Hypoxia-activated Ca2+ currents in pacemaker neurones of rat rostral ventrolateral medulla in vitro.

We examined the effects of brief periods of hypoxia or application of cyanide on the discharge and membrane properties of medullary pacemaker neurones in slices of the rostral ventrolateral reticular nucleus (RVL) of the medulla oblongata of rats. Stable intracellular recordings were obtained from seventy-nine neurones within the RVL which exhibited spontaneous rhythmic discharge in the absence of excitatory postsynaptic potentials (EPSPs). The membrane potential cycles of these neurones could be reset with an evoked spike without eliciting EPSPs or inhibitory postsynaptic potentials and hence met criteria of RVL pacemaker neurones. Hypoxia, produced by reducing O2 from 95 to 20% for 40 s or exposure to cyanide (30-300 microM for 40 s), reversibly increased neuronal discharge 1.6-fold (20% O2) or 2.6-fold (300 microM cyanide), respectively, in association with membrane depolarization and a significant fall in membrane resistance. The membrane responses to hypoxia and cyanide were observed in the presence of tetrodotoxin (TTX) at a concentration (10 microM) which eliminated spontaneous spikes or spikes evoked by intracellular depolarization. When recorded at a holding potential of -70 mV by single-electrode voltage clamp, hypoxia or cyanide (300 microM) elicited inward currents of 0.44 +/- 0.06 and 0.58 +/- 0.08 nA, respectively, which are attenuated by reducing the concentration of extracellular Ca2+ ions, and abolished by 2 mM CoCl2 and 100 microM NiCl2, but not affected by 50 microM CdCl2, replacement of 83% extracellular Na+, or adenosine deaminase (2U ml-1). We conclude that hypoxia and cyanide directly excite RVL pacemaker neurones in vitro by a common mechanism: activation of Ca2+ channel conductance.     

Monoclonal antibodies against selected peanut allergens: Production and use as affinity agents

An extract of dry‐roasted commercial peanut mix (CPE) was examined for allergenic activity in peanut‐sensitive individuals, using skin tests and radioallergosorbent (RAST) assays. Proteins in the extract were characterized by sodium dodecyl sulfate polyacry‐lamide gel electrophoresis (SDS‐PAGE) and immunoblotting. The proteins were electro‐eluted in three fractions in the ranges 15–25, 26–58 and 65 kDa. The 15–25 kDa molecular weight fraction produced the most reactive skin tests in peanut‐sensitive subjects and was chosen for monoclonal antibody production. Six hybridoma cell lines secreting peanut‐specific antibodies of the IgM isotype (kappa light chain) were produced. Immunopurified CPE proteins were then subjected to SDS‐PAGE, resulting in five major bands with approximate molecular weights of 14, 25, 38, 40 and 44 kDa. Immunoblotting of these separated proteins revealed: (1) three bands with approximate molecular weights of 38, 44 and 65 kDa, which bound IgE from peanut‐sensitive patients; and (2) that the monoclonal antibodies recognized epitopes in bands at approximate molecular weights of 12, 14, 23 and 25 kDa. RAST inhibition assays showed that the affinity‐purified proteins were able to inhibit the binding of serum IgE from peanut‐allergic individuals to solid‐phase CPE.     

人不同来源T细胞对丝裂原、PMA和A23187的反应性比较

本文比较了胎儿胸腺细胞、正常成人外周血Ｔ细胞及儿童扁桃体Ｔ细胞对丝裂原ＰＷＭ、ＰＨＡ和ＣｏｎＡ、ＰＫＣ激活剂ＰＭＡ和Ｃａ２＋导入剂Ａ２３１８７的反应性。结果表明：胎儿胸腺细胞对ＰＷＭ的反应对ＰＨＡ和ＣｏｎＡ的反应性很弱；正常成人外周血Ｔ细胞对ＰＨＡ的反应性最强，对ＰＷＭ和ＣｏｎＡ的反应性较弱；儿童扁桃体Ｔ细胞对３种丝裂原的反应性均很强；ＰＭＡ单独作用能强烈诱导儿童扁桃体Ｔ细对正常成人外周血Ｔ细胞及胸腺细胞作用较弱；ＰＷＭ和ＰＭＡ对胎儿胸腺细胞及正常成人外周血Ｔ细胞均有明显协同作用，但对儿童扁桃体Ｔ细胞无协同作用；Ａ２３１８７单独作用对３种Ｔ细胞的增殖作用均很弱，Ａ２３１８７对ＰＷＭ诱导的增殖均有部分抑制作用。这些结果对于认识处于不同发育阶段及不同部位的的活化途径和功能有一定意义。     

Is chemotherapy always necessary for patients with nonmetastatic gestational trophoblastic neoplasia with histopathological diagnosis of choriocarcinoma?

Objective

To evaluate expectant management versus immediate chemotherapy following pathological diagnosis of gestational choriocarcinoma (GCC) in patients with nonmetastatic disease.

Clinical characteristic and intraoperative findings of uterine perforation patients in using of intrauterine devices (IUDs)

Background

Intrauterine devices (IUDs) are the most popular form of contraception used worldwide; however, IUD is not risk-free. IUD migrations, especially uterine perforations, were frequently occurred in patients. The aim of this study was to investigate the clinical characteristics and intraoperative findings in patients with migrated IUDs.

Results

29 cases of uterine perforation associated with migrated IUDs and 69 control patients were followed between January 2008 to March 2015. Patients who used IUDs within first 6 months from the last delivery experienced a characteristically high rate of the perforation of the uterine wall. A significantly larger number of IUD insertion associated with uterine perforation were performed in rural hospitals or operated at a lower level health care system. There was no clear difference in the age and presented symptoms in patients between two groups. Majority of contraceptive intrauterine devices was the copper-releasing IUDs. Furthermore, patients who used V-shaped IUD showed significantly higher incidence of pelvic adhesions when compared with the users of O-shaped IUDs.

Conclusions

Unique clinical characteristics of IUD migration were identified in patients with uterine perforation. Hysteroscopy and/or laparoscopy were the effective approaches to remove the migrated IUDs. Improving operating skills is required at the lower level of health care system.

     

Preeclampsia and maternal risk of breast cancer: a meta-analysis of cohort studies

Background: Pregnancy-related hypertensive disorders, including preeclampsia (PE) and pregnancy-induced hypertension (PIH), may influence the maternal risk of breast cancer. However, results of the cohort studies were inconsistent.

Methods: An updated meta-analysis of cohort studies was performed to evaluate the association between PE, PIH and maternal breast cancer incidence. Relevant studies were identified via searching of PubMed and Embase databases. A random effect model was applied to synthesize the results. Stratified analyses were performed to evaluate the potential influence of parity, gender of offspring, and study design on the association between PE and maternal breast cancer incidence.

Results: Ten cohort studies with 2,417,899 pregnant women were included. Maternal risk of breast cancer was not significantly affected by PE (risk ration [RR]?=?0.93, 95% confidence interval [CI]: 0.82–1.06, p?=?.27), or PIH (RR?=?0.95, 95% CI: 0.81–1.12, p?=?.54). Interestingly, PE was associated with significantly lowered maternal incidence of breast cancer in women who give birth to male offspring (RR?=?0.79, p?p?Conclusions: Current evidence did not support a conclusive association between PE, PIH and the maternal risk of breast cancer. Gender of the offspring may influence the association between PE and maternal breast cancer incidence.     

Prenatal diagnosis of Wolf–Hirschhorn syndrome: from ultrasound findings,diagnostic technology to genetic counseling

Purpose

Wolf–Hirschhorn syndrome (WHS) is a contiguous gene syndrome due to terminal chromosome 4p deletions. We explored prenatal diagnosis of WHS by ultrasound as well as karyotype and single nucleotide polymorphism array (SNP array) to characterize the structural variants of WHS prenatally.

Methods

Ten prenatal cases of WHS were evaluated for the indication of the invasive testing, the ultrasound features, and cytogenetic and microarray results.

Results

Eight cases were diagnosed by karyotyping and SNP array, while two cases were detected only by SNP array. Combining our cases with 37 prenatal cases from the literature, the most common sonographic features were IUGR (97.7%) and typical facial appearance (82.9%). Other less common phenotypes included renal hypoplasia (36.2%), cardiac malformation (29.8%), cleft lip and palate (25.5%), cerebral abnormalities (25.5%), skeletal anomalies (21.3%), and increased nuchal translucency/nuchal fold thickness (NT/NF) (19%).

Conclusions

The most common intrauterine phenotypes of WHS were severe IUGR and typical facial appearance with other less consistent ultrasound findings. Noninvasive prenatal testing (NIPT) is one very promising screening tool for WHS. SNP array can improve diagnostic precision for detecting WHS, especially for the cryptic aberrations that cannot be identified by the traditional karyotyping. Ectopic kidney may be a previously unrecognized phenotype of WHS.

     

Ⅰ型星形胶质细胞条件培养液抑制H2O2诱导PC12细胞凋亡机制

目的: 探讨Ⅰ型星形胶质细胞条件培养液(ACM-1)对H2O2诱导PC12细胞凋亡的抑制作用与硫氧还蛋白(Trx)表达的关系.方法: 采用MTT法检测ACM-1预处理H2O2诱导PC12细胞的活力.流式细胞术,Real time-PCR法及免疫印迹法分别检测细胞凋亡率,Trx-1和Trx-2mRNA的变化及Trx和Bcl-2蛋白水平的变化.结果: 20%浓度的ACM-1能提高H2O2诱导PC12细胞的活力,细胞凋亡率下降,Trx-1的mRNA量升高,Trx和Bcl-2的蛋白量增加.结论: ACM-1对H2O2诱导PC12细胞凋亡的抑制作用可能是通过Trx上调介导的,而Trx的抗凋亡作用和Bcl-2的上调有关.     

A Novel Missense Mutation in the Nuclear Factor-κB Essential Modulator (NEMO) Gene Resulting in Impaired Activation of the NF-κB Pathway and a Unique Clinical Phenotype Presenting as MRSA Subdural Empyema

Introduction  

We describe a previously unreported 437 T→G missense mutation producing a V146G substitution in the first coiled-coil (CC1) domain of nuclear factor-κB essential modulator (NEMO) in a 9-month-old boy with ectodermal dysplasia with immunodeficiency who presented with methicillin-resistant Staphylococcus aureus subdural empyema. We performed in vitro experiments to determine if this novel mutation resulted in impaired NF-κB signaling.     

Functional evaluation of the role of reticuloendotheliosis virus long terminal repeat (LTR) integrated into the genome of a field strain of Marek's disease virus

MDV-GX0101 is a field strain of Marek's disease virus with a naturally occurring insertion of the reticuloendotheliosis virus (REV) LTR fragment. In order to study the biological properties of REV-LTR insertion in the MDV genome, we constructed a full-length infectious BAC clone of MDV-GX0101 strain and deleted the LTR sequences by BAC mutagenesis. The pathogenic properties of the LTR-deleted virus were evaluated in infected SPF birds. The study demonstrated that the LTR-deleted virus had a stronger inhibitory effect on the growth rates of the infected birds and induced stronger immunosuppressive effects. Surprisingly, however, the ability for horizontal transmission of the LTR-deleted virus appeared to be significantly weaker than its parental LTR-intact virus. Even though the precise molecular mechanisms are still not clear, the results of our studies demonstrate that the retention of the REV-LTR in the MDV genome decreases its pathogenic effects but increases its potential for horizontal transmission.