46, XY gonadal dysgenesis: new SRY point mutation in two siblings with paternal germ line mosaicism

Stoppa‐Vaucher S, Ayabe T, Paquette J, Patey N, Francoeur D, Vuissoz J‐M, Deladoëy J, Samuels ME, Ogata T, Deal CL. 46, XY gonadal dysgenesis: new SRY point mutation in two siblings with paternal germ line mosaicism. Familial recurrence risks are poorly understood in cases of de novo mutations. In the event of parental germ line mosaicism, recurrence risks can be higher than generally appreciated, with implications for genetic counseling and clinical practice. In the course of treating a female with pubertal delay and hypergonadotropic hypogonadism, we identified a new missense mutation in the SRY gene, leading to somatic feminization of this karyotypically normal XY individual. We tested a younger sister despite a normal onset of puberty, who also possessed an XY karyotype and the same SRY mutation. Imaging studies in the sister revealed an ovarian tumor, which was removed. DNA from the father's blood possessed the wild type SRY sequence, and paternity testing was consistent with the given family structure. A brother was 46, XY with a wild type SRY sequence strongly suggesting paternal Y‐chromosome germline mosaicism for the mutation. In disorders of sexual development (DSDs), early diagnosis is critical for optimal psychological development of the affected patients. In this case, preventive karyotypic screening allowed early diagnosis of a gonadal tumor in the sibling prior to the age of normal puberty. Our results suggest that cytological or molecular diagnosis should be applied for siblings of an affected DSD individual.     

4-苯基丁酸抗内质网应激诱导胰岛β细胞凋亡

目的 探讨4-苯基丁酸（4-PBA）对2型糖尿病（T2DM）大鼠胰岛β细胞内质网应激（ERS）诱导的细胞凋亡的保护作用。方法 高脂饮食喂养加小剂量链脲佐菌素（STZ）腹腔注射建立SD大鼠T2DM模型,造模成功后取其中10只给予4-PBA灌胃20d。放射免疫法检测各组大鼠游离脂肪酸（FFA）、血糖及胰岛素的变化。醛品红染色观察胰岛形态改变,RT-PCR检测内质网相关蛋白（Caspase-3、GRP78、CHOP）的mRNA表达变化。结果 高脂对照组（EM> n/EM>＝10）和T2DM组（ EM>n/EM>＝8）大鼠血清FFA比正常对照组（EM>n/EM>＝10）显著增加,4-PBA治疗后显著下降（ EM>n/EM>＝8）。4-PBA治疗升高了T2DM造成的     

Comparative Evaluation of Two Hemagglutinating Encephalomyelitis Coronavirus Vaccine Candidates in Mice

Porcine hemagglutinating encephalomyelitis (PHE) is caused by the coronavirus hemagglutinating encephalomyelitis virus (PHE-CoV), and the recent, rapid spread of PHE-CoV in piglets from many countries emphasizes the urgent need for a PHE-CoV vaccine. Here we use a murine model for evaluation of the induction of humoral and cellular immune responses by inactivated and PHE-CoV DNA vaccines in order to define the immune correlates for protection against PHE-CoV. The inactivated vaccine was composed of purified PHE-CoV and aluminum hydroxide gel (alum), which was chosen as an adjuvant because of its long history of safety for human use. The PHE-CoV DNA vaccine was constructed by subcloning the S1 gene of PHE-CoV into the pVAX1 vector to create the recombinant plasmid pV-S1. Our results showed that the inactivated PHE-CoV vaccine (IPV) elicited a high level of humoral immunity, resulting in good protection efficacy against PHE-CoV challenge. The IPV induced the IgG1 subclass of serum antibodies and expression of the cytokine interleukin-4 (IL-4), suggesting that the IPV generated a predominantly Th2-type immune response. The DNA vaccine was found to mediate primarily a cellular immune response with high levels of IgG2a and the cytokines IL-2 and gamma interferon (IFN-γ). However, mice that were vaccinated twice with the DNA vaccine and boosted with the IPV could mount a sufficient neutralizing antibody response against live PHE-CoV, with little variation in IgG1 and IgG2a levels, and showed high levels of IL-2 and IL-4. This response may activate both B and T cells to mount a specific humoral and cellular immune response that could, in turn, elicit a phagocyte-mediated defense against PHE-CoV infections to achieve viral clearance.     

云南苯丙氨酸羟化酶基因点突变及外显子5内新突变Y166X(C→G)的鉴定

目的研究云南苯丙氨酸羟化酶基因点突变分布概况,以提高该地区苯丙酮尿症（ＰＫＵ）的基因诊断率。方法应用ＰＣＲ－ＡＳＯ探针斑点杂交,ＰＣＲ－ＳＳＣＰ、ＡＳＰＣＲ和ＤＮＡ直接测序等技术,对１３／１４名云南籍ＰＫＵ患儿的苯丙氨酸羟化酶（ＰＡＨ）基因外显子４、５、６、７、１０、１１和１２进行了鉴定分析。结果共检出５种错义突变：Ｒ２４３Ｑ（５／２６）、Ｙ２０４Ｃ（３／２８）、Ｒ４１３Ｐ（２／２８）、Ｔ４１８Ｐ（１／２８）及Ｇ２４７Ｖ（１／２６）,３种无义突变Ｙ１６６Ｘ（Ｃ→Ｇ）（２／２６）、Ｗ３２６Ｘ（１／２８）及Ｙ３５６Ｘ（２／２６）和１种静止突变Ｖ３９９Ｖ（２／２６）。经检索国际ＰＡＨ基因突变数据库,其中Ｙ１６６Ｘ（Ｃ→Ｇ）为首次发现的新突变。结论云南藉ＰＫＵ与中国北方人群有相类似的最常见的ＰＡＨ基因突变类型（Ｒ２４３Ｑ、Ｙ２０４Ｃ、Ｙ３５６Ｘ、Ｖ３９９Ｖ和Ｒ４１３Ｐ）,但与南方人群ＰＡＨ基因突变特点则不同。该发现有助于提高云南ＰＫＵ的基因诊断率,并对我国ＰＡＨ基因突变不同地区及人群的分布、起源研究有参考价值。     

戈壁滩驻训官兵变应性鼻炎患者睡眠质量与应对方式研究

目的研究戈壁滩驻训官兵变应性鼻炎患者睡眠质量情况及应对方式。方法采用匹兹堡睡眠质量指数(PSQI)和特质应对方式问卷(TCSQ),对戈壁滩驻训官兵被确诊为变应性鼻炎(AR)70例进行测评调查。结果 1驻训官兵AR患者PSQI各因子分值较驻地健康官兵差异显著(P<0.01);2驻训官兵AR患者的应对方式与驻地健康官兵比较差异显著(P<0.01);3消极应对方式与PSQI呈显著负相关(P<0.05或P<0.01)。结论戈壁滩驻训官兵AR患者睡眠质量与消极应对方式呈显著负相关。     

人着色性干皮病D组基因对白细胞介素-6促进人血管平滑肌细胞增殖作用的影响

目的: 探讨人着色性干皮病D组基因(XPD)对白细胞介素-6(IL-6)促进人血管平滑肌细胞(VSMCs)增殖作用的影响。方法: 用脂质体转染法将重组质粒pEGFP-N2/XPD和空质粒pEGFP-N2稳定转染VSMCs,然后给予1×10⁵ U/L IL-6孵育48 h。实验分为6组:(1)空白对照组;(2)pEGFP-N2组;(3)pEGFP-N2/XPD组;(4)IL-6组;(5)IL-6 + pEGFP-N2组;(6)IL-6 + pEGFP-N2/XPD组。用荧光显微镜观察绿色荧光蛋白报告基因表达情况;用MTT法观察细胞增殖活力;用流式细胞仪检测细胞周期和凋亡率;用RT-PCR和Western blotting检测XPD、Bcl-2、Bax和野生型P53(wt-P53)表达量的变化。结果: 在荧光显微镜下,可在转染了重组质粒pEGFP-N2/XPD或空质粒pEGFP-N2的细胞中观察到绿色荧光,即转染成功;MTT结果显示,重组质粒pEGFP-N2/XPD的转染抑制了细胞增殖活力(P<0.05),并能抑制IL-6促进VSMCs增殖的作用(P<0.05);流式细胞仪结果显示,重组质粒pEGFP-N2/XPD的转染引起了细胞G₀/G₁期增加(P<0.05)、S期减少(P<0.05)、凋亡率增加(P<0.01),并能抑制IL-6促进VSMCs G₀/G₁期减少、S期增加、凋亡率降低的作用(P<0.01);RT-PCR和Western blotting检测发现,重组质粒pEGFP-N2/XPD的转染使得XPD表达增高(P<0.05或P<0.01),同时Bcl-2表达降低,Bax和wt-P53表达增高(P<0.05或P<0.01),并能抑制IL-6促进VSMCs的Bcl-2表达增高、Bax和wt-P53表达降低的作用(P<0.05或P<0.01)。结论: XPD能抑制VSMCs增殖并促进其凋亡,并能抑制IL-6促进VSMCs增殖和降低其凋亡的作用,有望成为治疗动脉粥样硬化的靶点。     

肾上腺髓质素真核表达载体对皮瓣缺血再灌注损伤的影响

背景：肾上腺髓质素可在心、脑、肾、肝等器官的缺血再灌注过程中起保护作用。 目的：观察肾上腺髓质素真核表达载体对大鼠腹部皮瓣缺血再灌注损伤的影响及其作用机制。 方法：构建大鼠肾上腺髓质素真核表达载体,将SD大鼠随机分假手术组、模型组、维拉帕米组、重组质粒组。重组质粒组腹部皮内注射肾上腺髓质素真核表达载体,其余各组注射等量生理盐水,各组注射4次后建立缺血再灌注模型,维拉帕米组于皮瓣内注射维拉帕米。 结果与结论：与模型组相比,重组质粒组皮瓣组织中丙二醛、血管内皮素1的含量分别降低了38.50%(P  < 0.01),54.73%  (P  < 0.01),超氧化物歧化酶的含量增加了50.67%(P < 0.05)。皮瓣组织学检查结果显示重组质粒组皮瓣炎性细胞浸润与水肿程度也较模型组明显减轻。结果证实肾上腺髓质素真核表达载体对大鼠皮瓣缺血再灌注损伤具有保护作用,其机制与减轻脂质过氧化及减少炎症细胞浸润有关。     

FCεRI gene promoter polymorphisms and total IgE levels in susceptibility to atopic dermatitis in Korea

IgE-dependent activation of mast cells and basophils through the high-affinity IgE receptor (FcεRI) is involved in the pathogenesis of allergen-induced immune responsiveness in atopic diseases like atopic dermatitis (AD). We sought to determine FcεRI gene polymorphisms are associated with AD in Korean patients, and analyzed the relevance of FcεRI gene polymorphisms and serum IgE levels. We conducted a case-control association analysis (175 patients and 56 controls) of Korean subjects. Genotyping was performed using the TaqMan fluorogenic 5' nuclease assay, and serum levels of IgE were measured using a fluorescence enzyme immunoassay. We found that there were no significant relationships between FcεRI and AD, although there were trends towards an association between the 66T>C (rs2251746) polymorphism and total serum IgE levels in the Korean AD patients. In conclusion, while the 66T>C (rs2251746) of the FcεRIα polymorphism may be linked to AD and higher serum IgE levels, polymorphisms in the FcεRIβ gene did not confer susceptibility to AD in our patient sample.     

Tamoxifen enhances therapeutic effects of gemcitabine on cholangiocarcinoma tumorigenesis

Cholangiocarcinoma is a highly malignant tumor with limited therapeutic options. We have previously reported that tamoxifen (TMX) induces apoptosis of cholangiocarcinoma cells and reduces cholangiocarcinoma tumorigenesis in mice. In the present studies, we determined the effect of combination therapy of TMX and gemcitabine (GMT), another chemotherapeutical reagent for many cancers, on cholangiocarcinoma tumorigenesis and investigated the responsible mechanisms. GMT inhibited cell growth and induced apoptosis of cholangiocarcinoma cells in a concentration-dependent manner. TMX enhanced GMT-induced apoptosis of cholangiocarcinoma cells. Consistently, GMT (15?mg/kg) inhibited cholangiocarcinoma tumorigenesis in nude mice by 50%. TMX (15?mg/kg) enhanced the inhibitory effect of GMT on tumorigenesis by 33%. The inhibition of tumor growth correlated with enhanced apoptosis in tumor tissues. To elucidate the mechanisms underlying the additive effects of TMX on GMT-induced apoptosis, we determined the activation of caspases in cholangiocarcinoma cells exposed to GMT, TMX, or both. Activation of caspases 9 and 3, as well as cytochrome c release to the cytosol, was demonstrated in cells exposed to both reagents. In contrast, TMX activated caspase 2, whereas GMT had no effect. Inhibition of caspase 2 activation decreased TMX-, but not GMT-, induced activation of caspase 3 and apoptosis of cholangiocarcinoma cells. Similarly, activation of caspase 2 was found in tumors from TMX-treated mice, but not GMT-treated mice. Therefore, the enhanced effect of TMX on GMT-induced cholangiocarcinoma cell death is partially mediated by activation of caspase 2. TMX and GMT both induce apoptosis and inhibit cholangiocarcinoma tumorigenesis, which may be attributed to the activation of distinct apoptosis signals by TMX and GMT. Our studies provide in vivo evidence and molecular insight to support the use of TMX and GMT in combination as an effective therapy for cholangiocarcinoma.