[Phenotype and genotype analysis of a pedigree affected with Joubert syndrome due to variant of TMEM237 gene].

OBJECTIVE: To explore the pathogenesis of two siblings (including a fetus) from a pedigree affected with Joubert syndrome. METHODS: Peripheral blood samples of the proband and his parents as well as amniotic fluid and abortion tissues of the fetus were collected. Part of the samples were used for the extraction of DNA, and whole exome sequencing (WES) was carried out to screen potential variants in the proband and his parents. Suspected variants were subjected to bioinformatics analysis with consideration of the clinical phenotype, and were verified by Sanger sequencing of the proband, fetus and their parents.The remainders were used for the extraction of RNA, and the mechanism of splicing variant was validated by reverse transcription-PCR (RT-PCR). RESULTS: WES showed that both patients have carried c.175C>T (p.R59X) and c.553+1G>A compound heterozygous variants of the TMEM237 gene. Among these, c.175C>T was a nonsense mutation inherited from the asymptomatic mother, while c.553+1G>A was an alternative splicing mutation inherited from the asymptomatic father. RT-PCR showed that this variant has resulted in aberrant splicing by exon skipping. CONCLUSION: The compound heterozygous variants of the TMEM237 gene probably underlay the etiology of Joubert syndrome in this pedigree. Above finding has enriched the phenotype and variant spectrum of the TMEM237 gene, and facilitated genetic counseling and prenatal diagnosis for the family.

Enrichment of adenosine using thermally responsive chromatographic materials under friendly pH conditions.

A thermally responsive boronate affinity chromatographic material, which showed thermal sensitivity, had been successfully applied for the enrichment and separation of cis-diol-containing compounds, and the capture and release process could be facilitated by adjusting the temperature. However, in this system, the pH of the mobile phase must be higher than 9.8, and alkaline media can lead to the degradation of labile compounds; the use of silica beads also limits its use. In this study, thermally responsive boronate affinity chromatographic material, namely poly(N-isopropylacrylamide-co-N-acryloyl-3-aminophenylboronic acid) grafted silica, was successfully prepared by atom transfer radical polymerization. Its structure was confirmed by IR spectroscopy and the graft ratio was 20.8%, determined by thermogravimetric analysis. Furthermore, the capture/release of adenosine, a cis-diol, was performed from pH 5.0-9.0 and 10-50°C. The elution of adenosine was remarkably retarded at decreased temperatures and adenosine could be captured completely at 10°C at pH values of 5.0-9.0. The enrichment of adenosine could be achieved by simply changing the temperature from 10 to 50°C. Therefore, this material not only improved the stability of the silica, but was also suitable for the capture of oxidation-sensitive biological analytes. Moreover, it could be used for the enrichment of cis-diol-containing compounds in LC with MS.

Effects of ß-conglycinin on growth performance, immunoglobulins and intestinal mucosal morphology in piglets.

One of the main causes of allergic reactions in young animals is ß-conglycinin, an antigenic glycoprotein found in soya beans. Therefore, the objective of the study was to investigate the effects of a prior immunisation with ß-conglycinin on growth performance, serum immunoglobulin levels and intestinal histology in piglets. Forty piglets (7 d of age) were randomly divided into four groups of ten piglets each. Piglets of Groups Im and Im+S were immunised twice by hypodermic injection with ß-conglycinin at 500 µg/kg body weight (BW) at day 7 and 21 of age. At day 23, Groups Im+S and S were intramuscularly injected with 5000 µg ß-conglycinin per kg BW. The piglets of Group C received a physiological saline solution by hypodermic injection. All piglets were weaned at the age of 23 d and blood samples were taken on days 7, 21 and 35. At the end of the trial, five piglets per group were slaughtered and the intestine was collected for evaluating mucosal histology. Compared to Group C, in Group S the average daily gain (ADG), feed intake and gain:feed ratio were decreased (p < 0.01), and serum levels of IgG and IgE were increased (p < 0.01). Furthermore, in this group the structure of duodenal and jejunal mucosa was severely damaged. But in Groups Im and Im+S the ADG was increased (p < 0.05), serum IgE levels were decreased (p < 0.01) and the intestinal mucosa was not damaged. The results suggest that prior immunisation with ß-conglycinin can increase ADG and serum IgG levels and decrease serum IgE levels. Therefore, this method is also potentially able to protect the structural integrity of the intestinal mucosal epithelia and alleviate allergic reactions in piglets.

Typical hemophagocytic syndrome associated with cytomegalovirus infection in an immunocompetent patient: a case report and literature review. / ä¸€ä¾‹èŠ¦å¯æ›¿å°¼æˆåŠŸæ•‘æ²»å ç–«åŠŸèƒ½æ­£å¸¸æ‚£è€ å› å·¨ç»†èƒžç— æ¯’æ„ŸæŸ“å¯¼è‡´å—œè¡€ç»†èƒžç»¼åˆå¾çš„ç— ä¾‹æŠ¥å‘ŠåŠæ–‡çŒ®å›žé¡¾.

Cytomegalovirus (CMV) infection is currently prevalent in populations throughout the world, and 56%|-94% of the global population is seropositive for CMV. CMV infection mainly affects immunocompromised hosts. In these cases, it can cause significant symptoms, tissue-invasive disease, and many sequelae including death (Dioverti and Razonable, 2016). The vast majority of healthy adults with CMV infection experience an asymptomatic course; when symptomatic, it manifests as a mononucleosis-like syndrome in approximately 10% of patients (Sridhar et al., 2018). The gastrointestinal tract and central nervous system appear to be the most frequent sites of severe CMV infection in immunocompetent individuals (Rafailidis et al., 2008). However, CMV infection is relatively rarely recorded in immunocompetent hosts.

Separation of peptides with an aqueous mobile phase by temperature-responsive chromatographic column.

Peptide separation technology is significant and is still an analytical challenge in proteomic studies. We report a simple preparation method for poly(N-isopropylacrylamide) grafted silica through the copolymerization of N-isopropylacrylamide with acetyl moieties immobilized on the silica surfaces. Differential scanning calorimetry results indicated that the prepared silica exhibited a sharp phase transition at 35.03°C. Silica grafted with poly(N-isopropylacrylamide) was evaluated as a temperature-responsive chromatography medium for the separation of peptides using 0.2 M NaCl solution as a mobile phase. Results indicated that at 10°C, the peptides were not resolved, but baseline separation with prolonged retention time at 50°C was attained. Particularly, a mixture of four peptides was efficiently separated within 8 min. The theoretical plate number of every peptide was more than 2500, and the resolutions were more than 3.40. The increased selectivity of the temperature-responsive column resulted from the temperature-modulated hydrophobic interaction with peptides. The retention times of these peptides were related to their hydrophobicities. This protocol provided a reliable set of chromatographic tool usable across all research and development applications that required isolation and analysis of peptides. It may represent a step forward in the complex analysis of hydrophobic and other proteins.

Let-7i-3p inhibits the cell cycle, proliferation, invasion, and migration of colorectal cancer cells via downregulating CCND1.

Dysregulated microRNAs are closely related to the malignant progression of colorectal cancer (CRC). Although abnormal let-7i-3p expression has been reported in various human cancers, its biological role and potential mechanism in CRC remain unclear. Therefore, the purpose of this study was to investigate the expression and regulation of let-7i-3p in CRC. Here, we demonstrated that let-7i-3p expression was significantly downregulated in three CRC cell lines while CyclinD1 (CCND1) was upregulated compared with the normal colon epithelial FHC cells. Moreover, bioinformatics and luciferase reporter assays revealed that CCND1 was a direct functional target of let-7i-3p. In addition, let-7i-3p overexpression or CCND1 silencing inhibited cell cycle, proliferation, invasion, and migration and diminished the activation of p-ERK in HCT116 cells. However, exogenously expressing CCND1 alleviated these effects. Taken together, our findings may provide new insight into the pathogenesis of CRC and let-7i-3p/CCND1 might function as new therapeutic targets for CRC.

Effects of deoxynivalenol exposure on cerebral lipid peroxidation, neurotransmitter and calcium homeostasis of chicks in vivo.

During current research, the effects of deoxynivalenol (DON) exposure on cerebral lipid peroxidation, neurotransmitter secretion and calcium homeostasis in chicks were evaluated. One hundred and twenty Hailan chicks (male, 1-day-old) were randomly divided into four groups. Chicks in low, medium and high dose groups were fed with 0.27, 1.68 and 12.21 mg/kg-1 DON respectively by gavage according to feed intake. Chicks in control group were fed with physiological saline by gavage. The trials were conducted for 36 d. At the end of the trials, twenty chicks per group were sacrificed, and the cerebra were collected for measuring the brain indices. Compared with the control group, the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase were significantly decreased in treatment groups (P < 0.05), the contents of malondialdehyde in high dose group were increased (P < 0.05), the catalase activities and nitric oxide contents in medium and high dose groups were decreased (P < 0.05), and the activities of T-AOC in high dose group were reduced (P < 0.05). Compared with the control group, the concentrations of norepinephrine and 5-hydroxytryptamine in high dose group were obviously increased (P < 0.05), while the concentrations of dopamine were decreased (P < 0.05). Meanwhile, the concentrations of calcium and calmodulin (CaM) in medium and high dose groups were lower than those of the control group (P < 0.05), and the gene relative expression of CaM mRNA in treatment groups were significantly reduced (P < 0.05), in a dose-dependent manner. These results suggested that DON exposure can affect the cerebral lipid peroxidation, neurotransmitters secretion and the balance of calcium homeostasis in chicks.

Silencing of the hTERT gene by shRNA inhibits colon cancer SW480 cell growth in vitro and in vivo.

Human telomerase reverse transcriptase (hTERT) is the key enzyme responsible for synthesizing and maintaining the telomeres on the ends of chromosomes, and it is essential for cell proliferation. This has made hTERT a focus of oncology research and an attractive target for anticancer drug development. In this study, we designed a small interfering RNA (siRNA) targeting the catalytic subunit of hTERT and tested its effects on the growth of telomerase-positive human colon carcinoma SW480 cells in vitro, as well as on the tumorigenicity of these cells in nude mice. Transient and stable transfection of hTERT siRNA into colon cancer SW480 cells suppressed hTERT expression, reduced telomerase activity and inhibited cell growth and proliferation. Knocking down hTERT expression in SW480 tumors xenografted into nude mice significantly slowed tumor growth and promoted tumor cell apoptosis. Our results suggest that hTERT is involved in carcinogenesis of human colon carcinoma, and they highlight the therapeutic potential of a hTERT knock-down approach.