Objective To investigate the biological effect of wild p16 gene on senescence of human keloid fibroblasts (HKFb) and discuss the feasibility of gene therapy on keloid. Methods Firstly, the HKFb was transfected with pcDNA3-p16 and the positive clones selected by G418. Then, the fibroblast changes were investigated by determining growth curve, cycle profile and ultrastructure of the transfected HKFb. Meanwhile, morphological changes of the transfected HKFb were identified by senescence associated-?-galactosidase staining. Results After HKFb was transfected by p16, the morphology of HKFb changed obviously from the third passage on. The cell proliferation was inhibited markedly and the cell cycle profile analysis showed that about 96.7% cells were trapped in stage G1. The electroscopic analysis of HKFb also showed that the lysosomes were overloaded. Senescence associated-?-galactosidase staining identified the senescence of HKFb. Conclusions The wild p16 gene can induce the HKFb into senescence in advance, as may provide experimental basis for gene therapy on human keloid.

Objective To explore the value of diagnostic and differential diagnostic with the Neutrophil‐lymphocyte count ratio (NLCR) and C‐reactive protein cmmunity‐acquired pneumonia(CAP) .Methods The NLCR ,as well as white blood cell counts , neutrophil count ,lymphocyte count and the concentration of CRP were measured in 40 patients with bacterial pneumonia ,40 pa‐tients with mycoplasma pneumonia ,40 patients with viral pneumonia and 40 healthy subjects ,and the results was analyzed by statis‐tical methods .Results In bacterial group ,the NLCR and the concentration of CRP were significantly higher than those in myco‐plasma group ,viral group and normal controls(P<0 .05) .According to the results of ROC curve analysis in the diagnosis of bacte‐rial CAP ,the areas of NLCR and CRP under ROC curve are 0 .911 and 0 .896 ,respectively ,which have good diagnostic sensitivity and specificity .Conclusion The NLCR and CRP in peripheral blood have great significance in diagnosis and differential diagnosis of bacterial community‐acquired pneumonia .

Background and purpose:Lung cancer is one of the leading causes of cancer death in the world. The aim of this study was to evaluate whether Fas-670 G>A and Fasl-844 T>C polymorphisms were associated with the risk of lung cancer.Methods:Data from 400 lung cancer patients with speciifc histological diagnosis were collected from 2010 to 2015. Meanwhile, data from matched healthy controls with the same gender and ±5 years were also collected. The genotypes of Fas-670 G>A and Fasl-844 T>C polymorphisms were determined by TaqMan lfuorescent probe method, and the results were analyzed using SPSS 16.0 software.Results:A total number of 386 cases and 394 controls were successfully genotyped. Compared with AA genotypeofFasgene, the GA and GG genotype carriers had no signiifcantly increased risk of lung cancer.The OR values were 1.05 (95%CI: 0.77-1.44) and 0.77 (95%CI: 0.81-1.99) respectively. Compared with TT genotype ofFasl gene, the CT and CC genotype carriers had signiifcantly increased risk of lung cancer. The OR values were 1.37 (95%CI: 1.01-1.86) and 1.74 (95%CI: 1.09-2.77), respectively. Conclusion:Fasl-844 T>C polymorphism may be involved in lung cancer risk but not Fas-670 G>A polymorphism.

Objective:To investigate the clinical efficacy of amoxicillin combined with metronidazole in the treatment of peri-implantitis and its effect on serum matrix metalloproteinase-8 (MMP-8), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels.Methods:Sixty-four patients with peri-implantitis who received treatment from January 2017 to December 2019 in Xiasha Hospital, SiR Run Run Shaw Hospital, Zhejiang University School of Medicine were included in this study. They were randomly assigned to receive treatment with metronidazole (control group, n = 32) or amoxicillin combined with metronidazole (observation group, n = 32). Clinical efficacy, periodontal pocket depth (PD), clinical attachment level (CAL), sulcus bleeding index (SBI), plaque index (PLI), matrix metalloproteinase-8 (MMP-8), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) levels and the incidence of adverse reactions were compared between the control and observation groups. PD, CAL, SBI and PLI were correlated with MMP-8, TNF-α and IL-1β levels. Results:The clinical efficacy in the observation group was significantly higher than that in the control group [93.75% (30/32) vs. 75.00% (24/32), χ2 = 4.267, P < 0.05]. After treatment, PD [(3.02 ± 0.24) mm], CAL [(1.70 ± 0.14) mm], SBI [(11.54 ± 3.26)%], PLI (0.87 ± 0.11) in the observation group were significantly lower than those in the control group [(4.47 ± 1.37) mm, (3.77 ± 1.06) mm, (18.84 ± 5.11)%, (1.12 ± 0.15), t = 5.897, 10.952, 6.813, 7.603, all P < 0.01]. After treatment, serum MMP-8 [(0.78 ± 0.23) μg/L], TNF-α [(71.24 ± 10.33) μg/L], IL-1β [(1.20 ± 0.14) μg/L] levels in the observation group were significantly lower compared with the control group [(1.12 ± 0.35) μg/L, (113.28 ± 15.73) μg/L, (2.55 ± 1.02) μg/L, t = 4.592, 12.637, 7.417, all P < 0.01]. Spearman correlation coefficient analysis showed that PD, CAL, SBI and PLI were positively correlated with serum MMP-8, TNF-α and IL-1β levels (all P < 0.01). Conclusion:Amoxicillin combined with metronidazole is highly effective in the treatment of peri-implantitis because it can promote plaque regression, reduce inflammatory reaction, and has a certain clinical promotion value.

Background and purpose:Studies have shown that DOC-2 could work as a potential tumor suppressor geue,and the role of DOC-2 in terms of the inhibition of cell growth and its mechanism remain unknown.Our paper is to investigate the effect and mechanism of DOC-2 expression on the tumorigenesis viability of ovarian cancer cell line HO-8910 from the aspects of clone efficiency,cell cycle and animal model test.Methods:Three cell lines were used including HO-8910,8910-P93(transfected with DOC-2 gene) and 8910-pcDNA3.1(transfected with the vector pcDNA3.1).Firstly,soft agar method was used to measure the clone efficiency.The cell cycle were analyzed by flow cytometer.The tumorigenesis viability was compared by athymic mouse test.Results:After being transfected with DOC-2 gene,the clone efficiency of 8910-P93 was markedly reduced.There was no difference between the 8910-pcDNA3.1 and HO-8910.G1 and G2 arrest were observed for 8910-P93.The athymic mouse test showed that the neoplasm derived from 8910-P93 was much smaller than that in the controls.Conclusions:DOC-2 could iniibit the tumorigenesis viability of human ovarian cancer line HO-8910.

Background and purpose:DOC-2 serves as one of the tumor suppressing genes in human ovarian cancer and plays a role in the process of cell growth and differentiation.This study was to investigate the role of DOC-2 in the TGF? signal pathway and verification of the interaction between DOC-2 and TGF?Ⅲ receptor.Methods:The bait vector was constructed by inserting the PID domain of DOC-2(nDOC-2)into yeast express vector pGBKT7.pGBKT7-nDOC2 was transformed into the yeast AH109 and confirmed to be expressed.After the human foetus brain cDNA library had been transformed,the positive clones was screened by both nutrition defect medium and X-?-gal.The putative positive clones were sequenced and analyzed to get the DOC-2 interactive proteins.Furthermore,after the DOC-2 cDNA and TGF?Ⅲ receptor cDNA had been co-transfected into the human ovarian cancer cell line HO-8910 together,the interaction between DOC-2 and TGF?Ⅲ receptor was investigated by immunoprecipitation and Western blot.Results:21 putative positive clones were picked after being screened and sequenced.Three of them were identified as Homo sapiens partial mRNA for betaglycan(TBR Ⅲ gene),homo sapiens protocadherin gamma subfamily C3(PCDHGC3)and APLP1(amyloid beta precursor-like protein 1).The analysis by immunoprecipitation and Western blot showed that the interaction between DOC-2 and TGF?Ⅲ receptor could form protein complex.Conclusions:The three encoding proteins might participate in the DOC-2 signal pathway.DOC-2 might play as an essential role in the TGF?signal pathway by interacting with TGF?Ⅲ receptor.

Objective To investigate a new method of measuring serum total protein by improving biuret double reagent through two point terminal method in order to dissolve the sample's interference of hemolysis, icterus, lipid , polymeric glucose.Methods The precision, recovery rate, the examination limit, the linear scope and the interference experiments were done by improving biuret reagent in order to ensure the method's basic function. The method was compared to Doumas method in order to ensure its reliability and the effection of dissolving the interference.Results The proportion of sample and reagent was 5∶220 ?l and the linear scope was 140 g/L. Regress equation:Y=484.0+48.06X,r=0.999 3.The within run CV(n=20) was 0.24% and the day to day CV(n=20) was 1.07%.The average recovery rate was 99.5%(95.6%~103%).The examination limit was 2.36 g/L. The interference was not significant when Hb concen tration was lower than 2.1 g/L, Bil lower than 148 ?mol/L,TG lower than 28.2mmol/L and polymeric glucose lower than 30 g/L. While measuring normal sample, improving method and Doumas method were highly correlation. Regress equation:Y=-2.435 4+1.037 4X,r=0.988 2. There wasn't a significant reference between results.Conclusion It could dissolve effectively the sample's interference of hemolysis, icterus, lipid, polymeric glucose and increase the result's accuracy while adopting improving biuret double reagent through two point terminal method.

Objective To investigate the application and needs' survey of mobile App health software in the patients with respiratory chronic disease. To make mobile App health software development more targeted and practical in the near future. To provide the foundation for continuous nursing service. Methods A survey research method was adopted in which self-designed questionnaires were administered to 163 patients with respiratory chronic disease from respiratory ward and outpatient clinic. Results Totally 119(73.0%) patients used smartphone, the mean score of experience in using mobile phones was (32.4 ± 14.1), which was in the medium range. There were 80 patients had ever installed and used this kind of App. The frequency of use of mobile App health software was Breathing Exercises App, Healthy Exercise App, Healthy Diet App and Medicine Remind App. The major reason for using of these App was disease prevention, self health care, disease and health monitoring and rehabilitation exercises. There were 120 patients would like to install App health software specific to respiratory chronic disease. They hope the characters of these App were simple operated, practical and without product placement. These App health software should include following function:timely feedback and guidance, providing practical method and help to register. Conclusion Most patients with respiratory chronic disease had the hardware basis and experience basis of using mobile App health software. More targeted and practical mobile App health software for the patients with respiratory chronic diseases should be developed to provide the foundation for continuous nursing service and patient′s self-management.

Objective To analyze the mutations of causal genes in sixteen Chinese patients with suspicious Bartter syndrome,and follow up their treatment results.Methods Mutations were identified by the next generation sequencing and the multiplex ligation-dependent probe amplification (MLPA).Clinical and biochemical features at the first presentation as well as follow-up results were reviewed.Results 15 different CLCNKB gene mutations were identified in sixteen patients with BS,including 11 novel ones.A novel missense mutation and a novel small deletion were found from SLC12A1 gene.A novel gross deletion was found in CLCNKA gene.A recurrent missense mutation was identified from BSND gene.The whole gene deletion mutation of CLCNKB gene was the most frequent mutation (32％),and the rate of gross deletion was up to 50 percent in this group of Chinese patients.The most common clinical manifestations were development retardation (15/16),polydipsia and polyuria (15/16).All of the patients were detected with hypokalemia,hypochloremia and metabolic alkalosis.Indomethacin treatment had significant improvement to the stature and weight restoration.Conclusion The present study has found 19 mutations,including 14 novel ones,which enriches the human gene mutation database (HGMD) and provides valuable references to the genetic counseling and diagnosis of Chinese population.

Objective To investigate the CT features of hepatic echinococcosis in children.Methods Clinical and CT data of 27 cases with hepatic echinococcosis confirmed by pathology and/or serology were retrospectively analyzed.Results Totally there were 26 cases of cystic echinococcosis and 1 case of alveolar echinococcosis.Ten cases were with simple hepatic echinococcosis and 17 cases with other organic echinococcosis,18 cases of intrahepatic single hydatid and 9 cases of intrahepatic multiple hydatid.A total of 1 alveolar lesion and 47 cystic lesions were found,including unilocular cyst accounted for 51.06％ (24/47),cysts with collapse of the internal capsule accounted for 42.55％ (20/47),cysts with wall calcification accounted for 4.26％ (2/47) and multiple cysts accounted for 2.13％ (1/47).There was no solid lesions.The cyst diameters which were larger than 5 cm accounted for 74.47％ (35/47).Conclusion The CT can qualitatively diagnose hepatic echinococcosis in children.It is a reliable method of examination.