Objective To study the morphology and the effect of bovine lactoferricin (LfcinB) for Jurkat T leukemia cells and HFL-I cell and further validate and analyze the signal transduction passage way of LfcinB-induced apoptosis. Method The Jurkat T leukemia cells and fibroblasts stained with Hoechst 33258 after in vitro treatment with LfcinB were observed through fluorescence microscope, compared with positive and negative control groups to analyze the difference between the effect of LfcinB on Jurkat T leukemia cells and HFL-I cell with gel electrophoresis analysis of DNA fragmentation. Early apoptosis and mitochondrial membrane potential of Jurkat T leukemia cells were analyzed by flow cytometry, and the changes of caspase-3, caspase-8, caspase-9 and cytochrome C in endochylema of Jurkat T leukemia cells after exposed to LfcinB during 4, 6 and 8 h were detected by Western blotting. Results DNA Ladder was shown by gel electrophoresis analysis of DNA fragmentation in Jurkat T leukemia cells following treatment with LfcinB, which was not found in HFL-I cell. Through fluorescence microscope, we found that nucleus of LfcinB-exposed Jurkat T leukemia cells stained with Hoechst 3325 became condense, but the nucleus of HFL-I cell was not changed. The results of flow cytometry analysis indicated that apoptosis rates of Jurkat T leukemia cells exposed to LfcinB for 2, 4 h were 11.5% and 17.8% respectively, with decline of mitochondria membrane potential. Immunoblot analysis showed that LfcinB could increase the content of cytochrome C, with the activated caspase-3 and caspase-9 increase gradually in endochylema of Jurkat T leukemia cells, however there was no effect of LfcinB on caspase-8. Conclusion LfcinB induced apoptosis of Jurkat T lenkemia cells, but not affected fibroblasts. After Jurkat T leukemia cells, contacted with LfcinB for more than 2 h, the contents of caspase-3, caspase-9 and cytochrome C in the cells were cumulatively increasing, which further validated that LfcinB induced Jurkat apoptosis by intra-cellular signal pathway depending on caspase family.

Background and purpose:Lung cancer is one of most common diseases,The number of new cases of lung cancer are more than 500 000 per year in the world and it has been ranked as number one in terms of incidence of malignant tumors in China.Unfortunately about 40 percent of the patients were diagnosed as locally advanced unresectable non-small cell lung cancer(NSCLC).Elderly patients with NSCLC also showed an increasing trend in the past years.The purpose of this study was to evaluate the efficacy and safety of gemcitabine(GEM) and carboplatin(CBP) used as induction regimen in the treatment of elderly patients with locally advanced unresectable NSCLC.Methods:42 cases of elderly patients have been cytologically and pathologically confirmed with locally advanced unresectable NSCLC,the age of the patients ranged from 65 to 75.The patients were treated with the combined regimen of gemcitabine and cisplatin.GEM 1 000 mg/m2 intravenously injected by drip on the 1st,8th day and the dosage of CBP was AUC 5 that was used on the 1st day,21 days apart to each cycle,most patients received 2-3 cycles.Treatment response was evaluated according to the criteria of RECIST(Response Evaluation Criteria in Solid Tumor),the side effect of the regimen was judged based on WHO criteria.Results:42 patients were evaluable and received a total of 89 cycles chemotherapy.There were no complete regression that could be observed,but 17 cases had partial regression(PR),22 cases with no change(NC) and 3 cases with progression disease(PD).The overall response rate was 40.5%.The main side effects were hematological toxicity.Conclusions:The GC regimen could be used as induction treatment for elderly patients with locally advanced unresectable NSCLC,and the regimen could be well tolerated and is safe in terms of side effects.

Objective To study expression of extracellular signal-regualated kinase in intraepithelial neoplasia,prostatic cancer and BPH tissue. Methods Expression of extracellular signal-regualated kinase (ERK) was determined in 12 prostatic intraepithelial neoplasia (PIN),11 prostate cancer and 16 benign prostatic hyperplasia specimens.ERK expression was determined by immunohistochemistry. Results ERK expression was positive in all the 12 PIN specimens,6 of which being overexpressed.In the 11 prostate cancer specimens,overexpression was observed in only one while the other 10 under expressed.Overexpreassion was not observed in all the BPH specimens whereas 12 of the 16 showed underexpression. Conclusions ERK expression is obviously higher in PIN tissue (P

BACKGROUND:Drug eluting stents and endothelium stents for clinical treatment of vascular stenosis can lead to delayed endothelialization and restenosis. A rapamycin eluting stent combined with CD34 antibody can play a synergistic role to offset delayed endothelialization and intimal hyperplasia due to antiproliferative drugs, but it is stil in the pilot phase. OBJECTIVE:To observe the ability of rapamycin eluting stent combined with CD34 antibody to capture endothelial progenitor cels, and to observe the differentiation characteristics of the captured cels. METHODS:Scanning electron microscope and indirect immunofluorescence were used to observe the morphology and differentiation characteristics of captured endothelial progenitor cels. Under a fluorescence microscope, we observed the captured endothelial progenitor cels and the degree of endothelialization after implantation of the rapamycin eluting stent combined with CD34 antibody into rabbit ear vein. RESULTS AND CONCLUSION:Under the scanning electron microscope, fusiform-like cels with a diameter of 6-8 μm were captured by the composite stent, and 24 hours later, the cels became ful-shaped. The captured cels had the appearance characteristics of endothelial progenitor cels. Results from indirect immunofluorescence observation showed that there were a lot of red fluorescent spots on the coating which represented adherent cels positive for vascular endothelial growth factor receptor-2; the composite stent was largely covered with vascular endothelial cels at 24 hours after stent implantation, and fuly covered at 48 hours, but there was no abnormal cel cluster. These findings indicate that the rapamycin eluting stent combined with CD34 antibody can be specific to rapidly capture endothelial progenitor cels in the peripheral blood, and the stent can be completely covered with vascular endothelial cels at 48 hours after stent implantation, thereby achieving rapid endothelialization and promoting the repair of endothelial cels.

Objective To analyze the clinical features,diagnosis and treatment of thrombotic thrombocytopenic purpura (TTP) in patients with systemic lupus erythematosus (SLE).Methods Clinical manifestations,laboratory findings,diagnosis,treatment and prognosis of 14 SLE patients with TTP were retrospectively analyzed.Results Of the 14 patients diagnosed with SLE and TTP,4 were men and 10 were women.The median age at diagnosis was 23 (17-69) years old.In five patients,the onset of SLE preceded TTP,and in nine patients SLE and TTP occurred simultaneouslv.All the 14 patients had thrombocytopenia and hemolytic anemia,12 had fever,11 had neurologic abnormalities,and 11 had renal dysfunction.Eight patients presented with the classic pentad of symptoms.Six patients were given steroids (alone or in combination with intravenous immunoglobulin and cyclophosphamide),and eight patients were treated with steroids in combination with plasmapheresis,with response rates of 2/6 and 6/8,respectively.Six patients died,with overall mortality rate of 6/14.No patients relapsed during the follow-up period.Conclusions SLE and TTP share some similar clinical symptoms.As a result,repeated examinations of peripheral blood smears are very important for early diagnosis.The renal damage in patients of co-existing diseases is more serious than those with TTP alone or SLE alone.Early diagnosis and prompt treatment with plasma exchange and steroids may improve the prognosis in SLE patients with TTP.

Objective To observe and compare the curative effects of zoledronic acid (ZA) combined with radiotherapy and ZA combined with chemotherapy in the treatment of bone metastasis of non-small cell lung cancer (NSCLC). Methods Seventy-eight patients with NSCLC bone metastasis treated by radiotherapy or chemotherapy were taken in our hospital from January 2010 to June 2014, including 39 cases of ZA combined conventional fractionation radiotherapy (radiotherapy combined group), and 39 cases of ZA combined with chemotherapy (paclitaxel liposome + cisplatin) (chemotherapy combined group). Each group had 39 cases. WHO objective evaluation standard, efficacy evalulation of solid tumor metastasis and curative effect standard grading of pain, anticancer agent toxicity classification standard, Karnofsky standard were used for evaluating and analyzing the patients with primary lung tumor, bone metastasis, degree of pain, adverse reactions and functional status (once before and after the treatment). Results The efficiency rate of primary lung tumor, the efficiency rate of bone metastasis, the total effective rate of pain relief and the improvement rate of functional status (Karnofsky score increased by 10 points or more)in the radiotherapy combined group and chemotherapy combined group were 82.05 % (32/39) vs. 79.49 % (31/39), 48.72 % (19/39) vs. 51.28 %(20/39), 82.05 % (32/39) vs. 84.62 % (33/39), 66.67 % (26/39) vs. 71.79 % (28/39) respectively, and the differences were not statistically significant (the values of x2 were 0.224, 0.237, 0.195, 0.259 respectively, all P> 0.05); Although the two groups showed low-grade fever, bone marrow suppression, esophagitis, liver and kidney damage, gastrointestinal reactions and other adverse reactions, the adverse reactions of two groups were close to [28.21 % (11/39) vs. 30.77 % (12/39)] (x2 = 0.314, P> 0.05). Fortunately, these reactions were controlled well after symptomatic treatment. Conclusion ZA combined with radiotherapy or chemotherapy is a safe and effective way for bone metastasis of NSCLC, which should be taken based on the individual condition of the patients.

Objective To investigate the liver protective effects of Xiaoyaosan in carbon tetrachloride-induced acute liver injury in mice.Methods Thirty mice were randomly divided into a control group, a model group, and aXiaoyaosan group, with 10 mice in each group. The mice in theXiaoyaosan group were intragastrically administrated withXiaoyaosan, the mice in the remaining two groups were fed with an equal volume of distilled water. After 7 days, acute liver injury were inducedvia intraperitoneal injection of carbon tetrachloride peanut oil solution. The serum levels of alanine transaminase(ALT), aspartate transaminase (AST), malondialdehyde(MDA)and superoxide dismutase(SOD)were measured, and pathological changes of liver tissue was tested after 16 hours.Results The serum levels of ALT(136.46±15.75 U/Lvs. 22.96±6.23 U/L), AST(145.37±16.39 U/Lvs. 31.89±7.26 U/L), and MDA level in the liver tissue(17.48±3.45 nmol/mgvs. 4.22±1.08 nmol/mg)in the model group were significantly higher than those in the control group(allP<0.01), SOD level in the liver tissue significantly lower than that in the control group(261.60±20.29 U/mgvs. 336.73±25.34 U/mg,P<0.01). The serum levels of ALT(89.38±6.96 U/L,P<0.01), AST(119.04±20.44 U/L, P<0.05), MDA level(10.30±2.22 nmol/mg,P<0.01) in the liver tissue in theXiaoyaosan group levels were significantly lower than those in the model group, and SOD level(304.77±31.71 U/mg,P<0.01) in the liver tissue were significantly higher than that in the model group.ConclusionXiaoyaosan has liver protective effects in carbon tetrachloride-induced acute liver injury in mice.

ObjectiveTo observe the effect of estrogenic-like effects ofWujia-Shenghua capsule and effective medication site.MethodsThrough D-101 macroporous resin column methods,Wujia-Shenghua capsule with 60% ethanol extraction was separated for water elution part, 20% ethanol elution part, 40% ethanol elution part, 60% ethanol elution part. Then each elution part was respectively mixed into concentration of 10mg/ml,1mg/ml, 0.1mg/ml, 0.01 mg/ml and acted on the MCF-7 cell to have, MTT test and rate of PR calculation.Results Compared with the blank control group(100%), when the concentration was 0.1 mg/ml, water elution part, 20% ethanol elution part and 60% ethanol elution part(98.10%, 101.06%, 106.04%)had no effect on the proliferation of MCF-7 cells, there was not statistically significant(P>0.05), while the 40% ethanol elution part(108.22%)can promote the proliferation of MCF-7 cells,there was a statistically significant(P<0.05). When the concentration was 1 mg/ml, 20%, 40% and 60% ethanol elution part(111.72%, 122.48%, 115.35%)can distinctly promote the proliferation of MCF-7 cells, there was a significant difference between four groups(P<0.01).ConclusionThe 40% ethanol elution part ofWujia-shenghua capsule has the strongest estrogen activity on plant.

Objective To observe the effect of astaxanthin on radiotherapy sensitivity of lung cancer A549 cells transplanted in nude mice. Methods Twenty BALB/c nude mice were divided into four groups:control group (mice were gavaged with pure water containing with 10% DMSO), astaxanthin group (mice were gavaged with astaxanthin suspension containing with 10%DMSO, astaxanthin was given to mice with the dose of 50 mg/kg on the first day, and every other day in the following days with a total of 7 times), radiotherapy group (mice were gavaged with pure water containing with 10%DMSO, the tumor site was given local radiotherapy with a dose of 5 Gy per time and the total dose was 15 Gy) and combination group (mice were given 50 mg/kg astaxanthin and radiotherapy with 15 Gy total irradiated dose). When the minor axis of the tumor reached 5 mm we began experiment. Tumor growth curve was measured by detecting the line of apsides every other day. Mice were killed on the second day after the last time of astaxanthin administration. Weights of tumor were measured by a balance and then tumor mass was processed into paraffin sections. Expressions of proliferating tumor cell antigen Ki-67, phosphorylated-signal transducers and activators of transcription (p-STAT3), and cell apoptosis (measured by terminal deoxynucleotidyl transferase mediated dUTP nick- end labeling, Tunnel) were detected by immunohistochemistry. Results Compared with control group, the transplanted tumor growth rate slowed down in other three groups (P<0.05), and tumor growth was the most slowly in the combination group. Tumor weight, Ki-67 and p-STAT3 expressions were decreased gradually in turn in control group, astaxanthin group, radiotherapy group and combination group. The anti-tumor rate and percentage of cell apoptosis were increased gradually in turn. There was significant difference between groups by multiple comparison statistics(P<0.05). Conclusion Astaxanthin enhances radiotherapy sensitivity of human lung cancer A549 cells in nude mice by down-regulating the expression of p-STAT3.

Objective Through investigating the clinicians’ scientific research level and weakness,we can carry out targeted clinical research training systematically,and then strengthen their utilization of clinical resources and data,and finally promote the transformation of scientific research resuits.Methods Filling and submitting the online questionnaires through WeChat,a popular chatting tool in China,physicians from 13 affiliated hospitals of a university in Shanghai have joined this research.Results 507 valid questionnaires have been received online:In terms of clinical research capacity,there seemed to be more barriers in the following issues:comprehension of the types and applications to design a clinical trial,formulation of the details including PICO elements(Patient-Intervention-Comparison-Outcome),methodological application including the category and principle of randomization,the category and principle of blind method and how to control the confounding elements and probable bias,and statistical problems in scientific research including discrimination of the different definition between FAS(Full Analysis Set) based on the principle of ITT(Intention-To-Treat) and PPS(Per-Protocol Set),how to write a standardized SAP(Statistical Analysis Plan) and how to calculate the sample size of a trial),and various management of clinical trials including data management,follow-up management,adverse event management and so on,and writing a protocol and a CRF (Case Report Form) in a standard and professional way.As for the needs for training,the top three topics were how to design a clinical research in a standardized style,how to practice the methodology and how to utilize the statistical skills into clinical trials.Conclusions Standardized design and writing of clinical trial protocols according to the CONSORT (Consolidated Standards of Reporting Trials) and SPIRIT (Standard Protocol Items:Recommendations for Interventional Trials),and the application of epidemiological and statistical methods are still short boards that most clinicians need to improve urgently,also are what they most concerned about at the same time.So it is necessary for physicians to receive systematic clinical research training to enhance their scientific research capacity.