Objective To investigate the effects of interferon-α (IFN-α) and gefitinib on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods Colon cancer cell line HCT116 was selected as research objective. The biological effects of IFN-α and gefitinib alone or combined on the cells were observed at different time point (after worked for 24, 48 and 72 hours). The proliferation inhibition of the medicine on the HCT116 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Morphologic changes were observed under optical microscope. Apoptosis was measured by flow cytometry (FCM). The results were analyzed with SPSS 13.0 software, two groups compare was tested by t test, and single factor variance analysis was for multiple group data compare. Results IFN-α and gefitinib alone or combined could significantly inhibit the proliferation of HCT116 cells (P<0.05), and there was a time and dose-dependent manner between the degree of inhibition and the working time and concentration of the medicine. With the work of the medicine, apoptosis morphologic changes were observed in the cells. And FCM result indicated that the apoptosis rate significantly increased. After treated with IFN-α and gefitinib alone or combined for 72h, the cell apoptosis rate were 15.6%±0.6%, 13.6%±0.4% and 31.2%±0.3% respectively, which was obviously higher than control group (6.8%±0.3%, P<0.05). Conclusion Both IFN-α and gefitinib were able to inhibit the proliferation and induce apoptosis of HCT116 cells moreover, and a synergistic effect was observed while combine used there two medicines.

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

Objective Based on the theory of preventive treatment and constitution regulation, to observe the clinical efficacy of point application for stable chronic obstructive pulmonary disease (COPD).Methods Seventy-nine qi deficiency or yang deficiency patients in stable COPD were randomized into a treatment group of 40 cases and a control group of 39 cases. The control group received conventional medication and the treatment group additionally received point application in canicular days or the three-nine days in winter. The frequency of catching cold, the situation of acute exacerbation of chronic obstructive pulmonary disease (AECOPD), constitution types and scores of traditional Chinese medicine (TCM), pulmonary function [the level of forced expiratory volume in one second (FEV1), the percentage of FEV1 in predicted value, FEV1’s ratio to forced vital capacity (FVC)], BODE index (body mass index, airflow obstruction, dyspnea and exercise capacity index) and therapeutic effects were recorded in the two groups before and after the treatment. The correlation analysis was performed between the scores of constitution and the frequency of catching cold, AECOPD, syndrome scores of TCM, BODE index.Results The cold times, AECOPD attack times, times of hospitalization for AECOPD and syndrome scores of TCM were significantly changed in the treatment group after treatment (P<0.01,P<0.05). There were statistically significant changes in the cold times, AECOPD attack times, and syndrome scores of TCM in the two groups after treatment (P<0.05). The total effective rate was 85.7% in the treatment group versus 70.3% in the control group, and the difference was statistically significant (P<0.05). The scores of the main abnormal constitution (qi deficiency or yang deficiency) and balanced constitution were significantly changed in the treatment group after treatment (P<0.01,P<0.05). There were statistically significant differences in the scores of the main abnormal constitution and balanced constitution between the two groups after treatment (P<0.01,P<0.05). There were no statistically significant changes in each item of pulmonary function in the two groups after treatment (P>0.05). After the treatment, the change of the BODE score in the treatment group was significantly different from that in the control group (P<0.01). A significant positive correlation was found between the main abnormal constitution scores and the cold times, AECOPD attack times, times of hospitalization for AECOPD, syndrome scores of TCM in the treatment group after treatment (P<0.05,P<0.01). A significant negative correlation was found between the balanced constitution scores and the cold times, AECOPD attack times, times of hospitalization for AECOPD, syndrome scores of TCM in the treatment group after treatment (P<0.05,P<0.01).Conclusion Point application is an effective method for stable COPD.

Objective To investigate the pathologic changes in the pancreas of rats after intraperitoneal injection of DETC,a kind of superoxide dismutase (SOD) inhibitor,and to compared that with another model of chronic pancreatitis by pancreatic duct injection of TNBS.Methods The rats were randomly divided into DETC group,DETC control group,TNBS group,TNBS control group,normal control group.Rats in DETC group received an intra-peritoneal injection of DETC twice a week,and rats in DETC control group received an intra-peritoneal injection of same amount of normal saline.Rats in TNBS group was injected with 2％ TNBS ethanol phosphate buffer into the pancreatic duct,while rats in TNBS control group was treated with injection of same amount of ethanol phosphate buffer,and rats in normal control group received no treatment.The rats were sacrificed after 2 w,4 w,6 w and 8 w.The serum levels of amylase were determined,and pathological and ultrastructure changes of the pancreas were measured.The levels of SOD,GSH-PX activity and MDA content were detected.The expressions of α-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN in tissue were detected by immunohistochemical assay.The TGF-β1 mRNA expression was detected by RTPCR.Results No rat died in DETC group.The mortality rate of TNBS group was 15％.The serum levels of amylase were not statistically different between the 2 groups.The fibrosis scores of rat in DETC group at 4 w was 3.4 ± 1.l,which was significantly higher than that in TNBS group (3.0 ± 1.3,t =3.462,P ＜ 0.05).At 6 w,the damage scores of rat in DETC group was 9.1 ± 1.8,which was significantly higher than that in TNBS group (8.4 ± 1.8,t =2.943,P ＜ 0.05).Scores of vacuolar degeneration and fatty infiltration of rat in DETC group were higher than those in TNBS group,but the difference between the two groups was not statistically significant.Two weeks later,ultrastructure changes of pancreas could be observed,and large amounts of regenerative or mature collagen could be seen at 4 w.The SOD activity of DETC group was significantly decreased when compared with those in TNBS group (t =5.468,P ＜ 0.01).The GSH-PX activity of DETC group at 2 w,6w was significantly decreased when compared with those in TNBS group (t =6.497,10.125,P＜0.01).While the activity of MDA at 6 w,8 w was significantly increased when compared with those in TNBS group (t =3.350,5.407,P ＜0.05).The differences at other time points were not statistically significant.The expressions of (a)-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN,and TGF-β1 mRNA were not statistically significant between the 2 groups.Conclusions Sustained suppression of SOD activity can successfully induce chronic pancreatitis.Fatty infiltration and fibrosis in pancreas in DETC group occurs earlier with more severe presentation than that in TNBS group.Intraperitoneal injection of DETC is easy with low mortality rate,which is an ideal method for chronic pancreatitis model induction.

The paper analyzed legally the following predicaments of legitimization of rural doctors in China:no legitimacy protection for their practice,vague criteria for practicing medicine,conflicts between legal regulations for medical practice and rural realities,and lack of a liability insurance system.The authors,based on an exploration of the legislative framework for rural doctors,made four corresponding legislative proposals as follows:to clearly define the legal scope and identity of rural doctors,to establish a qualification and employment system for rural doctors,to develop practicing norms applicable for rural doctors,and to set standards for their income.

Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.

Objective To evaluate the relationship between neoadjuvant chemotherapy (combination of taxanes and anthracyclines ) induced-neutropenia and the efficacy of neoadjuvant chemotherapy and long-term survival in operable breast cancer patients. Methods Two hundred and eleven patients received 4 cycles of neoadjuvant chemotherapy (combination of taxanes and anthracyclines).Clinicopathological characteristics were compared between patients with neoadjuvant chemotherapy-induced neutropenia and patients without neutropenia. The efficacy of neoadjuvant chemotheray and long-term survival rate were analyzed. Results Among 211 patients there were 51 (24. 2% ) cases suffering from neutropenia and 160 (75.8%) cases were of no-neutropenia. The response to chemotherapy in patients with neutropenia were more effective than in no- neutropenia ones ( P ＜ 0. 05 ). The 5-year disease-free survival (DFS) in patients with neutropenia was 82. 4%, while the 5-year disease-free survival ( DFS) with nonneutropenia was 60% ( P ＜ 0. 01 ). Additionally, the 5-year overall survival ( OS ) in patients with neutropenia was 90. 2% and in patients with non-neutropenia patients was 67. 5% ( P ＜ 0. 01 ).Conclusions Chemotherapy-induced neutropenia during neoadjuvant chemotherapy combination of taxanes and anthracyclines in patients with operable breast cancer has a better prognosis. The sensitivity of tumors given to chemotherapeutic drugs could be evaluated by chemotherapy-induced neutropenia.

The aim of this project is to establish a fibroblast growth factor-21 (FGF-21) signaling pathway targeted cell model, for screening a class of FGF-21 receptor agonists as anti-diabetic candidates. FGF-21 requires beta klotho transmembrane proteins as co-receptor for the activation of tyrosine kinase FGF receptor (FGFR) signaling, thereby activating a series of intracellular signaling pathways and regulating gene transcription for glucose metabolism. Firstly a recombinant plasmid expressing co-receptor beta klotho and EGFP reporter genes was constructed. After introducing the recombinant plasmid into package cells, the cell culture supernatant was used to infect 3T3-L1 cells, which were then screened for stably expressing beta klotho gene. Administration of FGF-21 increased the expression of GLUT1 and stimulated GLUT1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.

Objective To evaluate the diagnostic value of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) in combination with flow cytometry (FCM) in lymphoma.Methods From January 2011 to December 2011,the cases of suspicious lymphoma with EUS-FNA examination at Shanghai Ruijin Hospital were retrospectively analyzed.The final diagnosis was according to pathological diagnosis of specimen from the surgery and follow up results.The sensitivity,specificity and accuracy of EUS-FNA combined with FCM in lymphoma diagnosis were initially analyzed.Results A total of 14 suspicious lymphoma patients were collected,eight cases were diagnosed as lymphoma by pathological examination of specimen from the surgery or.tissue from aspiration,four cases were non-lymphoma lesions and two cases still had no final diagnosis.The sensitivity and specificity of FCM alone in lymphoma diagnosis were 4/8 and 4/4 respectively.Six cases of lymphoma were detected by EUS-FNA with FCM.The sensitivity,specificity and accuracy of EUS FNA combined with FCM were 6/8,6/6 and 10/12 respectively.Conclusion EUS-FNA combined with FCM has better diagnostic value in lymphoma,especially for gastrointestinal lymphoma and those surrounding deep lesions.

ObjectiveTo investigate the expression of protein inhibitor of activated STAT-1 ( PIAS1 )in rat with acute pancreatitis (AP) and study its prediction value for severity and prognosis.MethodsSD rats were randomly divided into control,acute edematous pancreatitis (AEP),and acute necrotizing pancreatitis (ANP) groups with 15 rats in each group.The rats were sacrificed at 0.5,6,16,24,48,72 h postoperatively.Serum CRP and amylase levels were determined.Water content of pancreas was measured.The pancreatic tissue was routinely harvested and underwent pathologic examination and was scored.The PIAS1protein expression was measured by Western blot and immunohistochemistry method.The survival rates at 72h were recorded and the risk analysis of multiple factors was investigated by Cóx regression method.ResultsThe pathologic score,water content of pancreas,serum CRP and amylase levels at 16h in ANP group were (7.70 ±2.55),(2.91 ±0.57)％,(0.36 ±0.05)mg/L,(3141 ±625)U/L,which were significandy higher than those in AEP group [(2.60 ±1.04),(2.04 ±0.47)％,(0.24 ±0.05)mg/L,(1984 ± 637)U/L)],and also significantly higher than those in control group [ (0.80 ±0.42),( 1.21 ±0.27)％,(0.14 ±0.03)mg/L,(978 ± 353) U/L,(P ＜ 0.05or 0.01 ) ].The survival time of ANP rats was [ (26.36 ± 3.41 ) h,95％ CI:19.67～33.06 h],which were significantly shorter than that in AEP group [ (57.26 ±4.16)h,95％ CI:49.11 ～ 65.42) ],and also significantly shorter than that in control group [ (71.33 ±0.54) h,95％ CI:70.26 ～71.40,P ＜0.01 ].The expression of PIAS1 at 16h in ANP group was significantly lower than those in AEP group and control group ( 0.10 ± 0.01 vs.0.80 ± 0.07,0.87 ± 0.05,P ＜ 0.01 ).The Cox analysis suggested that PIAS1 was an independent prognosis factor for the survival time of AP rats.ConclusionsPIAS1 is lowly to moderately expressed in ANP,and is negatively correlated with AP severity,and may be an independent risk factor for the prediction of severity and prognosis of AP.