Objective To explore the inhibitory effects of endogenous hydrogen sulfide, a novel and important gas-eous transmitter generated in mammalian tissues mainly by cystathionine β-synthase (CBS) or cystathionineγ-lyase (CSE) on the apoptosis of the rat hepatic BRL cell line in physiological condition. Methods BRL cells were cultured, and divid-ed randomly into several groups in different phases of the experiment, including negative-siRNA (control) group, CBS siRNA (CBS 1~3) group and CSE siRNA (CSE 1~3) group, which were used to select the most efficient sequences of siRNAs at 48 or 24-hour transfection. Solution group and (CBS+CSE) siRNA group were added to detect the variation of apoptosis. The BRL cell line was observed and evaluated at 0, 4, 8, 12, and 24 hrs after siRNA transfection. When the mechanisms of the apoptosis were detected, CBS/CSE siRNAs were transfected individually or jointly into BRL cells, and compared with nega-tive-siRNA group to examine the variation. The genic and protein expression of CBS/CSE were detected by RT-PCR and Western blot assay. After transfection of CBS/CSE siRNA, the apoptosis of BRL cells was detected by Hoechst stain and flow cytometry (FCM). The mitochondrial membrane potential (MMP) changes were observed by fluorescent staining. Western blot assay was used to examine the protein expression of intracytoplasm cytochrome C (Cyt C) and cleaved-caspase 3. Re-sults CBS and CSE were observed in BRL cells. After transfection of CBS/CSE siRNA, endogenous H2S generation de-creased and the apoptosis of BRL cells increased. Accordingly, the expression of intracytoplasm-Cyt C and cleaved-caspase 3 increased. Conclusion The inhibition of endogenous H2S synthesis induced the apoptosis of BRL cells under physiologi-cal condition, which may be involved in mitochondrial pathway of apoptosis.

Objective:To investigate the effect of LINC00261 on the radiosensitivity of nasopharyngeal carcinoma and tumor formation and its underlying mechanism in nude mice.Methods:qRT-PCR was used to detect the relative expression levels of miR-620 and LINC00261 in radiosensitive and radioresistant nasopharyngeal carcinoma tissues. After the 6-10B and HNE-3 cells were irradiated with 0, 2, 4, 6, and 8 Gy 60Coγ-ray, the relative expression levels of miR-620 and LINC00261 were measured by qRT-PCR. After over-expression or silencing of LINC00261 and inhibition of miR-620 expression, the cells were irradiated with 4 Gy 60Coγ-ray. Clone formation assay was performed to detect the radiosensitivity of nasopharyngeal carcinoma cells. Flow cytometry was used to detect cell apoptosis. Western blot was utilized to detect the expression levels of Cleaved caspase-3 and Cleaved caspase-9 proteins. Luciferase reporter assay was adopted to analyze the targeting relationship between LINC00261 and miR-620. The changes in tumor formation were observed in tumor-bearing nude mice. Results:Compared with the radiosensitive tissues, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in radioresistant tissues (both P<0.05). After different doses of irradiation, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in 6-10B and HNE-3 cells (both P<0.05). After overexpression of LINC00261 and interference with miR-620 expression, the expression levels of Cleaved caspase-3 and Cleaved caspase-9 were significantly up-regulated (both P<0.05), the cell apoptosis rate was remarkably increased ( P<0.05) and the cell survival fraction was significantly enhanced in 6-10B and HNE-3 cells ( P<0.05). LINC00261 targetedly regulated the expression of miR-620. Overexpression of miR-620 could attenuate the radiosensitization and pro-apoptotic effects of LINC00261 overexpression on nasopharyngeal carcinoma cells. LINC00261 overexpression could significantly reduce the tumor formation weight of nasopharyngeal carcinoma in nude mice ( P<0.05). Conclusion:Overexpression of LINC00261 can increase the radiosensitivity of nasopharyngeal carcinoma cells probably by targeted regulation of miR-620 expression.

Objective To determine the levels of serum visfatin and HbA1c in patients with coronary heart disease and ex-plore the correlation with the severity of coronary atherosclerosis .Methods Totally 264 patients were enrolled who performed cor-onary angiography totally ,visfatin and HbA1c levels were detected respectively of 33 cases of control group ,51 cases of atheroscle-rosis group ,75 cases of single-vessel disease group ,72 cases of double-vessel disease group ,33 cases of triple-vessel disease group . Gensini score was used for evaluation of coronary artery lesion ,and to establish a multiple linear regression analysis of the relation-ship between each risk factor for coronary heart disease .According to the degree of coronary artery stenosis ,the patients also could be divided into the control group(33 cases) ,the non severe stenosis group (174 cases) and the severe stenosis group (57 cases) ,the changes of visfatin and HbA1c levels were analyzed in the three groups .Results In the groups by the coronary lesion count ,HbA1c levels increased with the degree of coronary artery lesions in the control group [(4 .98 ± 0 .21)% ] ,the atherosclerosis group [(5 .58 ± 0 .36)% ] ,the single-vessel disease group[(6 .17 ± 0 .48)% ] ,the double-vessel disease group[(6 .63 ± 0 .80)% ] ,the tri-ple-vessel disease group[(7 .97 ± 1 .49)% ] ,and comparisons had significant difference between any two groups(P< 0 .05) ;Visfatin level in the control group ,the atherosclerosis group ,the single-vessel disease group ,the double-vessel disease group ,the triple-ves-sel disease group were(0 .73 ± 0 .42)μg/L ,(1 .50 ± 0 .87)μg/L ,(3 .45 ± 2 .50)μg/L ,(5 .45 ± 2 .96)μg/L ,(9 .21 ± 6 .35)μg/L ,a-mong them coronary heart disease group (the single-vessel disease group ,the double-vessel disease group ,the triple-vessel disease group) is higher than the atherosclerosis group and the control group(P< 0 .05) ;the atherosclerosis group is higher than the con-trol group ,but there was no statistically significant difference(P> 0 .05) ;In according to the degree of coronary artery diameter ste-nosis ,the levels of visfatin ,HbA1c in severe stenosis group [(8 .25 ± 4 .86)μg/L ,(7 .35 ± 1 .43)% ] is significantly higher than the non severe stenosis group [(3 .22 ± 2 .74)μg/L ,(6 .14 ± 0 .70)% ] and the control group [(0 .73 ± 0 .42)μg/L ,(4 .98 ± 0 .21)% ] , P< 0 .01 ;The non severe stenosis group is significantly higher than the control group (P< 0 .01) .The levels of visfatin ,HbA1c ,hs-CRP ,LDL and TC had positive correlation with Gensini score(P< 0 .01) .The level of HDL was negatively correlated with Gensini score(r= - 0 .535 ,P < 0 .01) .The levels of visfatin ,HbA1c ,hs-CRP ,LDL and TC had positive correlation with visfatin ( P <0 .01) ,and the level of HDL and TG were negatively correlated with visfatin(P< 0 .01) .In multiple linear regression analysis ,the factors which finally entered the regression equation were HbA1c ,LDL ,hs-CRP ,visfatin and HDL .Conclusion The levels of visfa-tin and HbA1c is closely related to the severity of coronary atherosclerosis .Combined detection of visfatin and HbA1c can be used as important indicators for evaluating the severity of coronary atherosclerosis .

Objective To observe the effect of pressure-protective brace with pressure-sensitive device in the early stage rehabilitation training enhance bone healing, shorten the treatment course and reduce complications,a kind of independently developed pressure-protective brace with pressure-sensitive device was utilized with quantified discontinuous longitudinal stress stimulation under doctors' regulation according to procedure. Methods The pressure-protective brace with pressure sensitive device for rehabilitation training was developed in May 2008 ,and was applied in clinics during January 2009 to June 2010. Forty elder patients,with complete clinical data, underwent Dynamic Hip Screw (DHS) internal fixation of femoral intertrochanteric fracture were were enrolled into this study. These cases were assigned into experimental group and control group with 20 patients respectively. The patients of experimental group performed lower extremity rehabilitation training wearing the pressure-protective brace. The load training of lower extremities with double crutches was modulated by doctors through regulating the threshold value of pressure in different time and different condition after operation according to the prearranged rehabilitative plan of individuation. The controls were instructed to performed lower extremity rehabilitation training in traditional way. Both the clinical healing and bone union time in all cases were evaluated according to the uniform standard. Results Total 40 patients were followed up for 13.0 - 24. 0 weeks ( average, 17.6 weeks ). Clinical healing time was 7.0 - 12. 0 weeks ( average,9. 1 weeks ) and bone healing time was 12. 0 - 16.0 weeks(average,13. 7 weeks)in experimental group. While in control group,the clinic healing time and bone union time was 9. 0 - 13.0 weeks( average, 11.3 weeks) and 14. 0 -20. 0 weeks (average, 16. 6 weeks)respectively. The Independent T-test results showed that whether clinic healing time or bone healing time presented significant differences between experimental group and the controls( P＜0. 01 ). All of the fractures in these two groups were healed at the end time of follow up without adverse complications,including fracture displacement, implant break, implant loose and failure. Conclusion The pressure-protective brace with pressure sensitive device used for quantifying rehabilitation training can enhance bone union, shorten the treatment course and reduce complications. This method further proves that discontinuous compressive stress in a certain range can stimulate fracture healing.

Objective To analyze the risk factors of BKV infection and compare the real-time PCR procedure and urinary sediment smears of patients checked for decoy cells. Methods The peripheral blood samples of 129 renal recipients were collected. According to the result of PCR, 129 patients were divided into 2 groups:①BKV-DNA(+);②BKV-DNA(-). The sex, age, cold ischemia time, hemotodialysis duration, immunosuppressive agent and other clinical parameters were compared between the 2 groups and a Logistic regression was performed to analyze the risk factors of BKV infection. Results There were 20(15. 5%) patients in BKV-DNA(+), 109(84. 5%)patients in BKV-DNA(-)group. Logistic regression found that the cold ischemia time, hematodialysis duration, living donor were significantly related to the BKV-DNA. The results of the real-time PCR procedure and urinary sediment smears of patients checked for decoy cells were related. Conclusion Real-time fluorescent quantitative PCR and urine decoy cell are good way for detection of BKV infection after renal transplantation. The cold ischemia time and hematodialysis duration and brain death donor were the risk factors of BKV infection post renal transplantation.

OBJECTIVETo investigate the changes in methylation levels of the promoters of the tumor suppressor gene Wilms' tumor gene on the X-chromosome (WTX) and its possible role in gastric cancer.

METHODSWTX promoter methylation levels were detected in 20 pairs of specimens of gastric cancer and matched normal tissues and in 3 gastric cancer cell lines (MGC803, SCG7901, and BGC823) using the Sequenom MassARRAY quantitative analysis system. The gastric cancer cell line BGC823 was treated with 5-aza-2'-deoxycytidine (5-aza-dC) for demethylation and the changes in the level of WTX promoter methylation were investigated.

RESULTSWTX promoter methylation levels were very low and showed no significant differences among normal gastric tissues, gastric cancer tissues and the 3 gastric cancer cell lines. In BGC823 cells, treatment with 5-aza-dC did not obviously affect the promoter methylation levels of WTX.

CONCLUSIONHigh methylation levels of WTX promoters are rare in gastric cancer.

Cell Line, Tumor ; Chromosomes, Human, X ; DNA Methylation ; Genes, Wilms Tumor ; Humans ; Promoter Regions, Genetic ; Stomach Neoplasms ; genetics ; metabolism

Objective The purpose of this study was to explore common complications and their clinicopathological features in pediatric liver transplantation.Methods Clinical and pathological data of 240 liver biopsies from 168 children that conducted liver puncture from January 2015 to May 2018 in Tianjin First Central Hospital was retrospectively analyzed.We comprehensively analyzed incidence rate and pathological features of various complications,and correlations between acute rejection and C4d staining result or Banff score.Results A total of 86.67％ (208/240) liver biopsies could be definitely diagnosed with incidence rate of main complications in descending order as follows:T cell mediated rejection (TCMR) 60.57％ (126/208),drug-induced liver injury (DILI) 17.31％ (36/208),biliary complication 8.17％ (17/208),vascular complication 3.37％ (7/208),ischemia/reperfusion injury (IRI) 2.88％ (6/208),antibody mediated acute rejection (AMR) 1.92％ (4/208),HBV infection 1.92％ (4/208),non-alcoholic fatty liver disease (NAFLD) 1.44％ (3/208),chronic rejection (CR) 0.96 ％ (2/208) and HCV infection 0.48 ％ (1/208).TCMR and AMR in acute rejection (AR) accounted for 96.92％ (126/130) and 3.08％ (4/160),and into(portal-based,PB)type TCMR accounted for 96.03％(121/126) with the detectable rate of BP type subtype TCMR of 26.45％(32/121)within 30 d.There were 65.87％ (83/126)、25.40％ (32/126) 和4.76％ (6/126) of BP TCMR samples with "Banff ACR RAI" score within 3-5,6-7 and 8-9,and RAI score was negatively correlated with postoperative time (r =0.127,P =0.084).The incidence rate of central perivenulitis (CP) and portal eosinophils infiltration (PEI) in BP TCMR was 63.63％ (77/121) 和43.80％ (53/ 121),respectively,additionally,the PEI level was positively correlate with RAI score (P＜0.05).CP TCMR and AMR occurred within 30d-365 d and 8 d-180 d,respectively postoperative,while,the two CR occurred at 1095 d and 1335 d postoperative,and significant correlation was strikingly observed between rejection subtype and postoperative time (Z =9.231,P =0.026).C4d positive rate was 10％ (24/240),which was associated with Banff score and postoperative time,besides,C4d score was also correlated with rejection subtype and RAI score.The occurrence of DILI was mainly at time of ＜90 d or ＞180 d postoperative,and the detectable rate of biliary complication within 180 d postoperative was 82.35％ (14/17),IRI Appear in ＜30d.Hepatic artery complication account for nearly 57.14％ (4/7),occurrence time is ≤90 d.Occurrence of HBV infection,CMV infection and NAFLD were mainly at ＞365 d,＜90 d and ＜365 d,respectively.Conclusion There were lots of differences in clinical and pathological features among multi pediatric liver transplantation complications.Liver puncture plays an important role in rejection subtype classification and grading,as well as in non-rejection complications identification.

OBJECTIVETo investigate the prognostic factors relevant to sudden sensorineural hearing loss.

METHODSThe internationally accepted standardized clinical research methods, unified design, and unified program were adopted to conduct the prospective clinical multi-center study. The sudden deafness patients between 18 to 65 years old, with the course of this disorder less than two weeks, and without any medical treatments were collected, and then, divided into four types according to the hearing curve: type A, acute sensorineural hearing loss in low tone frequencies; type B, acute sensorineural hearing loss in high tone frequencies; type C, acute sensorineural hearing loss in all frequencies; and type D, total deafness. The factors, in terms of age, gender, type of initial audiogram, time delay before the first visit, and severity of hearing loss, were included in the analyses.

RESULTSA total of 1 024 cases with single side sudden deafness were collected in the study from 33 hospitals in China from August 2007 to October 2011, inclusive of for 492 males (48.05%) and 532 females (51.95%). The average age was (41.2 ± 12.8) years old. There were 553 cases (54.00%) in left ear, and 471 cases (46.00%) in right ear. The curative effects of different types were shown as follows: the type in low tone frequencies had the highest rate of 90.73%, the type in all frequencies was 82.59%; the type of total deafness was 70.29%; and the type in high tone frequencies had the lowest rate of 65.96%. It had significant difference of the effective rate between different types (χ(2) = 231.58, P = 0.000). Age, time delay before first visit, and severity of initial hearing loss were significantly correlated with hearing improvement.

CONCLUSIONSInitial audiogram of SSNHL might predict hearing recovery. The young in age and a short time delay before starting treatment are positive prognostic factors for hearing recovery in SSNHL. The initial severity of hearing loss is negative prognostic factor of hearing recovery.

Adult ; China ; Female ; Hearing Loss, Sensorineural ; diagnosis ; Hearing Loss, Sudden ; diagnosis ; Hearing Tests ; Humans ; Male ; Middle Aged ; Prognosis ; Prospective Studies