Common molecular etiologies are rare in nonsyndromic Tibetan Chinese patients with hearing impairment

Background

Thirty thousand infants are born every year with congenital hearing impairment in mainland China. Racial and regional factors are important in clinical diagnosis of genetic deafness. However, molecular etiology of hearing impairment in the Tibetan Chinese population living in the Tibetan Plateau has not been investigated. To provide appropriate genetic testing and counseling to Tibetan families, we investigated molecular etiology of nonsyndromic deafness in this population.

Methods

A total of 114 unrelated deaf Tibetan children from the Tibet Autonomous Region were enrolled. Five prominent deafness-related genes, GJB2, SLC26A4, GJB6, POU3F4, and mtDNA 12S rRNA, were analyzed. Inner ear development was evaluated by temporal CT. A total of 106 Tibetan hearing normal individuals were included as genetic controls. For radiological comparison, 120 patients, mainly of Han ethnicity, with sensorineural hearing loss were analyzed by temporal CT.

Results

None of the Tibetan patients carried diallelic GJB2 or SLC26A4 mutations. Two patients with a history of aminoglycoside usage carried homogeneous mtDNA 12S rRNA A1555G mutation. Two controls were homozygous for 12S rRNA A1555G. There were no mutations in GJB6 or POU3F4. A diagnosis of inner ear malformation was made in 20.18% of the Tibetan patients and 21.67% of the Han deaf group. Enlarged vestibular aqueduct, the most common inner ear deformity, was not found in theTibetan patients, but was seen in 18.33% of the Han patients. Common molecular etiologies, GJB2 and SLC26A4 mutations, were rare in the Tibetan Chinese deaf population.

Conclusion

The mutation spectrum of hearing loss differs significantly between Chinese Tibetan patients and Han patients. The incidence of inner ear malformation in Tibetans is almost as high as that in Han deaf patients, but the types of malformation vary greatly. Hypoxia and special environment in plateau may be one cause of developmental inner ear deformity in this population.     

Use of human amniotic epithelial cells as a feeder layer to support undifferentiated growth of mouse spermatogonial stem cells via epigenetic regulation of the Nanog and Oct-4 promoters

Spermatogonial stem cells (SSCs) are defined by unique properties like other stem cells. However, there are two major challenges: long-term cultivation of normal SSCs into stable cell lines and maintaining the SSCs as undifferentiated and capable of self-renewal. Here, we compared different culture methods for mouse SSCs isolated and cultured from testicular tissue. We found that human amniotic epithelial cells (hAECs) can behave as feeder cells, allowing mouse SSCs to maintain a high level of alkaline phosphatase (AP) activity when cultured long-term. Also, we observed that expression of Nanog, Oct-4 and other important stem cells markers were higher in mouse SSCs cultured on hAECs compared to those cultured on MEF or without any feeder cells. Furthermore, we demonstrated that the CpG islands of the Nanog and Oct-4 promoters were hypomethylated in cells cultured on hAECs. In addition, mouse SSCs cultured on hAECs exhibited higher levels of H3AC and H3K4Me3 in the Nanog and Oct-4 promoters than those cultured on MEF or without feeder cells. Taken together, these results suggest that the hAEC-induced epigenetic modifications at the Nanog and Oct-4 locus could be a key mechanism for maintaining mouse SSCs in an undifferentiated state capable of self-renewal.     

Transverse False Tendons in the Left Ventricular Cavity Are Associated with Early Repolarization

BackgroundLeft ventricular false tendons (LVFTs) are related to precordial murmurs, ventricular arrhythmias and some repolarization abnormalities. Early repolarization (ER) is a specific type of repolarization abnormality.ObjectiveThe aim of the present study was to investigate the relationship between LVFTs and ER.MethodsThis study retrospectively included 99 consecutive healthy subjects and 33 patients with ER. Early repolarization was defined as an elevation of the QRS-ST junction of >0.1 mV from baseline in at least 2 inferior or lateral leads, manifested as QRS slurring or notching. Each participant was examined using echocardiography with second harmonic imaging, and the attachments of the LVFTs were recorded.ResultsA total of 93 LVFTs were present in 82 (83%) of the 99 healthy subjects. Of these 93 LVFTs, the majority (79/93, or 84.9%) were longitudinal-type LVFTs, which originated from the basal interventricular septum (IVS) and progressed toward the apical segment of the left ventricular free wall. There were significant differences in the positioning of the LVFTs between the ER patients and control (P < 0.0001). LVFTs between mid-IVS to the middle of the LV free wall were found more common in patients with ER compared with control subjects (47.5% vs. 6.5%, P < 0.0001). In the ER group, LVFTs between the basal IVS to the apical segment of LV free wall were only identified in 21% of the LVFTs, compared to a value of 84.9% for the control group (P < 0.0001). The distribution of LVFT trends in the ER group was also significantly different from that in the control group (P < 0.05).ConclusionsLVFTs are commonly visualized using echocardiography. An LVFT from the basal IVS to the apical segment of the left ventricular free wall may be a normal anatomical structure in the left ventricular cavity. On the contrary, transverse false tendons in the left ventricular cavity may be associated with ER.     

Effects of 17α‐methyltestosterone on the growth performance,development, and reproduction of gynogenetic topmouth culter (Culter alburnus Basilewsky)

This study investigated the effects of 17α‐methyltestosterone (17α‐MT) on the physiological properties of gynogenetic topmouth culter (Culter alburnus). Five 17α‐MT concentration (0, 10, 25, 40, and 60 μg/g) groups were set up in Experiment 1, and three treatment and recovery methods of the above five concentrations were studied in Experiment 2. In Experiment 3, fish were fed with increasing concentrations of 17α‐MT (from 40 μg/g to 140 μg/g) in an outdoor pond. In Experiment 1, growth was promoted during the initial 60 days in the 10 μg/g group, whereas inhibition was first observed at 20 days after treatment in the 60 μg/g group. Increased concentration (40–60 μg/g) of 17α‐MT significantly inhibited the expression of FOXL2 and CYP19a gene but significantly activated the expression of AMH and DMRT1 gene. The levels of superoxide dismutase, estradiol, testosterone, growth hormone, and tetraiodothyronine in the 40 and 60 μg/g groups were significantly higher than those in the other concentration groups at 40 days after treatment. In Experiment 2, almost all indexes detected besides the expression of reproduction‐related genes in the 17α‐MT treatment groups were recovered to those in the control groups. All gonads observed in Experiments 1 and 2 remained in a primordial state after treatment with different concentrations of 17α‐MT. In Experiment 3, 100% of the detected gonads developed into testes at 126 and 460 days post‐fertilization after three‐month outdoor 17α‐MT induction. These results suggest that 17α‐MT affects the physiological processes related to sexual differentiation in topmouth culter and that feeding 17α‐MT with increasing dose in an outdoor pond provides an effective protocol for producing neo‐male topmouth culter.     

油茶果生炭疽菌小分子GTP酶Rab7的功能研究

【目的】炭疽病是油茶的一种重要病害,果生炭疽菌是油茶炭疽病的主要致病菌。本文对果生炭疽菌小分子GTP酶Rab7进行研究,为油茶炭疽病的防控治理提供依据。【方法】构建CfRAB7基因敲除载体,通过PEG介导的原生质体转化、抗性筛选和PCR电泳验证获得果生炭疽菌突变体菌株△Cfrab7和互补菌株△Cfrab7/CfRAB7。进一步分析CfRAB7基因敲除突变体△Cfrab7的生长、产孢、附着孢的形成、胁迫应答、液泡融合和致病力等生物学表型。【结果】在PDA和MM培养基上,突变体△Cfrab7的菌落直径显著减小,产孢量和附着孢形成率显著降低,且不能穿透玻璃纸;在10mmol/LH2O2条件下,△Cfrab7生长受到明显抑制;进一步研究发现突变体△Cfrab7液泡无法正常融合,在油茶有伤和无伤的幼叶上均不发病。【结论】CfRAB7基因参与调控果生炭疽菌生长产孢、附着孢形成、H2O2胁迫应答、液泡融合和致病力。     

基于水分控制的切花百合生长预测模型

光合作用与干物质生产是观赏植物外观品质形成的基础。水分是影响植物光合作用与干物质生产的重要因子。为定量研究水分对切花百合光合作用与干物质生产的影响,以切花百合品种‘索邦’(Lilium‘Sorbonne’)为试验材料,于2009年3月至2010年1月在南京的连栋温室内开展了不同定植期和不同水分处理的栽培试验,以基于光温的温室花卉生长动态预测模型为基础,定量分析了不同定植期和不同水分处理条件下切花百合叶面积指数、光合速率和干物质生产的动态影响,并确定了切花百合正常生长的临界基质水势,建立了基质水势对切花百合光合速率和干物质生产影响的动态预测模型。结果表明,本文所建模型对切花百合叶片最大总光合速率和植株总干质量的预测效果较好,模型对叶面积指数、叶片最大总光合速率和植株干物质量的预测值与实测值之间的决定系数(r2)分别为0.97,0.96,0.94,相对根均方差(rRMSE)分别为7.12%、4.37%、11.14%。该模型能较好地预测水分对切花百合叶片最大总光合速率和植株总干质量的动态影响,可为进一步优化切花百合生产的水分管理提供决策支持。     

Doxorubicin hydrochloride enhanced antitumour effect of CEA‐regulated oncolytic virotherapy in live cancer cells and a mouse model

Oncolytic adenovirus (OA) has attracted increasing attention due to their specific proliferation in tumour cells and resulting in lysis of tumour cells. To further improve the antitumour effect of OA, in this study, we combined CD55‐TRAIL‐IETD‐MnSOD (CD55‐TMn), a CEA‐controlled OA constructed previously, and chemotherapy to investigate their synergistic effect and possible mechanisms. MTT assay was performed to detect antitumour effects. Hoechst 33 342 and flow cytometric analysis were used to examine cell apoptosis. Western blotting was performed to examine cell pyroptosis and apoptosis mechanism. Animal experiment was used to detect antitumour effect of doxorubicin hydrochloride (Dox) combined with CD55‐TMn in vivo. We firstly found that Dox promotes gene expression mediated by CEA‐regulated OA and virus progeny replication by activating phosphorylation of Smad3, and Dox can enhance antitumour effect of CEA‐regulated CD55‐TMn by promoting cell apotopsis and cell pyroptosis. Thus, our results provide an experimental and theoretical basis on tumour therapy by combination treatment of the oncolytic virotherapy and chemotherapy and it is expected to become a novel strategy for liver cancer therapy.     

The Motilin Gene Evolved a New Function in Kangaroo Rats and Kangaroo Mice (Dipodomyinae)

The motilin receptor gene was lost in the ancestral lineage of rodents. Subsequently, the gene encoding its ligand, motilin, has experienced different evolutionary fates. Previous genomic analyses had shown that the motilin gene (MLN) became a pseudogene independently in the lineages leading to the guinea pig and the common ancestor of the mouse and rat, yet an intact, and thus potentially functional, open reading frame for the MLN was preserved in the Dipodomys ordii genome. As only a single MLN haplotype from D. ordii was available, and this sequence is from a low coverage draft genome, it is possible that the intact MLN found in the draft kangaroo rat genome is an artifact, or represents an intermediate in the process of becoming a pseudogene. In order to establish whether an intact MLN is retained in kangaroo rats despite the loss of its specific receptor, and to investigate the evolutionary mechanisms underlying the retention of this gene sequence, we isolated MLN sequences from species that represent the diversity of the Dipodomyinae [the monophyletic Dipodomyinae subfamily consists of two genera: Dipodomys (kangaroo rats) and Microdipodops (kangaroo mice)]. The results demonstrate that the MLN sequence is well conserved in Dipodomyinae, and it codes for a predicted motilin peptide sequence possessing a conserved N-terminal pharmacophore and the potential to be processed and secreted as a hormone. The observations that the MLN evolved as a functional gene during the radiation of the Dipodomyinae, species that have lost their original motilin receptor, suggest that the MLN has undergone a lineage-specific physiological adaptation to a new function.