Altered microRNA expression profiles of human spermatozoa in normal fertile men of different ages.

MicroRNAs (miRNAs) are mediators of the aging process. The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility. Twenty-seven donors were divided into three groups by age (Group A, n = 8, age: 20-30 years; Group B, n = 10, age: 31-40 years; and Group C, n = 9, age: 41-55 years) for high-throughput sequencing analysis. Samples from 65 individuals (22, 22, and 21 in Groups A, B, and C, respectively) were used for validation by quantitative real-time polymerase chain reaction (qRT-PCR). A total of 2160 miRNAs were detected: 1223 were known, 937 were newly discovered and unnamed, of which 191 were expressed in all donors. A total of 7, 5, and 17 differentially expressed microRNAs (DEMs) were found in Group A vs B, Group B vs C, and Group A vs C comparisons, respectively. Twenty-two miRNAs were statistically correlated with age. Twelve miRNAs were identified as age-associated miRNAs, including hsa-miR-127-3p, mmu-miR-5100_L+2R-1, efu-miR-9226_L-2_1ss22GA, cgr-miR-1260_L+1, hsa-miR-652-3p_R+1, pal-miR-9993a-3p_L+2R-1, hsa-miR-7977_1ss6AG, hsa-miR-106b-3p_R-1, hsa-miR-186-5p, PC-3p-59611_111, hsa-miR-93-3p_R+1, and aeca-mir-8986a-p5_1ss1GA. There were 9165 target genes of age-associated miRNAs. Gene Ontology (GO) analysis of the target genes identified revealed enrichment of protein binding, membrane, cell cycle, and so on. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of age-related miRNAs for target genes revealed 139 enriched pathways, such as signaling pathways regulating stem cell pluripotency, metabolic pathways, and the Hippo signaling pathway. This suggests that miRNAs play a key role in male fertility changes with increasing age and provides new evidence for the study of the mechanism of age-related male fertility decline.

Identification of Frailty and Its Risk Factors in Elderly Hospitalized Patients from Different Wards: A Cross-Sectional Study in China.

OBJECTIVES: To survey the difference of frailty prevalence in elderly inpatients amongdifferent wards; to compare the diagnostic performance of five frailty measurements (Clinical Frailty Scale [CFS], FRAIL, Fried, Edmonton, Frailty Index [FI]) in identifying frailty; and to explore the risk factors of frailty in elderly inpatients. PARTICIPANTS AND METHODS: This was a cross-sectional study including 1000 inpatients (mean age 75.2±6.7 years, 51.5% male; 542, 229, and 229 patients from cardiology, non-surgical, and surgical wards, respectively) in a tertiary hospital from September 2018 to February 2019. We applied the combined index to integrate the five frailty measurements mentioned above as the gold standard of frailty diagnosis. Multivariate logistic regression models were used to determine the independent risk factors of frailty. RESULTS: Frailty prevalence was 32.3% (Fried), 36.2% (CFS), 19.2% (FRAIL), 25.2% (Edmonton), 35.1% (FI) in all patients. The frailty was more common in non-surgical wards, regardless of the frailty assessment tools used (non-surgical wards: 27.5% to 51.5%; cardiology ward: 14.9% to 29.3%; surgical wards: 18.8% to 41.9%). CFS≥5 showed a sensitivity of 94.1% and a specificity of 85.2% for all patients. FI≥0.25 showed a sensitivity of 94.8% and a specificity of 87.0% for all patients. Age [odds ratio (OR) = 1.089, P<0.001], education level (OR = 0.782, P=0.001), heart rate (OR = 1.025, P<0.001), albumin (OR = 0.911, P=0.002), log D-dimer (OR = 2.940, P<0.001), ≥5 comorbidities (OR = 2.164, P=0.002), and ≥5 medications (OR = 2.819, P<0.001) were independently associated with frailty in all participants. CONCLUSION: Frailty is common among elderly inpatients, especially in non-surgical wards. CFS is a preferred screening tool and FI may be an optimal assessment tool. Old age, low educational level, fast heart rate, low albumin, high D-dimer, ≥5 comorbidities, and polypharmacy are independent risk factors of frailty in elderly hospitalized patients.

[ABCC8, KCNJ11 and GLUD1 gene mutation analysis in congenital hyperinsulinism pedigree].

OBJECTIVE: To explore the ABCC8, KCNJ11, and GLUD1 gene mutations of the 11 patients diagnosed as congenital hyperinsulinism (CHI). METHODS: A total of 11 CHI children hospitalized in Beijing Children's Hospital from November 2008 to February 2012 and their parents were chosen as the study subjects. Direct sequencing of PCR-DNA was used to analyze the 39 exons of ABCC8 gene, non-translational region and exon of KCNJ11 gene and 6, 7, 10, 11 and 12 exons of GLUD1 gene. RESULTS: An P629PfsX17 heterozygous mutation of ABCC8 gene was detected in case 1 and his father, an W288X heterozygous mutation of ABCC8 gene was detected in case 4 and his father, A640V and Q1196X mutations in ABCC8 gene in case 5 whose father only carried the Q1196X mutation. In case 6 and his father, an R269H mutation was found in GLUD1 gene. The genotype of 4 children's mothers was normal. No mutations were found in other 7 patients and their parents. CONCLUSIONS: The ABCC8 gene mutations are the main pathogenic mechanisms of Chinese children with CHI. In Chinese, P629PfsX17, W288X, A640V and Q1196X heterozygous mutation of ABCC8 gene and R269H heterozygous mutation of GLUD1 gene may lead to CHI. The inheritance mode of the mutations may be paternally or de novo.

Correlation between DNase I hypersensitive site distribution and gene expression in HeLa S3 cells.

Mapping DNase I hypersensitive sites (DHSs) within nuclear chromatin is a traditional and powerful method of identifying genetic regulatory elements. DHSs have been mapped by capturing the ends of long DNase I-cut fragments (>100,000 bp), or 100-1200 bp DNase I-double cleavage fragments (also called double-hit fragments). But next generation sequencing requires a DNA library containing DNA fragments of 100-500 bp. Therefore, we used short DNA fragments released by DNase I digestion to generate DNA libraries for next generation sequencing. The short segments are 100-300 bp and can be directly cloned and used for high-throughput sequencing. We identified 83,897 DHSs in 2,343,479 tags across the human genome. Our results indicate that the DHSs identified by this DHS assay are consistent with those identified by longer fragments in previous studies. We also found: (1) the distribution of DHSs in promoter and other gene regions of similarly expressed genes differs among different chromosomes; (2) silenced genes had a more open chromatin structure than previously thought; (3) DHSs in 3'untranslated regions (3'UTRs) are negatively correlated with level of gene expression.

The tyrosine kinase c-Src directly mediates growth factor-induced Notch-1 and Furin interaction and Notch-1 activation in pancreatic cancer cells.

The proteolytic activity of Furin responsible for processing full length Notch-1 (p300) plays a critical role in Notch signaling. The amplitude and duration of Notch activity can be regulated at various points in the pathway, but there has been no report regarding regulation of the Notch-1-Furin interaction, despite its importance. In the present study, we found that the Notch-1-Furin interaction is regulated by the non-receptor tyrosine kinase, c-Src. c-Src and Notch-1 are physically associated, and this association is responsible for Notch-1 processing and activation. We also found that growth factor TGF-α, an EGFR ligand, and PDGF-BB, a PDGFR ligand, induce the Notch-1-Furin interaction mediated by c-Src. Our results support three new and provocative conclusions: (1) The association between Notch-1 and Furin is a well-regulated process; (2) Extracellular growth factor signals regulate this interaction, which is mediated by c-Src; (3) There is cross-talk between the plasma growth factor receptor-c-Src and Notch pathways. Co-localization of Notch-1 and c-Src was confirmed in xenograft tumor tissues and in the tissues of pancreatic cancer patients. Our findings have implications for the mechanism by which the Notch and growth factor receptor-c-Src signaling pathways regulate carcinogenesis and cancer cell growth.

[Comparative analysis of modified Dewar method versus arthroscopic double Endobutton fixation technique for the treatment of acute acromioclavicular joint dislocation].

OBJECTIVE: To compare the clinical outcomes of modified Dewar method versus arthroscopic double Endobutton fixation technique for the treatment of acute acromioclavicular joint dislocation (Rockwood types III-V). METHODS: All cases with acute acromioclavicular joint dislocation (Rockwood types III-V) were treated at our department from October 1997 to October 2009. Among them, 28 cases undergoing modified Dewar method were followed up. There were 20 males and 8 females aged 18 - 68 years old with a mean follow-up period of 6.8 years. And the arthroscopic technique of Endobutton fixation was employed for another 24 cases. There were 19 males and 5 females aged 19 - 65 years old with a mean follow-up period of 1.5 years. The radiographic findings, clinical outcomes and complications of two groups were compared. RESULTS: The good/excellent rate of modified Dewar group and arthroscopic double Endobutton group were 92.8% and 95.8% respectively. There was no significant difference between two groups. No significant difference existed between two groups as to the VAS (visual analogue scale) pain score and UCLA (University of California at Los Angeles) score. The modified Dewar group had a higher rate of ectopic ossification in coracoclavicular ligament than that of the arthroscopic double Endobutton group (25% vs 8.33%). Yet there was no statistical significance. However, the distance between clavicle and coracoid process was larger in the modified Dewar group (11.96 vs 8.54 mm, P < 0.05). CONCLUSION: Both modified Dewar method and arthroscopic double Endobutton fixation technique are both efficient therapies for acute acromioclavicular dislocation (Rockwood types III-V). The former tends to be more invasive while the latter can better maintain the relationship of coracoid process and clavicle.

MicroRNA miR-886-5p inhibits apoptosis by down-regulating Bax expression in human cervical carcinoma cells.

OBJECTIVE: MicroRNA (miRNA) plays an essential role in the progression of a variety of cancers, but its role in cervical cancer progression is not well defined. We aimed to test whether special miRNAs and their target mRNAs contribute to cervical cancer progression. METHODS: The expression profiles of 1145 microRNAs in cervical squamous cell carcinomas (CSCC) and adjacent non-tumor tissues were investigated using an Illumina microRNA microarray platform. Differentially expressed miRNAs were validated by RT-PCR. Downstream target validation was performed for miR-886-5p. RESULTS: We found that the expression levels of seven miRNAs differed significantly between CSCC tissues and adjacent non-tumor tissues. Forced expression of one miRNA, miR-886-5p, over-expressed in CSCC tissues lowered expression of the pro-apoptotic protein Bax, reduced apoptosis and promoted cell proliferation in H8, an HPV16-immortalized human cervical squamous epithelial cell line. Knockdown of miR-886-5p increased Bax protein and apoptotic cell death in cells of the cervical squamous carcinoma cell line, SiHa. CONCLUSION: MicroRNA miR-886-5p inhibits apoptosis of cervical cancer cells by down-regulating the production of Bax.

[Effects of different repair methods in treatment of supraspinatus tendon tear: experiment with rabbits].

OBJECTIVE: To compare the effects of different repair methods in treatment of supraspinatus tendon tear. METHODS: Forty-five New Zealand rabbits aged 4 approximately 6 months underwent cutting off of the left supraspinatus tendon for the width of 8 mm, and then were randomly divided into 3 equal groups: Group A, undergoing suture anchor simple suture; Group B, undergoing suture anchor mattress suture; and Group C, undergoing transosseous mattress suture. Eight weeks after the operation the rabbits were killed. The shoulder girdles of 12 rabbits randomly selected from each group were used for mechanical test to determine the intensity strength of the supraspinatus tendon, and the left shoulder girdler of the other rabbits underwent decalcification, HE staining, and histological study to observe the healing of the end point of the supraspinatus tendon. RESULTS: Two rabbits. One in Group A and on in Group B underwent infection. The healing rate was 93.33% for both Groups A and B, and was 100% for Group C, however without significant differences among them (all>0.05). The mean maximum tension loads of the supraspinatus tendon of Groups A, B, and C were 82.39+/-20.70 N, 81.80+/-20.31 N, and 88.58+/-17.24 N respectively, however without significant differences among them (F=0.44, >0.05). Histological examination showed that at the end point the supraspinatus tendons of Groups A and B connected to the lamellar bone and that three-zone structure (tendon, cartilage, and bone) could be found at the end point of the supraspinatus tendon in Group C, but compared with the normal end point, the number of cartilage cells decreased. CONCLUSION: The effects of the 3 suture methods are not significantly different from each other for the repair of supraspinatus tendon in respect to healing rate and healing strength, but compared with the suture anchor simple suture method and suture anchor mattress suture method, the transosseous mattress suture method shows greater post-healing strength of tension and the tendon healing situation is closer to the normal tendon structure.