Neural cell adhesion molecule (NCAM) is a glycoprotein expressing on the surface of neurons, glial cells, bone cells and natural killer cells. NCAM plays an important role in the process of cell - cell adhesion and cell migration, and is also a model protein to study polysialic acid. In this paper, NCAM gene from mouse mammary gland cells (NMuMG) was cloned into eukaryotic expression vectors pcDNA3.1(+) and transfected into mutant Chinese hamster ovary cells ldlD-14. The stable transfection over-expressing NCAM was obtained through the G418 selection and confirmed by Western blotting. Due to unique characters of ldlD-14 cells, carbohydrate chain of NCAM molecule can be easily manipulated with or without adding galactose in the serum free medium, and this modification can provide the basis for further studies on the effect of glycosylation on NCAM molecular function.
Animals ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Female ; Galactose ; Glycosylation ; Mammary Glands, Animal ; cytology ; Mice ; Neural Cell Adhesion Molecules ; biosynthesis ; Polysaccharides ; chemistry ; Sialic Acids ; chemistry ; Transfection

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

OBJECTIVETo investigate the methods and effects of secondary intervention for mid-long term complications of endovascular repair (EVR) in aortic diseases.

METHODSFrom May 1999 to Jun 2007, 21 patients with mid-long term complications after EVR were treated in our center. Of these cases, 15 cases received first EVR for abdominal aortic aneurysm (AAA), 3 cases for thoracic aortic aneurysm (TAA) and 3 cases for aortic dissection (TAD). The mid-long term complications included 11 cases of type I endoleak, 4 cases of type II endoleak, 2 cases of type III endoleak and 4 cases of migration of stent grafts. Proximal or distal extensions were used for type I and III endoleak in 9 cases. Fenestrated, scallop and bifurcated stent grafts were used to reconstruct the aortic arch in 3 cases. Emboli technique was used in treating type II endoleak. Thrombectomy and bypass technique were used in 4 cases with stent graft limb occlusion. One ruptured AAA accepted open surgery.

RESULTSSecondary endovascular technique were undergone in 20 (95.2%) cases. One case died in 30 days after the secondary intervention and endoleak remained after the secondary operation in 5 cases. Three cases died of the secondary intervention.

CONCLUSIONSEndoleak and limb occlusion were the chief mid-long complications after EVR. Secondary endovascular technique can be used in most cases and carries great challenges in aortic arch lesions.

Adult ; Aged ; Aged, 80 and over ; Aortic Diseases ; surgery ; Blood Vessel Prosthesis Implantation ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; therapy ; Retrospective Studies ; Stents ; adverse effects ; Treatment Outcome

OBJECTIVESTo explorer the change of several signal pathway and their signal after liver transplantation.

METHODSClassified 34 punctured donor liver samples and 10 normal liver samples as A (no rejection) groups, B (mild/moderate acute rejection) groups, C (serious acute rejection) groups, D (chronic rejection/fibrosis) groups and E (control) groups, MAPK, Ras and p53 were performed immunohistochemistry analysis and image analysis. MAPK and Ras were performed in situ hybridizition. Then image analysis was performed.

RESULTSThe protein expression of MAPK, Ras, increase by turns of A, B and C groups (1.42+/-0.28, 3.88+/-0.87, 6.68+/-0.57 in MAPK; 1.27+/-0.12, 2.80+/-0.30, 3.93+/-0.20 in Ras; corresponding), and decrease by turns of D and E groups (1.49+/-0.37, 0.88+/-0.20 in MAPK; 1.47+/-0.21, 1.01+/-0.12 in Ras; corresponding, F=178.39 in MAPK and 320.59 in Ras, groups B, C vs groups A, D, E, P<0.001 in MAPK and Ras), The protein expression of p53 is higher in treated groups (The results of groups A to E are 2.09+/-0.13, 2.39+/-0.11, 2.03+/-0.19, 2.26+/-0.18 and 0.35+/-0.08, corresponding, F=360.08, groups E vs groups A, B, C, D, P<0.001). Expression of MAPK, Ras mRNA is as same as that of protein.

CONCLUSIONThe MAPKs pathway has role in rejection response after liver transplantation. And it seemed that the MAPKs and p53 are one regulation mechanism for protecting the hepatocyte from damage after liver transplantation.

Humans ; Immunohistochemistry ; In Situ Hybridization ; Liver Transplantation ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinases ; analysis ; Signal Transduction ; physiology ; Tumor Suppressor Protein p53 ; analysis ; ras Proteins ; analysis

OBJECTIVETo prevent and manage frequent complications after endovascular repair of infrarenal abdominal aortic aneurysm (AAA).

METHODSThe data of 71 cases of infrarenal abdominal aortic aneurysm (AAA) treated by endovascular repair were analysed retrospectively. The reasons, managements, results and prognosis of frequent complications were investigated.

RESULTSSeventy-one cases of infrarenal AAA were treated by endovascular repair with 100% success rate. There was no surgical conversion to open aneurysm repair. There were 8 cases of primary endoleak, 1 case of nervous complication and acute thrombosis. An average follow-up period was 26 +/- 5 months. Three persistent endoleaks and 4 secondary endoleaks were found during the follow-up period. The endoleak rate was 9.8% (7/71) within 1 month postoperatively and mortality rate was 1.3% (1/71). Total mortality rate was 4.2% (3/71). Two patients died from acute myocardial infarction and one from acute heart failure.

CONCLUSIONSEndovascular treatment of abdominal aortic aneurysm is technically feasible and can effectively exclude aortic aneurysms from the circulation. Endoleak is a chief complication after endovascular repair of infrarenal AAA.Additional procedures and follow up are very important. Endoleak with enlarged aneurysm should be treated actively.

Aged ; Aged, 80 and over ; Aortic Aneurysm, Abdominal ; surgery ; Blood Vessel Prosthesis Implantation ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; therapy ; Prognosis ; Retrospective Studies ; Stents ; Treatment Outcome ; Vascular Fistula ; etiology ; prevention & control ; therapy

OBJECTIVETo discuss the change of blood supply pattern in visceral arteries of Stanford B dissection. The visceral arteries include celiac trunk (CA), superior mesenteric artery (SMA) and renal artery (RA).

METHODSBy retrospectively analysing the clinical data of 52 cases with Stanford B dissection, the blood supply pattern of visceral arteries was confirmed by aortography and the changes before and after endovascular repair were compared.

RESULTSAfter repair: the stenosis lesions disappeared in 7 cases supported by true channel completely but one. Twenty-two visceral arteries supported by true and false channel simultaneously recovered true channel chiefly but one. One recovered true channel chiefly and one had no change in 2 visceral arteries supported by false channel completely. Four recovered true channel chiefly and one had no change in 5 visceral arteries without blood support. 88.9% blood support got better and 11.1% blood support had no change in 36 damaged visceral arteries after endovascular repair.

CONCLUSIONBlood support from true and false channel simultaneously is the chief pattern in the injured visceral arteries before repair; Endovascular repair technique is benefit to recovering the blood support of true channel.

Adult ; Aged ; Aneurysm, Dissecting ; physiopathology ; surgery ; Aortic Aneurysm ; physiopathology ; surgery ; Celiac Artery ; physiopathology ; Female ; Humans ; Male ; Mesenteric Artery, Superior ; physiopathology ; Middle Aged ; Regional Blood Flow ; Renal Artery ; physiopathology ; Retrospective Studies

To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.
ABO Blood-Group System ; genetics ; China ; DNA Mutational Analysis ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Phenotype ; Sequence Deletion

OBJECTIVETo investigate the resistance genes and antibiotic resistance patterns against beta-lactams in Pseudomonas aeruginosa prevalent in burn ward.

METHODSK-B method was performed to test bacterial resistance patterns against 9 species of beta-lactams in Pseudomonas aeruginosa isolated from wounds and dressings of the patient in burn wards. Seven species of resistance genes against beta-lactams were detected with PCR. Tazobactam-inhibited piperacillin resistance test was performed to study whether the above strains produce extended spectrum beta-lactams.

RESULTSAll 12 strains of bacteria with resistance genes detected were resistant to penicillin and cephalosporins (100%), among them 11 were resistant to all antibiotics. Tazobactam-inhibited piperacillin resistance test demonstrated that all strains with resistance genes were ESBLs.

CONCLUSIONHigh incidence of beta-lactams resistance genes is found in Pseudomonas aeruginosa isolated from burn ward, and they have close relationship with the occurrence of multiple drug-resistance.

Burn Units ; Burns ; microbiology ; Genes, Bacterial ; Humans ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification ; beta-Lactam Resistance ; genetics

BACKGROUND: Posterior lumbar interbody fusion is the main repair method for senile degenerative lumbar spondylolisthesis. For elderly patients with osteoporosis, single screw rod system fixation regularly results in dislocation. The incidence of implant loosening is high, and therefore, effective internal fixation is not achieved. OBJECTIVE: This study will use the modified posterior lumbar interbody fusion procedure combined with bone cement augmentation for the treatment of senile degenerative lumbar spondylolisthesis so as to increase the stability of the vertebral body, make the fixator firm, and to maximize the recovery of postoperative motor function. METHODS: Totally 113 patients with senile degenerative lumbar spondylolisthesis, aged 65-70 years, irrespective of sex, will be recruited from the Department of Orthopedics of Affiliated Hospital of Qinghai University of China. The patients will be treated with modified posterior lumbar interbody fusion combined with bone cement augmentation. Follow-up will be performed at 3 and 12 months. The primary outcome measure is recovery of motor function as indicated by the postoperative Oswestry Disability Index score at 12 months. The secondary outcome measures are the change in the rate of excellent and good Oswestry Disability Index scores (comparing preoperative scores with the 4-month postoperative scores), Visual Analogue Scale scores, intervertebral space height, foraminal height, the preoperative and 4 and 14 months postoperative slip distance and slip angle, incidence of adverse reactions, success rate of vertebral fusion, and incidence of secondary slip 4 and 14 months postoperatively. This trial has been approved by the Medical Ethics Committee of Affiliated Hospital of Qinghai University of China(approval number:QHY023G).The study protocol will be proformed in accordance with the Declaration of Helsinki, formulated by the World Medical Association. Written informed consent will be obtained from all participants. This trial was designed in October 2017. The recruitment of subjects and data collection will begin in June 2018. The recruitment will be finished in December 2018. Outcome measures will be analyzed in January 2020. This trial will be completed in February 2020. The results of the trial will be reported in a scientific conference or disseminated in a peer-reviewed journal. This trial had been registered in the Chinese Clinical Trial Registry (registration number: ChiCTR1800015335). Protocol version (1.0). DISCUSSION: This trial aims to observe the efficacy of modified posterior lumbar interbody fusion combined with bone cement augmentation in the treatment of senile degenerative lumbar spondylolisthesis and to validate whether the procedure is safe and reliable.