Genetic diseases that have shown clinical symptoms in neonatal period are often characterized with atypical symptoms and serious condition, which contributes to difficulties in diagnosis and treatment.With the progress of sequencing technology, the next-generation sequencing technology is gradually applied to the clinical field with its advantages of high throughput, low cost and rapid detection.As one of next-generation sequencing technologies, the whole exome sequencing technology(WES)captures, enriches and sequences the genomic exon regions, and then the large amount of WES data is analyzed by bioinformatics methods and screened to find variant site in gene that causes genetic disease.WES technology has gradually become an important means of diagnosis in neonatal genetic diseases because of its advantages of comprehensive results and short reporting period.

The aim of the present study is to investigate the effects of sequoyitol (Seq) on expression of eNOS and NOX4 in aortas of type 2 diabetic rats. Type 2 diabetic rats induced by high fat and high sugar diet and low dose of streptozotocin (STZ, 35 mg x kg(-1)) and were administered Seq (12.5, 25 and 50 mg x kg(-1) x d(-1)) for 6 weeks. The fasting blood glucose (FBG) and body weight were tested. Acetylcholine (Ach) induced endothelium-dependent relaxation and sodium nitroprusside (SNP) induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Aortic morphological change was observed with HE staining. The level of serum insulin was measured by radioimmunoassay. The total antioxidative capacity (T-AOC), malondialdehyde (MDA) and NO levels in aortas were determined according to the manufacturer's instructions. In addition, the expressions of eNOS and NOX4 in aortas were measured by immunohistochemisty, real-time PCR or Western blotting. The results showed that Seq significantly decreased FBG and insulin resistance, and improved aortic endothelium-dependent vasorelaxation function. The expressions of NOX4 and MDA content were obviously decreased, while the expression of eNOS, the levels of NO and T-AOC increased significantly in aortas of diabetic rats with Seq treatment. In conclusion, Seq protects against aortic endothelial dysfunction of type 2 diabetic rats through down-regulating expression of NOX4 and up-regulating eNOS expression.
Animals ; Aorta ; metabolism ; pathology ; Blood Glucose ; metabolism ; Body Weight ; Diabetes Mellitus, Experimental ; chemically induced ; metabolism ; physiopathology ; Diabetes Mellitus, Type 2 ; chemically induced ; metabolism ; physiopathology ; Hypoglycemic Agents ; pharmacology ; Inositol ; analogs & derivatives ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Male ; Malondialdehyde ; metabolism ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidation-Reduction ; drug effects ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Vasodilation ; drug effects

OBJECTIVETo investigate the protective effect and mechanism of sequoyitol (Sep) on high glucose-induced human umbilical vein endothelial cells (HUVECs) injury.

METHODSHUVECs were cultured with high glucose (30 mmol/L) in the presence or absence of sequoyitol (0.1, 1 and 10 micromol/L) for 24 h. Cell proliferation was measured by BrdU marking and cell cycle was detected by flow cytometry. 2', 7'-dichlorofluorescein diacetate was used to evaluate intracellular reactive oxygen species (ROS) levels. The NO, malonydialdehyde (MDA) and H2O2 levels were determined by colorimetric method according to the manufacturer's instructions. The expression of endothelial nitric oxide synthase (eNOS) and NADPH oxidase 4 (NOX4) were measured by real-time PCR and Western blot.

RESULTSIn the present study, we found that sequoyitol pretreatment for 1 h significantly decreased cell injury, promoted cell proliferation. Meanwhile sequoyitol significantly down-regulated NOX4 expression and decreased the level of ROS, MDA and H2O2 and obviously increased NO levels and up-regulated eNOS expression.

CONCLUSIONSequoyitol alleviates high glucose-induced cell injuries in HUVECs via inhibiting oxidative stress and up-regulating eNOS expression.

Cell Proliferation ; Cells, Cultured ; Glucose ; toxicity ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; metabolism ; Inositol ; analogs & derivatives ; pharmacology ; Malondialdehyde ; metabolism ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism

Acute Coronary Syndrome ; blood ; Aged ; Cell Membrane ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; metabolism

ObjectiveTo evaluate prognostic factors affecting local tumor progression after radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC).MethodsA total of 246 HCC patients (343 lesions) underwent RFA treatment in our department and were enrolled into this study.The average tumor size was 3.7 cm ( range 0.9 ～ 3.7 cm).Regular follow-up with enhanced CT was performed to evalutate the treatment results.Kaplan-Meier model and log-rank test were used in univariate analysis and COX regression model was used in multivariate analysis to identify risk factors for local tumor progression.ResultsThe local tumor progression rate was 11.4％ (39/343 lesions),and the average time from initial RFA to local tumor progression was 12.0 months.Univariate analysis indicated tumor size ( P ＜0.001 ),close to intrahepatic vessels ( P ＜0.001),tumor boundary ( P =0.020),pathological grade( P =0.010) and CEUS before RFA ( P =0.001) as risk factors for local progression.The following factors were identified as independent prognostic factors for local tumor progression by multivariate model:tumor size (P ＜ 0.001),isolated or close to intrahepatic vessels( P ＜0.001) and CEUS before RFA(P =0.018).ConclusionsTumor size,CEUS before RFA and close to intrahepatic vessels are the most important factors for local progression after RFA.Being awaring of possible risk factors for local tumor progression may increase the treatment efficacy and help to promote the use of RFA technique.

Objective To introduce the outcomes of tracheal resection combined with primary end-to-end anastomosis for benign cervical tracheal stenosis,and to discuss the strategy for prevention of surgical complications.Methods The clinical data of 22 patients due to different causes benign cervical tracheal stenosis surgery were analyzed retrospectively.Results The length of cervical tracheal stenosis ranged was 2.2-4.2 cm.Grade Ⅱ stenosis was present in 6 patients.Grade Ⅲ stenosis was present in 11 patients and grade Ⅳ stenosis in 5 patients.Successful extubation was achieved in all 22 cases.After surgery,temporary hoarseness occurred to 1 case;unilateral pulmonary atelectasis with pleural effusion occurred to 1 case; subcutaneous emphysema with infection occurred to 1 case; mild dysphagia occurred to 3 cases;a slight deepening of the tone of voice in 10 patients with women occurred to 5 cases,granulation tissue growth near the suture occurred to 3 cases,and suture dehiscence did not occur in any patient.The follow-up period ranged from 6-45 months,no patient developed restenosis.Conclusions It presents a high success rate and good functional result of tracheal resection combined with primary end-to-end anastomosis.Therefore,it is an effective and reliable approach for the management of benign cervical tracheal stenosis.To avoid complications,the preoperative assessment,patients selection and postoperative management should be emphasized.

The aim of the present study is to investigate the effects of sequoyitol (Seq) on expression of eNOS and NOX4 in aortas of type 2 diabetic rats. Type 2 diabetic rats induced by high fat and high sugar diet and low dose of streptozotocin (STZ, 35 mg x kg(-1)) and were administered Seq (12.5, 25 and 50 mg x kg(-1) x d(-1)) for 6 weeks. The fasting blood glucose (FBG) and body weight were tested. Acetylcholine (Ach) induced endothelium-dependent relaxation and sodium nitroprusside (SNP) induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Aortic morphological change was observed with HE staining. The level of serum insulin was measured by radioimmunoassay. The total antioxidative capacity (T-AOC), malondialdehyde (MDA) and NO levels in aortas were determined according to the manufacturer's instructions. In addition, the expressions of eNOS and NOX4 in aortas were measured by immunohistochemisty, real-time PCR or Western blotting. The results showed that Seq significantly decreased FBG and insulin resistance, and improved aortic endothelium-dependent vasorelaxation function. The expressions of NOX4 and MDA content were obviously decreased, while the expression of eNOS, the levels of NO and T-AOC increased significantly in aortas of diabetic rats with Seq treatment. In conclusion, Seq protects against aortic endothelial dysfunction of type 2 diabetic rats through down-regulating expression of NOX4 and up-regulating eNOS expression.

Objective To restore the incisal angle defect of anterior upper teeth with partial cov-erage porcelain veneer, and evaluate the clinical performance of the restoration. Methods A total of 12 patients were included, and 14 anterior upper teeth with small incisal angle defect were involved in this study, after tooth reduction of 2 mm wider than the defect of every teeth, 14 partial coverage IPS-Empress Ⅱ veneers were made and binded with Variolink Ⅱ adhesive system. During the observation period of 36-48 months after insertion, prosthodontic results were evaluated, using the CDA quality e-valuation system, and marginal integrity, anatomic form, surface and color of the veneers, and sec-ondary caries were examined. Results During the period of 36-48 observation, all of the 14 restora-tions were rated satisfactory. Conclusions With strict control of indications, the partial coverage porcelain veneer could function satisfactorily as an alternative restoration for small incisal defect of an-terior upper teeth.

Objective To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection. Methods Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2、IFN-?、TNF-?、IL-4、 IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d. Results Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL-2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-? was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-? reached a peak after 80 d p.i., and decreased slowly after 140 d p.i., The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i. Conclusion The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection.

Objective To investigate the role of nerve growth factor (NGF) in the pathogenesis of diabetic neuropathy (DNP) and the correlation of NGF level with,sensory nerve conduction velocity (SNCV) and duration of DNP.MethodsThe 41 patients with DNP,35 diabetics without DNP and 33 healthy controls were enrolled.And the serum level of NGF was measured with ELISA,the SNCV of left medial nerve,ulnar nerve,peroneal nerve and tibial nerve were measured too. Then the correlation analysis was completed. ResultsThe serum level of NGF was significantly lower in DNP patients [(665.18±188.32) ng/L] than in healthy controls [(976.44±159.07)ng/L] and in diabetics without DNP [(943.32±167.33) ng/L,F=9.316,P<0.001].The NGF level of DNP patients was positively correlated with SNCV of left medial nerve (r=0.810,P<0.001 ),left peroneal nerve (r=0.760,P<0.001) and duration of DNP (r=0.542,P<0.001).Conclusions The serum level of NGF is lower in DNP patients than in healthy controls.The decrease of the production of NGF may play a role in the pathogenesis of DNP.