IL2RG基因新突变致重症联合免疫缺陷病的临床表型和基因分析

目的探讨白细胞介素-2受体共同γ链(IL2RG)基因突变所致重症联合免疫缺陷病的临床特征和基因突变类型。方法回顾分析1例IL2RG基因突变所致重症联合免疫缺陷病患儿的临床资料。结果患儿,男,48天,生后早期多重感染,白细胞减少,抗生素治疗效果差。患儿IgG 2.93 g/L,IgA0.07 g/L,lgM 0.16 g/L,C3 0.67 g/L,C4 0.13 g/L;CD 3+CD 4+T淋巴细胞0. 03%、CD 3+CD 8+T淋巴细胞0. 1%、CD 3-/CD 16+CD 56+NK细胞2. 6%;CD 3-CD 19+B淋巴细胞96.76%。二代基因测序显示患儿IL2RG基因(HG19位置chrX:70328484)存在半合变异c.816_819delGATT(p.L273fs*20),为罕见移码突变。软件预测该突变导致蛋白质合成提前出现终止密码,为1类致病突变。患儿母亲为杂合状态,患儿父亲无此突变,患儿姨妈也有该杂合突变。结论发现1例由IL2RG基因[c.816-819delGATT (p.L273fs*20)]突变所致的重症联合免疫缺陷病患儿。基因测序分析结合性别鉴定可在先证者家系中筛查携带者及产前诊断。     

脑瘫精细运动功能测试量表在偏瘫型患儿中需要重新建立评估标准与程序

目的 分析脑瘫精细运动功能测试量表(FMFM)在偏瘫和非偏瘫型脑瘫间的项目功能差异(DIF)。方法 收集2001至2018年复旦大学附属儿科医院康复中心及其医联体儿童康复协作网诊断为脑瘫并行FMFM评估的患儿,从FMFM评估数据中提取偏瘫患儿对侧数据,随机选择30%样本作为DIF的焦点组样本;根据整体样本中非偏瘫型与偏瘫型的比例确定采用随机分层匹配对照组样本。采用Conquest软件把两组合并数据样本分别与FMFM中B~E区的全部56个项目以及经项目内容分析后划分出的30个单手项目和26个双手项目进行Rasch分析,以内聚拟合度(Infit)的均方来确定FMFM的单维性,标准为MnSq在0.6～1.4,并对全部56项、单双手项目组进行项目功能差异分析。结果 1 556例脑瘫患儿的3 442次FMFM评估作为整体样本和数据,焦点组198个和对照组792个FMFM评估数据进入本文分析,单独样本配对检验显示两组FMFM分值[112(96,146) vs 113(95,145)]差异无统计学意义。经过3轮项目内容分析,30个归入单手项目,26个归为双手项目。全部56个项目的难度尺度对数值之差的绝对值>0.5分别有41个(73.2%),30个单手和26个双手项目的难度尺度对数值之差的绝对值>0.5分别有23个(76.7%)和9个(34.6%)。结论 FMFM全部56个操作项目在偏瘫和非偏瘫型脑瘫间存在DIF,在单手项目中尤其显著,为了更为精准地评价偏瘫儿童的精细运动功能,需要重新建立评估标准与程序。     

Tenascin-C基因在子宫内膜癌中的表达及临床意义

目的：研究 Tenascin - C 在子宫内膜正常组织、子宫内膜增生组织及子宫内膜癌组织中的表达,探讨Tenascin - C 基因与子宫内膜癌发生、发展及预后的关系。方法：采用免疫组化 SP 法,分别检测14例正常子宫内膜组织、18例子宫内膜增生组织和78例子宫内膜癌组织中 Tenascin - C 的表达情况,并分析子宫内膜癌组织中 Tenascin - C 的表达和临床病理特征之间的关系。结果：Tenascin - C 在子宫内膜癌组织阳性率明显高于正常子宫内膜组织、子宫内膜增生组织,差异有统计学意义（P <0.05）。Tenascin - C 在子宫内膜癌中的表达与浸润肌层深度、分化程度、FIGO 分期和淋巴结转移密切相关（P <0.05）。结论：Tenascin - C 基因的表达与子宫内膜癌的发生、发展和转移密切相关,因此对 Tenascin - C 基因的检测可用于判断子宫内膜癌恶性程度及评估其预后的生物学指标。     

~（18）F-FDG SPECT/CT检查结合CA125检测在卵巢癌术后复发转移诊断中的价值

目的：探讨18F-FDG SPECT/CT检查结合CA125检测在卵巢癌术后复发病例的早期诊断价值。方法：对43例卵巢癌术后病例同时进行18F-FDG SPECT/CT检查和CA125检测,怀疑复发的病例进行CT/MRI检查和临床随访,以进一步确诊。结果：43例病人中,18F-FDG SPECT/CT检查发现核素异常浓聚24例,经CT/MRI检查和临床资料证实为转移灶19例,阳性率和假阳性率分别为44.2%、11.6%,特异性为83.8%。本组病例CA125升高13例,证实为转移灶病例中CA125升高12例,阳性率和假阳性率分别为27.9%、2.3%,特异性为96.9%。结论：18F-FDG SPECT/CT检查和CA125检测是卵巢癌术后复发的可靠检查方法。     

Immortalized mouse fetal liver stromal cells support growth and maintenance of human embryonic stem cells

Human embryonic stem cells (hESCs) are usually maintained in an undifferentiated state by co-culture with feeder cells. The feeder cells are important for the growth of hESCs. A novel spontaneously immortalizated mouse fetal liver stromal cell line, named KM3, was isolated from a?13.5?day mouse fetal liver. In this study, we examined whether KM3 cells could be used as feeders to support the growth of hESCs. hESCs cultured on KM3 cells showed a similar proliferation rate and characteristics to mouse embryonic fibroblasts?(MEFs) after prolonged culture, including morphology, unlimited and undifferentiated proliferative ability, maintenance of normal karyotypes, formation of embryoid bodies in?vitro and typically immature teratomas in?vivo. Our results indicate that the immortalized KM3 cell line has the potential to support the growth and maintenance of hESCs. The cell line may be used for the large-scale expansion of hESCs in a low-cost and less labor-intensive manner.     

RNA干扰环氧化酶-2基因表达对A549细胞体外恶性增殖的影响

目的研究不同靶点对环氧化酶-2(COX-2)基因表达的抑制及其对A549细胞体外恶性增殖的影响。方法以COX-2第3、7和10外显子为靶点,构建3个带有人U6启动子的COX-2小干扰RNA(siRNA)表达载体。用脂质体lipofectamine介导,分别将3个siRNA表达载体及2个空载体转染到COX-2表达阳性的A549细胞,建立转染细胞株。采用逆转录-聚合酶链反应(RT-PCR)和Western blot方法检测COX-2表达水平的变化。通过细胞生长曲线、集落形成实验研究COX-2不同干扰靶点对A549细胞体外增殖的影响。结果3个siRNA和U6启动子序列正确并准确克隆到pEGFP载体,转染细胞株分别命名为A549-3、A549-7、A549-10、A549-p和A549-pU6。转染后24、48和72 h,A549-p细胞均有绿色荧光表达,而A549-pU6、A549-3、A549-7和A549-10细胞均未观察到绿色荧光。RT-PCR和Western blot结果显示,以COX-2第3、7和10外显子作为靶点进行干扰,COX-2表达均受到抑制。与A549细胞比较,A549-3、A549-7、A549-10细胞的COX-2 mRNA表达量分别降低10.6%、33.4%和61.2%,COX-2蛋白表达量分别降低26.7%、44.7%和56.2%。细胞生长曲线、集落形成试验的结果显示,A549-10细胞生长减慢,集落形成率减少,而第3外显子和7外显子两个干扰靶点对A549细胞生长没有明显的影响。结论以COX-2第10外显子为靶点干扰效果最好,对A549细胞的体外恶性增殖有明显的影响。     

胸段食管癌术后预防性照射的预后分析

目的 探讨胸段食管癌根治术后预防性照射的范围对长期生存的影响。 方法 回顾分析2000— 2007年间 201例胸段食管癌根治术后行预防性照射患者资料,比较照射范围差异对生存影响,并对可能影响预后的因素行Cox模型多因素分析。Kaplan Meier法计算OS率,Logrank检验差异。结果 5年随访率为97.0%。全纵隔、全纵隔+胃左区、全纵隔+双锁骨上区、中上纵隔+双锁骨上区、全纵隔+双锁骨上+胃左区照射的 5年OS率分别为21.7%、37.1%、38.7%、34.8%、19.8%( P=0.406),多因素分析结果显示仅仅术后N分期为预后影响因素(P=0.009)。预防性照射后锁骨上淋巴结转移11例、中上纵隔淋巴结转移34例、腹腔淋巴结转移10例。结论 胸段食管癌根治术后预防性照射范围应包括中上纵隔和双锁骨上区。     

Over expression of aromatase protein is highly related to MMPs levels in human breast carcinomas

Estrogens can play a critical role in the development of breast cancer. Aromatase which catalyzes the formation of aromatic C18 estrogens from C19 androgens is regarded to be responsible for the cancer local production of estrogen. Studies not only from aromatase transfected breast cancer cells, but also transgenic mouse, which overexpressed aromatase, demonstrated that in situ produced estrogen plays more important roles than circulating estrogens in breast tumor promotion and progression. Matrix-metalloproteinases (MMPs) have been implicated in the proeolytic process, which play important roles in the aggressiveness of cancer cells including invasion of adjacent tissue and metastasis to distant sites. Expression of MMP2 and 9 may be stimulated by estrogens in hormonal dependent breast cancers, since tumor aromatase can stimulate breast cancer growth and progression in both an autocrine and a paracrine manner. Theoretically aromatase overexpression, that causes relatively high estrogen concentration in situ, may be positively related to MMP2 and 9 expression, indicate worse prognosis in breast cancers, and maybe insensitive to tamoxifen therapy. In the present study, we studied the expression of aromatase activity and MMPs in human breast carcinoma both in vitro and in vivo. In human breast carcinoma cell lines including MCF-7, MDA-MB-231 and MDA-MB-435, the expression of aromatase levels both in mRNA and protein activity was related to MMP2 and MMP9. In humam breast cancer samples, we demonstrated that aromatase expressions were strongly associated with MMP2 and MMP9 levels. It was interesting to observe that the positive relationship was only present in the ER and/or PR positive patients. This may indicate that both MMP2 and MMP9 were up regulated by estrogen produced by aromatase through ER. So in endocrine therapy, either blocking the ER by tamoxifen or inhibiting the aromatase by aromatase inhibitors for example letrozole, may both inhibit tumor growth and lower the metastatic potential especially in ER positive breast cancer patients by means of down-regulation of MMP2/9.     

Effects of salidroside on glioma formation and growth inhibition together with improvement of tumor microenvironment

Objective：To test the effects of salidroside on formation and growth of glioma together with tumor microenvironment.Methods：Salidroside extracted from Rhodiola rosea was purified and treated on human glioma cells U251 at the concentration of 20 μg/mL.3-（4,5-dimethylthiazol-2-yl）-2,5-dephenyltetrazolium bromide （MTT） assay for cytotoxicity and flow cytometry （FCM） for cell cycle analysis were performed.Then for in vivo study,xenotransplantation tumor model in nude mice was generated and treated with salidroside at the concentration of 50 mg/kg.d for totally 20 d.Body weight and tumor size were detected every 2 d after the treatment.The levels of 8-isoprostane,superoxide dismutase （SOD） and malondialdehyde （MDA）,special markers for oxidative stress,were detected while immunofluoresence staining was performed for astrocyte detection.Results：For in vitro study,salidroside could decrease the viability of human glioma cells U251 and the growth of U251 cells at G0/G1 checkpoint during the cell cycle.For in vivo study,salidroside could also inhibit the growth of human glioma tissue in nude mice.The body weight of these nude mice treated with salidroside did not decrease as quickly as control group.In the tumor xenotransplantation nude mice model,mice were found of inhibition of oxidative stress by detection of biomarkers.Furthermore,overgrowth of astrocytes due to the stimulation of oxidative stress in the cortex of brain was inhibited after the treatment of salidroside.Conclusions：Salidroside could inhibit the formation and growth of glioma both in vivo and in vitro and improve the tumor microenvironment via inhibition of oxidative stress and astrocytes.