Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P＜0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.

Animals ; China ; Elapidae ; Humans ; Male ; Snake Bites ; Snake Venoms ; poisoning ; Young Adult

Objective: To systemically review the safety and efficacy of Absorb bioresorbable vascular scaffold (Absorb BVS) versus cobalt chromium-everolimus eluting stent (CoCr-EES) for treating the patients with coronary artery disease (CAD) by percutaneous coronary intervention (PCI).
Methods: We searched relevant literatures in PubMed, Embase, Cochrane Library, CNKI and Wanfang database from 2008-01 to 2015-10, meanwhile, collected published data and randomized controlled trials from meeting abstracts and websites to compare Absorb BVS and CoCr-EES for treating CAD patients. The quality of literatures were assessed and extracted by modiifed Jadad score, Meta-analysis was conducted by STATA 12.0 soft ware.
Results: There were 4 eligible trials with 3,389 patients were enrolled. Absorb BVS group, n=2,164 and CoCr-EES group, n=1,225. During the mean of 1.1 years follow-up period, the following incidences were similar between 2 groups:target lesion failure (OR=1.29, 95%CI 0.95-1.74, P=0.10), all cause death (OR=1.31, 95%CI 0.60-2.87, P=0.50), cardiac death (OR=1.38, 95%CI 0.45-4.24, P=0.57), myocardial infarction (OR=1.30, 95%CI 0.93-1.80, P=0.12), deifnite or probable stent thrombosis (OR=2.08, 95%CI 0.95-4.54, P=0.07), re-vascularization (OR=1.03, 95%CI 0.80-1.33, P=0.81), target lesion re-vascularization (OR=1.06, 95%CI 0.67-1.66, P=0.81) and the patient-oriented composite endpoint (OR=0.95, 95%CI 0.66-1.35, P=0.76).
Conclusion: Absorb BVS and CoCr-EES had the similar safety and efifcacy for treating the low and moderate risk CAD patients who had angina, ischemia while without symptoms. The longer period of follow-up study and larger randomized, controlled clinical trials were needed for Absorb BVS application in clinical practice.

Objective The feasibility of predicting the B-cell epitopes of human Neutrophil Gelatinase-Associated Lipocalin (NGAL) was discussed by applicating bioinformatics technology.Linear epitope molecules that have diagnostic value were screened and these recombinant linear multi-epitope peptides were constructed,and expressed.The immunogenicity of the recombinant linear multi-epitope peptides were also identified.Methods NGAL amino acid sequence was got from GenBank in the Department of Clinical Laboratory of the Second Affiliated Hospital of Nanjing Medical University in July 2015,the Predicted,ABCpred,BepiPred,BcePred,and Lasergene softwares were used to predict the linear B cell epitope prediction.The predict epitopes were constructed and prokaryotic expressed,and then the single epitope antigens which could reacted with commercially available polyclonal NGAL antibody were screened out by Western blot.Finally,the multi-epitope peptide was constructed,expressed,and identified through immunoreactions.Results Eight possible epitopes were obtained after prediction.pET32a-N1-N8 prokaryotic expression vector were used to express the predict epitopes.After purification and Western blot analysis,three of the epitopes have strong antigenicity,and then a soluble fusion protein was expressed and obtained from the multi-epitope prokaryotic expression vector pET22b-Ngal_MEP1.The fusion protein was successfully purified by Ni2 + affinity column.Western blot analysis showed that the fusion protein had a strong antigenicity.Conclusions The constructed multi-epitope linear NGAL antigen peptides can obtain high soluble expression in prokaryotic expression system,and have a strong immunoreactivity,which can be used in subsequent antibody preparation.

Puerarin (PUE), as an isoflavone component, has a wide range of pharmacological activities, while its poorly aqueous solubility limits the development of solid oral dosage forms. In this study, PUE along with nicotinamide (NIC) were prepared into the coamorphous system by solvent-evaporation method and characterized by powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FT-IR). In addition, its dissolution behavior and solubilization mechanism were also investigated. PUE-NIC coamorphous was a single homogeneous binary system, with a single glass transition temperature at 35.1 ℃. In comparison to crystalline PUE, during the dissolution process, coamorphous PUE-NIC not only exhibited the "liquid-liquid phase separation" (LLPS) phenomenon, but the formation of A_p type complexation (1∶1 and 1∶2) between PUE and NIC molecules was also verified, which significantly improved the solubility of PUE and prolonged the supersaturation time, and would benefit its absorption.

Objective To investigate the clinical efficacy of hemorrhoids suppositories plus fumigation and hip bath with Chinese medicine on postoperative pain and edema of patients with mixed hemorrhoids.Methods A total of 62 cases of mixed hemorrhoids to be treated surgically were randomly divided into the trial group and the control group,with 31 cases in each group.Both groups of patients were given the surgery of mixed hemorrhoids(high ligation of hemorrhoids or Milligan-Morgan hemorrhoidectomy)and routine postoperative nursing.Additionally,the control group was treated with rectal hemorrhoids suppository,and the trial group was treated with rectal hemorrhoids suppository plus fumigation and hip bath with Chinese medicines of Carthami Flos,Persicae Semen,Corydalis Rhizoma,Talcum,Toosendan Fructus,Aurantii Fructus,Phellodendri Chinensis Cortex,Myrrha,and Olibanum.The course of treatment for the two groups covered 2 weeks.The changes of pain score and edema score in the two groups were observed before and after treatment,and time for the relief of the symptoms,clinical efficacy and therapeutic satisfaction were also compared.Results(1)After 2 weeks of treatment,the total effective rate of the trial group was 96.77%(30/31),and that of the control group was 80.65%(25/31).The intergroup comparison(tested by chi-square test)showed that the therapeutic efficacy of the trial group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,the pain scores and edema scores of patients in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the decrease of the scores in the trial group was significantly superior to that in the control group with statistically significant difference(P＜0.01).(3)The time for pain relief and time for edema subsidence in the trial group were significantly shorter than those of the control group,and the differences were statistically significant(P＜0.01).(4)The total satisfaction rate of the trial group was 96.77%(30/31),while that of the control group was 77.42%(24/31).The intergroup comparison(tested by chi-square test)showed that the therapeutic satisfaction of the trial group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion The therapeutic effect of hemorrhoids suppository plus fumigation and hip bath with Chinese medicine in treating postoperative pain and edema of mixed hemorrhoids is remarkably,and the therapy can effectively improve the degree of edema and pain,promote the recovery of the patients,and improve the patient's therapeutic satisfaction.

Objective: To systematically review the safety and efifciency for dual antiplatelet therapy (DAPT) ≤ 6 months and DAPT ≥ 12 months in patients after drug eluting stent (DES) implantation. Methods: We collected the data for randomized clinical trials for DAPT ≤ 6 months and DAPT ≥ 12 months in patients after DES implantation up to 2015-01 by searching the literatures of PubMed, EMBASE, Cochrane Library, Scopus and Chinese literature database, and meanwhile collected the relevant reporting cases from both domestic and international cardiovascular conferences for this study. There were 2 investigators independently conducted the literature screening, data extraction and quality evaluation, Meta analysis was performed with STATA 12.0 software. Results: A total of 15,378 patients from 7 eligible studies were enrolled and the patients were divided into 2 groups: DAPT ≤ 6 months group,n=7672 and DAPT ≥ 12 months group,n=7706. Meta analysis indicated that DAPT ≤ 6 months could effectively reduce the major bleeding (OR=0.58, 95% CI 0.37-0.91,P=0.017). While the other incidences between 2 groups were similar as all cause death (OR=0.90, 95% CI 0.73-1.11,P=0.314), cardiac death (OR=0.93, 95% CI 0.70-1.24,P=0.617), myocardial infarction (OR=1.13, 95% CI 0.91-1.41,P=0.275), in stent thrombosis (OR=1.21, 95% CI 0.79-1.85,P=0.382) and cerebrovascular accidents (OR=1.00, 95% CI 0.66-1.51,P=1.000). Conclusion: The incidence rates of cardiovascular and cerebrovascular events are similar in patients with DAPT ≤ 6 months and DAPT ≥ 12 months after DES implantation. DAPT ≤ 6 month had the lower risk of bleeding, which is rather suitable for the patients received new generation of DES, with higher risk of bleeding, lower risk of thrombosis and with poor compliance to medication; however, the large and randomized clinical trials are needed to make ifnal conclusion.

A GeXP based multiplex PCR assay was developed to simultaneously detect six different chicken respiratory viruses including H5, H7, H9 subtypes of avian influenza virus(AIV), new castle disease virus (NDV), infectious bronchitis virus(IBV) and infectious laryngotracheitis virus(ILTV). According to the conserved sequences of genes of each pathogen, seven pairs of specific primers were designed, and the reaction conditions were optimized. The specificity and accuracy of GeXP were examined using samples of single and mixed infections of virus. The sensitivity was evaluated by performing the assay on serial 10-fold dilutions of cloned plasmids. To further evaluate the reliability, thirty-four clinical samples were detected by GeXP. The corresponding specific fragments of genes were amplified. The detection limit of GeXP was 10(2) copies/microL when all of 7 pre-mixed plasmids containing target genes of six chicken respiratory viruses were present. In the detection of thirty-four clinical samples, the results of GeXP were accorded with the viral isolation completely. In conclusion, this GeXP assay is a rapid, specific, sensitive and high-throughput method for the detection of chicken respiratory virus infections. It can be applied in rapid differential diagnosis for clinical samples, and also provide an effective tool to prevent and control chicken respiratory diseases with similar clinical symptoms.
Animals ; Chickens ; Influenza A virus ; classification ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; diagnosis ; virology ; Multiplex Polymerase Chain Reaction ; methods ; Poultry Diseases ; diagnosis ; virology ; Respiratory Tract Infections ; diagnosis ; veterinary ; virology

BACKGROUNDIntratracheal instillation of blood induces self-repaired acute lung injury. However, the mechanism of repair has been unclear. Heme-oxygenase (HO)-1, which catalyzes heme breakdown, acts as an inducible defense against oxidative stress and plays an important role in inflammation. The objective of this study was to test the role of HO-1 in lung injury caused by intratracheal instillation of red cells.

METHODSForty healthy, male Sprague-Dawley rats were randomly divided into five groups: normal group, saline group, erythrocyte group, erythrocyte+zinc-protoporphyrin (ZnPP, HO-1 inhibitor) group and saline+ZnPP group. At 2 days after intratracheal instillation of red cells, lung tissues and lavage samples were isolated for biochemical determinations and histological measurements.

RESULTSHistological analysis revealed that administration of ZnPP worsened the acute lung injury induced by instilled erythrocytes. HO-1 was over-expressed in the erythrocyte group and in the erythrocyte + ZnPP group. Compared with the erythrocyte + ZnPP group, the levels of total protein, lactate dehydrogenase and tumor necrosis factor-alpha in the lavage were lower (P < 0.01), while the level of interleukin-10 was higher in the erythrocyte group (P < 0.01).

CONCLUSIONHO-1 protects against erythrocyte-induced inflammatory injury in lung.

Animals ; Erythrocytes ; physiology ; Heme Oxygenase (Decyclizing) ; analysis ; physiology ; Interleukin-10 ; analysis ; Lung ; pathology ; Lung Injury ; prevention & control ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

We investigated inhibition of Moloney leukemia virus 10 (MOV10) upon xenotropic murine leukemia virus-related virus (XMRV) and made a preliminary study of the mechanism of action. Using transfection, infection, western blotting and real-time polymerase chain reaction, we found that MOV10 inhibited XMRV replication. Using MOV10 overexpressed in viral producer cells, MOV10 was shown to reduce the infectivity of XMRV. MOV10 could be incorporated into XMRV, suggesting that MOV10 could undergo encapsidation by XMRV during viral assembly. MOV10 could also restrict the DNA production of XMRV in target cells. We found that the putative RNA-helicase domain of MOV10 maintained most of its XMRV inhibition. These results suggest that MOV10 could be required during the retroviral lifecycle. Perturbation of MOV10 disrupts the generation of infectious viral particles, suggesting that MOV10 has broad antiretroviral activity. Hence, MOV10 could be actively involved in host defense against retroviral infection.
Humans ; Moloney murine leukemia virus ; physiology ; RNA Helicases ; physiology ; Virus Replication