Objective To investigate the expression of macrophage migration inhibition factor (MIF) in pulmonary tissues of the smokers with and without chronic obstructive pulmonary disease (COPD).Methods The subjects were assigned into three groups:non-smokers without COPD (control group,n =12),smokers without COPD (smoker group,n =13) and smokers with COPD (COPD group,n =16).The specimens were obtained from lung tissues as far away from cancer focus as possible (＞ 5cm).Real-time quantitative PCR and immunohistochemistry were used to investigate the expression and distribution of MIF in pulmonary tissues.The relationship between the severity of airflow obstruction and the differential expressions of MIF in lung tissues of the smokers with or without COPD was analyzed.Results (1) MIF mRNA expression in COPD group (4.87 ± 1.79) was higher than that in the smoker group (2.16 ±0.72;P＜0.01),which was higher than that in the control group (1.09 ±0.48;P ＜0.01).(2)Immunohistochemistry analysis showed that MIF protein expression in lung tissues of the COPD group (0.277±0.025) was higher than that in the smokers group (0.199 ±0.034;P ＜0.01),which was significantly higher than that in control group (0.130 ±0.021 ;P ＜0.01).(3) Correlation analysis of MIF mRNA expression in the lung tissues and pulmonary function parameters of forced expired volume in one second (FEV1) percentage of predicted (FEV1 pred) and ratio of FEV1 to forced vital capacity (FEV1/FVC) suggested that MIF mRNA expression in the lung tissues was negatively related with FEV1 pred (r=-0.578,P ＜ 0.01) and FEV1/FVC (r =-0.607,P ＜ 0.01).Conclusions MIF expression significantly increases in the smokers with COPD,and MIF level in the lung is positively correlated with airflow limitation.The results suggest that MIF may play an important role in the pathogenesis of smokinginduced COPD.

The purpose of this study was to investigate the changes of protein kinase Calpha (PKCalpha) and cyclin D1 expressions in pulmonary arteries from smokers with normal lung function and smokers with mild to moderate chronic obstructive pulmonary disease (COPD). The peripheral lung tissues were obtained from 10 non-smokers with normal lung function (non-smoker group), 14 smokers with normal lung function (smoker group), 11 smokers with mild to moderate COPD (COPD group). The morphological changes of pulmonary arteries were observed by HE-staining. The expressions of alpha-smooth muscle actin (alpha-SMA), proliferating cell nuclear antigen (PCNA), PKCalpha and cyclin D1 proteins in pulmonary artery smooth muscle cells (PASMCs) were immunohistochemically determined. The percentages of PCNA-positive cells were taken as the smooth muscle cells proliferation index (PI). The mRNA expressions of PKCalpha and cyclin D1 in PASMCs were evaluated by real-time fluorescence PCR. Morphometrical analysis showed that the ratio of pulmonary artery wall area to total area (WA%) in smoker group and COPD group was significantly greater than that in non-smoker group (P<0.01). The PASMCs proliferation index in smoker group and COPD group was significantly higher than that in nonsmoker group (P<0.01). The protein levels of PKCalpha and cyclin D1 in PASMCs were significantly increased in smoker group and COPD group as compared with non-smoker group (P<0.01). The mRNA expressions of PKCalpha and cyclin D1 in PASMCs were significantly elevated in smoker group and COPD group as compared with non-smoker group (P<0.01). Significant correlations were found between PKCalpha protein and WA% or PI (P<0.01). Correlations between cyclin D1 protein and WA% or PI also existed (P<0.01). The expression of PKCalpha was positively correlated with the expression of cyclin D1 at both protein and mRNA levels (P<0.01). In conclusion, increased expressions of PKCalpha and cyclin D1 might be involved in the pathogenesis of abnormal proliferation of PASMCs in smokers with normal lung function and smokers with mild to moderate COPD.

Objective To explore the methods of diagnosis and appropriate treatment of pancreatic duct stone.Methods Clinical data of 16 patients with pancreatic duct stone from March,2005 to August,2009 were analyzed retrospectively.Results 15 patients presented with varying degrees of upper abdominal pain,another one with irregular diarrhea.Serum and urine amylase level was higher than the upper limit of normal level in 3 patients;serum glucose was elevated in 4 patients.The diagnostic accuracy by Bultrasonography,CT,MRI,ERCP and KUB was 93.8%(15/16),68.8% (11/16),57.1% (4/7);100%(2/2) and 50% (3/6),respectively.2 cases underwent endoscopic pancreatic sphincterotomy + pancreatic stent drainage,14 cases were treated with surgery,including transpancreatic duct lithotomy + pancreatic jejunal anastomosis in 12 cases,pancreatoduodenectomy in 1 ease,and pancreatic body and tail resection +pancreatic jejunal Roux-en-Y anastomosis in one case.All operations were successful without mortality,and abdominal pain was significantly improved.Follow up of 14 cases showed no stone recurrence.Two patients were lost in follow up,so the follow up rate was 87.5% with the duration ranging from 1 to 53 months.Conclusions B-ultrasonography was the best imaging examination for pancreatic duet stone,but the combined application of imaging tests could significantly improve the diagnostic yield,and imaging examination provided an important basis for the choice of treatment method.With the improvement of endoscopic techniques,ERCP will be as important as surgery for the treatment of pancreatic duct stone.

Objective To investigate the diagnosis and surgical treatment of pancreatic cystic tumor.Methods The clinical data of 19 cases of pancreatic cystic tumor from January 2000 to August 2009 was retrospectively analyzed. Results Patients with pancreatic cystic tumor has no specific clinical feature.Ultrasound and CT were main image examinations, but they could not distinguish the pathologic types, and the diagnostic accuracy when compared with postoperative pathologic results was 57.9% (11/19) and 68.4%(13/19) respectively. The tumors were located in the pancreatic head and neck in 5 cases, body and tail in 14 cases, the maxim diameter was between 3 ～ 15 cm. All patients underwent surgical treatment; the rate of curative resection was 84. 2% ( 16/19 ). The rate of intraoperative misdiagnosis was 21.0% ( 4/19 ).Pathologic examination results showed 6 cases of serous cystadenoma, 6 cases of mucinous cystadenoma, 5 cases of mucinous cystadenocarcinoma, and 2 cases of intraductal papillary mucinous adenoma. 15 ( 78.9% )patients were followed up. Among the 3 patients with mucinous cystadenocarcinoma, one patient who received curative resection survived for 4 years with no evidence of recurrence; the other 2 patients died 4 months and 7 months later. 12 cases of cystadenoma were alive without recurrence. Four patients, including 2 patients of cystadenoma and 2 patients of cystadenocarcinoma were lost in follow-up. Conclusions To be aware of pancreatic cystic tumors is the key to reduce misdiagnosis and mistreatment. Surgical treatment is the treatment of choice with excellent prognosis.

The regulative role of extracellular signal-regulated kinase(ERK)signaling pathway on proliferation and apoptosis in rats' airway smooth muscle cells(ASMCs)was investigated.Primary cultures of ASMCs were established and cells between passages 4 and 7 were used for experiments.ASMCs were U-eated with ERK activator epidermal growth factor(EGF)and inhibitor PD98059.The expressions of ERK mRNA and protein were detected by RT-PCR and immunofluorescence staining.Proliferation of ASMCs were detected by MTT eolorimetric assay and [3H] thymidine incorporation.Apoptosis of ASMCs were detected by Hoechst staining and Annexin-V FITC PI donble staining.The levels of ERK1/2,phosphorylated forms of ERK1/2(p-ERK1/2) and proeaspase-3 protein were detected by Western blotting.The expressions of ERK mRNA and ERK protein were obviously observed in ASMCs. Compared with control,the absorbanee(A490) value and DNA synthesis index of PD-treated ASMCs were significantly decreased(P＜0.05).The apoptotic index and the percentage of the early apoptotic cells were significantly increased(P＜0.05).The expressions of ERK1/2 and pERK1/2 protein were significantly down-regulated.The expressions of procaspase-3 protein was significantly increased.Compared with c,ontrol,the A490 value and DNA synthesis index were significantly increased(P＜0.05)in EGF-treated cells.Apoptotic index and the percentage of the early apoptotic cells were significantly decreased(P＜0.05).The levels of ERK1/2 and pERK1/2 protein were significantly increased,and the levels of procaspase-3 protein were significantly decreased, There were no significant differences between control and P+E group(P＞0.05).ERK signaling pathway may play an important role in regulating ASMCs proliferation and apoptosis and ERK regulating the apoptosis of ASMCs possibly relates to the expressions of procaspase-3 protein. The finding will help to understand the mechanisms of asthmatic ASMCs involved in abnormal proliferation.

Objective To systematically evaluate the safety and effectiveness of combined portal vein resection and reconstruction in the resection of hilar cholangiocarcinoma.Methods Literatures were researched using Cochrane Library,PubMed,Embase,China Biology Medicine disc,China National Knowledge Infrastructure,Wanfang database,VIP database from January 31,2006 to January 31,2016 with the key words including “hilar cholangiocarcinoma”“Klatskin tumor”“Bile duct neoplasm”“Vascular resection”“portal vein resection”“肝门部胆管癌”“血管切除”“门静脉切除”.The clinical studies of resection of hilar cholangiocarcinoma with portal vein resection and construction and without vascular resection and construction were received and enrolled.Two reviewers independently screened literatures,extracted data and assessed the risk of bias.Patients who underwent resection of hilar cholangiocarcinoma combined with portal vein resection and reconstruction were allocated into the portal vein resection group and patients who didn't undergo vascular resection were allocated into the no vascular resection group.Analysis indicators included (1) results of literature retrieval;(2) results of Meta-analysis:① incidence of postoperative complications (hepatic failure,biliary fistula,intra-abdominal hemorrhage),② postoperative mortality,③ patients' prognosis,④ related indicators of postoperative pathology (lymph node metastasis rate,moderate-and low-differentiated rate,nerve invasion rate,negative rate of resection margin).The heterogeneity of the studies was analyzed using the I2 test.The hazard ratio (HR) and 95％ confidence interval (CI) were used for assessing the prognostic indicators.The incidence of complications,mortality and pathological indicators were evaluated by the odds ratio (OR) and 95％ CI.Results (1) Results of literature retrieval:13 retrospective studies were eurolled in the meta-analysis,and the total sample size was 1 668 cases including 437 in the portal vein resection group and 1 231 in the no vascular resection group.(2) Results of Meta-analysis:① incidence of postoperative complications was respectively 39.86％ in the portal vein resection group and 35.27％ in the no vascular resection group,with no statistically significant difference between the 2 groups (OR =1.12,95％ CI:0.82-1.53,P ＞0.05).The results of subgroup analysis showed that hepatic failure,biliary fistula and intra-abdominal hemorrhage were postoperative main complications,and the incidences were 17.09％,8.79％,6.25％ in the portal vein resection group and 10.62％,9.69％,2.51％ in the no vascular resection group,respectively,with no statistically significant difference between the 2 groups (OR =0.48,1.13,0.82,95％ CI:0.23-1.02,0.45-2.83,0.21-3.12,P ＞ 0.05).② Postoperative mortality was respectively 5.38％ in the portal vein resection group and 3.88％ in the no vascular resection group,with no statistically significant difference between the 2 groups (OR =1.16,95％ CI:0.62-2.14,P ＞ 0.05).③ There was statistically significant difference in patients' prognosis between the 2 groups (HR =1.81,95％ CI:1.52-2.16,P ＜ 0.05).④ The related indicators of postoperative pathology:lymph node metastasis rate,moderate-and low-differentiated rate and negative rate of resection margin were 41.55％,76.42％,63.74％ in the portal vein resection group and 33.42％,66.75％,64.29％ in the no vascular resection group,respectively,with no statistically significant difference between the 2 groups (OR =1.45,1.59,0.67,95％ CI:0.95-2.21,0.97-2.61,0.37-1.20,P ＞ 0.05).The nerve invasion rate was 83.47％ in the portal vein resection group and 64.90％ in the no vascular resection group,with a statistically significant difference between the 2 groups (OR =2.61,95 ％ CI:1.45-4.70,P ＜ 0.05).Conclusion Combined portal vein resection and reconstruction is safe and feasible in the treatment of hilar cholangiocarcinoma,and the prognosis of patients with portal vein invasion is worse than that without portal vein invasion.

Objective To compare the safety and effectiveness of duodenum-preserving pancreatic head resection (DPPHR) with pylorus-preserving pancreaticoduodenectomy (PPPD) in the treatment of chronic pancreatitis with a pancreatic head mass.Methods Medline,Biosis,Cochrane Library,Science Citation Index Database,CBM Database,Wan Fang and CNKI were searched systematically.The bias risk of the included trials was assessed according to the assessing tools as suggested by the Cochrane Handbook.Review Manage 5.2 was used to perform the statistical analysis.Results 7 RCTs with 226 patients were included in the meta-analysis which showed that there were no significant differences between PPPD and DPPHR in overall postoperative morbidity,postoperative hospital stay,complete pain relief,pancreatic fistula,exocrine insufficiency,symptom score at 5 to 7-year follow-up,and quality of life score at 14 to 15-year follow-up (P ＞ 0.05).While DPPHR had significant superiorities in operation time,blood replacement,delayed gastric emptying,occupational rehabilitation after the operations,weight gain,quality of life score at 1 to 2-year follow-up,symptom score at 5 to 7-year follow-up,and physical functioning score at 14 to 15-year follow-up.Conclusions DPPHR is more favourable than PPPD in reducing the use of blood replacement,shortening operation time,delayed gastric emptying,occupational rehabilitation after the operations,weight gain,physical functioning,and in improving quality of life of patients.

AIM: To investigate whether Ligustrazine(LTZ) has an effect on the changes of protein kinase C(PKC) signaling pathway induced by inflammatory mediators involved in asthma in normal human peripheral blood lymphocytes (PBL). METHODS: 10 mL peripheral venous blood was obtained from each of 63 health humans and treated as follows. The activities of PKC from cytosolic and membrane fractions in PBL were measured by -ATP-catalyzing assay, after PBL had been isolated and performed by following processes: (1) First: three groups treated with 5 g/L LTZ(n=6) or 5 ?mol/L Ro31-8220 (n=6); Paired untreated PBL served as control of this group, as well as the negative controls of the following groups(n=6); (2)Second : three groups treated with 100 nmol/L Methacholine (Mch, n=5), 5 g/L LTZ+100 nmol/L Mch(n=5)or 5 ?mol/L Ro31-8220(a PKC inhibitor)+100 nmol/L Mch(n=5); (3)Third: three groups treated with 100 nmol/L histamine, 5 g/L LTZ+100 nmol/L histamine(n=5) or 5 ?mol/L Ro31-8220+100 nmol/L histamine(n=5); (4)Fourth: three groups treated respectively with 100 nmol/L PMA(a PKC activator, n=5), 5 g/L LTZ+100 nmol/L PMA(n=5) or 5 ?mol/L Ro31-8220+100 nmol/L PMA(n=5). RESULTS: (1)LTZ had no effect on the activities of PKC in inactive PBL in normal humans; (2) Methacholine or histamine resulted in an increase in membrane PKC activity of normal human PBL, which was partly suppressed by LTZ (all P

AIM:To investigate the activity change of voltage-dependent delayed rectifier potassium channel(Kv) on human airway smooth muscle cells(HASMCs) after transfection of Kv1.5 antisense oligonucleotides(AsOND),and to discuss the regulating mechanism of Kv1.5.METHODS: The mRNA and protein expressions of Kv1.5 in liposome-mediated Kv1.5 AsOND transfected HASMCs were measured with techniques of reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blotting.Kv activities in transfected HASMCs were investigated with whole-cell patch clamp.RESULTS: After HASMC were transfected by liposome-mediated Kv1.5 AsOND,the mRNA and protein expressions of Kv1.5 were decreased,and Kv activity was inhibited,which made the cell membrane potential(Em) inclined to depolarization.CONCLUSION: Transfection of Kv1.5 AsOND made the function of Kv in HASMCs decreased.Kv1.5 may play a critical role in the regulation of Kv activity.

AIM: To explore the change of delayed rectifier potassium channel (K_V) activity in alveolar macrophages (AM) in chronic obstructive pulmonary disease (COPD) rats. METHODS: COPD model was established by exposure of the animals to cigarette smoke. With whole-cell voltage- or current-clamp techniques, K_V activity, membrane capacitance and resting membrane potential (Em) in AM from COPD model and control rats were compared. RESULTS: (1) Significant increases in total mononuclear cells and AM in bronchoal aveolar lavage fluid (BALF) were found in COPD group compared with in control group. (2) The AM K_V current altitude in COPD group [(520.5?38.7)pA, (+50) mV, n=30] was significantly lower than that in control group [(713.6?44.4)pA, (+50) mV, n=30, P0.05), but had more positive Em (P