The mutation of lysosomal trafficking regulator (LYST) gene and the clinical data of an adult patient who showed an abnormal gait with Chediak-Higashi syndrome were analyzed retrospectively. The whole exon sequencing was applied, and Sanger sequencing was used to verify the results. All members of the family were genetically verified for the same mutation site. The sequencing revealed the presence of c.421C>T(p.Arg141 *) mutation in LYST gene in the proband, which was inherited from his parents. The mutation was found in the homozygous state for the proband, both his parents being heterozygous for the same mutation. This mutation type was not reported in the human gene mutation database. According to the American Society of Medical Genetics and Genomic Society′s guide to the interpretation of genetic variation, the mutation of c.421C>T was identified to be pathogenic.

The effects of vascular endothelial growth factor (VEGF) on neural differentiation of human embryonic stem cells (hESCs) in vitro and the possible mechanism were observed. The hESCs lines, TJMU1 and TJMU2, were established and stored by our laboratory. hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups: group A, routine induction; group B, routine induction+10 ng/mL VEGF; group C, routine induction+10 ng/mL VEGF+10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were detected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence. The percentage of Nestin positive cells in group B was significantly higher than in groups A and C, while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C (P<0.01). There was no significant difference between groups A and C (P>0.05). It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

Aim To evaluate the vasorelaxant effect of two new chemical entities, J35242 and J35243, on iso-lated rat thoracic aorta rings as Rho-kinase inhibitors, and further to explore the underlying mechanisms of these two compounds. Methods Isolated rat thoracic aorta rings pre-contracted by KCl or norepinephrine ( NE) were used to evaluate the vasodilatory effect of J35242 and J35243 . Through the interventions of sev-eral tool drugs, the mechanisms of compounds concern-ing endothelium, K+ channels and Ca2+ were studied. Results J35242 and J35243 showed potent relaxant effect on both KCl and NE pre-contracted vessels, and exhibited partial endothelium dependency. L-NAME and Methylene Blue( MB) could influence the relaxant effect of these compounds. Meanwhile, the compounds could inhibit intracellular Ca2+ release and extracellu-lar Ca2+ influx, which indicated that the compounds might block the calcium channels to relax the vessels. In addition, the two compounds probably did not dilate the aorta rings through opening potassium channels. Conclusions J35242 and J35243 have vasorelaxant effects on vessels in vitro and the potency of J35242 is stronger than that of J35243 . The underlying mecha-nisms might be endothelium-dependent. Also the com-pounds might block Ca2+ channels, lowering intracel-lular Ca2+ concentration to relax the vessels.

Objective To evaluate the values of prenatal surveillances of ultrasonography in twin pregnancies with amniotic fluid discordance. Methods Two hundred and seventy cases of diamniotic twins were included. Both postnatal outcomes and prenatal amniotic fluid discordant variations were analyzed,and the incidences of amniotic fluid discordance were compared between the monochorionic-diamniotic(MCDA)and dichorionic-diamniotic (DCDA) gruop. Results Twenty four cases of twins with amniotic fluid discordance were found in the study. The incidence of amniotic fluid discordance in MCDA group was much higher than that in DCDA group (28.9% vs 5.6%, P ＜0. 001 ) ,and 24 cases of twin-to-twin transfusion syndrome (TTTS) were diagnosed in the former group and none were found in the latter group. Downward tendency of amniotic fluid discordance was shown in non-TTTS cases of MCDA group. Compared with TTTS cases, the postnatal outcomes of non-TTTS cases with amniotic fluid discordance were much better in MCDA group ( P ＜0.001 ). Conclusions TTTS and MCDA twins with amniotic fluid discordance may overlap each other in the early stage. Serial surveillances of ultrasonography are necessary for prenatal differentiating twin pregnancies complicated by amniotic fluid discordance,and providing strong support to clinical treament.

OBJECTIVE: To explore the genetic basis for a patient with intellectual disability. METHODS: Whole exome sequencing and Sanger sequencing were carried out for the patient. The result was verified in her family. RESULTS: DNA sequencing revealed that the patient has carried a heterozygous nonsense c.40C>T (p.Arg14X) variant of the TRIP12 gene, which was de novo in origin. The variant was unrecorded in the Human Gene Mutation Database. Based on the American College of Medical Genetics and Genomics standards and guidelines, the variant was predicted to be pathogenic (PVS1+ PS2+ PP3). CONCLUSION The patient was diagnosed with autosomal dominant intellectual disability due to heterozygous c.40C>T variant of the TRIP12 gene.
Carrier Proteins/genetics* ; Codon, Nonsense ; Female ; Heterozygote ; Humans ; Intellectual Disability/genetics* ; Ubiquitin-Protein Ligases/genetics* ; Whole Exome Sequencing

BACKGROUNDProtein kinase C(PKC) is a potentially important target for can-cer therapeutics due to its potential role in carcinogenesis.Abnormal expression and increasing activity of PKC-α are present in non-small cell lung cancer(NSCLC).PKC inhibitor can show anti-tumor effects through inducing tumor cell apoptosis,enhancing cytotoxic effects and down-regulating expressions of multidrug resistance gene.By observing the effects of PKC inhibitor chelerythrine chloride(CH) on drug-sensitivity to cisplatin of four NSCLC cell lines its mechanism of effect initially is explored.

METHODSNSCLC cell lines(H1299,H460,A549 and cisplatin-resistant A549) were dealed with PKC inhibitor CH respectively.The expressions of PKC-α mRNA and protein in NSCLC cell lines were examined by reverse transcription polymerase chain reaction(RT-PCR) and Western blot.The apoptosis rates of NSCLC cells lines were detected by flow cytometry.The drug-sensitivity to cisplatin of NSCLC cells lines was measured by methabenzthiazuron(MTT) assay.

RESULTSThe expression levels of PKC-α mRNA and protein in cisplatin-resistant A549 cell lines were significantly higher than H1299,H460 and parent A549 cell lines(P < 0.05).The expression levels of PKC-α mRNA and protein in four NSCLC cell lines decreased at different extent.The apoptosis rates of cisplatin-resistant A549 cell lines increased obviously after treating with CH for 4 and 24 hours,but it was not seen in H1299,H460 and parent A549 cell lines.The IC50 value of cisplatin of NSCLC cell lines decreased at different degree after treating with CH and it was more obvious in cisplatin-resistant A549 cell lines(P < 0.05).

CONCLUSIONSHigh expressions of PKC-α mRNA and protein exist in all four NSCLC cell lines.PKC inhibitor CH can enhance the drug-sensitivity of NSCLC cell lines to cisplatin by inhibiting their expression of PKC-α mRNA and protein.When compared with parent A549 cell lines,cisplatin-resistant A549 cell line's drug-sensitivity to cisplatin is increasing more efficiently by PKC inhibitor CH through inhibition of PKC-α protein's expression and elevation of tumor cell apoptosis rates.

Objective? To investigate and analyze the relationship between relocation stress,family function and coping strategy in parents of children transferred from Intensive Care Uni(t ICU). Methods? Using the method of cross-sectional study, the parents of children transferred from ICU to general ward in Children's Hospital Affiliated to Zhengzhou University from October 2018 to March 2019 were selected as the subjects of study by convenience sampling. General Information Questionnaire, Family Relocation Stress Scale (FRSS), Feetham Family Functioning Survey(FFFS),and the Coping Health Inventory for Parents(CHIP) were used to evaluate the relocation stress level and analyze the relationship between scores of FRSS, FFFS, and CHIP. A total of 200 questionnaires were distributed and 179 valid ones were retrieved, yielding an effective recovery rate of 89.5%. Results? The total score of FRSS of the 179 ICU-transferred children's parents was (34.52±8.46), which was in a high level of stress; the total average score of FFFS was (1.27±1.14); the total score of "coping style use frequency" was (144.57±13.57) in CHIP, and the total score of "coping style effect"was (78.20±14.83); correlation analysis showed that each dimension and total score of FFFS was negatively correlated with the level of relocation stress of parents (P＜0.05), and positively correlated with the "frequency and role of coping styles" (P＜0.05). Conclusions? The relocation stress of ICU-transferred children's parents is serious. Family function and coping style are closely related to the level of relocation stress. Medical staff should pay attention to the adaptation of families in the process of transfer, encourage parents to adopt more positive ways to cope with diseases and improve the overall health of children and families.

The effects of vascular endothelial growth factor (VEGF) on neural differentiation of human embryonic stem cells (hESCs) in vitro and the possible mechanism were observed. The hESCs lines,TJMU1 and TJMU2, were established and stored by our laboratory, hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups: group A, routine induction; group B, routine induction+10 ng/mL VEGF; group C, routine in-duction+10 ng/mL VEGF+10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were de-tected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence.The percentage of Nestin positive cells in group B was significantly higher than in groups A and C,while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C (P<0.01). There was no significant difference between groups A and C (P>0.05). It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

PURPOSE: c-Met and its ligand, hepatocyte growth factor (HGF), play a critical role in oncogenesis and metastatic progression. The aim of this study was to identify inhibited enzymogram and to test the antitumor activity of SIM-89 (a c-Met receptor tyrosine kinase inhibitor) in non-small cell lung cancer. MATERIALS AND METHODS: Z′-LYTE kinase assay was employed to screen the kinase enzymogram, and mechanism of action (MOA) analysis was used to identify the inhibited kinases. Cell proliferation was then analyzed by CCK8 assay, and cell migration was determined by transwell assay. The gene expression and the phosphorylation of c-Met were examined by realtime-PCR and western blotting, respectively. Finally, the secretion of HGF was detected by ELISA assay. RESULTS: c-Met, activated protein kinase (AMPK), and tyrosine kinase A (TRKA) were inhibited by SIM-89 with the IC₅₀ values of 297 nmol/L, 1.31 µmol/L, and 150.2 nmol/L, respectively. SIM-89 exerted adenosine triphosphate (ATP) competitive inhibition on c-Met. Moreover, the expressions of STAT1, JAK1, and c-Met in H460 cells were decreased by SIM-89 treatment, and c-Met phosphorylation was suppressed in A549, H441, H1299, and B16F10 cells by the treatment. In addition, SIM-89 treatment significantly decreased the level of HGF, which accounted for the activation of c-Met receptor tyrosine kinase. Finally, we showed cell proliferation inhibition and cell migration suppression in H460 and H1299 cells after SIM-89 treatment. CONCLUSION: In conclusion, SIM-89 inhibits tumor cell proliferation, migration and HGF autocrine, suggesting it's potential antitumor activity.
Adenosine Triphosphate ; Blotting, Western ; Carcinogenesis ; Carcinoma, Non-Small-Cell Lung* ; Cell Movement ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hepatocyte Growth Factor ; Lung Neoplasms ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Protein-Tyrosine Kinases

OBJECTIVETo identify novel common mutations among patients with non-syndromic hearing loss (NSHL).

METHODSHigh-throughput gene capture technology was used to analyze 18 patients with NSHL in whom common mutations of deafness genes including GJB2, SLC26A4, GJB3, and mtDNA were excluded. Suspected mutation was verified with Sanger sequencing.

RESULTSNext generation sequencing has identified 62 mutations in 29 genes associated with hearing loss, which included 54 missense mutations, 4 splicing mutations, 3 deletional mutations, and 1 nonsense mutation. Mutations occurring more than twice in the 18 patients were verified by Sanger sequencing. This has confirmed 15 mutations in 8 genes, including 3 missense mutations (p.C2184G, p.L2825P, p.H1888Y) which have not been reported previously. Meanwhile, p.L445W, p.D866N, and IVS919-2A>G were common causative mutations.

CONCLUSIONA number of common causative mutations, e.g., p.L445W, p.D866N, IVS919-2A>G, have been identified by high-throughput capture technology, which may facilitate the research and genetic diagnosis for hearing loss.

DNA, Mitochondrial ; genetics ; Deafness ; genetics ; Female ; Hearing Loss ; genetics ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Male ; Mutation ; genetics