[Overexpression of Numb gene can inhibit the progression of biliary liver fibrosis in adult liver].

Objective: To investigate whether the overexpression of Numb gene can effectively intervene the progression of cholestatic liver fibrosis (CLF) in adult liver. Methods: Twenty-four SD rats were randomly divided into sham operation (Sham, n=6), common bile duct ligation (BDL, n=6), empty vector plasmid (Numb-EV, n=6) and numb gene overexpression group (Numb-OE, n=6). The CLF model was prepared by common bile duct ligation. Simultaneously, the model was established, and the adeno-associated virus (AAV) carrying the cloned numb gene was injected into the rats' spleens. Samples were collected at the end of four weeks. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), serum total bilirubin (TBil), serum total bile acid (TBA), liver histopathology, liver tissue hydroxyproline (Hyp) content, and alpha smooth muscle actin (α-SMA), cytokeratin (CK) 7, and CK19 expression conditions were determined in liver tissue. An analysis of variance was used to compare the means of multiple groups. Results: Compared with the Sham group, the Numb mRNA level in the rat liver tissue was significantly decreased in the BDL group (0.872±0.237 vs. 0.452±0.147, P=0.003). Compared with the Numb-EV group, the Numb mRNA level in the liver tissue was significantly increased in the Numb-OE group (0.487±0.122 vs. 1.094±0.345, P＜0.01). Compared with the Sham group, the Hyp content (µg/L) (288.46±49.49 vs. 901.98±271.85, P＜0.01) and the α-SMA mRNA level (0.858±0.234 vs. 8.976±1.398, P＜0.01) were significantly higher in the BDL group. Compared with the Numb-EV group, the Hyp content (864.32±113.54 vs. 580.44±171.77, P=0.039), the α-SMA mRNA level (6.138±1.443 vs. 1.322±0.859, P＜0.01) and the protein levels were significantly reduced in the Numb-OE group. Compared with the Sham group, the serum ALT, AST, TBil, and TBA levels were significantly increased in the BDL group (P＜0.01), and the ALB content was significantly decreased (P＜0.01). Compared with the Numb-EV group, AST and TBil levels were significantly reduced in the Numb-OE group (P＜0.01), as were the ALT and TBA levels (P＜0.05); however, the ALB content was significantly increased (P＜0.01), and the differences were statistically significant. Compared with the Sham group, the mRNA expression levels of CK7 and CK19 were significantly increased in the BDL group (1.40±0.42 vs. 43.78±7.56; 1.11±0.51 vs. 363.81±134.84, P＜0.01). The mRNA expression levels of CK7 and CK19 were significantly reduced in the OE group (343.19±81.22 vs. 3.22±2.34; 40.53±14.02 vs. 15.68±9.36,P＜0.01). Conclusion: Overexpression of the Numb gene can inhibit CLF progression in the adult liver, which may become a new target for CLF therapy.

[What else should surgeons do when ERAS is in-depth development].

Since the concept of enhanced recovery after surgery(ERAS) has been proposed,it is valued,enriched and developed along with controversy and discussion. Although the content of ERAS involves multidisciplinary fields and is not an independent branch of surgery,surgeons,as an indispensable part,play an important role. At present,surgeons should pay more attention to post-operation complication rate,unplanned readmission rate and reoperation rate,and avoid these problems to the most extent by combining excellent surgical techniques with solid non-technical skills. As consequence,we would implement ERAS much better and be of great benefit to patients.

Balloon sizing during transcatheter aortic valve implantation : Comparison of different valve morphologies.

BACKGROUND: Although balloon sizing has been found to be useful during transcatheter aortic valve implantation (TAVI), its effectiveness in patients with bicuspid aortic valve (BiAV) remains unknown. METHODS: Patients who underwent balloon sizing were retrospectively identified. The study comprised 67 patients (61.2% with BiAV). Preprocedural hypothetical transcatheter heart valve (THV) sizing was based on multislice computed tomography (MSCT) measurements at the annulus. Changes in valve size after balloon sizing were reviewed. Postprocedural MSCT measurements and the grade of paravalvular aortic regurgitation (PAR) were compared. RESULTS: When comparing patients with a BiAV and those with a tricuspid aortic valve (TiAV), there was no significant difference (p = 0.97) in the proportion of decreased (43.9% vs. 46.2%), unchanged (51.2% vs. 50.0%), or increased (4.9% vs. 3.8%) valve sizes chosen on the basis of MSCT findings. The anticipated annular sizing ratio for patients who received a smaller valve was 7.2% (3.5-10.5%) while it was 15.7% (12.5-19.0) for the others (p < 0.01), and no significant difference in the proportion of mild (or more severe) PAR cases was found between the groups (37.9% vs. 30.6%, p = 0.53 at the 1­month follow-up). Stent frame expansion and eccentricity index were comparable between the BiAV and TiAV subgroups among patients who received a smaller THV after balloon sizing. CONCLUSION: Balloon sizing is a useful tool that is complementary to the current gold standard of MSCT for THV size selection as well as for BiAV morphology assessment.

[The effect and mechanism of 2-methoxyestradiol on human laryngeal papilloma cell line].

Objective:To explore the signal pathway that mediates the effect of 2-methoxyestradiol（2ME2） on human laryngeal papilloma cell line, in terms of cell proliferation and neovascularization. Method:HIF-1α expression of human laryngeal papilloma cell line（Hs840. T） was interfered using siRNA, and the cells were then processed by 2ME2 in two concentrations. RT-PCR and ELISA were performed to detect the difference of HIF-1α in cells with normal or lower HIF-1α mRNA level, with ELISA test of excretory VEGF level and CCK8 test of cell viability. Result:The IC50of 2ME2 in Hs840. T was 0.309 µmol/L in terms of the inhibition effect of cell proliferation（P<0.01）. Baseline level of intracellular HIF-1α was detectable, and procession of Hs840. T cells by 2ME2 of 0.4 µmol/L inhibited the transcription and expression of HIF-1α by（76.8±2.0）% and（68.6±3.5）% [vs blank group（100.0±2.7）% and（100.0±6.9）%, P<0.01]. VEGF excretion decreased to（50.8±2.1） and（28.1±4.0）% [vs blank group（100.0±3.1）%, P<0.01]after procession by 2ME2 of 0.2 µmol/L and 0.4 µmol/L. After the successful interference of HIF-1α by siRNA, the inhibition effect on cell proliferation by 2ME2 of 0.4 µmol/L decreased to（51.5±3.8）% [vs control group（65.7±1.7）%, P<0.01]. siRNA interference of HIF-1α lead to a decrease of HIF-α mRNA and protein level to（16.3±0.9）% and（7.4±0.8）% [vs cells not interfered（76.8±2.0）% vs（68.6±3.5）%, P<0.01]. Secretory VEGF dropped to（41.0±2.9）% [vs cells not interfered（28.1±4.0）%, P<0.05]. Conclusion:2ME2 has a significant inhibitory effect on human laryngeal cell line. The inhibition of cell proliferation was mediated by a lower level of HIF-1α and therefore lower VEGF. 2ME2 might serve as a novel potential therapy for patients of recurrent respiratory papillomatosis.

[The expression and clinical progression of androgen, estrogen and prolactin receptor in laryngeal squamous cell carcinoma].

Objective:The aim of this study is to analyze the different expression and function of androgen receptors (AR) and estrogen receptor (ER-α) in laryngeal squamous cell carcinoma (LSCC). In combination with the expression of prolactin receptor (PRLR), we analyzed the prognostic impact of three receptors on the laryngeal carcinoma. Method:In this study, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PC) analysis were performed on the tumor tissues and adjacent normal tissues in 112 LSCC patients (95 males and 17 females). We found that hormone receptor expression is closely related to the clinical tumor lesions and overall survival data. Result:The expression of AR, ER-α and PRLR in tumor tissues were much higher than those in adjacent normal tissues (P>0.05) at both protein and mRNA levels. The higher PRLR level indicate poor survival in LSCC patients (P=0.02), while higher ER-α expression could influence the survival with considerable trend toward significance (P=0.06). Furthermore, the higher expression of ER-α in tumours were corresponding with PRLR cytoplasmic higher level expression (r=0.802, P=0.04). This mutual promoted effect between ER-α and PRLR possibly suggests potential mechanisms among those sex related hormone receptors in laryngeal cancer. Conclusion:It has become increasingly credible that the sex related hormone receptors play an important role in the development of LSCC.

[Factors for lymph node metastasis and survival rate in papillary thyroid microcarcinoma: from a large cohort of 3 607 cases series].

Objective: To investigate high-risk factors for lymph node metastasis (LNM) and 10-year survival rate in patients with papillary thyroid microcarcinoma (PTMC). Methods: We retrospectively analyzed the clinical data of 3 607 patients with PTMC received surgery in our hospital from January 2005 to December 2015. Univariate and multivariate analyses were performed to identify the factors associated with metastasis and 10-year survival rate of the patients was calculated. Results: The patients accepted an average follow-up of 68 months, with follow-up from 18 to 138 months. Total LNM rate in the 3 607 patients with PTMC was 17.13%. Central lymph node metastasis rate was 16.86%, which was related to gender (males 21.89% vs females 15.26%, P<0.01), age (<45 years old 24.44% vs ≥45 years old 11.96%, P<0.01), tumor diameter (P<0.01) and multifocality (P<0.01). Lateral lymph node metastasis rate was 1.05%, which was related to age (<45 vs ≥45, P<0.01) and tumor diameter (P<0.01). The 10-year-survival rate was 93.0% and 10-year recurrence-free survival rate was 87.5%. Conclusion: PTMC may metastasize to cervical lymph node, especially in males, with age younger than 45, multifocal tumors and maximal tumor diameter larger than 0.7 cm. There was no significant correlation between these clinical features associated with LNM and the 10-year survival rate.

[Effect of obstructive sleep apnea on sleep architecture of acute ischemic stroke patients].

Objective: To investigate the effect of obstructive sleep apnea (OSA) on sleep architecture in acute ischemic stroke (AIS) patients. Methods: Seventy AIS patients with polysomnography examination from June 2014 to April 2016 were included in the Second Affiliated Hospital of Soochow University. Twenty-seven healthy controls during the same period were chosen as control group. According to apnea-hypopnea index (AHI), AIS patients were divided into AIS group (AHI<5/h) and AIS+ OSA group (AHI≥5/h). Their general conditions, Neurological function defect (national institutes of health stroke scale, NIHSS) on admission and at discharge and self-care ability (modified Rankin Scale, mRS) at discharge and 3 months later (mRS score <2 reflected good prognosis, and mRS score≥2 reflected unfavorable prognosis) were collected. Results: Fifty-two patients (74.3%) had OSA in total seventy AIS patients. Total sleep time (TST) was significantly shorter and sleep efficiency (SE) was lower in AIS group than the control group (P=0.007, 0.008, respectively). AIS+ OSA group had longer non-rapid eye movement (NREM)1 than control group [24.9(21.3) vs 14.3(10.6), P=0.044]. Compared with AIS group, AIS+ OSA group had shorter NREM3 [13.0(13.2) vs 19.6(12.8), P=0.039]. There was no significant difference between the infarct location of AIS group and AIS+ OSA group. However, AIS+ OSA group had higher mRS score observed at 3 months through follow-up visit than AIS group (P=0.027). Spearman correlation analysis showed a positive correlation between unfavorable prognosis of stroke at 3 months and atrial fibrillation, the oxygen desaturation index (ODI), percentage of oxygen saturation <90% of total recording time (TS90%), AHI, the longest time of apnea and hypopnea, respiratory related microarousal and NIHSS, while a negative correlation between unfavorable prognosis of stroke and NREM3, lowest nocturnal oxygen saturation (L-SaO(2)). On Logistic analysis, the unfavorable prognosis was independently predicted by AHI and NIHSS. Conclusions: Sleep architecture of cerebral infarction patients are disturbed with its characteristic of shorter total sleep time and lower sleep efficiency. Cerebral infarction patients with OSA have longer NREM1 and shorter NREM3.

[Preliminary findings for metabolite profiles of papillary thyroid carcinoma and laryngeal squamous cell carcinoma].

Objective:To identify distinct metabolite profiles of papillary thyroid cancer (PTC) and laryngeal squamous cell carcinoma (LSCC).Method:Tumor and adjacent non-tumor specimens were collected from 57 PTC and 33 LSCC patients. Distinct metabolite profiles of tissues were examined using a combination of gas chromatography-time-of-flight mass spectrometry and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. The data were analyzed with multivariate statistical analysis to compare the distinct metabolite profiles and related pathways of these three tumors.Result:A panel of 46 and 41 differentially expressed metabolites were identified in tumor and adjacent non-tumor tissues of PTC and LSCC, respectively. Increased glycolysis, amino acids metabolism, one carbon metabolism and tryptophan metabolism were found in these two types of tumor tissues compared to adjacent non tumor tissues. Purine and pyrimidine metabolism was significantly elevated in PTC and LSCC tumor tissues, while taurine and hypotaurine were only higher in PTC tumor tissues. The fatty acid metabolism was detected at lower level in both PTC and LSCC tumor tissue.Conclusion:PTC and LSCC tumor tissues not only have common metabolic signatures characterized by increased glycolysis, amino acids metabolism, one carbon metabolism and tryptophan metabolism, but also have distinct metabolic signatures. It is helpful to understand the nature of these two tumors.

Elevated Plasma Level of Interferon-λ1 in Chronic Spontaneous Urticaria: Upregulated Expression in CD8(+) and Epithelial Cells and Induction of Inflammatory Cell Accumulation.

Interferon- (IFN-) λ1 is regarded as a potent bio-active molecule in innate immunity. However, little is known about its role in chronic spontaneous urticaria (CSU). We therefore investigated expression of IFN-λ1 in CSU, its cellular location, and its influence on inflammatory cell accumulation by using flow cytometry analysis, skin tissue dispersion, immunohistochemical stain, and a mouse peritoneal inflammation model. The results showed that level of IFN-λ1 was 2.0-fold higher in plasma of the patients with CSU than the level in healthy control (HC) subjects. Among leukocytes examined, only CD8(+) T cells expressed more IFN-λ1 in CSU blood. Double labeling immunohistochemical staining revealed that IFN-λ1(+) inflammatory cells such as mast cells, eosinophils, B cells, neutrophils, and macrophages were mainly located in dermis, whereas epidermis tissue highly expressed IFN-λ1. IFN-λ1 induced a dose-dependent increase in number of eosinophils, lymphocytes, mast cells, macrophages, and neutrophils in the peritoneum of mice at 6 h following injection, which was inhibited by pretreatment of the animals with anti-intercellular adhesion molecule- (ICAM-) 1 and/or anti-L-selectin antibodies. In conclusion, IFN-λ1 is likely to play a role in the pathogenesis of CSU. Blocking IFN-λ1 production may help to reduce the accumulation of inflammatory cells in the involved CSU skin.

Identification of LCK mutation in a family with atypical epidermodysplasia verruciformis with T-cell defects and virus-induced squamous cell carcinoma.

BACKGROUND: Inherited epidermodysplasia verruciformis (EV) is a rare skin disorder characterized by susceptibility to specific types of human papilloma virus (HPV) and is strongly associated with skin carcinomas. Inactivating mutations in EVER1/EVER2 account for most cases of EV. However, more phenotypes related to but distinct from EV have been reported with an immunodeficiency state but without EVER1/EVER2 mutation, and the genetic basis for these atypical EV cases is poorly understood. OBJECTIVES: To identify the causative gene responsible for three siblings affected by atypical EV but without EVER1/EVER2 mutation. METHODS: Whole-exome sequencing followed by Sanger sequencing was performed to identify the gene responsible for the patients with atypical EV enrolled in our study. RESULTS: A homozygous splicing mutation was detected in LCK (c.188-2A>G). This mutation resulted in an exon 3 deletion T lymphocyte-specific protein tyrosine kinase isoform, which further led to frameshift mutation and subsequent mRNA decay. CONCLUSIONS: We demonstrate a novel mutation in LCK in a family affected by atypical EV with T-cell defects, HPV infection and virus-induced malignancy, providing new clues in the understanding of host defences against HPV and better genetic counselling of patients with the EV phenotype.

[The characteristic of metabolics of female papillary thyroid carcinoma and its relationship with primary tumor size and cervical lymph node metastases].

OBJECTIVE: To analyze the metabolic profiles of the female papillary thyroid carcinoma (PTC) and the relationship between the metabolic profiles and primary tumor size and cervical lymph node metastasis using a metabolomics approach. METHODS: Forty-three cases of female PTC were enrolled in this study. Gas chromatography-time-of-flight mass spectrometry and ultra performance liquid chromatography-time of flight-mass spectrometry were employed for analyzing metabolic profiles of tumor tissues and adjacent normal tissues in the female PTC. Cases were divided into Group T1 (tumor size≤2.0 cm) and Group T2 (tumor size>2.0 cm) according to the tumor size, and divided into Group N- (with negative cervical lymph node) and Group N+ (with positive cervical lymph node) according to the cervical lymph node conditions. We compared the metabolomic profiles between these groups. RESULTS: A panel of 46 differentially expressed metabolites was identified in the PTC specimens, compared with normal tissues. Increased metabolism of amino acid, purine and pyrimidine, tryptophan acid, one carbon, glycolysis, taurine and hypotaurine, and fatty acid were found in PTC tumors tissues. Amino acids, purine and pyridine, tryptophan, and carbon metabolism increased significantly in the tumor tissues of Group T2 compared with Group T1, while glycolysis, amino acid, purine and pyridine, tryptophan, and carbon metabolism increased significantly in the specimens of Group N+ . CONCLUSION: Distinct metabolic profiles were identified in the female PTC tissues, which were related to the primary tumor size and cervical lymph node metastases.

[Quality of life analysis of teachers in a primary school in Chengdu].

Objective:To investigate the prevalence of voice disorder and quality of life in the primary school teachers in a certain district of Chengduand examine the correlation between two voice-specific questionnaires.Method:Random sampling survey included 389 teachers from a certain district of Chengdu.Twelve item general health questionnaire(GHQ-12),voice handicap index 10(VHI-10) and voice-related quality of life (V-RQOL) were investigated and routine examination of throat with electronic fiber laryngoscopy were carried out for teachers.Result:There were 189 teachers who had voice disorders(voice disorders group),and 200 teachers who did not have voice disorders(control group).②The total score's difference of V-RQOL and VHI-10,functional(F) and emotion(E) dimensions in VHI-10 between two groups were statistically significant(P<0.05),however,there were no statistically significant difference of GHQ-12 between two groups. The total score of GHQ-12 was positively correlated with VHI-10(r=0.194,P <0.05),while the total score of V-RQOL was negatively correlated with GHQ-12(r=-0.209,P<0.05) and VHI-10 (r=-0.586, P <0.05).Conclusion:Voice disorders can affect teachers' psychology and voice related quality of life.Although the VHI-10 and V-RQOL had correlation,the correlation is not strong.So it is recommended combine VHI-10 and V-RQOL when the quality of life of voice disorders is evaluated.

Effects of growth differentiation factor 9 and bone morphogenetic protein 15 on the in vitro maturation of porcine oocytes.

Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are members of the transforming growth factor-ß (TGF-ß) family, and their roles in oocyte maturation and cumulus expansion are well known in the mouse and human, but not in the pig. We investigated GDF9 and BMP15 expressions in porcine oocytes during in vitro maturation. A significant increase in the mRNA levels of GDF9 and BMP15 was observed at germinal vesicle breakdown, with expression levels peaking at metaphase I (MI), but decreasing at metaphase II (MII). GDF9 and BMP15 protein localized to the oocyte cytoplasm. While treatment with GDF9 and BMP15 increased the expression of genes involved in both oocyte maturation (c-mos, cyclinb1 and cdc2) and cumulus expansion (has2, ptgs2, ptx3 and tnfaip6), SB431542 (a TGFß-GDF9 inhibitor) decreased meiotic maturation at MII. Following parthenogenetic activation, the percentage of blastocysts in SB431542 treatment was lower than in the control (41.3% and 74.4%, respectively). Treatment with GDF9 and BMP15 also increased the mRNA levels of maternal genes such as c-mos [a regulatory subunit of mitogen-activated protein kinase (MAPK)], and cyclinb1 and cdc2 [regulatory subunits of maturation/M-phase-promoting factor (MPF)]; however, SB431542 significantly decreased their mRNA levels. These data were supported by poly (A)-test PCR and protein activity analyses. Our results show that GDF9 and BMP15 participate in cumulus expansion and that they stimulate MPF and MAPK activities in porcine oocytes during in vitro maturation.

[Western blotting of RStV gene products in rice and insects].

The NS3 and NC protein genes encoded by RNA3 of RStV, the NCP and NSvc4 protein genes encoded by RNA4 were subcloned into the E. coli expression vector pGEX3X to express four groups of fusion protein under IPTG induction. These fusion proteins were used to immunize rabbits to raise antisera. The antisera against the E. coli-expressed proteins were available for probing the presence of the viral gene products in both rice plant and insect hosts. The expected gene products can be probed only in diseased rice plant with NCP antiserum and the corresponding products detected in both plant and RStV particle preparation with NC antiserum. The viral gene products probed by NS3 and NSvc4 antisera were different from the expected ones in size.

Production of animal model of traumatic pseudoaneurysm.

Three microsurgical techniques, i.e., electric cauterized vein patch (ECVP), electric cauterized vein segment (ECVS), and electric cauterized muscular cavity (ECMC), were employed to produce an ideal animal model of traumatic pseudoaneurysm (TPA). In this paper, 108 cases of TPAs were produced in rabbit femoral arteries. The successful rates of these microsurgical techniques, proved by angiography, autopsy in vivo, and histological examination, were 5.5, 16.7, and 88.9%, respectively. The successful rate of ECMC, with the merits of simple operation, a narrow neck but giant size of TPA which rarely ruptured, and the pulsatile mass occurring as early as 4 to 11 days after operation, was much higher than that of ECVP and ECVS (P < 0.01). By this reliable method of ECMC, the pathogenesis of TPA was observed as the following: the arterial wall was injured and partly ruptured, then the escaped blood was confined by fibrous connective tissue near the arterial opening. As a result, a located hematoma without any pulsatile mass and vascular systolic murmur formed. When a cavity occurred in the hematoma a few days later it gradually enlarged till the clot which plugged the arterial opening became so thin that arterial blood flowed into the cavity to form a pulsating hematoma. So, a pulsatile mass with vascular systolic murmur can be observed.

[Hyperbaric oxygen treatment for cystoid macular edema secondary to retinal vein occlusion].

A prospective study was made to estimate the effect of hyperbaric oxygen (HBO) on 14 eyes of cystoid macular edema (CME) secondary to retinal vein occlusion. The visual acuity was improved 2-6 lines (average 3.6 lines) after the treatment, while the control group of 12 cases showed little improvement. The therapeutic effect was stable, however, fluorescein angiography showed no leakage reduction from the perifoveal capillaries, or visual function was enhanced prior to eventual improvement in the fundus. Since HBO therapy has few contraindications nor side effects, it is an effective modality in the treatment of CME.