Objective To study the expression of STIM 1 gene in human hypopharyngeal carcinoma cell line FaDu and its effect on FaDu cell apoptosis .Methods Lentivirus infection was used to knock STIM 1 down in FaDu cells .Group STIM1-siRNA: the expression of STIM1 in FaDu cell was inhibited by STIM 1-siRNA lentivirus .Group control:FaDu cells were infected by negative control siRNA lentivirus . Real-Time PCR was applied to identify the efficacy of lenticirus infection and the expression of STIM 1 in FaDu cells.Western blot was used to identify the expression of STIM 1 protein after lenticirus infection .Flow cytometry assay was performed to detect the apoptosis of FaDu cells in the two groups.The data were statistically analyzed with SPSS 17.0 software.Results Compared with GAPDH (Ct=12.08 ±0.05),the expression of STIM1 in FaDu cells was significant expressed (Ct=22.21 ±0.05,P<0.001).Real-Time PCR analysis the relative mRNA expression of STIM1 in FaDu cells of control group and STIM 1-siRNA group were (1.00 ±0.08) and (0.12 ±0.01) respectively (P<0.001). Western blot showed that the expression of STIM 1 gene and protein in FaDu cells were inhibited significantly after STIM 1-siRNA lentiviral in-fection,which was in accordance with the results of Real-Time PCR analysis.Flow cytometry assay showed that the siRNA-mRNA group had a higher apoptosis percentage (9.81 ±0.56)% compared to the control group (4.36 ±1.32)%,with statistically significant difference (P<0.05).Conclusion STIM1 gene correlated significantly with FaDu cell apoptosis .It inhibits apoptosis of FaDu cells ,and it may be a potential diagnostic and therapeutic target for the hypopharyngeal carcinoma .

BACKGROUND:The pathogenesis of knee intraarticular adhesion is yet unknown. Excessive proliferation of fibroblasts is considered to cause knee intraarticular adhesion. OBJECTIVE:To study the preventive effects of methotrexate on knee intraarticular adhesion through fibroblast apoptosis induced by endoplasmic reticulum stress. METHODS:The viability of the cultured fibroblasts treated with methotrexate(10-5-10-9mol/L)or PBSwas determined after 24 hours. Fibroblast apoptosis was detected by Hoechst33342 staining. Endoplasmic reticulum stress-and apoptosis-related proteins, including cleaved-PARP, CHOP, Bax and Bcl-2, were determined by western blotassay. Eighteen healthy male New Zealand white rabbits were used to establish the knee intraarticular adhesion models, and equaly randomized into three groups, and received topical application of 2 or 1 g/L methotrexate, or normal saline (control). The preventive effects of methotrexate on knee intraarticular adhesion and CHOP expression in scar tissue were observed. RESULTS AND CONCLUSION:Methotrexate inhibited the proliferation and viability of fibroblasts in a dose-dependent manner. The number of apoptotic fibroblasts was significantly increased compared with control group. Protein expression of cleaved-PARP, CHOP, and bax was increased, while protein expression of bcl-2 was decreased with time. The animal experiment showed that preventive effects of 2 g/L methotrexate on knee intraarticular adhesion were superior to 1 g/L methotrexate treatment. CHOP expression in the scar tissue in the methotrexate groups was higher than the control group and that was higher in high-dose methotrexate group. Our results suggest that methotrexate prevents knee intraarticular adhesionviaendoplasmic reticulum stress-induced fibroblast apoptosis.

OBJECTIVETo evaluate the inhibition effect of STIM1 gene silencing on tumor growth of human hypopharyngeal carcinoma cell lines FaDu in nude mice.

METHODSSTIM1 gene in FaDu was silenced by lentiviral infection, and the effect of inhibition was detected by Real-time PCR and Western blot after lentiviral infection. Nude mice were divided into 2 groups, 5 mice in each group. Inhibition group: subcutaneous inject FaDu cells which STIM1 expression was inhibited.

CONTROL GROUPsubcutaneous inject FaDu cells infected with negative control siRNA-expressing lentivirus. Tumor volumes were measured by calipers, and small animal imaging was detected by NightOWL system on the day 10, 14, 18 and 22 after tumor inoculated. Tumor weights were evaluated in the day 22 after tumor inoculated. Statistical analysis was performed using standard student test(P value threshold was 0.05).

RESULTSThe expressions of human STIM1 gene and protein in FaDu cells were suppressed effectively after STIM1-siRNA lentiviral infection. The mean tumor volumes of control group and inhibition group were (51±25) mm3 and (40±35) mm3, respectively, on the day 10, (262±107) and (106±41) mm3 on the day 14, (716±226) and (340±158) mm3 on the day, (1 682±592) mm3 and (917±252)mm3 on the day 22 (P<0.05). On the day 22, the tumor weight was (1.22±0.41) g in control group and (0.66±0.26) g in STIM1-siRNA group (P<0.05). Small animal imaging showed that the tumors had a smaller fluorescence range with lower signal intensity in STIM1-siRNA group than in control group on the day 14, 18 and 22.

CONCLUSIONThe expression of STIM1 in human hypopharyngeal carcinoma cell lines FaDu can be inhibited effectively by lentiviral infection, causing the inhibition of tumor formation and growth.

Animals ; Cell Line, Tumor ; Gene Silencing ; Humans ; Hypopharyngeal Neoplasms ; pathology ; Lentivirus ; Membrane Proteins ; genetics ; Mice ; Mice, Nude ; Neoplasm Proteins ; genetics ; Neoplasm Transplantation ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Stromal Interaction Molecule 1

Objective:To explore the predictive value of abnormal ECG J waves for arrhythmias occurred during short term in patients with acute ST elevation myocardial infarction (ASTEMI ) .Methods :ECG and echocardio‐graphic monitoring recordings of 204 ASTEMI patients who received emergency percutaneous coronary intervention (PCI) in our hospital from Jan 2007 to Dec 2012 were retrospectively analyzed .Univariate and multivariate Logistic regression analyses were performed to analyze the influencing factors of arrhythmias within one week after myocar‐dial infarction .Results:Abnormal J waves detected by ECG were found in 82 cases (40.2% ) among the 204 pa‐tients ,most of which were distributed on inferior leads of ECG .Multivariate Logistic regression analysis indicated that abnormal J waves (OR=14.05 , P=0.01 ,95% CI 1.70~116.40) ,J waves distributed across ≥ two locations (OR=13.38 ,P=0.01 ,95% CI 1.53~38.68) and J wave amplitude≥0.2 mV (OR=4.28 ,P=0.02 ,95% CI 1.82~16.72) were independent predictors for sustained ventricular tachycardia/fibrillation (VT/VF) ,but they cannot be used as predictors for occurrence of all ventricular arrhythmias ,including non‐sustained VT ,sustained VT and VF (P>0.05 all) ,nor the occurrence of atrial arrhythmias , P>0.05 all .Multivariate Logistic regression analysis indicated that diabetes mellitus was an independent predictor for occurrence of ≥30s atrial tachycardia/fibrillation in ASTEMI patients (OR=2.29 ,P=0.047 ,95% CI 1.01~5.18) .Conclusion:Abnormal ECG J wave is an inde‐pendent predictor for occurrence of sustained ventricular tachycardia/fibrillation during short term after ASTEMI .

Objective To investigate clinical efficacy and prognostic factors of hilar cholangiocarcinoma.Methods The retrospective case-control study was conducted.The clinicopathological data of 322 patients with hilar cholangiocarcinoma who were admitted to the Xiangya Hospital of Central South University between December 2005 and November 2015 were collected.Preoperative staging and classification of tumor and treatment planning were carried out according to the results of laboratory and imaging examinations.Observation indexes:(1) clinical features and results of assisted examinations;(2) treatments and results of pathological examination;(3) followup and survival;(4) prognostic factors analysis:gender,age,preoperative highest total bilirubin (TBil),preoperative carcinoembryonic antigen (CEA),preoperative CA19-9,preoperative CA242,preoperative CA125,treatment methods and TNM staging.The follow-up of outpatient examination and telephone interview was perfornmed to detect patients' survival up to November 2016.Survival curve was drawn using the Kaplan-Meier method.Survival and univariate analyses were done using the Log-rank test,and multivariate analysis was done using the Cox proportional hazard model.Results (1) Clinical features and results of assisted examinations:among the 322 patients,there were 301 patients with a chief complaint of jaundice.Of the 322 patients,the preoperative highest levels of TBil,DBil,ALT and AST in 322 patients were 3.9-785.2 μmol/L,1.6-410.2 μ mol/L,14.8-484.5 U/L and 21.4-539.8 U/L,respectively.Levels of ALP and GGT in 272 patients were 93.8-1 890.0 U/L and 2.0-1 832.8 U/L,respectively.Seventy-seven of 292 patients had an elevated CEA level,272 of 298 patients had an elevated CA19-9 level,153 of 260 patients had an elevated CA242 level and 86 of 260 patients had an elevated CA125 level.According to Bismuth-Corlette type,24 patients were detected in type Ⅰ,115 in type Ⅱ,55 in type Ⅲa,63 in type Ⅲb and 65 in type Ⅳ.(2) Treatments and results of pathological examination:Of the 322 patients,104 patients underwent radical resection,including 79 with hilar bile duct resection (9 combined with vascular resection and reconstruction) and 25 with extended hepatic lobectomy (16 combined with caudate lobectomy),and 218 patients underwent palliative treatments,including 134 with external biliary drainage and 84 with internal biliary drainage.Five patients were dead in the perioperative period,of which 2 died of acute liver failure,1 died of systemic infection and multiple organ failure,1 died of acute renal failure and 1 died of acute suppurative cholangitis,septic shock and disseminated intravascular coagulation.Of 263 patients receiving pathological examination,adenocarcinoma was detected in 253 patients (12 with high-differentiated adenocarcinoma,85 with moderate-differentiated adenocarcinoma,33 with low-differentiated adenocarcinoma and 123 with indefinite differentiation),mucinous adenocarcinoma in 5 patients,cholangiocarcinoma in 3 patients and neuroendocrine carcinoma in 2 patients.TNM staging of 322 patients:stage Ⅰ was detected in 8 patients,stage Ⅱ in 53 patients,stage Ⅲ in 132 patients,stage Ⅳ in 96 patients and indefinite stage in 33 patients.(3) Follow up and survival:among the 322 patients,296 were followed up for 12-132 months,with a median follow-up time of 65 months,including 94 with radical resection and 202 with palliative treatments.Among the 296 patients,the median survival time and 1-,3-,5-year survival rates were 10 months,47.1％,20.2％ and 9.5％,respectively.0f296 patients with follow-up,median survival time and 1-,3-,5-year survival rates were 31 months,84.0％,46.2％,25.0％ in 94 patients receiving radical resection and 7 months,29.9％,8.1％ and 2.3％ in 202 patients receiving palliative treatment,respectively,with a statistically significant difference between the 2 groups (x2=78.777,P＜ 0.05).Among the 94 patients receiving follow-up and radical resection,the median survival time and 1-,3-,5-year survival rates were 31 months,82.1％,45.1％,25.7％ in 73 patients undergoing hilar bile duct resection and 35 months,90.5％,49.8％,22.1％ in 21 patients undergoing hepatic lobectomy,respectively,with no statistically significant difference (x2=0.186,P＞0.05).Among the 73 patients undergoing hilar bile duct resection,median survival time and 1-,3-,5-year survival rates were 16 months,57.1％,0,0 in 7 patients combined with vascular resection and reconstruction and 34 months,84.6％,49.5％,27.5％ in 66 patients undergoing simplex hilar bile duct resection,respectively,showing a statistically significant difference (x2 =11.977,P＜ 0.05).(4) Prognostic factors analysis:results of univariate analysis showed that preoperative highest TBil,preoperative CEA,preoperative CA242,preoperative CA125,treatment methods and TNM staging were related factors affecting prognosis of patients with hilar cholangiocarcinoma (x2=25.009,18.671,9.359,33.628,94.729,77.136,P＜0.05).Multivariate analysis showed that preoperative highest TBil ≥ 342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ and Ⅳ were the independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma (HR =2.270,2.147,3.166,2.351,95％ confidence interval:1.587-3.247,1.446-3.188,2.117-4.734,1.489-3.712,P＜0.05).Conclusions Prognosis of hilar cholangiocarcinoma is still unsatisfactory.The R0 resection is the key in radical surgery.Preoperative highest TBil≥342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ-Ⅳ are independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma.

Objective To assess the accuracy of puncture guided by intelligent positioning (IP) system using magnetic navigation.Methods Five prepared targeted models at three certain depth (<50 mm, samll depth;50-100 mm,medium depth;>100 mm,large depth) underwent puncture guided by intelligent positioning system using IP and conventional ultrasound (US),respectively.Puncture errors,the number of attempt and spent time were recorded and compared .Results For the targets at small,medium and large depth,the errors of IP was (1.88 ±1.18),(1.56 ±0.56) and (3.99 ±1.10) cm,and the errors of conventional US was (4.52 ±2.23),(4.49 ±1.73) and (3.93 ±2.19) cm respectively.The errors of IP were significantly less than those of conventional US at small(t=-2.345,P=0.047) and medium(t=-3.608,P=0.007) depth,but there was no statistically significant difference at large depth (t=0.058,P=0.955). In the IP group,there were statistically significant differences for puncture errors between the small and large depth,as well as between medium and large depth ( F =8.923,P =0.010).There was no statistically significant difference for the errors of IP between the small and medium depth (t=-1.927,P=0.501).For the targets at small,medium and large depth,each puncture was performed in single attempt when guided by IP and in 2,1 and 2 attempt when guided by conventional US .At small and large depth,the numbers of attempt of IP were significantly less than those of conventional US (U=-2.372,P=0.018;U=-2.39, P=0.032).Whereas at medium depth,there was no significant difference (U=-1.000,P=0.690).For the targets at small,medium and large depth,each puncture spent (21.20 ±2.39)s, (27.00 ±4.00)s and (31.80 ±3.83)s when guided by IP,and(45.20 ±9.68),(26.80 ±4.21) and (54.60 ±13.48)s when guided by conventional US.The spent time of IP was less than that with conventional US for small and large depth targets(t =-5.383, P =0.001;t =-3.637, P =0.007).Whereas no statistically significant difference was found for the medium depth target (t=0.077,P=0.916).Conclusion In comparison with conventional US,IP system guided puncture is more accurate and the number of attempt and spent time is less .

Objective:To explore the relationship between abnormal ECG J wave and in‐hospital prognosis in patients with acute ST -segment elevation myocardial infarction (ASTEMI) .Methods :ECG and related clinical data of 204 ASTEMI patients ,who received emergency percutaneous coronary intervention (PCI) in our hospital from Jan 2007 to Dec 2012 ,were retrospectively analyzed .According to the presence of abnormal J wave or not ,patients were di‐vided into abnormal J wave group (n= 82 ,occupied 40.2% ,82/204) and no abnormal J wave group (n= 122 , 59.8% ,122/204) .Single‐and multiple‐factor Logistic regression analysis were used to analyze risk factors of in -hospital death .Results:During hospitalization ,incidence rate of sustained ventricular tachycardia/fibrillation in pa‐tients with abnormal ECG J waves was significantly higher than those without abnormal J waves (9.8% vs .1.6% , P=0.008) , but among the 12 cases (5.9% ) of cardiac deaths ,only six cases had abnormal J waves .Logistic regres‐sion analysis indicated that abnormal ECG J wave cannot predict in‐hospital death of ASTEMI patients (OR=0.99 , 95% CI :0.34~ 2.90 , P= 0.987) ,while age can be regarded as an independent predictor factor for in‐hospital prognosis of these patients (OR = 1.08 ,95% CI :1.01 ~ 1.15 , P= 0.02) .Conclusion:For ASTEMI patients , though the incidence rate of sustained ventricular tachycardia/fibrillation in abnormal J wave group is significantly higher than those of without abnormal J waves group during hospitalization ,but abnormal ECG J waves cannot pre‐dict short-term prognosis of these patients .

Objective To investigate the high risk factors and emergency treatments to the patients with intraoperative acute encephalocele caused by delayed intracranial hematomas at non operating region. Methods The clinical data of 40 patients with intraoperative acute encephalocele caused by delayed intracranial hematomas at non operating region from January 2000 to December 2013 were analyzed retrospectively. Summarized the high risk factors and emergency treatments in this case. Results The 22 cases with contrecoup injury. 14 cases with extensive contusion of brain,16 cases with skull fracture and 10 cases with small intracranial hematoma (Volume <15mL).The delayed intracranial hematomas to the acute encephalocele include,32 cases at the offside. And 24 cases with epidural hematoma. 32 cases had to undergo reoperation, survived 16 cases. The total death rate was 60%. Conclusion The high risk factors are ex-tensive contusion of brain, offside skull fracture and the delayed intracranial hematomas at non operating region. Under-standing Its high risk factors in clinical, with much more foreseeability, and sufficient preoperative preparation, right in-traoperative treatment, a scientific system comprehensive treatment postoperation can save the patients' life in maximum.

Objective To observe the effects of Langchuangqing granule on the Forssman cutaneous vasculitis reaction in guinea pigs , the antibo-dies, complements and immunoglobulin levels in MRL/lpr mice and the antibodies levels in the spleen lymphocytes.Methods In the forssman cutaneous vasculitis reaction , the immune serums were intradermally in-jected to the back skin of guinea pigs followed by tail vein injection of 0.5%evans blue.The skins dyed evans blue showed the model estab-lished.The guinea pigs were divided into five groups:model group , con-trol group and high-dose, middle-dose, low-dose ( Langchuangqing granule,18.27, 9.13, 4.56 g· kg -1) of test groups.The optical densi-ty( OD) of the skin was tested after continuously intragastric administra-tion for 8 days.In the lupus experiment in MRL/lpr mice, the models were established naturally with the mice growing up.The mice were divided into six groups: blank group , model group , control group and high-dose, middle-dose, low-dose(Langchuangqing granule,45.67, 22.83, 11.42 g· kg -1) of test groups.The blood was taken in all groups after intragastric administration for 6 weeks.Then the anti -double stranded DNA anti-body(dsDNA), antinuclear antibody (ANA), complement 3(C3), complement 4(C4), immunoglobulin M(IgM), immunoglobulin G(IgG) levels were measured by enzyme -linked immunosorbent assay (ELISA)method.In the spleen lymphocytes experiment , the dsDNA and ANA levels were measured in the lymphocyte supernatant after the effects of serum containing Langchuangqing granule on spleen lymphocytes.Results In the forssman cutaneous vasculitis reac-tion, compared with model group , the OD of the skin was significantly reduced in high dose test group in guinea pigs ( P<0.05 ).In the lupus experiment in MRL/lpr mice , compared with model group , high dose of langchuangqing granule significantly reduced dsDNA[(47.25 ±6.05) vs (37.51 ±3.26) ng· L-1] and IgM levels, improved C3 lev-els(P<0.05, P<0.01).Also in the middle dose group, IgM levels notably reduced and C3 increased(P<0.05,P<0.01 ).In the spleen lymphocytes experiment , the dsDNA levels in lymphocyte supernatant significantly reduced af-ter treated with the 20%drug-containing serum ( P<0.05 ).Conclusion The effects of Langchuangqing granule on the systemic lupus erythematosus may be related to the immunological functions of Autoantibodies , complement , immunoglobulin.

OBJECTIVETo investigate the chemical constituents of red alga Corallina pilulifera.

METHODCompounds were isolated by normal phase silica gel and Sephadex LH - 20 gel column chromatography, reverse phase HPLC and recrystallization. Their structures were elucidated by spectroscopic methods including MS, 1H-NMR, 13C-NMR, HSQC, HMBC. Cytotoxicity of the compounds was screened by using standard MTT method.

RESULTSeven compounds were isolated from red alga C. pilulifera, their structures were identified as (E) -phytol epoxide (1), phytenal (2), phytol (3), dehydrovomifoliol (4), loliolide (5), 3beta-hydroxy-5alpha, 6alpha-epoxy-7-megastigmene-9-one (6), 4-hydroxybenzaldehyde (7).

CONCLUSIONAll of the compounds were obtained from this species for the first time. These compounds were inactive (IC50 > 10 microg x mL(-1)) in the MTT assay.

Benzaldehydes ; chemistry ; isolation & purification ; pharmacology ; Benzofurans ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; drug effects ; Humans ; Phytol ; chemistry ; isolation & purification ; pharmacology ; Rhodophyta ; chemistry