Objective To observe the effects of VEGF antisense oligonucleotide on growth of micrangium of xenograft in nude mice.Methods VEGF-ASODN was synthesised.After transfection with VEGF-ASODN,RNA copy was detected by real-time PT-PCR,VEGF protein was examined by ELESA,Survival of cells was tested by MTT.Growth of cells was shown by growth curve.After the model of human gastic cancer xenografts in nude mice was developed,model animals were divided randomly into three groups:antisense group、scrambled group and saline group.ResultsVEGF-ASODN can reduce the VEGF mRNA and VEGF protein in cells and supernate remarkably.Survival and growth of cells were suppressed.It can significantly reduce VEGF protein in serum of nude mice.It also reduced the volum and weight of xenografts.Density of micrangium decreased in xenografts.Conclusion VEGF-ASODN can suppress the growth of micrangium in xenografts.

10.3969/j.issn.1007-3969.2013.05.005

OBJECTIVETo compare the difference in the clinical efficacy on infantile Tourette syndrome between the integrated therapy of acupuncture and pingganjianpi decoction and haloperidol tablets.

METHODSForty-seven children were randomized into an observation group (25 cases) and a control group (22 cases). In the observation group, acupuncture was applied to Taichong (LR 3), Baihui (GV 20), Zhongwan (CV 12), Zusanli (ST 36), etc. The needles were retained for 30 min. Acupuncture was given once a day and there were 5 days at intervals after 10 times of acupuncture. Additionally, pinggan jianpi decoction was prescribed. In the control group, haloperidol tablets were prescribed, starting from the small dose, 0. 05 mg/kg per day, twice a day. The treatment of 30 days made one session and 3 sessions were required. Yale global tic severity scale (YGTSS) was adopted to observe tic time, tic frequency and tic severity score before treatment and in 30 days, 60 days and 90 days after treatment in the two groups. The efficacy and adverse reactions were compared between the two groups.

RESULTSThe total effective rates were 40. 0% (10/25), 64.0% (16/25) and 76.0% (19/25) in the observation group and were 59.1% (13/22), 68.2% (15/22) and 77.3% (17/22) in the control group in 30 days, 60 days and 90 days after treatment respectively. The effect in 30 days after treatment in the control group was better than that in the observation group (P<0. 05). The differences at the other time points were not significant between the two groups (all P>0. 05). The tic time, tic frequency and tic severity score at the each time point after treatment were reduced obviously as compared with those before treatment (all P<0. 05). Each item score in the control group was reduced obviously as compared with that in the observation group in 30 days after treatment (all P<0. 05). The differences at the other time points were not significant between the two groups (all P>0.05). The probability of adverse reaction in the observation group was less than that in the control group.

CONCLUSIONThe integrated therapy of acupuncture and medicine achieves the similar effect on infantile Tourette syndrome to haloperidol tablets and the side effects of it are less.

Acupuncture Therapy ; Child ; Child, Preschool ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Tourette Syndrome ; drug therapy ; therapy ; Treatment Outcome

OBJECTIVE:To evaluate the effectiveness,safety and economy about Wenxin granules in the treatment of cardio-vascular diseases based on Meta-analysis. METHODS:Using“Wenxin granules”“Wenxinkeli”“Wenxin”as the search terms, searching PubMed,ProQuest,Springer,The Cochrane Library,CNKI,VIP and Wanfang data(Jan. 2000-Oct. 2015)through com-puter,relevant journals and conference papers by hand,randomized controlled trials about Wenxin granules in the treatment of car-diovascular diseases were screened and classified according to indications. Meta-analysis was performed by using RevMan 5.3 soft-ware,while cost-effectiveness analysis and minimum cost analysis were used for economic evaluation. RESULTS:Totally,38 liter-atures were involved,including 20 literatures of arhythmia,9 of premature ventricualr contraction,4 of coronary disease angina pectoris,3 of unstable angina pectoris,2 of pediatric viral myocarditis. Meta-analysis showed that compared with control drug, Wenxin granules showed better clinical efficacy for arrhythmia with less ADR;showed better clinical efficacy and ECG efficacy for premature ventricualr contraction with less ADR;showed better clinical efficacy for angina pectoris of coronary disease,unstable angina pectoris and pediatric viral myocarditis,with statistical significance (P<0.05). The incremental cost-effectiveness analysis showed that the additional cost of Wenxin granules in the treatment of above indications was worth it all,compared to control drug. Single factor sensitivity analysis supported the stability of results. CONCLUSIONS:Wenxin granules have a better effectiveness, safety and economy in the treatment of cardiovascular diseases,but still need high-quality evidence to support.

Background and purpose: Current colorectal cancer patient-derived xenografts (PDXs) models were established by samples taken during surgery. However, metastatic colorectal cancer (mCRC) patients have less surgical opportunities, and it was difcult to obtain enough tumor fragment. The aim of the present study was to es-tablish mCRC PDXs by image-guided biopsy. Methods: A total of 12 patients with colorectal cancer who underwent surgery were included. All patients had recurrent lesions or metastatic lesions needed to be histologically confirmed, and none of them had contraindication to biopsy. Tumor tissues not required for clinical diagnosis were used to establish mCRC PDXs. Results: Seven PDXs grew sufficiently for transfer into mice. The success rate was 77.8%. Conclusion:The PDXs established by image-guided biopsy had the advantage of convenient operation, good reproducibility, high achievement ratio, short experimental periodicity and reliably retain specific genetic and morphological features of the primary patient tumors.

Objective To evaluate the clinical value of ultrasound breast imaging reporting and data system (BI‐RADS) in small solid breast masses with diameter ≤1 cm. Methods The ultrasound features of 230 solid breast masses with diameter ≤ 1 cm were described by ultrasound BI‐RADS, the relationship between ultrasound features, BI‐RADS final assesment and pathology were analyzed. Results Of these 230 masses, 72 (31 3.% ) were pathologically confirmed to be malignant and 158 (68 7.% ) to be benign. The ultrasound BI‐RADS features of mass shape, margin, orientation, posterior acoustic features, and microcalcificaition were significantly different between malignant and benign masses( P < 0.05). Irregular shape, noncircumscribed, nonparallel orientation, postrior shadowing, microcalcifications were regarded as malignant ultrasound features, their positive predictive values(PPV), sensitivity, specificity and accuracy for malignancy were 53 3.% -100%, 2 8.% -75 0.%, 82 3.% -100%, 69 6.% -80 9.%, respectively. One hundred and fifty‐two(66.1% ), 62(27 0.% ), 16(7 0.% ) masses were classified into grade 3, 4, and 5, respectively. The PPV for grade 3, 4 and 5 were 10 5.%, 64 5.%, 100% respectively. Among BI‐RADS grade 3 cases, 87 5.%malignant masses were intraductal carcinoma in situ and special type of invasive cancer, among pathological benign BI‐RADS grade 4 masses, 90 9.% were hyperplasia and intraductal papilloma. Conclusions In small breast masses with diameter ≤ 1 cm, due to the sensitivity of malignant signs are not high, the overlap between signs of benign and malignant lesions, pathological type and other factors, the positive predictive value of BI‐RADS grade 3 is higher than criteria of American College of Radiology, so BI‐RADS classification requires further detailed study.

The incorporation of site-specific recombination systems can help to overcome bottlenecks in livestock transgenic technology. For evaluating the efficiency of Cre/lox mediated DNA recombination in embryos and somatic cells, a working model was established using rat mammary carcinoma cell line SHZ-88, aimed at creation of and use repeatedly of selected "friendly loci" in transgenic livestock. An integration vector pTE-lox2272-DsRed-loxP-GFP-loxP, which red fluorescence gene DsRed served as the first target gene and green fluorescence gene GFP as marker gene, was constructed for introduction of acceptor loci in genome. At the same time a replacement vector pT-lox2272-neo-loxP in which Neo coding sequence served as the second target gene was also constructed for replacing DsRed gene. Transgenic cell clones were produced by electroporating SHZ-88 cell with the integration vector. Cells from three transgenic clones selected randomly were further amplified and were then co-electroporated with the replacement vector as well as cre gene. Analysis of the expression patterns of DsRed and GFP indicated that among the 1 070 cell colonies the efficiency on marker GFP deletion was 91.1% and the efficiency on gene replacement was 29.3%. Molecular analysis by PCR and Southern blotting confirmed that the color patterns as expressed by cell colonies could represent the actual molecular events. This working model mediated by Cre/lox system should be useful for the improvement of the present animal transgenic technology.

Background and purpose:Risk of invasive cervical carcinoma (ICC) and its precancerous lesions following high risk human papillomavirus (hrHPV) infection may vary according to HPV types and geographic regions. Analyzing HPV-type distribution in cervical samples from local women aged 30 years and older, this study aimed to identify HPV types with higher risk of developing CIN2+, and to compare diagnostic performance for CIN2+using these types and type combinations. Methods:Cervical samples with histology follow-up from patients of a tertiary cancer center in Shanghai were collected for HPV genotyping by PCR-RDB. The risk associations of HPV types with CIN2+were estimated by logistic regression analysis, and ROC curves were plotted for diagnostic performance evaluation. Results:A total number of 413 specimens were obtained, including 38 CIN1, 184 CIN2/3, 126 ICC patients and 65 negative control people. The 4 most common HPV types in CIN2+were HPV16, 58, 33 and 18, in descending order. And only HPV16 (P<0.000 1), 58 (P=0.002), 33 (P=0.015) were signiifcantly associated with CIN2+lesions. Besides, the area under the ROC curve of the HPV16/18/33/58 test scored statistically higher than the HPV16/18 test did (P=0.006 6). Conclusion:A combined test of HPV16/18/33/58 may offer better performance for detecting CIN2+lesions in our geographic region.

Objective To explore the subcellular localization of Galectin-9 and its effect on allogeneic immune response.Methods The plasmid pEGFP-N1 was inserted with Galectin-9 fragment which was amplified from pBKCMV-Galectin-9 by PCR.The recombinant plagmid wag then transfected into CHO cells using JetPEI in vitro.The cells were cultured in G418 selecting mediam to obtain the stably-transfected cells.The transcription and expression of Galectin-9 gene were verified by immunohistochemical staining and RT-PCR.The solid-phase transgenic CHO cells or freshly-cultured supernatant wag added into the mixed lymphocyte response system to detect the inhibitory effect of Galectin-9.Galectin-9 protein wag administered intraperitoneally for 7d consecutively.Results The expression of Galectin-9 wag localized in the cytosol of CHO.The allogeneic mix lymphocyte proliferation was dose-dependently inhibited by the freshly-cultured supernatant from stably-transfected CHO cells.Furthermore,the supernatant from stably-transfected CHO cells dose-dependently inhibited the level of IL-2.The inhibitory effect could be reversed by Tim-3-Fc blocking.Administration of Galectin-9 significantly prolonged the survival of allogeneic cardiac transplants[(22.7±1.2)d vs(7.2±0.4)d)].Conclusion Galectin-9 may be secreted in physical situation to exert its immunomodulatory function on allogeneic immune response.Furthermore.Galectin-9 may be a novel therapeutic drug in transplant medicine.

To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejection (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.