Objective To construct a replication-deficient recombinant adenovirus expression vector of human heparanase (hpa). Methods The hpa gene was cloned at the downstream of CMV promoter of the adenoviral shuttle plasmid pDC315 in sense direction, and the resultant recombinant plasmid pDC315-hpa was transfected into HEK293 cell together with plasmid pBHGlox (deltaE1,3) containing adenoviral genome, then the replication-deficient recombinant adenovirus expression vector of hpa (Ad-hpa) was obtained, and it was identified by infection test, electronic microscope observation and PCR amplification. Results After purification and concintration,the titer of Ad-hpa reached 5?10 10pfu/ml. Virus particles could be found in virus concintration solution, and replication of virus was observed in HEK293 cells was observed under transmission electron microscope. Both adenovirus and hpa special fragment could be amplified from Ad-hpa by PCR, whereas hpa special fragment could not be amplified from the control. Conclusion The replication-deficient recombinant adenovirus expression vector of hpa was constructed successfully. This study established a foundation for further study on hpa vaccines and gene therapy for carcinoma.

Objective To compare the procedural difficulty index (PDI) and immediate outcome (IM) of percutaneous coronary intervention (PCI) in patients with various stages of myocardial infarction.Methods Ninety-four patients with myocardial infarction were divided into three groups, direct PCI(n=38), delayed PCI(n=22) and late PCI(n=34). The characteristics of infarct-related coronary artery, PDI and IM of PCI were evaluated angiographically, and severe procedural complications (SPC) and major adverse cardiac events (MACE) during hospitalstay were documented. Results In the three groups, PDI was 1.47 ?1.79, 1.82 ?1.72 and 2.85 ?2.83, respectively (P

Transcriptional regulation is one of the major regulations in plant adventious shoot regeneration, but the exact mechanism remains unclear. In our study, the RNA-seq technology based on the IlluminaHiSeq 2000 sequencing platform was used to identify differentially expressed transcription factor (TF) encoding genes during callus formation stage and adventious shoot regeneration stage between wild type and adventious shoot formation defective mutant be1-3 and during the transition from dedifferentiation to redifferentiation stage in wildtype WS. Results show that 155 TFs were differentially expressed between be1-3 mutant and wild type during callus formation, of which 97 genes were up-regulated, and 58 genes were down-regulated; and that 68 genes were differentially expressed during redifferentiation stage, with 40 genes up-regulated and 28 genes down-regulated; whereas at the transition stage from dedifferentiation to redifferention in WS wild type explants, a total of 231 differentially expressed TF genes were identified, including 160 up-regualted genes and 71 down-regulated genes. Among these TF genes, the adventious shoot related transcription factor 1 (ART1) gene encoding a MYB-related (v-myb avian myeloblastosis viral oncogene homolog) TF, was up-regulated 3 217 folds, and was the highest up-regulated gene during be1-3 callus formation. Over expression of the ART1 gene caused defects in callus formation and shoot regeneration and inhibited seedling growth, indicating that the ART1 gene is a negative regulator of callus formation and shoot regeneration. This work not only enriches our knowledge about the transcriptional regulation mechanism of adventious shoot regeneration, but also provides valuable information on candidate TF genes associated with adventious shoot regeneration for future research.
Arabidopsis ; growth & development ; Arabidopsis Proteins ; physiology ; Gene Expression Regulation, Plant ; Genes, Plant ; Plant Shoots ; growth & development ; RNA ; Regeneration ; Seedlings ; growth & development ; Transcription Factors ; physiology ; Up-Regulation

Objective To prepare the standard molecular weight fragment mixtures. Methods Primers were designed to prepare clones which contained different sizes of standard molecular weight fragments. The template used for amplification of insert fragments was the pMD18-T vector. Bacteria culture and plasmid extraction were used to obtain abundant target fragment. Unlabeled DNA fragments were prepared by double digestion of the recombinant plasmids, and the fluorescent adaptor was prepared by annealing with two partial reverse complimentary DNA fragments. The unlabeled fragments and fluorescent adaptor were connected by DNA ligation reaction assisted with T4 DNA ligase. In this way, different sizes of standard molecular weight fragments were prepared. Standard molecular weight fragment mixture was finally prepared by mixing all the fragments together before purification. Results Ten standard molecular weight fragments of different sizes were prepared. The sizes of each fragment are 80bp, 124bp, 194bp, 224bp, 254bp, 304bp, 349bp, 399bp, 424bp and 454bp. The internal standard could accurately determine the size of PCR products amplified with the DNATyper15 kit. Conclusion Using this method, the standard molecular weight fragment mixture which meet the requirements of research and laboratory use was prepared, perfectly providing a new method for preparation of the DNA molecular weight standards. The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.

Objective This study aimed to explore clinical characteristics of four types of obesity based on metabolic classification. Methods Forty-eight obese patients were divided according to their clinical characteristics into 4 groups including metabolic healthy obesity (MHO), hypometabolic obesity (LMO), hypermetabolic obesity (HMO), and metabolic obesity with inflammation (IMO). 20 normal weight individuals were also recruited as a control group. Body fat, body weight, visceral index, and basal metabolism were measured by Omron body fat meter. Fat content and its distribution were measured by dual energy X-ray absorptiometry. All participating patients underwent various tests for 75 g oral glucose tolerance, blood glucose, insulin, C peptide. Lipid profile, thyroid function and sex hormones levels, and inflammation factors were also measured. Results (1)Patients in MHO group had higher body fat content, but had no metabolic disorder and inflammation. Their hormones levels were normal. (2) Lower metabolic rate and lower hormones levels were found in the patients in LMO group with increasing visceral fat. Trunk/subcutaneous fat mass was significantly higher than that in MHO group(1. 19 ± 0. 25 vs 0. 97 ± 0. 32, P<0. 05). There were abnormal lipid and glucose metabolism in LMO group. The insulin action index was significantly lower than that in MHO group(0. 006 6 ± 0. 002 7 vs 0. 012 1 ± 0. 009 5, P<0. 05). The area under the curve of glucoseconcentrationwassignificantlyhigherinLMOgroupthanthatinMHOgroup[(18.71±8.68vs12.70±4.63) mmol/L, P<0. 05]. (3)Heart rate and blood pressure were higher in HMO group. The heart rate was significantly increased compared with that in MHO group [(90. 50 ± 8. 24 vs 73. 20 ± 14. 11) beat/min, P<0. 05]. The waist circumference was significantly larger than that in MHO group [(111. 88 ± 10. 54 vs 98. 05 ± 15. 56) cm, P<0. 05]. (4) In IMO group, insulin action index was significantly lower than MHO group (0. 007 0 ± 0. 003 3 vs 0.0121±0.0095,P<0.05). ThetrunkfatmassanduricacidlevelsweresignificantlyhigherthanMHOgroup [(17236.38±4610.60vs15816.10±5453.42)gand(468.28±121.32vs376.84±97.14) μmol/L,bothP<0. 05]. Patients in IMO group had acanthosis nigricans, but their glucose level was relatively normal. Conclusion The metabolic-based obese diagnosis is essential for understanding the obesity etiology and providing individualized treatment.

Objective To report our first clinical experience with a novel modified culotte technique for the treatment of true coronary bifurcation lesions. Methods The novel modified culotte technique (the mono-ring culotte) stenting was done in which the side branch (SB) stent was deployed firstly followed by ex vivo wiring of a most proximal cell of SB stent with the hard end of main branch (MB) wire. Secondly, the MB stent was deployed through the most proximal cell of SB stent. The procedure was ended with kissing balloon dilation. From June 2014 to March 2015, 15 patients with true coronary bifurcation lesion were treated with mono-ring culotte stenting in our center. Results The procedures were successful in all cases without procedural complication and in-hospital major adverse cardiovascular events. The procedural time was (34. 3 ± 9. 6) min, fluoroscopic time was (18. 1 ± 3. 8) min, and contrast volume was (112. 0 ± 24. 5) ml, respectively. Post-procedurally, the residual stenosis of the main and the side branch were (10. 0 ± 2. 5)% and (10. 2 ± 5. 3)% , respectively. Conclusions The mono-ring culotte stenting is safe and feasible for treatment of true coronary bifurcation lesions, and may be superior to the conventional culotte stenting.

Objective To develop and validate a model based on deep learning for automatic diagnosis of early gastric cancer ( EGC) to improve detection and diagnosis of EGC. Methods A total of 5159 images ( including 1000 images of EGC and 4159 images of other benign lesions or normal patients) obtained from May 2014 to December 2016 were collected from endoscopic database in changhai Hospital. Then 4449 images were selected randomly for a deep convolutional neural network ( CNN ) training, of which 768 were diagnosed as EGC and 3681 diagnosed as other benign lesions or normal. The remaining 710 images were used to test the model by comparing with diagnostic results of four endoscopists. Results The deep learning model showed accuracy of 89. 4％ ( 635/710 ) , sensitivity of 88. 8％ ( 206/232 ) and specificity of 89. 7％ ( 429/478) for EGC. The mean time required for diagnosis was 0. 30 ± 0. 02 s. The performance of the model was superior to that of four endoscopists. Conclusion The model based on deep learning has high accuracy,sensitivity and specificity for detecting EGC,which could assist endoscopists in real-time diagnosis.

Purpose: Bone destruction and pain caused by cancer is one of the most devastating complications of cancer patients with bone metastases, and it seriously affects the quality of patients’ life. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule with increased expression in a variety of tumors. This study focused to clarify the specific function of EMMPRIN in bone metastasis of breast cancer. Materials and Methods: Adenovirus with shRNA-EMMPRIN was transfected into MRMT-1 rat breast carcinoma cells, and the MRMT-1 cells with different expression levels of EMMPRIN were implanted into the bone marrow cavity of rat tibia. Next, the effect of down-regulation of EMMPRIN was evaluated as follows: bone damage was detected by X-ray radiological and tartrate-resistant acid phosphatase staining; the tumor burden was evaluated by hematoxylin and eosin staining; the test of pain-related behaviors was assessed used the bilateral paw withdrawal mechanical threshold; and the levels of secretory factors in tumor conditioned medium were determined by using enzyme-linked immunosorbent assay. Results: We found that down-regulation of EMMPRIN in tumor cells can simultaneously reduce tumor burden, relieve cancer-induced bone destruction and pain. Conclusion: Materials and Methods EMMPRIN is expected to be a therapeutic target for relieving bone metastasis of breast cancer and alleviating cancerinduced bone destruction and pain. The method of targeting EMMPRIN may be a promising strategy for the treatment of cancer in the future.

Purpose: Bone destruction and pain caused by cancer is one of the most devastating complications of cancer patients with bone metastases, and it seriously affects the quality of patients’ life. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule with increased expression in a variety of tumors. This study focused to clarify the specific function of EMMPRIN in bone metastasis of breast cancer. Materials and Methods: Adenovirus with shRNA-EMMPRIN was transfected into MRMT-1 rat breast carcinoma cells, and the MRMT-1 cells with different expression levels of EMMPRIN were implanted into the bone marrow cavity of rat tibia. Next, the effect of down-regulation of EMMPRIN was evaluated as follows: bone damage was detected by X-ray radiological and tartrate-resistant acid phosphatase staining; the tumor burden was evaluated by hematoxylin and eosin staining; the test of pain-related behaviors was assessed used the bilateral paw withdrawal mechanical threshold; and the levels of secretory factors in tumor conditioned medium were determined by using enzyme-linked immunosorbent assay. Results: We found that down-regulation of EMMPRIN in tumor cells can simultaneously reduce tumor burden, relieve cancer-induced bone destruction and pain. Conclusion: Materials and Methods EMMPRIN is expected to be a therapeutic target for relieving bone metastasis of breast cancer and alleviating cancerinduced bone destruction and pain. The method of targeting EMMPRIN may be a promising strategy for the treatment of cancer in the future.