mL, 取20 μL,加入99TcmO4- 185 MBq,标记率可达(90.0±3.0)%. 产物在体外稳定;细胞培养观察其抑制内皮细胞生物活性与AS无显著差异. 结论:用99Tcm直接法标记AS简单高效,且对AS生物活性无明显影响.     

导航下手术治疗脑血管疾病

目的：探讨无框架脑立体定位手术在治疗脑血管性疾病中的应用价值。方法：术前将MRI数据传入无框架脑立体定向导航系统（SealthStation)计算机工作站,进行三维建模,标记点定位,手术入路设计,开颅前行参考环及探针确认,标记点注册,设计手术切口,术中实时定位,1999年11月至2001年6月行无框架脑立体定向手术117例,其中动静脉畸形46例,动脉瘤42例,海绵状血管瘤29例,结果：颅内病灶或重要解剖定位结构准确,机显病灶定位误差均在2mm以内,术后神经功能损害4例,占3．4％,无手术死亡,结论：无框架脑立体定向手术能准确进行病灶定位,有效保护脑组织,减少神经损伤,切除病灶,改变了传统开颅手术模式,是微创神经外科的保证。     

Viral shedding in Chinese young adults with mild 2009 H1N1 influenza

Background  The duration of viral shedding and the transmission of 2009 H1N1 influenza among individuals, especially among the younger population with mild illness, are not well understood now. The aim of this study was to determine the viral shedding of the young adult patients with mild 2009 H1N1 influenza in China.

Methods  From September 2009 to January 2010, the clinical data and serial nasopharyngeal swabs of 67 patients with 2009 H1N1 influenza and 37 patients with seasonal influenza aged from 18 years to 35 years were collected. The nasopharyngeal swab samples were detected by real time RT-PCR to determine the viral shedding. All the patients did not receive the antiviral therapy but Chinese medicine for detoxicating.

Results  Among the patients with H1N1 virus infection, 82.1% (55/67) patients presented with fever symptom, while more patients with high fever (≥39ºC) were found in seasonal influenza patients (P <0.05). For the H1N1 patients, the median interval between the symptom onset and the undetectable RNA was six days (4–10 days). But viral shedding was still found in 31.3% patients after 7 days following illness onset. The median interval between disappearance of fever and an undetectable viral RNA level was three days (2–8 days), and 17.9% patients were found to be viral shedding 6 days later after normalization of body temperature. For the seasonal influenza patients, 94.6% patients were detected out viral RNA within 7 days. The median interval of seasonal influenza between the symptom onset and the undetectable RNA was four days (3–8 days). The median interval between disappearance of fever and an undetectable viral RNA level was three days (2–6 days).

Conclusion  It suggests that 7 days isolation period from the illness onset or 24 hours after the resolution of fever and respiratory symptoms are not long enough to cut off the transmission among Chinese young adults with mild illness.

     

The Effect of Different Treatments of (–)-Epigallocatechin-3-Gallate on Colorectal Carcinoma Cell Lines

Backgroud: (-)-Epigallocatechin-3-gallate (EGCG), the major component of green tea, is well documented to induce apoptosis and cell cycle arrest in cancer by targeting multiple signal transduction pathways. However, EGCG is extremely unstable in general culture conditions and rapidly degraded. So, to what extent EGCG or which degradation products of EGCG play a role in anti-tumor is still unknown. In this study, we evaluated the effect of different treatments of EGCG on HCT116 cells.

Design: MTT assay was applied to evaluated the inhibitory effect of different treatments of EGCG on HCT116 cells. Cell cycle and apoptosis were performed by flow cytometry. Finally, western blot analysis was used to elucidate the molecular mechanism associated with cell cycle arrest and apoptosis.

Results: Compared with control, both EGCG and O-EGCG (i.e., EGCG being pre-incubated at 37°C for 3 h) significantly inhibited HCT116 cells growth. Surprisingly, we found that the inhibitory effect of O-EGCG was stronger than that of EGCG. The IC₅₀ values of EGCG and O-EGCG were 8.75 and 5.40 μM, respectively. Cell cycle analysis showed that 20 μM of EGCG simultaneously caused cell cycle arrest at G1 and G2 phase in HCT116 cells, differing from O-EGCG which exclusively caused cell cycle arrest at G2. This result suggested that parent EGCG at the early treatment might cause cell cycle arrest at G1. As time went on, EGCG disappeared and degraded products of EGCG were formed which might cause cell cycle arrest at G2. Further studies revealed that EGCG induced cell cycle arrest at G1 by downregulation of cyclin E and cyclin D1 and upregulation of p21. On the other hand, O-EGCG induced HCT116 cells apoptosis mainly by increasing the expression of p53 and cleaved caspase-3, which might be the underlying reason why O-EGCG had stronger inhibitory effect on HCT116 cells line than EGCG.

Conclusions: The pretreatment of EGCG may be an effective way to enhance its antitumor effect.     

藏药牦牛角中的主要化学成分

采用180／80AAS仪、WYD－2型氢化物原子荧光光谱仪以及Wates-AAA型氨基酸分析仪等分别测定了传统藏药牦牛角的K、Na、 Ca、Mg、Al、P、Cu、Zn、Fe、Mn、Co、Se、As、Hg、Pb、Cd等16种矿物质常量、微量元素和17种氨基酸的含量。结果表明,在青藏高原这一独特生态环境条件下生活的牦牛其藏医入药的牦牛角,含有丰富的矿物质常量、微量元素以及人体所必需的氨酸基等化学成分。     

2～5岁儿童贫血状况及其与生长发育的关系

目的了解某市城区2～5岁在园儿童贫血状况,分析相关影响因素,为改善本地区儿童贫血状况提供依据。方法采用分层整群抽样的方法,在该市城区随机抽取4所公立幼儿园和8所私立幼儿园,对每所幼儿园2～5岁在园儿童进行一般情况调查,进行体格发育测量和口腔检查,并采末梢静脉血检测血红蛋白。结果共调查2 993名儿童,男1 424,女1 569,平均月龄为(51.23±11.73)月,公立幼儿园儿童占64.0%,单因素分析显示,私立幼儿园儿童贫血高于公立幼儿园,月龄越大儿童贫血检出率越低,非贫血儿童生长发育状况好于贫血儿童;多因素Logistic回归结果显示,月龄高[0.985(0.978～0.992)]为儿童贫血的保护因素,龋齿数多[1.034(1.007～1.062)]和私立幼儿园[3.276(2.782～3.857)]为儿童贫血的危险因素。结论该地区2～5岁在园儿童贫血检出率高,针对该地区儿童采取预防贫血的干预措施势在必行;贫血检出率随月龄的增高而降低,在研究儿童贫血与生长发育的关系时需考虑到月龄。     

腹腔镜、胆管镜、扩张导管、T管支架治疗肝左外叶及胆总管结石17例

目的探讨腹腔镜、胆管镜、扩张导管(逐级扩张导管、球囊导管)、T管支架(T形管＋塑料胆管支架,LCDTS)治疗肝左外叶及胆总管结石的应用体会。 方法回顾性分析2002年4月至2017年9月间符合入选标准的17例患者的临床资料。 结果腹腔镜下切除肝左外叶和探查胆总管17例,切除胆囊12例。胆管镜取石、逐级导管扩张左肝管和乳头并留置LCDTS 52.9% (9/17),逐级导管扩张左肝管和推挤左肝管结石出总管切口并留置LCDTS 23.5% (4/17),逐级导管联合球囊导管扩张左肝管并留置LCDTS 11.8%(2/17),因肝内胆管残石和狭窄扩张左肝管并留置LCDTS 11.8% (2/17)。术后残石2例(11.8%),胆汁漏1例(5.9%)。无肠穿孔、胆管穿孔、大出血、急性胰腺炎等并发症,无围手术期再手术和死亡患者。术后总并发症发生率为17.6%(3/17)。 结论只要病例选择合适,LCDTS治疗肝左外叶及胆总管结石是可行、有效和安全的。