scRNA-seq reveals that origin recognition complex subunit 6 regulates mouse spermatogonial cell proliferation and apoptosis via activation of Wnt/β-catenin signaling / 亚洲男科学杂志(英文版)

The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

Neonatal Immune State Is Influenced by Maternal Allergic Rhinitis and Associated With Regulatory T cells

PURPOSE: Maternal influences contribute to the origin of allergic diseases, but the mechanisms are not clear. The current literature prompted the role of epigenetics in the development of allergic diseases. We sought to investigate the roles of regulatory T (Treg) cells and Forkhead box p3 (Foxp3) DNA methylation in the process of maternal transmission of allergic rhinitis (AR) susceptibility. METHODS: BALB/c female mice (AR mother) were sensitized by intraperitoneal injection of Dermatophagoides pteronyssinus (Der p) 1 on day 1 and 7. Then they mated with normal male mice on day 8. From day 21 to 28, the female mice were intranasal challenged with Der p 1 continuously. The normal controls were given with normal saline in the same way. On postnatal day 3, Female mice and their offspring were sacrificed to detect their histopathology in nasal mucosae, cytokines in sera of mother and spleen homogenates of offspring, Treg cells count, Foxp3 mRNA expressions, and Foxp3 DNA methylation levels in spleens. RESULTS: Compared with the normal controls, neonatal offspring of Der p 1-stimulated female mice (AR offspring) showed the elevation of interleukin (IL)-4 (P<0.01) and IL-17 (P<0.01), the submission of IL-10 (P<0.01) in spleen homogenates. Further, Treg cells count in AR offspring decreased remarkably compared with the normal offspring (P<0.01). Though the difference of Foxp3 DNA methylation level between AR offspring and normal control offspring was not obvious, correlation analysis demonstrated that there was significantly positive correlation between Foxp3 DNA methylation level of mother and that of offspring (r=0.803, P<0.01). CONCLUSIONS: Under the influence of Maternal AR, their neonatal offspring develop into T-helper type 2 (Th2) dominant immune state, which is closely associated with the recession of Treg cells. Foxp3 DNA methylation may be a mechanism responsible for that maternal effect but still need more studies to ensure.

Application of nasopore and budesonide suspension on tamping after endoscopic sinus surgery / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the clinical effect of nasal packing of pulmicort respules combined withnasopore after endoscopic sinus surgery.@*METHOD@#A total of 30 CRSwNP and CRSsNP patients with bilateral functional endoscopic sinus surgery and finished following up visit were randomly choosed, conventionally select the left nasal cavity as the experimental group, the right nasal cavity as the control group. Experimental group to pack the nasal cavity with pulmicort respules union nasopore after surgery and control group to pack the nasal cavity with only nasopore after surgery. The differences were observed in patients with subjective symptoms and recovery of mucosa of operative cavity between the two groups after two weeks, one month and three months.@*RESULT@#(1) The postoperative VAS symptoms score about nasal obstruction, nasal secretion, headache, dizziness and distending pain after two weeks,one month and three months in the experiment group were significantly better than those in the control group(P0.05).@*CONCLUSION@#The application of nasal packing of pulmicort respules combined with nasopore after functional endoscopic sinus surgery can effectively relieve postoperative uncomfortable symptoms, promote recovery of mucosa of perative cavity, which deserves clinical promotion.

Proton Pump Inhibitor and Prokinetic Drug on the Treatment of Laryngopharyngeal Reflex Diseases:A Meta-analysis / 听力学及言语疾病杂志

Objective To evaluate the efficacy and safety of proton pump inhibitor (PPI)plus prokinetic drug on the treatment of laryngopharyngeal reflex .Methods According to predetermined criteria of inclusion and exclu-sion ,Cochrane Central Register of Controlled Trails(CENTRAL) ,Embase ,Elsevier ,Pubmed ,VIP ,Wanfang Data , CNKI and CBM were searched for the randomized controlled trails of proton pump inhibitor and prokinetic drug on the treatment of laryngopharyngeal reflex diseases from the date of establishment of the datebases to November 2012 .Revman 5 .1 software and GRAED profiler 3 .6 software were applied to analysis the data ,which was extrac-ted by three independent evaluators .Results A total of 346 related articles were obtained ,7 trails were included , 614 patients in total .The results of Meta -analysis showed that :the overall effictive rate of combination therapy with proton pump inhibitors plus prokinetic drug in patients with laryngopharyngeal reflex was higher than those in the control group [RR=1 .18 ,95% CI(1 .10 ,1 .28) ,P<0 .0001] .The rate of improvement of laryngeal signs exam-ined by fiberoptic laryngoscope was higher than those in the control group [RR= 1 .38 ,95% CI(1 .16 ,1 .65) ,P<0 .001] .The average relief time of dysphonia symptoms was shorter than those in the control group [MD = -1 .70 , 95 CI (-2 .95 ,-0 .45) ,P<0 .001] .Conclusion The effect of proton pump inhibitor plus prokinetic drug on the treatment of laryngopharyngeal reflex is superior to those of proton pump inhibitors or prokinetic drug only ,but in terms of safety ,more large-sample ,high-quality randomized controlled trails are required .

A case-control study on the duration of sleeping and cerebral infarction / 中华流行病学杂志

Objective To explore the relationship between duration of sleeping and cerebral infarction.Methods A case-control study involved 1037 cerebral infarction patients admitted by the Second Affiliated Hospital of Harbin Medical University,December 2011-December 2012 as cases.Another 1205 adults free from cerebro-vascular diseases who had undergone physical examination in the hospital at the same period,were served as controls.All the subjects were interviewed with unified questionnaire.Chi-square test,u-test and multivariate logistic regression analysis were performed.Results After adjustment for potential confounding factors including age,sex,body mass index,wrist-hip ratio,smoking,alcohol intake,hypertension,diabetes mellitus,coronary artery disease and lipid parameters,data from the multivariate logistic regression analysis showed that the risk of cerebral infarction was greater in people who slept less than 6 hours per night than those who slept between 6 hours and 8 hours per night,with an odds ratio (95％ CI) as 2.81 (95％ CI:1.68-4.70).There was no significant association between factor as ‘sleeping longer than 8 hours/pre day' and cerebral infarction.Through the subgroup analysis,data showed that the association between ‘ shorter than 6 hour sleep/night' and cerebral infarction consistently exsited,across the categories of sex,and the degree of association was greater in women than in men,with the odds ratio as 5.58 (95％CI:1.78-17.52) and 2.00(95％CI:1.10-3.64) respectively.Conclusion Short sleeping duration might increase the risk of developing cerebral infarction.

Receptor-binding ability of fragments 260-600 and 397-796 of SARS-associated coronavirus spike protein / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To investigate the interaction between the host cell and the truncated S fragments to identify the receptor-binding domain of the spike (S) protein of SARS-associated coronavirus (SARS-CoV).</p><p><b>METHODS</b>Two different fragments S260-600 and S397-796 of the SARS-CoV S protein were expressed in Escherichia coli (E.coli) using a pET expression vector, respectively. The two recombinant proteins were separately verified by Western blot, purified by nickel-affinity chromatography, and incubated with Vero cells, a susceptible cell line of SARS-CoV infection, for cell binding assay. After the sequential probing with sera from convalescent SARS-patients and FITC-labeled anti-human IgG, the cells were analyzed by flow cytometry. The NIH 3T3 cell, a non-permissive cell line of SARS-CoV infection, was used as controls.</p><p><b>RESULTS</b>The recombinant proteins S260-600 and S397-796 were efficiently expressed in an insoluble form in E.coli. The appropriate expression of the proteins was confirmed by Western blotting using both SARS patients' sera and anti-6 x histidine antibody. The flow cytometry results showed that the both proteins were able to bind Vero cells, but the binding ability of S260-600 was somewhat stronger than that of S397-796. In contrast, the S260-600 protein did not bind NIH3T3 cells.</p><p><b>CONCLUSION</b>Both S260-600 and S397-796 exhibited different receptor binding activity. The S260-600 fragment probably contains the important receptor binding domain and could be a potential candidate for the development of SARS vaccine and anti-SARS therapeutics.</p>