Objective To study the influence and mechanism of hepatocyte growth factor (HGF) on myotube phenotype by myotube transdifferentiation induced by transforming growth factor-β1 (TGF-β1). Methods C2C12 cells were cultured in differentiation medium to induce myotubes formation. The cells were randomly devided into 3 groups. The control group without growth factor interruption. The induction group was supplemented with TGF-β1 (5 ng/mL) while the inhibition group was supplenmented with both TGF-β1 (5 ng/mL) and HGF (30 ng/mL). After 12 hours, the expressions of connective tissue growth factor (CTGF) protein in myotubes were detected by Western blot, the levels of CTGF mRNA were measured by RT-PCR. Results Compared to the control group, the protein and mRNA levels of CTGF significantly increased in TGF-β1 treated group , whereas the protein and mRNA levels of CTGF were significantly lower in inhibition group than those in induction group (P < 0.05). Conclusion HGF can inhibit the effect of TGF-β1 on the expression of CTGF in myotubes , which provides the evidences on the study of skeletal muscle cell transdifferentiation.

Objective To explore molecular mechanisms underlying the in vitro counteracting effect of curcumin on malignant melanoma.Methods Cultured A375 and C8161 human melanoma cells were cultivated in vitro,and randomly divided into several test groups and a control group to be treated with different concentrations of curcumin and dimethyl sulfoxide respectively for different durations.Then,methyl thiazolyl tetrazolium (MTT) assay,Transwell assay,flow cytometry and Western blot were performed to evaluate the effect of curcumin on the proliferation,invasion and cell cycle of,as well as expressions of AKT/mTOR signaling pathway-related proteins in A375 and C8161 cells respectively.Statistical analysis was carried out by using t test.Results MTT assay showed that the treatment with curcumin of 5-35 mg/L for 24-96 hours significantly inhibited the proliferation of both A375 and C8161 cells compared with that with dimethyl sulfoxide (all P ＜ 0.001),and the inhibitory effect was in a dose-dependent manner within the range of 5-15 mg/L for A375 cells and within the range of 5-10 mg/L for C8161 cells,and in a time-dependent manner from 0 to 48 hours for both cells.After treatment for 24 hours,the 50％ inhibitory concentration (IC50) of curcumin against A375 cells and C 8161 cells was 10 mg/L and 5 mg/L respectively.Transwell assay demonstrated that the invasion of A375 and C8161 cells was significantly suppressed by 72-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively (both P ＜ 0.001).Flow cytometry showed that the cell cycle of A375 and C8161 cells was arrested at G2/M phase after 24-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively,with significant differences in the proportion of A375 cells and C8161 cells in G2/M phase between the test group and control group (A375 cells:35.00％ ± 3.54％ vs.120.80％ ± 7.46％,P＜ 0.001;C8161 cells:19.33％ ± 4.04％ vs.85.00％ ± 9.53％,P ＜ 0.001).Western blot revealed that the expressions of AKT/mTOR signaling pathway-related proteins were decreased in A375 and C8161 cells after 24-hour treatment with 10 mg/L and 5 mg/L curcumin respectively.Conclusion Curcumin can inhibit the proliferation and invasion of A375 and C8161 cells,likely by blocking cell cycle and inhibiting activation of the AKT/mTOR signaling pathway.

Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

Objective To assess the feasibility and curative effects of laser surgery in treatment of early-stage glottic carcinoma under self-retaining laryngoscope with nasal endoscopic assistance.Methods Thirteen cases of glottic carcinoma(TisN0M0:2 cases;T1aN0M0:7 cases;T1bN0M0:3 cases;T2N0M0:1 case) extirpated with Nd:YAG laser or Diomed laser from June 1999 to March 2003 were retrospectively analyzed.Because of the poor glottic exposure with self-retaining laryngoscope under general anesthesia,various angular nasal endoscopes were used.The tumor was distinctly displayed on monitor screen with nasal endoscopic assistance and were completely removed through vapourization or removed with Nd:YAG laser or Diomed laser.Results The follow-up period was three to five years.Two patients were found to have local recurrence.One patient received treatment with Nd:YAG laser surgery under electronic laryngoscope with local and superfacial anesthesia,and the patient was followed up for three years without showing recurrence.The other patient underwent vertical hemilaryngectomy and was followed up for three and half years without showing recurrence.All 13 cases were followed up for three years,and all of them were alive.Three-year survival rate was 100%(13/13).When they were followed up for five years,one patient lost track(counted as death),therefore the five-year survival rate was 92.3%(12/13).Conclusions With the assistance of various angular nasal endoscope,the glottic tumors can be exposed distinctly under self-retaining laryngoscope and removed completely.Nd:YAG laser or Diomed laser is an effective and safe procedure with minimal invasion for the treatment of early-stage glottic carcinoma.The advantages of this technique include lower complication rate and high rate of preservation of laryngeal functions,avoidance of tracheotomy and the sequela of laryngeal fissure.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

The editorial board and publication ethics committee of the CEO decided to retract this paper from our journal.

OBJECTIVE: To investigate the influence of denervation on myofiber morphology of the adductor and the abductor in patients with recurrent laryngeal nerve (RLN) paralysis and to provide experimental evidence for the clinical feasibility of RLN repair. METHOD: Adductor muscles were acquired from the lateral cricoarytenoid muscle (LCAM) and abductor muscles from the posterior cricoarytenoid muscle(PCAM). Normal human PCAM and LCAM are treated as control group (n = 7). Thirty-eight cases of PCAM with damaged RLN were divided into five groups according to the duration of their RLN damage: 0.5-1 year (7 cases), > 1-2 years (10 cases), > 2-3 years (8 cases), > 3-6 years (8 cases) and > 6 years (5 cases); twenty-nine cases of LCAM were also divided into five groups: 0.5-1 year (7 cases), > 1-2 years (6 cases); > 2-3 years (6 cases), > 3-6 years (6 cases) and > 6 years group(4 cases). They were all stained with HE and Masson three-color staining, the fiber cross-sectional area of muscle tissue and collagen connective tissue were quantitative analyzed. The changes of myofiber morphology of adductor and abductor muscles after the loss of the RLN were analyzed with image analysis system. RESULT: The transverse areas of myofibers gradually decreased and those of collagen fibers gradually increased with the prolongation of denervation. (1) Difference between the denervated groups of LCAM of 0.5-1 year, > 1-2 years and > 2-3 years groups were not significant (P > 0.05). Fiber cross-sectional area of > 3-6 years group decreased most obviously with significantly difference compared with > 2-3 years group (P < 0.05); (2) There were obvious difference between the control group, 0.5-1 year group, > 1-2 years group, > 2-3 years group and > 3-6 years of PCAM(P < 0.05); (3) There was no significant difference between the group of > 3-6 years and > 6 years of two kinds of laryngeal intrinsic muscle (P > 0.05); (4) Fiber cross-sectional area of each group of the LCAM after 1 year denervation were significantly greater than that of the PCAM under same conditions (P < 0.05). CONCLUSION The influence of denervation on myofiber morphology following denervation is different between the abductor and adductor owing to the different fiber type composition and functional properties. The rate of muscle atrophy of the adductor is slower than that of the abductor. To restore the structure and function of denervated laryngeal muscles better, the recurrent laryngeal nerve injury repair surgery for PCA muscle function recovery should be carried out within 1 year after denervation while the surgery for LCA muscle function recovery should be carried out within 3 years after denervation.
Case-Control Studies ; Denervation ; Humans ; Laryngeal Muscles ; innervation ; pathology ; Myofibrils ; pathology ; Neurosurgical Procedures ; Recurrent Laryngeal Nerve ; pathology ; Staining and Labeling ; Vocal Cord Paralysis ; pathology ; surgery

OBJECTIVES: To investigate the surgical outcomes of different uvulopalatopharyngoplasty (UPPP). METHODS: All subjects underwent overnight polysomnography and were evaluated using the Epworth sleepiness scale (ESS), the Quebec sleep questionnaire and the snoring scale at the baseline and 3 and 12 months following operation. The primary endpoint was the overall effective rate representing the sum of the surgical success rate and effective rate. RESULTS: The overall effective rate at 12 months post surgery was 55.6% for simple UPPP, 95.8% for UPPP+GA, and 92.3% for UPPP+TBA. The surgical success rate at 3 and 12 months postoperation for UPPP+GA or UPPP+TBA was significantly higher than simple UPPP (P<0.05). Marked improvement was observed in all patients in the snoring scale score and the ESS score 3 and 12 months following surgery compared to the baseline (P<0.05 in all). CONCLUSION: UPPP, UPPP+GA, and UPPP+TBA are all effective in improving the surgical outcome of obstructive sleep apnea hypopnea syndrome (OSAHS) patients with multilevel obstruction. UPPP+TBA appears to be the most effective in treating OSAHS patients.
Humans ; Polysomnography ; Prospective Studies* ; Quebec ; Sleep Apnea Syndromes ; Sleep Apnea, Obstructive* ; Snoring ; Tongue* ; Surveys and Questionnaires

Objective To investigate the clinicopathologic features of Chinese patients with early gastric cancer (EGC) according to the World Health Organization(WHO) diagnostic criteria,and to explore the risk factors of lymph node metastasis (LNM) in EGC.Methods From 2002 to 2017,at the Second Affiliated Hospital of Nantong University,and from 2014 to 2017,at the People's Hospital of Qidong City,the People's Hospital of Haimen City and the People's Hospital of Rugao City,315 EGC patients with complete clinicopathological data were enrolled.The clinicopathologic features were analyzed including gender,age,tumor location,tumor size,macroscopic type,histological type (WHO),differentiation degree,depth of tumor invasion,Lauren subtype,infiltration pattern,ulceration,lymphoid follicles,lymphovascular invasion and perineural invasion.Chi-square test and Fisher's exact test were performed for univariate analysis and the logistic regression was used for multivariate analysis.Results The ratio of EGC was 10.0％ (315/3 140) of patients with gastric cancer and received surgery during the same period.Among the 315 EGC,the ratio of well-differentiated tubular adenocarcinoma was 11.7％ (37/315),which was lower than 24.8％(2 752/11 104) in Japan and 19.9％(41/206) in South Korea,and the differences were statistically significant (x2 =28.208,P＜0.01;x2 =6.51 0,P =0.011),however there was no statistically significant difference when compared with Western countries (11.9 ％,8/67;x2 =0.002,P=0.964).Among the 276 patients who underwent radical gastrectomy,49 (17.8 ％) patients had with LNM.The results of univariate analysis showed that tumor size,macroscopic type,differentiation degree,depth of invasion,infiltration pattern,ulceration and lymphovascular invasion were related with LNM(x2=9.327,6.038,6.381,34.983,19.309,52.297 and 5.058;all P＜0.05).The results of multivariate analysis revealed that lymphovascular invasion and ulceration were the independent risk factors of LNM (odd ratio (OR)=7.028 and 2.566,both P＜0.05).Conclusions There is obvious difference in pathological diagnostic standard of well-differentiated tubular adenocarcinoma between China,Japan and South Korea,which may influence the therapeutic strategy of EGC.Lymphovascular invasion and ulceration are independent risk factors of LNM in EGC.

OBJECTIVE: To investigate the expression of myosin heavy chain (MHC) in normal laryngeal muscle and the difference between the adductor and abductor. METHOD: Seven patients with total laryngectomy were enrolled in this study. The adductor muscles were acquired from the lateral cricoarytenoid (LCA) muscle and the abductor muscles were acquired from the posterior cricoarytenoid (PCA) muscle. The expression of myosin heavy chain were detected with fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining respectively. RESULT: (1) MHC-II b was expressed in laryngeal muscles at mRNA levels, and not expressed at the protein level; (2) At both mRNA level and protein level, the expression of MHC-I was higher in the PCA muscles than in the LCA muscles while MHC-II level was higher in the LCA muscles than in the PCA muscles. CONCLUSION (1) MHC-II b protein was not expressed in human laryngeal muscles; (2) Phenotypic differences were significant in laryngeal adductor and abductor muscles based on their different functions. PCA contained larger percentage of MHC-I fibers, while LCA contained more MHC-II fibers.
Aged ; Female ; Humans ; Laryngeal Muscles ; chemistry ; metabolism ; Male ; Middle Aged ; Myosin Heavy Chains ; genetics ; metabolism ; Phenotype ; Protein Isoforms