Apoptosis disorders have an important function in the development of gastrointestinal cancer. BNIP3 is a member of the BH3-only subfamily of the bcl-2 family, which contains a BH3 domain and a transmembrane domain, and belongs to the mitochon-drial pro-apoptotic proteins. BNIP3 induces cell death via the caspase-independent mitochondrial apoptotic pathway and mediates au-tophagic cell death. BNIP3 expression is regulated by hypoxia and other factors. BNIP3 expression in tumors exhibits tissue specificity;BNIP3 is highly expressed in some tumors, including breast, lung, and cervical tumors. In pancreatic, gastric, and colorectal cancers, BNIP3 is epigenetically silenced. The absence of BNIP3 in the tumors can cause tumor cells to tolerate hypoxia and may be associated with chemotherapy and radiotherapy resistance. A comprehensive understanding of the spectrum of BNIP3 expression in a various tu-mors is necessary to use BNIP3 as a marker in clinical applications to treat tumors and as a new target in tumor prognosis.

Objective To discuss the effect of somatostain SS on metabolism of free radicals in animal models of osteoarthritis,so to discuss the treatment mechanism of intra- articular injection of somatostatin on osteoarthritis.MethodsForty experimental Japanese big-ear rabbits were randomly divided into four groups:normal control group,blank model group,normal saline group and SS group,n =10 for each group.After modeling,the rats in SS group and normal saline group received intra- articular injection of somatostain and saline respectively on the right knee.Ten weeks after the modeling,we measured the movement of right knee joint in rabbits,the levels of nitric oxide ( NO ) and hyaluronic acid (HA) in serum from the heart blood,and the activity of SOD and the content of MDA in synovial tissue.Results Compared with the blank model group,the movement of the knee joint was significantly increased in SS group,(96.01 ± 1.06)vs (50.21 ± 1.80) (P ＜ 0.01 ).In the blank model group,the levels of NO and HA (μmoL/mL) in serum were ( 111.60 ± 2.76) and ( 309.11 ± 1.89 ),which was significantly decreased in SS group,which was( 80.14 ± 1.67 ) and ( 133.73 ± 2.75 ) ( P1 =0.0049,P2 =0.0052,P ＜ 0.01 ).Compared with the blank model group,the SOD activity was (15.77 ± 2.76) and the MDA in serum nmol/mg prot was( 1.33 ±1.03),while was significantly increased in SS group,(24.74 ± 1.67) ( P ＜ 0.01 ),and the levels of MDA in serum was significantly decreased in the SS group,(0.89 ±1.46) (P＜0.01).Conclusion Intra-articular injection of somatostatin in knee osteoarthritis can improve the oxidative stress,enhance the activity of the knee joint.

Objective To investigate the correlation between different clinicopathological factors and the recurrence and metastasis of advanced adenocarcinoma of the esophagogastric junction after curative resection,and to analyze the effects of the factors on the prognosis of these patients.Methods The clinical data of 385 patients with advanced adenocarcinoma of the esophagogastric junction who received curative resection at the Affiliated Hospital of Tianjin Medical University from January 2000 to January 2007 were retrospectively analyzed.There were 228 patients did not have tumor recurrence and metastasis (non-recurrence and metastasis group) and 157 patients had tumor recurrence and metastasis (recurrence and metastasis group).Risk factors which might influence postoperative recurrence and metastasis were analyzed using univariate analysis (chi-square test) and multivariate analysis (Logistic regression model).All patients were followed up via out-patient examination or phone call.The survival curve was drawn by Kaplan-Meier method,and the survival analysis was done by Log-rank test.Results The median time for follow-up was 36 months (range,3-108 months).A total of 157 patients had postoperative tumor recurrence and metastasis,and the mean time of tumor recurrence was 17.9 mouths.The results of univariate analysis showed that tumor type,differentiation degree,invasion depth,number of positive and negative lymph nodes,TNM staging were risk factors for the postoperative recurrence and metastasis of adenocarcinoma of the esophagogastric junction after curative resection (x2=5.248,13.493,12.319,18.315,9.704,10.281,P ＜ 0.05).The results of multivariate analysis showed that differentiation degree,invasion depth,number of positive and negative lymph nodes were the independent risk factors influencing the recurrence and metastasis of adenocarcinoma of the esophagogastric junction after curative resection (OR =1.805,1.809,1.520,0.763,P ＜0.05).The numbers of positive lymph nodes in the non-recurrence and metastasis group and the recurrence and metastasis group were 3.86 ± 0.28 and 6.89 ± 0.58,with a significant difference (t =5.118,P ＜ 0.05).The number of negative lymph nodes in the non-recurrence and metastasis group and the recurrence and metastasis group were 14.04 ±0.54 and 10.53 ±0.56,with a significant difference between the 2 groups (t =4.386,P ＜0.05).The 5-year survival rates of patients with the numbers of positive lymph nodes of 0,1-2,3-6 and more than 7 were 46.4％,43.8％,27.1％ and 7.2％,respectively,and the corresponding median survival time were 53,47,35 and 26 months.There was a significant difference in the 5-year survival rate among patients with different numbers of positive lymph nodes (x2 =54.783,P ＜ 0.05).The 5-year survival rates of patients with the number of negative lymph nodes under 9,between 10 and 15 and more than 16 were 22.1％,21.5％ and 45.5％,respectively,and the corresponding median survival time were 28,34,47 months.There was a significant difference in the 5-year survival rate among patients with different numbers of negative lymph nodes (x2=22.814,P ＜ 0.05).Conclusions Tumor type,invasion depth,number of positive and negative lymph nodes are independent risk factors of postoperative recurrence and metastasis of adenocarcinoma of the esophagogastric junction,and the number of positive and negative lymph nodes are important for the prognosis of patients with adenocarcinoma of the esophagogastric junction.

Objective To investigate the tumor diameter on the prognosis of patients with advanced esophageal squamous cell carcinoma(ESCC) after Ivor-Lewis surgical resection.Methods The clinical data of 254 patients with advanced ESCC who received Ivor-Lewis surgical resection at the Affiliated Hospital of Tianjin Medical University from January 2005 to December 2008 were retrospectively analyzed.All the patients were followed up via outpatient examination,telephone interview and correspondence till December 2013.Survival curve was drawn by the Kaplan-Meier method,and survival rate was analyzed using the Log-rank test.Receiver-operating-characteristic (ROC) curve analysis was used to determine the appropriate cutoff value of tumor size.Univariate and multivariate analysis were done using the chi-square test and COX regression model.Results Of 254 patients,223 patients were followed up for a median time of 30 months (range,3-108 months) with a follow-up rate of 87.80％ (223/254).The median total survival time was 27 months,and the 1,3,5-year overall survival rates were 72.7％,42.2％ and 31.3％,respectively.ROC analysis showed that the appropriate cutoff value of tumor diameter was 3.5 cm.The median survival time and 5-year survival rate were 36 months and 39.3％ in patients with tumor diameter ≤ 3.5 cm and 18 months and 25.4％ in patients with tumor diameter ＞ 3.5 cm,respectively,with a significant difference (x2 =9.494,P ＜ 0.05).The results of univariate analysis showed that the age,tumor diameter,depth of tumor invasion,lymph node metastasis and postoperative adjuvant therapy were related factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection (x2=4.459,9.494,6.993,10.382,5.507,P ＜ 0.05).The results of multivariate analysis showed that tumor diameter ＞ 3.5 cm,lymph node metastasis and no postoperative adjuvant therapy were the independent factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection (HR =1.631,1.681,0.677,95％ confidence interval:1.151-2.312,1.198-2.358,0.487-0.942,P ＜ 0.05).Of 159 patients without postoperative lymph node metastasis,median survival time and 5-year accumulated survival rate were 49 months and 46.4％ in patients with tumor diameter ≤ 3.5 cm and 23 months and 32.0％ in patients with tumor diameter ＞ 3.5 cm,respectively,with a significant difference (x2 =6.412,P ＜ 0.05).Conclusions The tumor diameter ＞ 3.5 cm,lymph node metastasis and no postoperative adjuvant therapy are the independent factors affecting the prognosis of patients with advanced ESCC after Ivor-Lewis surgical resection,meanwhile there is an assessed value of tumor diameter on the prognosis of patients without lymph node metastasis.

Objective To investigate the expression of Krüppel-like factor 4 (KLF4) protein in esophageal squamous cell carcinoma (ESCC) tissues and to explore its correlation with clinical pathological features as well as prognosis.Methods The expression of KLF4 protein in cancer tissues and normal esophageal tissues from surgical paraffin specimens of 98 thoracic ESCC cases with complete clinical,pathological and follow-up date were detected by immunohistochemistry.The expression of KLF4 at protein level in 20 freshly surgical esophageal cancer tissues and normal esophageal mucous tissues were examined by Western blot.The relation between the expression of KLF4 protein,clinicopathological characteristics and prognosis was analyzed,t-test was used for measurement data analysis.Chi-square test was performed to analyze the correlation between KLF4 protein expression and clinicopathological features.Survival analysis was analyzed by the Kaplan-Meier method.The comparisons of survival rates were analyzed by Log-rank test.Results The positive rate of KLF4 protein expression in normal esophageal tissues and ESCC tissues was 82.7％ (81/98) and 43.9％ (43/98),respectively,the difference was statistically significant (x2=31.701,P＜0.01).The expression of KLF4 at protein level in 20 cases of fresh esophageal cancer tissues and normal esophageal mucosa tissues was 0.576±0.050 and 0.684 ± 0.095,respectively,the difference was statistically significant (t =4.932,P＜0.01).The expression of KLF4 at protein level was correlated with lymph node metastasis and TNM stage (x2 =10.871 and 6.482,P=0.001 and 0.039),however not correlated with gender,age,location,tumor size,degree of differentiation and the depth of invasion (x2=0.214,3.442,5.748,0.891,0.013 and 1.479,P=0.644,0.064,0.056,0.345,0.911 and 0.477).In 98 patients,the 5-year survival rate of cases with KLF4 protein positive expression and negative expression was 48.8％ and 25.5％ and the median survival period was 55 months and 26 months,the differences were statistically significant (x2 =5.747 and 4.493,P=0.017 and 0.034).Conclusion KLF4 as a tumor suppressor gene may play an important role in the genesis,development and metastasis of ESCC,and may become a biological indicator of the severity and prognosis in ESCC.

Object To isolate and identify the chemical constituents from the rhizome of Cimicifuga dahurica (Turcz ) Maxim Methods The different chromatographic techniques were used to isolate ten constituents, and the spectral methods, such as IR, MS, 1HNMR, 13 CNMR and 2DNMR, were used to identify the structures Results Their structures were identified as cimigenol (Ⅰ), 24 epi 7, 8 didehydrocimigenol 3 O ? D xylopyranoside (Ⅱ), 7, 8 didehydrocimigenol 3 O ? D xylopyranoside (Ⅲ), 25 O acetyl 7, 8 didehydrocimigenol 3 O ? D xylopyranoside (Ⅳ), 3 arabinosyl 24 O acetylhydroshengmanol 15 glucoside (Ⅴ), isoferulic acid (Ⅵ), (E) 3 (3′ methyl 2′ butenylidene) 2 indolinone (Ⅶ), sucrose (Ⅷ), ? sitosterol (Ⅸ) and stigmastenol 3 O ? D glucopyranoside (Ⅹ), respectively Conclusion Compounds Ⅱ Ⅳ, Ⅹ are isolated for the first time from the title plant

Objective To investigate the role of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in rotenone-mediated dopaminergic cell damage. Methods Neuronal rat adrenal pheochromecytoma(PC12) cells treated with rotenone were used as the cell model of Parkinson' s disease (PD). Cell viability of PC12 cells after exposure to rotenone was detected by MTT (methyl thiazolyl tetrazolium) method. Immanohistechemistry was used to detect the phosphorylation of ERK1/2 in cells exposed to rotenone. Western blotting was used to verify the phosphorylation of ERK1/2 and to observe the effect of PD98059, an inhibitor of the upstream mitogen activated protein kinase kinase (MEK) that phosphorylates and activates ERK1/2, on rotenone-induced ERK1/2 phosphorylation and cell viability.Results The viability (represented by A570 of PC12 cells exposed to 5 μmoL/L rotenone) declined with the increase of exposure time from 1 h to 24 h (%, 1 h :75.46±5.47, 2 h : 70.42±1.94, 4 h : 65.23± 0.96, 8 h : 59.04 ± 2.85, 24 h :29.64 ± 1.63, comparison between different time points(F=143.014, P=0.000) ; compared with control groups(100.00±2.89), q value: 17.07, 20.58, 24. 19, 28.50, 48.95 respectively, all P <0.01). After exposure to rotenone, phosphorylated ERK1/2 aggregated in the PC12 cells. Western blotting indicated that rotenone induced a biphasic phosphorylation of ERK1/2, which increased from 30 min after rotenone treatment, reached the peak at 1-2 h, decreased at 4 h, and increased again at 8 h, and disappeared after 16 h; PD98059 significantly inhibited ERK1/2 phosphorylation induced by rotenone, and attenuated cell injury examined at 1, 2 and 8 h. Conclusions Our study suggested that ERK1/2 activation plays a detrimental role in rotenone toxicity, and raised the possibility that abnormal patterns of ERK1/2 activation may contribute to dopaminergic neuronal cell death in PD.

10.3969/j.issn.2095-4344.2013.25.020

Objective To investigate the effects of constant magnetic field (CMF) on proliferation, apopto-sis and nitric oxide (NO) secretion of rat bone marrow-derived endothelial progenitor cells (EPCs) intervened by C-reactive protein (CRP). Methods EPCs were isolated from rat bone marrow by density gradient centrifugation and cultured on fibronectin-coated dishes. The cells were divided into five groups, i. e., control group, CRP (12 μg/ml) group, CRP plus CMF (0.1, 0. 5, 1.0 mT) groups. Samples were collected 24 hours after incubation. Cell proliferation was measured by MTT chromatometry. Apoptosis rate was detected by flow-cytometry. NO content of culture medium was measured by nitrate reductase method. Results As compared with control group, cell prolifer-ation in CRP group reduced significantly (0. 265±0. 008 vs 0. 316±0. 011, P < 0.05), NO secretion also de-creased significantly [(22.7±4.5) μmol/L vs (37.6±3.8) μmol/L, P < 0.05], cell apoptosis rate elevated sig-nificantly [(10.8±0. 8) % vs (4.2±0.5)% ,P < 0.05]. Cell proliferation in CRP plus 0. 5 mT or 1.0 mT CMF group (0. 295±0. 009,0. 302±0. 010) were much more than those in CRP group (P<0.05), NO secretion contents [(28.3±4.9) μmol/L, (29.2±5.6) μmol/L]were also much more than those in CRP group (P < 0.05) , apopto-sis rate [(7.4±0.5)% ,(6.9±0.6)%]was significantly lower than that in CRP group (P <0.05). Conclusion CMF at intensity of 0.5 mT and 1.0 mT can antagonize the effects of CR, promote proliferation of EPCs and secretion of NO and inhibit apoptosis rate of EPCs.