WRKY transcription factors are novel transcriptional regulatory factors, which play an important role in regulating plant development, metabolism and other physiological processes. In this study, a new Dendrobium officinale WRKY transcription factor, designated as DoWRKY1 was cloned by using RT-PCR and RACE (GenBank Accession No. KF953910). Bioinformatic analysis demonstrated that, the full-length cDNA of DoWRKY1 was 1,704 bp. And DoWRKY1 contained a 1,629 bp open reading frame (ORF) that encoding a peptide of 542 amino acid residues. The putative DoWRKY1 protein contained two conserved WRKY domains and it belonged to the group I WRKY family protein. Yeast one-hybrid experiment showed that DoWRKY1 had transcriptional activation ability in yeast, and it could activate the expression of downstream report genes (His3 and Ade2). Semi-quantitative RT-PCR experiment showed that DoWRKY1 expressed in roots, stems, leaves and protocorm-like bodies. Real-time qRT-PCR proved that DoWRKY1 could be induced by methyl jasmonate (MeJA) and chitosan (Chitosan), and the expression level of this gene can reach the expression peak at 2 h and 1 h, respectively. These results are useful for further determination of the regulation function of this gene in secondary metabolism of D. officinale.
Cloning, Molecular ; Dendrobium ; genetics ; Gene Expression Regulation, Plant ; Plant Proteins ; genetics ; Transcription Factors ; genetics

Objiective To evaluate the effect of pearventricular device closure of non-muscular ventricular defercts(VSDs) in infants.Methods From April 2007 to February 2008,39 patients with non-muscular VSDs were received off pump surgical treatment,perventricular device closure were for all the patients.There were 16 males and 23 females with mean age of(14.5±7.8) months(12 to 36 months and mean weight of(12.4±2.3)kg(8.5 to 18.0 kg).There were 34 perimembranous and 5 subateria VSD.The diameter of defects were 3.0 to 11 mm[mean(6.1±2.0)mm].Results Thirty-seven(94.9%)VSDs ere successfully closed,while two were repaired under cardiopulmonary bypass(CPB)instead of device closure because of the complication of moderat aortic reurgitation.The diameter of occluders were 4 to 12mm[mean(8.2±2.0)mm].The tricuspid regurgitations decareasded after operation in 3 patients with perimenmbranous VSDs,while the closure caused new mild or trace tricuspid regurgitations in 8 patients.Six patients with perimembranours VSDs acquired the incomplete right bundle branch blocks affter device closure.The length of hospital stay was 3 to 5 days[mean(3.4±0.4)]after operation and no petient had blood transfusion.Conclustion Perventricular device closure is probably an effective and safe treatment for non-muscular VSDs in unfants.

Adolescent ; Adult ; Blood Transfusion ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Poisoning ; therapy ; Treatment Outcome

OBJECTIVETo investigate the feasibility of using new tracheal prosthesis made of biomaterials to replace extensive circumferential tracheal defects in mongrel dogs.

METHODSThree types of tracheal prostheses were developed, whose basic skeleton of tubular mesh was knitted with polypropylene monofilament and poly (lactic-co-glycolic acid) fiber. The inner side of type-I tubular mesh was first coated with polyurethane solution and then with collagen. The exterior of type-I was then immobilized with collagen-hydroxyapatite composites. In contrast, the internal and external walls of type-II were coated with polyurethane solution, which produced a prosthesis similar to a nonporous one, while type-III was coated only with collagen solution. Surgical resection and replacement of a segment of the cervical trachea was performed in 16 adult mongrel dogs. The efficacy of the implanted prosthesis periodically evaluated postoperatively.

RESULTSIn group A, only one died from prosthetic dehiscence, another from anastomotic leakage, and the others had uneventful postoperative courses. The implanted prosthesis was completely incorporated with the recipient trachea, where different length of reepithelialization occurred on the luminal surface of the reconstructed trachea. Macroscopic examination showed scattered and different sizes of neo-ossification surrounding the implanted prosthesis. The prosthesis was roentgenopaque when exposed to routine X rays. In contrast, a relatively high number of complications occurred postoperatively in group B and C.

CONCLUSIONType-I tracheal prosthesis may be used effectively for long-segment circumferential tracheal replacement, and appears very promising for clinical application, with further improvements in promoting the epithelialization.

Animals ; Biocompatible Materials ; Collagen ; Dogs ; Female ; Male ; Polyglycolic Acid ; Polypropylenes ; Polyurethanes ; Prostheses and Implants ; adverse effects ; Prosthesis Design ; Prosthesis Implantation ; Trachea ; surgery

OBJECTIVETo evaluate diffusion tensor tractography (DTT) of white matter in prognosis of acute lacunar infarctions.

METHODSTwenty-eight patients of pyramidal tract strokes at the acute phase (<3 days) with a marked motor defect were examined. Lesions were identified on diffusion weighted imaging and isotropic imaging of DTI, and infarctions and the tract were shown on the DTT images simultaneously. The anatomic location and pattern of the lesions were visualized on DTT, with regard of the corticospinal tract (CST), all patients were divided into three clinical subgroups: in Group 1 infarction lesions were close to CST, in Group 2 CST was partial involved, in Group 3 lesions centered in the pyramidal tract. Subsequently, they were compared with the National Institutes of Health Stroke Scale (NIHSS) scores at acute phase (<3 days), early chronic phase (8 approximately 14 days), and outcome(30 approximately 60 days).

RESULTNIHSS scores of Group 1(12/28) were not different with those of Group 2 (11/28) at the acute phase (U=-1.430, P>0.05), and NIHSS scores in Group 2 were significantly lower than those of Group 3(5/28) (U= -2.676, P <0.01). In the outcome, NIHSS scores of Group 1 were significantly lower than those of Group 2 (U= -2.501, P<0.05), NIHSS scores of Group 2 were significantly lower than those of Group 3 (U= -2.948, P<0.01). Among these three groups,Group 1 all had good recovery, Group 2 also had good recovery but sometimes with some mild motor disfunction, and Group 3 always had marked defect and minor improvement. Both rADC value and rFA value were induced in the acute lacunar infarctions.

CONCLUSIONDTT is helpful in prognestic valuation of acute lacunar infarction by providing visualized stereo localization of CST and infarction lesions.

Brain ; pathology ; Brain Infarction ; diagnosis ; pathology ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Nerve Fibers, Myelinated ; pathology ; Prognosis

OBJECTIVETo design and develop a novel esophageal prosthesis by selecting appropriate biomaterials, developing special manufacturing techniques, and investigating the feasibility of replacement of cervical esophagus in mongrel dogs.

METHODSIn accordance with the requirements of ideal esophageal substitutes, we designed a new type of esophageal prostheses. The inner stent were made with polyurethane of medical grade, and the outer surface of the prosthesis was coated with collagen-chitosan sponge. The silicone tube was used as a control. Thirteen adult mongrel dogs that were divided into two groups were used to establish the experimental models.

RESULTSIn the experimental group (n = 8), the esophageal prostheses were completely incorporated with the native esophagus and adherent to the surrounding host connective tissues. Epithelial linings of varying degrees were formed on the luminal surface, and complete epithelization was seen in 1 month postoperatively. The granulation at the sites of the anastomosis in this group was less significant than that of the control group. One dog has been surviving for 12 months up to now without any complications. In the control group (n = 5), esophageal epithelial was not observed on the luminal surface, constriction of the regenerated esophagus progressed and all the dogs died within 2 months after operation.

CONCLUSIONThese observations suggest that this esophageal prosthesis made of composite biomaterials has high biocompatibility and potential for long-segment esophageal reconstruction, which is promising for the clinical repair of esophageal defects.

Absorbable Implants ; Animals ; Artificial Organs ; Biocompatible Materials ; Chitosan ; Collagen ; Dogs ; Esophagus ; Implants, Experimental ; Models, Animal ; Polyurethanes ; Prosthesis Design ; methods ; Prosthesis Implantation

To investigate the transgenic expressing efficacy of helper-dependent adenoviral vector (HDAd) in vitro, we constructed a HDAd encoding enhanced green fluorescent protein (EGFP), denominated as HDAd/EGFP, performed large scale preparation and purification, and then identified the purified HDAd/EGFP under fluorescent microscope and electron microscope. After the concentration of HDAd/EGFP was determined by spectrophotometer, the transgenic expression efficiency of HDAd/EGFP was compared with first generation adenoviral vector encoding EGFP (FGAd/EGFP) in vitro. Therefore, we infected A549 cells with 2000 virus particles (vp) per cell by HDAd/EGFP and FGAd/EGFP respectively and analyzed EGFP expressing level by flow cytometry. Consequently, the fluorescent expression rate and fluorescent intensity of EGFP were higher in early infected A549 cells by HDAd/EGFP than by FGAd/EGFP. HDAd, capable of expressing transgene instantly and efficiently in vitro, is a potential vaccine vector.
Adenoviridae ; genetics ; metabolism ; Cell Line, Tumor ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Helper Viruses ; genetics ; metabolism ; Humans ; Transgenes ; Viral Fusion Proteins ; genetics ; metabolism

BACKGROUND: With the development of 3D printing technology, organ and tissue construction can be achieved by constructing a three-dimensional scaffold that is conducive to cell growth. OBJECTIVE: To solve the scaffold over-accumulation during 3D printing.METHODS: Fluent, a finite element analysis software developed by ANSYS Company in the United States, was used to analyze the extrusion process of print heads and to obtain suitable viscosity and extrusion pressure of materials for the 3D printing of cellulose gel composites. We then compared simulation results with experimental results. RESULTS AND CONCLUSION: The error between simulation results and experimental results was less than 5%. The simulated values at a kinetic viscosity of 45 and a pressure of 0.10-0.12 MPa solved the phenomenon of over-accumulation of cellulose gel composites during the 3D printing process, ensuring enough space for the 3D printed scaffold.

Background:Surgical treatment of both-column acetabular fractures is challenging because of the complex acetabular fracture patterns and the curved surface of the acetabulum.Seldom study has compared the application of three-dimensional (3D) printing technology and traditional methods of contouring plates intra-operatively for the surgical treatment of both-column acetabular fractures.We presented the use of both 3D printing technology and a virtual simulation in pre-operative planning for both-column acetabular fractures.We hypothesized that 3D printing technology will assist orthopedic surgeons in shortening the surgical time and improving the clinical outcomes.Methods:Forty patients with both-column acetabular fractures were recruited in the randomized prospective case-control study from September 2013 to September 2017 for this prospective study (No.ChiCTR1900028230).We allocated the patients to two groups using block randomization (3D printing group,n =20;conventional method group,n =20).For the 3D printing group,1:1 scaled pelvic models were created using 3D printing,and the plates were pre-contoured according to the pelvic models.The plates for the conventional method group were contoured during the operation without 3D printed pelvic models.The operation time,instrumentation time,time of intra-operative fluoroscopy,blood loss,number of times the approach was performed,blood transfusion,post-operative fracture reduction quality,hip joint function,and complications were recorded and compared between the two groups.Results:The operation and instrumentation times in the 3D printing group were significantly shorter (130.8 ± 29.2 min,t =-7.5,P ＜ 0.001 and 32.1 ± 9.5 min,t =-6.5,P ＜ 0.001,respectively) than those in the conventional method group.The amount of blood loss and blood transfusion in the 3D printing group were significandy lower (500 [400,800] mL,Mann-Whitney U=74.5,P ＜ 0.001 and 0 [0,400] mL,Mann-Whitney U =59.5,P ＜ 0.001,respectively) than those in the conventional method group.The number of the approach performed in the 3D printing group was significantly smaller than that in the conventional method group (pararectus + Kocher-Langenbeck [K-L] approach rate:35％ vs.85％;X2 =10.4,P ＜ 0.05).The time of intra-operative fluoroscopy in the 3D printing group was significantly shorter than that in the conventional method group (4.2 ± 1.8 vs.7.7 ± 2.6 s;t =-5.0,P ＜ 0.001).The post-operative fracture reduction quality in the 3D printing group was significantly better than that in the conventional method group (good reduction rate:80％ vs.30％;X2 =10.1,P ＜ 0.05).The hip joint function (based on the Harris score 1 year after the operation) in the 3D printing group was significantly better than that in the conventional method group (excellengood rate:75％ vs.30％;x2 =8.1,P ＜ 0.05).The complication was similar in both groups (5.0 ％ vs.25 ％;x2=3.1,P =0.182).Conclusions:The use of a pre-operative virtual simulation and 3D printing technology is a more effective method for treating bothcolumn acetabular fractures.This method can shorten the operation and instrumentation times,reduce Mood loss,blood transfusion and the time of intra-operative fluoroscopy,and improve the post-operative fracture reduction quality.

OBJECTIVETo construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma.

METHODSRestriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR).

RESULTSTwo gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24.

CONCLUSIONThe construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.

Carcinoma, Adenoid Cystic ; enzymology ; genetics ; pathology ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Matrix Metalloproteinase 15 ; genetics ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 7 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; Matrix Metalloproteinases ; genetics ; Neoplasm Metastasis ; Reverse Transcriptase Polymerase Chain Reaction ; methods