Objective:To investigate the effects of methyltransferase-like protein 14 (METTL14)-mediated long-chain non-coding RNA EIF3J antisense RNA1 (Inc EIF3J-AS1) on the migration and invasion of cholangiocarcinoma cells and its mechanism.Methods:From September 2017 to December 2018, 10 pairs of cholangiocarcinoma and adjacent normal tissues were collected from the First Affiliated Hospital of Naval Medical University, which were surgically resected and pathologically confirmed. The expression of METTL14 mRNA and Inc EIF3J-AS1 in cholangiocarcinoma tissues was detected by fluorescence quantitative PCR, and the protein expression of METTL14 was detected by Western blotting. Cholangiocarcinoma cell lines HUCCTI and RBE were divided into control group and METTL14 or Inc EIF3J-AS1 knockdown group. The corresponding normal lentivirus was transfected in the control group, and METTL14 or Inc EIF3J-AS1 knockdown group was transfected with lentivirus that interfered with the expression of METTL14 or Inc EIF3J-AS1, respectively. Transwell assay was used to detect the ability of cell migration and invasion, and Western blotting was used to detect the expression of epidermal growth factor receptor (EGFR) and AKT protein.Results:The expressions of METTL14 mRNA and lnc EIF3J-AS1 in cholangiocarcinoma tissues were significantly higher than those in adjacent normal tissues (0.075±0.012 vs 0.031±0.006, 0.140±0.032 vs 0.064±0.012), and there was a positive correlation between expression of METTL4 mRNA and expression of lnc EIF3J-AS1 ( r=0.883, P=0.0007). The expression of METTL14 protein in cholangiocarcinoma tissues was higher than that in adjacent normal tissues (0.354±0.131 vs 0.187±0.183). Compared with the control group, the expression of lnc EIF3J-AS1 was significantly lower in METTL14 or Inc EIF3J-AS1 knockdown group (0.217±0.020 vs 1.000±0.052, 0.149±0.066 vs 1.000±0.045). The migration and invasion ability of cell lines HUCCTI and RBE decreased significantly in lnc EIF3J-AS1 knockout group (5.00±0.58 vs 23.33±0.33, 20.33±0.67 vs 70.67±0.33; 12.00±0.58 vs 25.00±2.52, 22.33±0.89 vs 43.67±0.33). The expression of EGFR and p-AKT/AKT protein were also significantly decreased (0.109±0.015 vs 1.000±0.018, 0.226±0.036 vs 1.000±0.051; 0.118±0.052 vs 1.000±0.069, 0.132±0.098 vs 1.000±0.023). The above differences were statistically significant (all P<0.05). Conclusions:Abnormal expression of lnc EIF3J-AS1 in cholangiocarcinoma mediated by METTL14 can promote tumor cell migration and invasion.

Objective:To investigate the influence of echinoside on the expressions of inducible nitric oxide synthase (iNOS) and transforming growth factor-β1 (TGF-β1) in renal tissues of diabetic nephropathy rats.Methods:A total of 60 male SD rats were randomly divided into the normal group, model group, glibenclamide group and low-, medium-, high-dose echinoside treatment group. the rat model of diabetic nephropathy was established through the streptozocin induction and the glibenclamide treatment was applied as control. After molding, glibenclamide group was given 5 mg/(kg?d) of glibenclamide, and low-, medium-, high-dose echinoside treatment group was given 50, 100 and 200 mg/(kg?d) of echinacoside solution. The normal group and model group were intragastrically given equal volume of normal saline. After 4 weeks, the change rate of body weight, kidney weight/body weight, blood glucose, serum TC, TG, BUN, SCR and 24 h urinary protein levels were measured, and the NO release concentration in renal tissue cell culture medium was measured, the renal pathology was observed by HE staining, the expression of iNOS and TGF-β1 proteins in renal tissues was detected by Western blotting.Results:Compared with the model group, the body weight of rats in the medium-dose group significantly increased ( P<0.05); the weight change rate and kidney weight/body weight of rat in the medium-, high-dose group significantly decreased ( P<0.05), and the NO release concentration in the medium-, high-dose group decreased ( P<0.05); the level of blood glucose, TC, TG, BUN, SCr, and 24-hour urinary protein in the low-, medium- and high-dose groups significantly decreased ( P<0.05), and iNOS in renal tissue of rats in the low-, medium- and high-dose groups was significantly lower ( P<0.05). Compared with the model group, the relative expression levels of TGF-β1 (1.35 ± 0.06, 1.15 ± 0.05, 0.91 ± 0.04 vs. 4.58 ± 0.09) and TGF-β1 (1.24 ± 0.04, 1.41 ± 0.05, 0.89 ± 0.04 vs. 3.04 ± 0.05) in the low-, medium- and high-dose groups significantly decreased ( P<0.05). Conclusions:The echinoside can prevent weight loss, reduce blood glucose and triglyceride, inhibit the production of NO in renal tissues, reduce the expression of iNOS and TGF-β1 among diabetic nephropathy rats. It has been proved to play a protective role in diabetic nephropathy pathological tissues.

OBJECTIVE: To analyze the incidence and risk factors of benign liver space-occupying mass in patients with chronic hepatitis B (CHB) and the ultrasound features that differentiate these masses from small hepatocellular carcinoma. METHODS: We retrospectively analyzed the color Doppler and clinical data of 17 721 patients with CHB treated in the Hepatology Unit of Nanfang Hospital between January, 2016 and December, 2017. The data were compared with those of 21629 healthy control subjects undergoing routine physical examination in the Center of Heath Management of Nanfang Hospital during the same period. RESULTS: Compared with the control subjects, the patients with CHB had significantly higher incidences of hepatic cysts (11.8% 8.7%, < 0.05), hepatic hemangioma (8.2% 1.6%, < 0.05) and hepatic cirrhosis nodules (20.6% 2.4%, < 0.05). The incidences of hepatic cysts and cirrhosis nodules increased with age and was significantly higher in male than in female patients ( < 0.001). The highest incidence of hepatic hemangioma was found in CHB patients aged 30-49 years without a gender difference (>0.05). Sonographically, the benign liver masses commonly showed homogeneous echo within the lesion with clear boundaries and regular shape. Hepatic hemangioma was distinctively hyperechoic in 83.32% (1579/1895) of the patients, while small hepatocellular carcinoma presented with weaker peripheral and internal blood flow signals with a lower flow velocity in the arteries and a higher flow velocity in the portal vein. Liver cirrhosis nodules mostly showed a mixture of strong and weak echoes (79.60%; 7637/9595) without blood flow signal within or around the nodule; an increased volume of the nodule accompanied by heterogeneous echoes within the nodule indicated an increased probability of malignant lesion. Hepatic cysts often displayed no echo within the lesion, but the echo could be enhanced posteriorly. CONCLUSIONS The patients with CHB are at a significantly higher risk of developing hepatic cysts, hepatic hemangiomas and hepatic cirrhosis nodules than the control population, and an older age and the male gender are associated with a higher incidence of hepatic cysts or cirrhosis. The differences in the sonographic and hemodynamic features can help to differentiate hepatic benign mass from malignant lesions, and kinetic changes in sonography can be used to monitor potential malignant transformation of the cirrhotic lesions.

Objective To investigate the predictive value of HIT-antibodies(HIT-Ab) detection for new thrombus in suspected Heparin-Induced thrombocytopenia (HIT). Methods Retrospective cohort study. 472 suspected HIT patients were collected from July 2016 to November 2018, and all subjects under-went a 4Ts score and were sent for HIT-Ab tests. According to the results of HIT-Ab, there were four groups:412 cases of negative HIT-Ab (0-0.9 U/ml), 45 cases of weak-positive HIT-Ab (1.0-4.9 U/ml), 12 cases of moderate-positive HIT-Ab (5.0-15.9 U/ml), and 3 cases of strong-positive HIT-Ab (≥16.0 U/ml) respective-ly. Ultrasound or CT examination was used to confirm new thrombosis as a standard to evaluate the value of HIT-Ab for predicting new thrombus. The diagnostic efficacy of HIT-Ab for HIT was evaluated in clinically confirmed HIT. Results The incidence rates of new thrombus in each group were: 15.8％ in Negative HIT-Ab group (62/412), 48.9％in Weak-positiveHIT-Ab group (22/45), 75.0％in Moderate-positive HIT-Ab group (9/12), and100％in Strong-positive HIT-Ab group (3/3)(P<0.00). When HIT-Ab≥1.0 U/ml, the speci-ficity for diagnosing new thrombus was 93.0％, the sensitivity was 34.2％, the negative predictive value (NPV) was 84.2％, and the positive predictive value (PPV) was 56.5％. The diagnostic rates of HIT in each group were:negative 0％(0/412), weak-positive 62.2％(28/45), moderate-positive (12/12) and strong-positive (3/3) were 100％. When HIT-Ab≥ 1.0 U/ml, the specificity for HIT diagnosis was 96.0％, the sensitivity was 100％, NPV was 100％, and PPV was 71.5％. Conclusions In suspected HIT patients, the incidence of new thrombosis increases with the elevated HIT-Ab level. HIT-Ab detection can be used as a crucial tool for new thrombosis prediction and HIT diagnosis in suspected HIT patients. Clinicians can develop treatment strategies based on HIT-Ab levels.

Objective To explore the emergency strategy of microsurgical replantation to open degloving injuries in plantar soft tissue.Methods From July,2013 to July,2017,5 patients with plantar degloving injury were treated by micro-replantation,3 of which were injuried by traffic accident and 2 caused by crashed injury.After early debridement,4 cases received anastomosis of blood vessels,and 1 treated with Venous Autograft to get recovery of blood circulation.The avulsion flaps were sutured without tension.Results Four flaps survived after plantar replantation,and 1 developed a partial necrosis and secondery repaird with skin graft.The plantar repair made sensory function preserved in the maximum extent.All patients were followed-up for 0.5-2.0 years.The appearance,elasticity and feel of flaps was good.Conclusion Complete debridement,early recovery of blood supply and maximum preservation of plantar tissue function should be the key to successful replantation of open plantar degloving injuries.

Objective To further improve the value of clinical application of a quantitative analysis method called LF-index (LFI) which based on real-time tissue elastography (RTE). Methods We prospectively enrolled 116 consecutive patients with chronic hepatitis B (CHB) and all patients underwent a liver biopsy and RTE between January 2015 and December 2015 at the First affiliated hospital of Wenzhou Medical University. Univariate and multivariate analyses were performed, and the prediction models for predicting significant fibrosis and cirrhosis were derived from independent predictors. Results (1) In multivariate analyses, spleno-portal index (SPI) (OR=13.956, P=0.002) and LFI (OR=6.283, P=0.023) were confirmed as independent predictors of significant fibrosis. In multivariate analyses of patients with and without cirrhosis, we found significant differences in the γ-Glutamyl transferase (GGT) (OR=1.012, P=0.049), SPI (OR=5.676, P=0.002) and LFI (OR=14.102, P=0.001). (2)A novel model called LFI-SPI score (LSPS) for prediction of significant fibrosis was developed (area under receiver operating characteristic curve [AUROC]=0.87), showing the superiority of diagnostic accuracy than LFI (AUROC=0.76, P=0.0109), aspartate aminotransferase to platelet ratio index (APRI) (AUROC=0.64, P=0.0031), fibrosis-4 index (FIB-4) (AUROC=0.67, P= 0.0044) and Fibroscan (AUROC=0.68, P=0.0021). (3) We also developed a LFI-SPI-GGT score (LSPGS) for predicting cirrhosis, with an AUROC of 0.93. The diagnostic accuracy of LSPGS was similar to that of Fibroscan (AUROC=0.85, P=0.134), and was superior to LFI (AUROC=0.81, P=0.0113), APRI (AUROC=0.67, P<0.0001), and FIB-4 (AUROC=0.72, P=0.0005). Conclusions We developed new formulas, LSPS and LSPGS for predicting significant fibrosis and cirrhosis in this prospective study. LSP score was mainly used for screening of significant liver fibrosis, and LSPG score was mainly used to exclude cirrhosis.

Objective To investigate the effects of ghrelin on proliferation of vascular smooth muscle cells(VSMC)and the expression of mitochondrial fusion 2(Mfn-2)in cultured human aortic smooth muscle cells(HASMCs).Methods HASMCs were cultured in vitro,treated with different concentrations(10-9,10-8,10-7,10-6,10-5 mol/L)ghrelin or 10-6 mol/L ghrelin for different time(0,6,12,18,24 h).Subconfluent HASMCs at passage 4-6 were used in experiments.MTT essay was used to investigate the effect on proliferation of HASMCs.RT-PCR and Western blot were used to analyse the expression of Mfn-2.Results 10-7-10-5 mol/L ghrelin inhibited the proliferation of HASMCs,and the inhibitory effect of concentration of 10-6 mol/L was the most obvious(P<0.01).Ghrelin inhibited the proliferation of HASMCs in 6-24 h,and it reached the peak at 24 h(P<0.01).10-6 mol/L ghrelin significantly increased the expression of Mfn-2 mRNA and protein(P<0.01).The up-regulation of 10-6 mol/L ghrelin on Mfn-2 mRNA and protein expression reached the peak at 18 h(P<0.01).Conclusion Ghrelin might inhibit the proliferation of HASMC by up-regulating the expression of Mfn-2.

Objective:To investigate the site and characteristic ofp53 gene mutations in familial or early-onset breast cancer patients in part population of southern China.Methods:A total of 150 patients with familial and early-onset breast cancer in parts population of southern China were enrolled.Genomic DNA was isolated from each peripheral blood sample,and the entire coding sequence and exon and intron splicing region of p53 gene were amplificated by PCR in the 150 patients.The mutation analysis were detected by denaturing high performance liquid chromatography (DHPLC) and confirmed by DNA sequence analysis.Results:In the 150 patients with familial and early-onset breast cancer,6 mutations including one novel pathogenic mutation 869_888 ins20 (insert mutation) and 5 previously reported pathogenic mutations (deletion mutation 643_660de118 and 4 missense mutation 91G＞A,215C＞G,537T＞G,743G＞A) were identified in p53 gene encoding region in 9 patients of breast cancer.Moreover,one same sense mutation 141G＞A in exon 4,one 16 bases deletion in intron 3,and 9 single nucleotide polymorphisms in p53 gene introns were also identified.The total mutation frequency ofp53 gene in 150 patients with familial breast cancer and early-onset breast cancer from part population of southern China was 6.00％,and the mutation frequency of familial breast cancer and early-onset breast cancer was 6.81％ and 6.25％,respectively.Conclusion:The total mutation frequency ofp53 gene in 150 patients with familial breast cancer and early-onset breast cancer from partpopulation of southern China is higher than the frequency previously reported.The pathogenicity of the novel mutations (insert mutation) 869_888ins20 will be confirmed by function analysis in the future study.The deletion mutation 643_660de118 enriches the p53 gene mutation database among Chinese population,which is probably the specific mutation of breast cancer in Chinese population.

Objective:To observe the clinical effect of TOMY self-ligation appliance technique in the treatment of anterior crossbite and moderate crowding, and to explore its clinical application.Methods:A patient, manifested as anterior crossbite and moderate dentition crowing, diagnosed as Angle class Ⅲ subclass malocclusion, MaoⅡ1+Ⅰ1 malocclusion, high angle,and facial asymmetry, was selected.This patient was treated with TOMY self-ligation appliance technique without tooth extraction.The length and width of the dental arch, the width of the alveolar bone and the width of the basal arch were measured before and after treatment;the lateral radiographs of the patient were taken too.Results:After treatment,the upper and lower dentition arranged in neat rows, reached a neutral occlusal relationship;SNA, SNB and ANB had no obvious changes;the lip inclination of anterior teeth was increased by 10 degrees, and reached the normal value;the upper and lower dental arch width, alveolar bone width, basal bone arch width and dental arch length were lower than before treatment.Conclusion:For the patient with anterior crossbite and moderate dental crowding treated with non-extraction, the TOMY self-ligation technique can be considered to use, by increasing the arch width and length to improve the anterior crossbite and moderate crowding, in order to achieve the effective treatment.

Objective To evaluate the effect of long term home non-invasive positive pressure ventilation (HNIPPV ) in stable severe chronic obstructive pulmonary disease (COPD) .Methods Forty-two patients with stable severe COPD after hospital dis-charge were divided into 2 groups :The observation group (conventional treatment + HNIPPV ,22 patients) and control group (con-ventional treatment ,20 patients) .Parameters before and after one year follow-up observation were compared ,which includes arterial bloods gases ,lung function test ,6-min walking distance(6MWD) ,dyspnea grade ,scoring for emotional disorders ,the hospitalization rates .Results PaCO2 ,PaO2 ,6MWD ,dyspnea grade ,scoring for emotional disorders ,the hospitalization rates improved after one year in the observation group (P < 0 .05) .There were no significance of FVC and FEV1 between the two groups after one year . There were no patients who were dead or exited with other reason after one year in the two groups .Conclusion Long term HNIP-PV could decrease exacerbations ,respiratory failure and increase life quality ,this therapy is effective and safe for patients with sta-ble severe COPD .