Identification of potential candidate genes for the Huoyan trait in developing Wulong goose embryos by transcriptomic analysis.

1. The Wulong goose is a Chinese breed and a source of high-quality meat and eggs. A characteristic of the Wulong goose is that a proportion of the birds do not have eyelids, known as the Huoyon trait.2. Wulong geese exhibiting the Huoyan trait at embryonic stages of 9 days (E9), 12 days (E12) and 14 days (E14) were selected alongside those with normal eyelids for comprehensive transcriptome sequencing. Differentially expressed gene (DEG) and functional enrichment analyses were performed and finally, eight DEG were chosen to verify the accuracy of qPCR sequencing.3. Overall, 466, 962 and 550 DEG were obtained from the three control groups, D9 vs. N9, D12 vs. N12 and D14 vs. N14, respectively, by differential analysis (p < 0.05). CDKN1C, CRH, CROCC and TYSND1 were significantly expressed in the three groups. Enrichment analysis revealed the enrichment of CROCC and TYSND1 in pathways of cell cycle process, endocytosis, microtubule-based process, microtubule organising centre organisation, protein processing and protein maturation. CDKN1C and CRH were enriched in the cell cycle and cAMP signalling pathway.4. Some collagen family genes were detected among the DEGs, including COL3A1, COL4A5, COL4A2 and COL4A1. FREM1 and FREM2 genes were detected in both Huoyan and normal eyelids. There was a significant difference (p < 0.01) in FREM1 expression between ED9 and ED14 in female embryos, but this difference was not observed in male embryos.

Gga-let-7a-3p inhibits the proliferation and differentiation of chicken intramuscular preadipocytes.

1. Intramuscular fat (IMF) is a key parameter for chicken meat quality. IMF deposition is driven by genetic, nutritional and management factors, with genetics being the determining factor. Previous whole transcriptome sequencing revealed that microRNA gga-let-7a-3p was related to lipid metabolism in breast muscle. This study further investigated the potential role of gga-let-7a-3p in IMF deposition.2. The mimic and inhibitor of gga-let-7a-3p were individually transfected into chicken intramuscular preadipocytes. Subsequently, the proliferation and differentiation states of the cells were detected. Transcriptome sequencing was performed on cells transfected with gga-let-7a-3p mimic.3. The results indicated that gga-let-7a-3p suppressed the mRNA levels of proliferation and differentiation-related genes, as well as the protein levels. EdU and Oil Red O assays revealed that gga-let-7a-3p restrained preadipocyte proliferation and differentiation. In addition, a total of 333 up-regulated genes and 807 down-regulated genes were identified in cells transfected with gga-let-7a-3p mimic. Using Kyoto Encyclopaedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis, differential genes were found to be enriched in processes such as the peroxisome proliferator activated receptor (PPAR) signalling pathway and oxidative phosphorylation.4. The study demonstrated that gga-let-7a-3p inhibits the proliferation and differentiation of chicken intramuscular preadipocytes, which provides new understanding to further unravel the function of gga-let-7a-3p.

[Preliminary report on the use of total lumpectomyconical remnant gastric - esophagus side overlap anastomosis in radical resection of Siewert type II proximal gastric cancer].

Objective: There is no standard method for esophageal remnant gastric reconstruction for proximal gastrectomy. Reflux esophagitis caused by esophagogastrostomy remains a difficult surgical problem. To report the preliminary surgical results of novel esophagus-conical remnant gastric side overlap anastomosis (CGEO) , with particular emphasis on postoperative esophageal reflux. Methods: In June 2022, we developed a novel CGEO for laparoscopic proximal gastrectomy on two patients with Siewert type II esophagogastric junction adenocarcinoma. Surgical procedures for CGEO: (1) Laparoscopic proximal gastrectomy and preparation of conically shaped gastric remnant; (2) Determining anastomotic site of residual stomach and esophagus; (3) Side-to-side anastomosis of right esophageal wall to anterior of conical gastric remnant; (4) Valvuloplasty of esophageal stump. Results: Case 1 was a 71-year-old man with an operation time of 305 minutes and was successfully discharged from the hospital on the 9th day after surgery, and the postoperative pathology was T3N0M0. Case 2 was an 82-year-old man with an operation time of 325 minutes. He was discharged on the 10th day after surgery. In both cases, only mild esophageal mucosal changes were seen in gastroscopy, there were no obvious symptoms of esophageal reflux. There was also no significant weight change at half a year after operation. Conclusion: CGEO is moderately safe in radical surgery for proximal gastric cancer, and may have a preventive effect on the occurrence of postoperative esophageal reflux, but long-term results need to be confirmed by further studies with follow-up.

Tumor purity-associated genes influence hepatocellular carcinoma prognosis and tumor microenvironment.

Introduction: Tumor purity takes on critical significance to the progression of solid tumors. The aim of this study was at exploring potential prognostic genes correlated with tumor purity in hepatocellular carcinoma (HCC) by bioinformatics analysis. Methods: The ESTIMATE algorithm was applied for determining the tumor purity of HCC samples from The Cancer Genome Atlas (TCGA). The tumor purity-associated genes with differential expression (DEGs) were identified based on overlap analysis, weighted gene co-expression network analysis (WGCNA), and differential expression analysis. The prognostic genes were identified in terms of the prognostic model construction based on the Kaplan-Meier (K-M) survival analysis and Least Absolute Shrinkage and Selection Operator (LASSO) regression analyses. The expression of the above-described genes was further validated by the GSE105130 dataset from the Gene Expression Omnibus (GEO) database. We also characterized the clinical and immunophenotypes of prognostic genes. Gene set enrichment analysis (GSEA) was carried out for exploring the biological signaling pathway. Results: A total of 26 tumor purity-associated DEGs were identified, which were involved in biological processes such as immune/inflammatory responses and fatty acid elongation. Ultimately, we identified ADCK3, HK3, and PPT1 as the prognostic genes for HCC. Moreover, HCC patients exhibiting higher ADCK3 expression and lower HK3 and PPT1 expressions had a better prognosis. Furthermore, high HK3 and PPT1 expressions and low ADCK3 expression resulted in high tumor purity, high immune score, high stromal score, and high ESTIMATE score. GSEA showed that the abovementioned prognostic genes showed a significant correlation with immune-inflammatory response, tumor growth, and fatty acid production/degradation. Discussion: In conclusion, this study identified novel predictive biomarkers (ADCK3, HK3, and PPT1) and studied the underlying molecular mechanisms of HCC pathology initially.

[Related factors and prognosis analysis of esophagorespiratory fistula after esophageal cancer surgery].

Objective: To explore the factors associated with the development of esophagorespiratory fistula (ERF) after esophageal cancer surgery and its relationship with patient survival. Methods: A total of 241 patients with esophageal cancer after surgery, who received postoperative sputum suction through bronchoscope from West China Hospital of Sichuan University between January and December 2021 were included. The clinical data and airway features under bronchoscope of these patients were collected. Of the 241 patients, 203 were males (84.2%) and 38 were females (15.8%), aged (63.63±8.05) years. The related factors of ERF were analyzed by multivariate logistic regression analysis, and Kaplan-meier was used to analyze the relationship between bronchoscopic specific manifestations, treatment modality and patient survival. Results: Of the 241 postoperative patients with esophageal cancer, 21 (8.7%) developed ERF. There were 39 (16.2%) patients with bronchoscopic specific manifestations, including 16 cases (6.6%) of hyperemia, 13 cases (5.4%) of congestion, and 15 cases (6.2%) of erosion. Bronchoscopic specific manifestations of tracheal mucosa (OR=13.734, 95%CI: 3.535-29.074, P<0.001) and thoracotomy (OR=9.121, 95%CI 1.843-44.237, P=0.007) were independent risk factors for the development of ERF, and preoperative chemotherapy (OR=0.128, 95%CI: 0.052-0.607, P=0.006) was a protective factor in the occurrence of ERF. The median survival time was 224 (95%CI: 95-353)d in the stent-treated group (14 patients) after the onset of ERF, and the median survival time of patients in the supportive care group (7 patients) was 29 (95%CI: 8-50)d, and the survival difference was statistically significant (χ2=5.69, P=0.017). Conclusions: Bronchoscopic specific manifestations are independent risk factors for the development of ERF in postoperative patients with esophageal cancer and are useful in assessing the risk of developing ERF. After the occurrence of postoperative ERF, timely intervention by insertion of tracheal stents to seal the fistula may prolong the survival time of the patients.

[Clinical application of three-dimensional printed preformed titanium mesh combined with free latissimus dorsi muscle flap in the treatment of squamous cell carcinoma with skull defect in the vertex].

Objective: To explore the clinical effects of three-dimensional printed preformed titanium mesh combined with latissimus dorsi muscle flap free transplantation in the treatment of wounds with skull defect after radical surgery of squamous cell carcinoma in the vertex. Methods: A retrospective observational study was conducted. From January 2010 to December 2019, 5 patients with squamous cell carcinoma in the vertex accompanied with skull invasion who met the inclusion criteria were admitted to the Department of Burns and Plastic Surgery of the Second Affiliated Hospital of Air Force Medical University, including four males and one female, aged 50 to 65 years. The original lesion areas ranged from 5 cm×4 cm to 15 cm×8 cm. The titanium mesh was prefabricated via three-dimensional technic based on the result the scope of skull resection predicted with computerized tomography three-dimensional reconstruction before surgery. During the first stage, the soft tissue defect area of scalp (8 cm×7 cm to 18 cm×11 cm) after tumor enlargement resection was repaired with the preformed titanium mesh, and the titanium mesh was covered with latissimus dorsi muscle flap, with area of 10 cm×9 cm to 20 cm×13 cm. The thoracodorsal artery/vein was anastomosed with the superficial temporal artery/vein on one side. The muscle ends in the donor site were sutured together or performed with transfixion, and then the skin on the back were covered back to the donor site. On the 10th day after the first-stage surgery, the second-stage surgery was performed. The thin intermediate thickness skin graft was taken from the anterolateral thigh to cover the latissimus dorsi muscle flap. The duration and intraoperative blood loss of first-stage surgery were recorded. The postoperative muscle flap survival after the first-stage surgery and skin graft survival after the second-stage surgery was observed. The occurrence of complications, head appearance, and recurrence of tumor were followed up. Results: The average first-stage surgery duration of patients was 12.1 h, and the intraoperative blood loss was not more than 1 200 mL. The muscle flaps in the first-stage surgery and the skin grafts in the second-stage surgery all survived well. During the follow-up of 6-18 months, no complications such as exposure of titanium mesh or infection occurred, with good shape in the recipient sites in the vertex, and no recurrence of tumor. Conclusions: Three-dimensional printed preformed titanium mesh combined with latissimus dorsi muscle flap free transplantation and intermediate thickness skin graft cover is an effective and reliable method for repairing the wound with skull defect after extended resection of squamous cell carcinoma in the vertex. This method can cover the wound effectively as well as promote both recipient and donor sites to obtain good function and appearance.

[Research advances on application of botulinum toxin type A in scar prevention and treatment].

The wound healing time, tension of wound edge, proliferation of fibroblast, and extracellular matrix deposition are the important factors of scar formation, and botulinum toxin type A can regulate the above. Prevention and treatment of scar with botulinum toxin type A is one of the hot topics of clinical research in recent years. This paper briefly reviews researches by scholars at home and abroad on the mechanism, clinical application, complications, and adverse effects of botulinum toxin type A in scar prevention and treatment.

[Comparison and application of postprandial and fasting blood lipid levels in 839 physical examinees in Beijing].

Objective: To investigate the feasibility of application of non-fasting dyslipidemia cutoff values in community population. Methods: Self-control study was used. 839 physical examinees (292 males and 547 females) were recruited in clinical laboratory of Guang'an men Hospital from January to October 2018. The median (interquartile range) of age was 60 (54, 66) years. Blood samples were collected before and at 4 h after a standard breakfast. Comparison of fasting and postprandial lipoprotein levels was performed using Paired-Samples T Test or Two-Related-Samples Wilcoxon. The changes of 4-hour postprandial blood lipid levels and the percentages of postprandial dyslipidemia according to different stratification of fasting dyslipidemia were performed using one-way ANOVA and χ2 test, respectively. Results: Compared with fasting, 4-hour postprandial total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), non-high density lipoprotein cholesterol (non-HDL-C), apolipoprotein A1 (ApoA1) and apolipoprotein B (ApoB) decreased slightly, postprandial triglyceride (TG) increased by 0.72 mmol/L, and postprandial remnant-like lipoprotein cholesterol (RLP-C) increased by 0.27 mmol/L (t or Z values = 10.26,22.94,24.22,4.71,16.61,26.92,-23.58,-19.35, P<0.05, respectively). According to the non-fasting dyslipidemia cut-off values recommended by the European consensus, there were 10%, 16.6%, 10.1%, 12.3%, 30% and 34.9% of the population in the appropriate levels of fasting TC, LDL-C, HDL-C, non-HDL-C, TG and RLP-C distributed in elevated levels of postprandial, respectively. The changes of 4-hour postprandial TC, LDL-C, non-HDL-C and HDL-C increased with the elevation of fasting level (F=9.50,6.18,8.07,3.86,P<0.01), and the maximum changes of TC≤3.5%, LDL-C≤6.8%, non-HDL-C≤2.9%, HDL-C≤6.3%; the change of 4-hour postprandial TG increased slightly first and then decreased significantly (51.3% vs. 57.9% vs. 39.2%, F=19.05, P<0.01); the change of 4-hour postprandial RLP-C decreased (50.8% vs. 33.2%, F=10.40, P<0.01). The cut-off values of 4-hour postprandial dyslipidemia were TC ≥5.1 mmol/L, LDL-C ≥3.2 mmol/L, HDL-C ≤0.9 mmol/L, non-HDL-C ≥4.0 mmol/L and RLP-C ≥1.0 mmol/L. The cut-off values of borderline elevated and elevated TG levels were ≥2.2 mmol/L and ≥3.4 mmol/L, respectively. Conclusions: The cut-off values of postprandial dyslipidemia including TC, LDL-C, HDL-C, non-HDL-C and RLP-C were preliminarily established in community population, which could be applied to the routine lipid profile evaluation in the physical examination population. And it might be needed that postprandial TG was managed hierarchically according to different cut-off values.

[Application effects of free transplantation of autologous omentum in treating maxillofacial infection complicated with complex sinus tract].

Objective: To investigate the application effects of free transplantation of autologous omentum in treating maxillofacial infection complicated with complex sinus tract. Methods: The retrospective observational study method was used. Four patients with maxillofacial infection complicated with complex sinus tract were admitted to Department of Burns and Plastic Surgery of the Second Affiliated Hospital of Air Force Medical University from July 2017 to the December 2019, including 3 males and 1 female aged 36-60 years. Preoperative facial computed tomography (CT) was performed on patients for calculating the volume of sinus tract. During the operation, the sinus tract was thoroughly debrided, and the volume of the tissue defect was about 5 cm×3 cm×2 cm-10 cm×5 cm×3 cm after debridement. The tissue defect area was filled with omentum of 100-300 mL which was cut under laparoscopy. The artery and vein on the right side of the omentum were reserved as the vascular pedicle of the donor area, which were anastomosed with the facial artery and external jugular vein of the recipient area. The survival of omentum, and the occurrences of reinfection and complication were observed after operation, respectively. On the 10th day and in 1 month after the operation, the blood supply of omentum was examined by colored Doppler ultrasound and CT angiography, and the filling of tissue defect area was examined by head and face CT. During follow-up after the operation, the recoveries of face appearance and function and scar hyperplasia in the donor area. Results: The transplanted omentums in 4 patients survived after the operation with no reinfection and complication. On the 10th day and in 1 month after the operation, the transplanted omentums had good blood supply, and the filled area with omentum was in good shape, without formation of dead cavity. During follow-up of 6-10 months after surgery, the appearance and function of face recovered well, and there was no obvious scar hyperplasia in the donor area. Conclusions: After free transplantation of omentum in treating maxillofacial infection with complex sinus tract, the patients have good facial appearance and function, and the application of laparoscopy results in little damage to the patients and quick postoperative recovery.

[Changes and clinical significance of serum NSE and MBP levels in children with cerebral palsy at high altitude during comprehensive rehabilitation].

Objective: To explore the changes of serum neuron-specific enolase (NSE) and myelin basic protein (MBP) in children with cerebral palsy at high altitude during comprehensive rehabilitation and their clinical significance. Methods: A clinical randomized controlled study design was used to select 144 children with cerebral palsy who were diagnosed and treated in the Rehabilitation Center of Xining Traditional Chinese Medicine Hospital of Qinghai Province from June 2018 to October 2019, including 83 males and 61 females, aged 3-5 years old. According to the order of admission, the random number table was used to divide into a conventional treatment group (n=72, 40 males and 32 females) and a comprehensive treatment group (n=72, 43 males and 29 females). The conventional treatment group was treated with conventional rehabilitation. The comprehensive treatment group was treated with monosialotetrahexose ganglioside sodium on the basis of conventional rehabilitation. In addition, 30 healthy children aged 3-5 years, 16 males and 14 females, were selected as the control group during the physical examination of the Pediatrics Department of Xining Hospital of Traditional Chinese Medicine, Qinghai Province. The serum levels of NSE and MBP in each group were detected, and the children's GMFM-88 scores were evaluated before and after treatment. The SPSS19.0 software was used for statistical analysis, the count data was tested by χ2. Results: The serum NSE and MBP levels of the control group were (5.96±0.80), (0.71±0.15) µg/L. Before treatment, the serum NSE and MBP levels of children with severe, moderate, and mild cerebral palsy were [(21.63±1.92), (3.63±0.49) µg/L], [(17.86±1.43) µg/L, (2.21±0.07) µg/L] and [(15.14±0.95), (1.76±0.30) µg/L], respectively. After treatment, the serum NSE and MBP levels of the conventional treatment group and the comprehensive treatment group were [(13.54±2.41), (2.07±0.85) µg/L] and [(12.09±2.37), (1.81±0.69) µg/L], respectively, and the GMFM-88 score was (116.75±27.41) points and (125.94±24.93) points. The levels of NSE and MBP in the serum of children with cerebral palsy were significantly higher than those of normal children in the control group, and their levels increased with the degree of disease, and the corresponding gross motor function scores were lower. After treatment, the GMFM-88 scale assessment scores of the two groups of children were significantly improved (t values were 310.97 and 70.86, P values were both<0.05), and serum NSE and MBP levels decreased to varying degrees compared with before treatment. The decline in the comprehensive treatment group was greater than that in the conventional treatment group. Conclusions: Serum NSE and MBP levels in children with cerebral palsy at high altitude are significantly higher than those in healthy children, and their levels are closely related to the degree of impairment and GMFM-88 scores in children with cerebral palsy. Dynamic monitoring of changes in NSE and MBP levels may be responsible for the condition and treatment effects of children with cerebral palsy judgments based.

LncRNA NR2F1-AS1 promotes proliferation and metastasis of ESCC cells via regulating EMT.

OBJECTIVE: The aim of this study was to explore the expression of long non-coding ribonucleic acid (lncRNA) nuclear receptor subfamily 2 group F member 1-antisense RNA 1 (NR2F1-AS1) in esophageal squamous cell carcinoma (ESCC) tissues and cells and to investigate its effects on ESCC proliferation and metastasis. PATIENTS AND METHODS: The expression level of NR2F1-AS1 in 51 pairs of ESCC tissues and corresponding adjacent tissues was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Meanwhile, NR2F1-AS1 expression in ESCC cells was measured via qRT-PCR as well. Subsequently, specific interference sequences of NR2F1-AS1 were designed, synthesized, and transiently transfected into ESCC cells. 48 h later, qRT-PCR assay was performed to detect the interference efficiency. The effects of small interfering (si)-NR2F1-AS1 on the proliferation of ESCC cells were determined through cell counting kit-8 (CCK-8) and colony formation assay. Wound healing and transwell assays were conducted to investigate the influences of si-NR2F1-AS1 on the migration and invasion of ESCC cells. Additionally, the changes in the expressions of epithelial-mesenchymal transition (EMT) molecular markers were detected by Western blotting. RESULTS: QRT-PCR assay revealed that the expression level of NR2F1-AS1 was significantly up-regulated in 42 of 51 cases of ESCC tissues (42/51, 82.4%). Compared with esophageal mucosal epithelial HET-1A cells, NR2FA-AS1 was highly expressed in ESCC cells. CCK-8 and colony formation assay indicated that the proliferation of ESCC cells decreased remarkably after interference in NR2F1-AS1 expression. The results of wound healing and transwell assays showed that the migration and metastasis of cells were significantly lower in si-NR2F1-AS1 group than those in si-NC group. Western blotting demonstrated that the expressions of EMT molecular markers were changed after interfering with NR2F1-AS1 expression. CONCLUSIONS: NR2F1-AS1 was highly expressed in ESCC tissues and cells. Furthermore, high expression of NR2F1-AS1 promoted the proliferation and metastasis of ESCC cells by modulating EMT.

[Early diagnosis of subtype in early clinical stage lung adenocarcinoma by using an autoantibody panel and computed tomography].

Objective: To explore the value of CT and autoantibody panel to diagnosis the subtype in early clinical stage lung cancer,especially lepidic predominant adenocarcinoma (LPA),and to provide the correct information for the clinical and the prognosis evaluation. Methods: A retrospective study was conducted of 60 patients (total 63 subsolid nodules,which included 39 PSN and 24 pGGN) who underwent surgical resection or needle biopsy for stage Ⅰa or Ⅰb lung adenocarcinoma at Affiliated Tumor Hospital of Zhengzhou University between June 2017 and April 2018,age from 28 to 82 years old,and the average age was (52±7) years. All patients underwent a pretreatment chest LDCT and the peripheral blood of all patients were used to detect the tumor related antibody (including p53, GAGE7, PGP9.5, CAGE, MAGEA1, SOX2, GBU4-5) through enzyme linked immunosorbent assay. All the patients were divided into LPA group (43 subsolid nodules, which included AIS 10 subsolid nodules, MIA 11 subsolid nodules, LPA 22 subsolid nodules) and invasive predominant adenocarcinoma (IPA) group (20 subsolid nodules). The information of CT scanning was measured and assessed in lung and mediastinal windows with double blind method. The mean computed tomography (m-CT) value and the solid component/tumor ratio in three-dimensional (3D) and two-dimensional (2D) planes were measured and analyzed using computer-aided diagnosis (CAD) system. Results: There were 20 partial solid nodules in IPA group, 19 partial solid nodules in LPA group and 24 ground-glass nodules (χ(2)=19.278,P=0.000). There were 4 circular nodules, 16 irregular nodules in the IPA group, 21 circular nodules, 5 oval nodules and 7 irregular nodules in the LPA group χ(2)=8.587,P=0.003). The incidence of burr, vascular aggregation and bronchial truncation in IPA group was higher than that in LPA group (40.0% vs 16.3%, 70.0% vs 18.6%, 30.0% vs 2.3%, χ(2)=4.234,15.860,10.580, P=0.040,0.000, 0.001). The incidence of clear tumor lung interface in patients in LPA group was significantly higher than that in patients in IPA group (97.7% vs 65.0%, χ(2)=13.146,P=0.00). Of all the quantitative analysis of nodules,the m-CT value, the solid component/tumor ratios in three-dimensional (3D) and two-dimensional (2D) planes in IPA group were higher than those of LPA group ((-180±156) vs (-410±213) HU, 0.44±0.32 vs 0.14±0.26, 0.54±0.26 vs 0.18±0.26, t=-4.208, -3.951、-5.166, P=0.000, 0.000, 0.000). Among the 60 patients with lung cancer, there were 33 cases with positive antibody in peripheral blood, with a positive rate of 55.0%. The positive rate of 7-AABs was 70.0% in IPA group and 44.2% in LPA group, which had no statistical difference (χ(2)=3.647, P=0.056), the positive expression of tumor-associated antibodies was independent of the patient's age, CT value and nodular solid components and lung nodular volume ratio and area ratio, P>0.05, only in correlation with pleural traction (χ(2)=3.866, P=0.049). Conclusion: Compared with IPA, the imaging features of LDCT about the mGGN and PGGN appearance, clear tumor-lung interface, low m-CT and the solid component/tumor ratio in two-dimensional or three-dimensional (3D) planes are benefit for the diagnosis of the LPA; the expression of tumor-associated antibody group is independent of the age of the patient and the number of nodular solid components, and is only related to pleural depression, which is not conducive to the identification of LPA and IPA.

MiR-203a functions as a tumor suppressor in bladder cancer by targeting SIX4.

Increasing evidence indicates that microRNAs (miRNAs) have essential roles in various biological processes, including proliferation, migration, invasion, cell cycle progression and apoptosis. It is considered that miRNA de-regulation contributes to tumor progression and metastasis in various cancers, and MiR-203a has been identified as a tumor suppressor in cancers, such as glioma, gastric cancer and hepatocellular carcinoma. Herein, we established that miR-203a expression is significantly lower in bladder cancer tissues than in adjacent normal tissues, and that low miR-203a expression is associated with poor patient outcome. The over-expression of miR-203a inhibited bladder cancer cell proliferation, invasion, migration and EMT in vitro, and its up-regulation led to bladder cancer cell cycle arrest and apoptosis. This over-expression also inhibited the PI3K/Akt signaling pathway. Bioinformatics prediction software and luciferase reporter assay then confirmed that SIX4 is a direct target of miR-203a. We established negative correlation between SIX4 expression and miR-203a expression in bladder cancer tissues, and SIX4 silencing caused effects similar to miR-203a up-regulation Furthermore, SIX4 over-expression diminished the effects of miR-203a on bladder cancer cells in vitro. In summary, our study determined that miR-203a down-regulation is closely related to tumorigenesis in bladder cancer; thus suggesting that miR-203a is a potential prognostic marker and a potential target in bladder cancer treatment.

Evidence for prevalent Z = 6 magic number in neutron-rich carbon isotopes.

The nuclear shell structure, which originates in the nearly independent motion of nucleons in an average potential, provides an important guide for our understanding of nuclear structure and the underlying nuclear forces. Its most remarkable fingerprint is the existence of the so-called magic numbers of protons and neutrons associated with extra stability. Although the introduction of a phenomenological spin-orbit (SO) coupling force in 1949 helped in explaining the magic numbers, its origins are still open questions. Here, we present experimental evidence for the smallest SO-originated magic number (subshell closure) at the proton number six in 13-20C obtained from systematic analysis of point-proton distribution radii, electromagnetic transition rates and atomic masses of light nuclei. Performing ab initio calculations on 14,15C, we show that the observed proton distribution radii and subshell closure can be explained by the state-of-the-art nuclear theory with chiral nucleon-nucleon and three-nucleon forces, which are rooted in the quantum chromodynamics.

Effects of different levels of feed intake during four short periods of gestation and housing systems on sows and litter performance.

The current study investigated the effects of different levels of feed intake during 4 short periods of gestation and of housing systems on sow and litter performance. A total of 255 multiparous sows were allotted to 1-4 dietary treatments using a randomized complete block design blocking by initial body weight (BW), backfat (BF) and parity. Sows were housed either in individual stalls (n=129) or group pens (n=126) with 55 sows in each pen with electronic sow feeder during gestation. All sows were fed one common corn-soybean meal-based diet with the amount of 1.0×maintenance energy level of feed intake (106×BW0.75) throughout gestation except 4 periods of 7 d when dietary treatments were imposed on day 27, 55, 83 and 97 of gestation. During the 4 periods, sows were fed 1 of 4 different levels of feed intake: 0.5, 1.0, 1.5 and 2.0×maintenance energy level (0.5M, 1.0M, 1.5M and 2.0M, respectively). Results showed that both BW gain and BF change during gestation for sows on 1.5M (49.7kg and 3.1mm, respectively) and 2.0M (52.5kg and 3.7mm, respectively) levels of feed intake were significantly (P<0.01) greater than sows on 0.5M (26.1kg and -0.1mm, respectively) and 1.0M (35.6kg and 0.1, respectively) levels of feed intake. In contrast, lactation weight gain for sows on 1.5M (3.3kg) and 2.0M (3.4kg) levels of feed intake during 4 short periods of gestation were significantly (P<0.01) less than sows on 0.5M (18.4kg) and 1.0M (11.4kg) levels of feed intake during 4 short periods of gestation, whereas BF loss during lactation for sows on 1.5M (-3.6mm) level of feed intake during 4 short periods of gestation were significantly (P=0.03) higher than sows on 1.0M (-2.1mm) level of feed intake during 4 short periods of gestation. Additionally, average daily feed intake during lactation for sows on 0.5M (6.6kg/d) level of feed intake during gestation tended (P=0.06) to be greater than sows on 2.0M (5.9kg/d) level of feed intake. There were no differences (P>0.1) among 4 levels of feed intake in terms of numbers of total born and weaning piglets. However, both piglet weight at birth (1.46, 1.52, 1.53 and 1.51kg for piglets from sows on 0.5M, 1.0M, 1.5M and 2.0M levels of feed intake during gestation, respectively) and at weaning (6.37, 6.55, 6.64 and 6.38kg for piglets from sows on 0.5M, 1.0M, 1.5M and 2.0M levels of feed intake during gestation, respectively) were maximized at 1.5M level of feed intake. Sows housed in group pens had greater (P<0.01) net BW gain (24.7 vs. 19.2kg) from day 27 of gestation to weaning compared with sows housed in individual stalls. However, there were no differences (P>0.1) between the 2 housing systems in terms of litter size and piglet weight at birth and at weaning. In conclusion, increasing levels of feed intake during 4 short periods of gestation increased BW and BF gain during gestation and led to less BW gain and more BF loss during lactation. Piglet weight at birth and at weaning was maximized at 1.5M level of feed intake. However, housing systems did not affect reproductive performance. Group pen housing system may be beneficial in terms of increased overall BW gain during gestation and lactation.

[Comparison of the prognostic value of the seventh and eighth edition of The AJCC Esophageal Cancer Staging System for the patients with stage â ¡ and â ¢esophageal squamous cell carcinoma].

Objective: To compare and evaluate the prognostic value of the 7(th) and 8(th) edition of The AJCC Esophageal Cancer Staging System for patients with stage Ⅱ and Ⅲ esophageal squamous cell carcinoma. Methods: The clinical data of 328 esophageal cancer patients who received operation at Department of Esophageal Cancer, Tianjin Tumour Hospital from January 2006 to December 2010 were restrospectively analyzed. There were 63 female and 265 male patients. The mean age was 65 (range: 33 to 87) years. Univariate and multivariate analysis were performed to identify the prognosis factors. Results: The five years overall survival rates among patients with stage Ⅱ and Ⅲ were both significantly different (χ(2)=87.035, 84.730, all P=0.000) according to the 7(th) and 8(th) editions of the TNM staging systems. The five years overall survival rate among patients with stage ⅡB and ⅢA were significantly different (39.6% vs 23.4%, P=0.001) according to the 7(th) edition of the esophageal cancer staging systems.According to the 8(th) edition of the esophageal cancer staging system, the 5 years survival rate of patients with stage ⅡA and ⅡB, ⅢB and Ⅳ was statistically significant (58.5% vs. 35.5%, P=0.040; 18.9% vs. 0, P=0.000). In multivariate analysis, tumor size, T staging, N staging and tumor differentiation (HR=1.592, 95%CI: 1.185 to 2.139, P=0.002; HR=1.519, 95% CI: 1.236 to 1.867, P=0.000; HR=1.647, 95% CI: 1.448 to 1.874, P=0.000; HR=1.404, 95% CI: 1.059 to 1.861, P=0.018) were the main independent prognosis factors affecting the prognosis of esophageal squamous cell carcinoma patients. Conclusions: Both the 7(th) and the 8(th) editions of TNM staging systems are able to reflect the clinical prognosis of patients receiving radical resection of esophageal cancer, and the factors of tumor size, differentiaton, invasion depth and lymph node metastases are the independent predictors of prognosis. The 8(th) edition provides a more detailed and more reasonable for the staging of stage Ⅱ and Ⅲ for esophageal cancer patients than the 7(th) edition, and it is more accurate for the prognosis of patients with esophageal cancer after surgery.

Assessment of serum Golgi protein 73 as a biomarker for the diagnosis of significant fibrosis in patients with chronic HBV infection.

Transient elastography (TE) is accurate in staging fibrosis noninvasively. However, a reliable serum biomarker with comparable accuracy is also important, especially when TE is unreliable/unavailable. Therefore, we aimed to evaluate the diagnostic performance of serum Golgi protein 73 (GP73) for significant fibrosis in patients with chronic HBV infection. A total of 801 patients with chronic liver disease (CLD; 492 chronic HBV infection and 309 non-HBV liver disease) with liver biopsy performance were enrolled. Healthy controls (n = 180) and hepatocellular carcinoma (HCC) patients (n = 85) were included for comparisons. Liver biopsy was used as the reference method for fibrosis staging. Serum GP73 level was measured in duplicate in double-blind fashion. Serum GP73 was highest in HCC but also significantly higher in chronic hepatitis B than in healthy controls. The elevation of serum GP73 in non-HCC patients was significantly associated with the presence of significant fibrosis independently of ALT level, liver stiffness (LS) value, inflammation grade and other confounding factors. The diagnostic performance of serum GP73 was accurate in antiviral-naïve HBV patients (area under the receiver operating curve [AUROC], 0.76 95% CI: 0.72-0.81) but not in patients with ongoing antiviral treatment (AUROC, 0.60). The utility of serum GP73 was also confirmed in non-HBV CLD (AUROC, 0.80 95% CI: 0.75-0.85). Serum GP73 was comparable to LS (AUROC, 0.78 95% CI: 0.73-0.82) and significantly better than AST to platelet ratio index (APRI) (AUROC, 0.67 95% CI: 0.62-0.72) and FIB-4 (AUROC, 0.68 95% CI: 0.63-0.73). In conclusion, serum GP73 is an accurate serum marker for significant fibrosis in chronic HBV infection, with higher accuracy than APRI and FIB-4. Serum GP73 is potentially a complementary tool for TE when evaluating the necessity of antiviral treatment, particularly in patients without definite antiviral indication.

Epigenetic silencing of microRNA-137 enhances ASCT2 expression and tumor glutamine metabolism.

Tumor cells must activate specific transporters to meet their increased glutamine metabolic demands. Relative to other glutamine transporters, the ASC family transporter 2 (ASCT2, also called SLC1A5) is profoundly elevated in a wide spectrum of human cancers to coordinate metabolic reprogramming and malignant transformation. Understanding the molecular mechanisms whereby tumor cells frequently upregulate this transporter is therefore vital to develop potential strategies for transporter-targeted therapies. Combining in-silico algorithms with systemic experimental screening, we herein identify the tumor suppressor microRNA, miR-137, as an essential regulator that targets ASCT2 and cancer cell glutamine metabolism. Metabolic analysis shows that miR-137 derepression, similar to ASCT2 inactivation, significantly inhibits glutamine consumption and TCA cycle anaplerosis. Mechanistically, methyl-CpG-binding protein 2 (MeCP2) and DNA methyltransferases (DNMTs) cooperate to promote active methylation of the miR-137 promoter and inhibit its transcription, conversely reactivating ASCT2 expression and glutamine metabolism. Moreover, expression between miR-137 and ASCT2 is inversely correlated in tumor specimens from multiple cancer types, and ectopic ASCT2 expression markedly rescued miR-137 suppression of tumorigenesis. These findings thus elucidate a previously unreported mechanism responsible for ASCT2 deregulation in human cancers and identify ASCT2 as a critical downstream effector of miR-137, revealing a molecular link between DNA methylation, microRNA and tumor metabolism.

Effects of different feeding levels during three short periods of gestation on gilt and litter performance, nutrient digestibility, and energy homeostasis in gilts.

The present study investigated the effects of different feeding levels during 3 short periods of gestation on gilt and litter performance, apparent total tract digestibility (ATTD) of energy and nutrients, and energy homeostasis in gilts. A total of 18 gilts were allotted to 1 of 3 dietary treatments using a completely randomized design. All gilts were fed 1 common corn-soybean meal-based diet with the amount of 1.0 × maintenance energy intake (100 × BW (BW) kcal ME/d) throughout gestation except 3 periods of 7 d when dietary treatments were imposed on d 27, d 55, and d 83 of gestation. During the 3 short periods, gilts were fed 1 of 3 different feeding levels: 0.5, 1.0, and 2.0 × maintenance energy level (0.5M, 1.0M, and 2.0M, respectively). Results showed that gilts on 2.0M feeding level had higher ( < 0.05) weight gain from d 27 to 109 of gestation (37.05 vs. 15.34 kg) and greater ( < 0.05) BW change, average daily gain, and gain to feed ratio during gestation periods 1 (d 27-34) and 3 (d 83-90) when compared with gilts on 0.5M feeding level. No differences ( > 0.10) in litter performance were observed among the 3 feeding levels. Additionally, the slopes of BW change in response to feeding levels in period 1 were 4.32 kg/0.5M change from 0.5M to 1.0M feeding level and 3.72 kg/0.5M change from 1.0M to 2.0M feeding level, respectively. There were quadratic ( < 0.05) effects of feeding levels on ATTD of dry matter and gross energy during periods 1 and 2 (d 55-62). Furthermore, fasting plasma concentrations of acyl ghrelin and nonesterified fatty acid (NEFA) in period 1 were greater ( < 0.01) in gilts on 0.5M feeding level than those on 2.0M feeding level. In conclusion, increasing feeding levels during 3 short periods increased primiparous sow performance during these short periods but did not affect litter performance. ATTD of energy and nutrients, and BW change efficiency were maximized for gilts on 1.0 M feeding level. The data also indicated that sows on the lowest feeding level were exposed to negative energy balance as evidenced by the higher plasma acyl ghrelin and NEFA concentrations.