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Objective To explore the change rules of blood ethanol and blood acetaldehyde concentration, the impairment of psychomotor functions of different acetaldehyde dehydrogenase （ALDH） 2 genotype individuals after alcohol consumption and the relationship among them. Methods The ALDH2 genotypes in seventy-nine healthy volunteers were obtained by SNaPshotTM method, then divided into ALDH2*1/*1 （wild type） and ALDH2*1/*2 （mutant type） group. After volunteers consumed 1.0 g/kg of alcohol, blood ethanol concentration and blood acetaldehyde concentration at a series of time points before and after alcohol consumption and psychomotor functions, such as, visual selective response time, auditory simple response time and tracking experiment were detected. Biphasic alcohol response questionnaires were collected. Results After alcohol consumption, ALDH2*1/*2 group's blood ethanol and blood acetaldehyde concentration reached the peak earlier than ALDH2*1/*1 group. Its blood acetaldehyde concentration was higher than that of ALDH2*1/*1 group, 1-6 h after alcohol consumption. The psychomotor functions, such as visual selective response time and auditory simple response time in ALDH2*1/*2 group were more significantly impaired than those in ALDH2*1/*1 group after alcohol consumption. There was no statistical significance between the two groups in excitement or sedation reactions （P>0.05）. Pearson correlation coefficient test showed that blood acetaldehyde concentration was related with psychomotor function. Conclusion There are significant differences between the psychomotor function of ALDH2 wild type and mutant type individuals after alcohol consumption estimated to be related to the difference in blood acetaldehyde concentration after alcohol consumption.
Subject(s)
Humans , Acetaldehyde/metabolism , Alcohol Drinking/blood , Aldehyde Dehydrogenase/genetics , Aldehyde Dehydrogenase, Mitochondrial , Aldehyde Oxidoreductases , Ethanol/metabolism , Genotype , Polymorphism, Genetic/genetics , Psychomotor Performance/physiologyABSTRACT
OBJECTIVE To study the methodology of achieving stable co-expression of drug-metab?olizing enzymes in the HepG2 cells by the piggyBac (PB) transposon system. METHODS N-terminal attachment of enhanced green fluorscent protein plasmid (pEGFP- N2) and 2A peptide linked recombinant PB transposon plasmid containing dual-genes encoding drug metabolizing enzymes cyto?chrome P450 3A4 (CYP3A4) and CYP2C19 (pPB-CYP3A4-2A-2C19) were transfected into HepG2 cells respectively by Lipofectamine?LTX reagent, GenJetTM (Ver.Ⅱ) reagent and Neon?Transfection System reagent, which were widely used for large-sized DNA fragments transfection. 48 h later, the transfection efficiency and cell toxicity were detected and compared between the three methods so as to find a method with relatively high efficiency and low toxicity for later transfection.Then,three groups of recombinant PB transposons-single-gene transposon (PB-CYP3A4), 2A peptide linked dual-gene transposon (PB-CYP3A4-2A-2C19) and multiple single-gene transposon mixture〔PB-CYP3A4, PB-CYP2C8, PB-CYP2A6, organic anion transporting polypeptide 1B1 PB transposon (PB-OATP1B1)〕-were transfected into HepG2 cells respectively with the above established method.The puromycin (Puro)-resistant and GFP positive cell clones were picked up and further cultured. The mRNA, protein and metabolic levels of drug-metabolizing enzymes in monoclonal cell lines were detected by quantitative real-time PCR,Western blotting and high performance liquid chromatography-tandem mass spectrometry respectively after screening by Puro and green fluorescence. Comparisons of different groups were made using statistical analysis. RESULTS The comparison of three different transfection methods indi?cated that the transfection efficiency of GenJetTMwas up to(94.2±2.5)% and (89.3±3.3)%,significantly higher than those of the other two methods (P<0.01), along with lower cytotoxicity. Then GenJetTMwas chosen for later transfection. In the Puro-resistant monoclonal cell lines of single transposon PB-CYP3A4,PB-CYP3A4-2A-2C19 groups,the mRNA,protein and enzyme activity levels of drug-metabo?lizing enzymes were significantly increased respectively.The recombinant transposon (PB-CYP3A4-2A-2C19) containing 2A peptide could achieve stable and efficient co-expression of two metabolizing enzymes CYP3A4 and CYP2C19,while the expression of drug-metabolizing enzymes remained unbal?anced and random in those of multiple single-gene transposon mixture group (PB-CYP3A4, PB-CYP2C8,PB-CYP2A6,PB-OATP 1B1)(CYP3A4 was expressed in some cell clones only).CONCLUSION GenJetTM could be an effective method for the PB recombinant transposon transfection into HepG2 cells, by which the PB transposon could mediate stable expression of drug-metabolizing enzymes. In terms of multi-gene expression,a low and unbalanced expression is found by multiple transposons co-transfection method,which is different from that by virus mediated method.In contrast,mono-PB trans?poson linked by 2A peptide can achieve stable expression of multi-genes.
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Objective·To explore the value of applying CT/MRI image registration and fusion combined with 3D printing technique in pre-surgical planning of refractory pelvic tumors. Methods·A retrospective analysis was performed on the cases with refractory pelvic tumors admitted between March 2014 and December 2016 in Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. CT/MRI examinations were performed as a routine pre-surgical procedure, and the resulting images were subject to rapid registration and fusion with the Medraw software in order to accurately determine the tumor boundaries and to be used as references in designing and making patient-specific tumor models, prostheses, and implants by 3D printing technique. Results·A total of 13 cases with refractory pelvic tumors were included in this study (8 cases of chondrosarcoma, 1 case of osteosarcoma with aneurysmal bone cyst, 1 case of dedifferentiated sarcoma, 1 case of fibrous connective tissue hyperplasia, 1 case of solitary fibrous tumor/hemangiopericytoma, and 1 case of metastatic squamous cell carcinoma) with an average age of (50.0±8.5) years. According to the Enneking classification,there were 2 cases in zone Ⅰ , 2 in zones Ⅰ + Ⅱ , 1 in zones Ⅱ + Ⅲ , 3 in zones Ⅰ + Ⅱ + Ⅲ , 1 in zones Ⅰ + Ⅱ + Ⅳ , 1 in zonesⅠ + Ⅱ + Ⅲ + Ⅳ , 1 in zones Ⅰ + Ⅳ , 1 in zones Ⅱ + Ⅳ , and 1 in zone Ⅳ. The mean maximal tumor diameter was (15.15±4.81) cm without implicating the pelvic blood vessels. Results from intraoperative frozen section at resection boundaries and the installation of prosthesis showed that the actual surgical procedure was consistent with the surgical planning based on the reported image fusion technique. Conclusion·CT/MRI image registration and fusion technique combined with 3D printing technique can accurately determine the resection boundary of pelvic tumors and assist in designing of individualized prosthesis model and surgical guide, which has a great value in pre-surgical planning and clinical treatment of refractory pelvic tumors.
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Objective·To explore the value of applying CT/MRI image registration and fusion combined with 3D printing technique in pre-surgical planning of refractory pelvic tumors. Methods·A retrospective analysis was performed on the cases with refractory pelvic tumors admitted between March 2014 and December 2016 in Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. CT/MRI examinations were performed as a routine pre-surgical procedure, and the resulting images were subject to rapid registration and fusion with the Medraw software in order to accurately determine the tumor boundaries and to be used as references in designing and making patient-specific tumor models, prostheses, and implants by 3D printing technique. Results·A total of 13 cases with refractory pelvic tumors were included in this study (8 cases of chondrosarcoma, 1 case of osteosarcoma with aneurysmal bone cyst, 1 case of dedifferentiated sarcoma, 1 case of fibrous connective tissue hyperplasia, 1 case of solitary fibrous tumor/hemangiopericytoma, and 1 case of metastatic squamous cell carcinoma) with an average age of (50.0±8.5) years. According to the Enneking classification,there were 2 cases in zone Ⅰ , 2 in zones Ⅰ + Ⅱ , 1 in zones Ⅱ + Ⅲ , 3 in zones Ⅰ + Ⅱ + Ⅲ , 1 in zones Ⅰ + Ⅱ + Ⅳ , 1 in zonesⅠ + Ⅱ + Ⅲ + Ⅳ , 1 in zones Ⅰ + Ⅳ , 1 in zones Ⅱ + Ⅳ , and 1 in zone Ⅳ. The mean maximal tumor diameter was (15.15±4.81) cm without implicating the pelvic blood vessels. Results from intraoperative frozen section at resection boundaries and the installation of prosthesis showed that the actual surgical procedure was consistent with the surgical planning based on the reported image fusion technique. Conclusion·CT/MRI image registration and fusion technique combined with 3D printing technique can accurately determine the resection boundary of pelvic tumors and assist in designing of individualized prosthesis model and surgical guide, which has a great value in pre-surgical planning and clinical treatment of refractory pelvic tumors.
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GATA6 transcription factor belongs to the GATA family and contains 2 conserved zinc ifnger DNA binding domains. GATA6 not only presents in embryonic tissues but also found in heart, lung and pancreas and is essential for the maintenance of their function.The present review focuses on the critical roles of GATA6 in heart development and atrial septal defect to provide theoretical basis for diagnosis and treatment of atrial septal defect.
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ObjectiveTo identify mutations ofGATA4 andGATA6 genes in children with isolated congenital atrial septal defect (ASD).Methods From November 2012 to November 2013, 101 patients with ASD (99 unrelated patients and one twin) who were submitted to catheter-based intervention and 100 ethnicity-matched children without congenital heart disease, blood disorders and chromosomal abnormalities were enrolled. The blood was collected. The coding regions and lfanking regions of theGATA4 andGATA6 genes were ampliifed by polymerase chain reaction and sequenced using the dideoxvnucleotide chain termination technique, and then compared with the normal sequence in the Genbank.Results Two novel heterozygous missense GATA6mutations, c. G145A and c. G151A, were identiifed in 2 unrelated ASD patients, which were not present in the controls. These two mutations predicted the conversion of glycine into serine at amino acid residue 49 (G49S) and glutamate into lysine at amino acid residue 52 (K52E). A heterozygous missenseGATA6 mutation c.43 G>C, which caused a conversion from glycine to arginine, was found in 9 ASD patients and 7 controls. A single nucleotide polymorphism c.99G>T, which did not cause amino acid conversion inGATA4 gene, was found.ConclusionsGATA6 gene is an important transcription factor in heart development. The mutation ofGATA6 gene may cause the change of its transcriptional activity, and lead to ASD.
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Objective To observe the clinical effects of coronoid process of ulna osteotomy and bilateral anatomic locking plate on humerus condyle comminuted fracture in the elderly.Methods 17 elderly patients with humerus condyle comminuted fracture were collected in department of orthopaedics in our hospital from July 2005 to December 2013 and graded into C2 (6 cases) and C3 (11 cases) according to the standard of the American society for the prevention of bone trauma.All patients were treated by coronoid process of ulna osteotomy and bilateral anatomic locking plate and accessed by Mayo elbow performance score (MEPS) at the last follow-up.Results Postoperative incision in all patients were healed,and X-ray showed fracture healing occurred at 10 to 24 weeks after coronoid process of ulna osteotomy (average of 14 weeks).No iatrogenic ulnar nerve injury and nonunion were found at the follow-up.The MEPS were excellent in 13 cases,well in 3 cases,ordinary in 1 case,and the excellent and good rate was 94.1％.Conclusions Coronoid process of ulna osteotomy and bilateral anatomic locking plate in the treatment of humerus condyle comminuted fracture in the elderly have good clinical effects with many advantages such as exposure clearly,early elbow joint function exercise and avoiding the aggravation of elbow joint osteoarthritis.
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HuaFu Shengji is the primary traditional Chinese medicine (TCM) therapy for treating chronic skin ulcer. The high activities of the protein enzyme in the wound fluids is one of the main cause of healing delay. In order to investigate the effect of TCM Zhuhong ointment for promoting wound healing. This research focused on its influence on matrix metalloproteinase (MMP) activities in wound fluids with TCM Yang syndromes, directly on the activated MMP-1,2 activities in vitro and on MMP-1,-2,-9 production by HSF. 8 wound fluid samples were collected, which were diagnosed Yang Syndromes in TCM. Wound fluid activities of MMP-2 and MMP-9 were measured by gelatin zymogram assay. MMP-1 and MMP-2 activities in vitro were measured by substrate cleavage. CCK-8 was used to observe the toxicity of Zhuhong ointment on HSF. MMP-1,-2,-9 production by HSF were detected by confocal microscope. Zhuhong ointment from 1 to 25 g x L(-1) obviously inhibited MMP-2 activity in wound fluid. When Zhuhong ointment was over 5 g x L(-1), it showed significantly inhibitory effect on wound fluid MMP-9 activity. In vitro study, when the mercury concentration was 320 mg x L(-1), Zhuhong ointment solution directly inhibited both MMP-1 activity and MMP-2. But mercury concentration from 0.51-2.56 mg x L(-1), it could activate MMP-1 activity, and from 0.51-64 mg x L(-1), activate MMP-2 activity instead. The mercury concentration when Zhuhong ointment saturated in DMEM was 39.6 mg x L(-1). When the mercury concentration was over 1.23 mg x L(-1), Zhuhong ointment showed toxicity to HSF. At 1.23, 0.62, 0.31 mg x L(-1) of mercury concentration, it increased MMP-1 expression by HSF, and at 1.23, 0.62 mg x L(-1), decreased MMP-2 expression. However, at 1.23, 0.62, 0.31 mg x L(-1), it decreased MMP-9 expression. At higher concentration, Zhuhong ointment can inhibit MMP-2, MMP-9 activities in wound fluid with dose-dependent way and show a direct inhibitory effect on activated MMP-1 and MMP-2 in vitro. But at a lower concentration, it showed two-way adjustment, with increased MMP-1, MMP-2 activities and its expression by HSF and decreased MMP-9 activity.
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Humans , Body Fluids , Cells, Cultured , Dermatitis , Drug Therapy , Drugs, Chinese Herbal , Pharmacology , Fibroblasts , Physiology , Matrix Metalloproteinase 1 , Metabolism , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Wound HealingABSTRACT
<p><b>BACKGROUND</b>The elevated matrix metalloproteinase (MMP) activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide (As2O3), is a common traditional Chinese medicine wildly used in treating chronic wounds; it can remove necrotic tissue and promote tissue regeneration. This research was designed to evaluate the effects of As2O3 on production and activities of MMP-1, MMP-2 and MMP-9, and on regulation of its signal transduction pathway in human skin fibroblasts (HSFb) and human monocyte line (THP-1 cells) that were in an inflammatory state.</p><p><b>METHODS</b>We established three cell models; HSFb activated by TNF-α, THP-1 cells activated by phorbol 12-myristate 13-acetate (PMA) and an HSFb-THP-1 co-culture system. Three cell models was cultured with As2O3 for 24 hours. The levels of MMP-1, MMP-2, MMP-9, TNF-α and IL-1β in the cell culture supernatants were assayed by ELISA. The mRNA expressions of MMP-1, MMP-2 and MMP-9 were determined by RT-PCR. The activities of MMP-2 and MMP-9 were tested by Gelatin zymography assays. The phosphorylation levels of ERK1/2 and p38MAPK were assayed by Western blotting.</p><p><b>RESULTS</b>As2O3 inhibited the expression of MMP-1, MMP-2 and MMP-9 mRNA, the secretion and activity of MMP-1, MMP-2 and MMP-9 in HSFb and THP-1 cells in the inflammatory state (P < 0.05 and P < 0.01 respectively). It also inhibited the secretion of TNF-α and IL-1β in THP-1 cells and in the co-culture system (P < 0.05 and P < 0.01, respectively). It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFb and THP-1 cells (P < 0.05 and P < 0.01, respectively).</p><p><b>CONCLUSIONS</b>As2O3, as a main component of arsenolite, can inhibit the production of MMPs by HSFb and THP-1 cells in an inflammatory state through inhibiting the release of inflammatory factors and the activation of the MAPK cascade pathway. This may be a possible mechanism for arsenolite healing chronic wounds.</p>
Subject(s)
Humans , Arsenicals , Pharmacology , Cell Line , Cells, Cultured , Electrophoresis, Polyacrylamide Gel , Fibroblasts , Interleukin-1 , Metabolism , Matrix Metalloproteinase 1 , Metabolism , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Monocytes , Oxides , Pharmacology , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To summarize our experience in adult-to-infant living donor liver transplantation (A-ILDLT) and to analyze the efficacy and complications of A-ILDLT.</p><p><b>METHODS</b>The clinical data, surgical strategies and complications of 28 adult donors and infantile recipients who underwent A-ILDLT from April 2006 to December 2009 were retrospectively analyzed. These 28 patients (14 boys and 14 girls) aged from 80 days to 11.5 months with body weights of 3.08 to 10.3 kg at the time of operation . They suffered from biliary atresia with decompensated cirrhosis. The living donors were 15 mothers, 9 fathers, 3 grandma and 1 elder brother with ABO compatible with the infantile recipients. 27 Donor organs were the left lateral lobe grafts (segment II, III) and 1 graft was segment II. All patients were followed up for 5 to 24 months.</p><p><b>RESULTS</b>These grafts were orthotopically transplanted into the infantile recipients. The average length of stay was 9.3 days for the donor group without any complications. Postoperative immunosuppression included prednisone, Cyclosporin and mycophenolate mofetil (MMF). A total of 24 postoperative complications occurred in 20 recipients, including 5 vascular complications, 4 bleeding, 7 pneumonia, 2 bowel obstruction, 4 intestinal perforation and 3 rejection. Three recipients died of hepatic arterial thrombosis (HAT). The perioperative mortality rate of recipients was 10.7% (3/28) and the survival rate was 89.3% in peroperative period. One died of stricture of hepatic vein and 1 of accidental asphyxia during follow-up term. At present, 23 cases are still alive.</p><p><b>CONCLUSION</b>A-ILDLT has become an effective method to infants with end-stage liver disease. The postoperative vascular complication is the predominant cause of death.</p>
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Female , Humans , Infant , Male , Liver Diseases , General Surgery , Liver Transplantation , Methods , Living Donors , Retrospective Studies , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of Gamma knife surgery (GKS) in treating cavernous malformation (CM).</p><p><b>METHODS</b>From 1994 to 2001, 92 patients with 114 CMs were treated by GKS and then followed up for 2-8 years (mean 4.1+/-1.9). We analyzed the MRI features of CMs bleeding, efficacy of GKS, and the complications of treatment. Six pathological specimens after radiosurgery (1 from our group, 5 from other centers) were also assayed.</p><p><b>RESULTS</b>Among 43 patients who were treated by GKS to control their epilepsy, epileptic paroxysm was alleviated in 36 patients (83.7%), including 12 (27.9%) seizure-free. Rebleeding was confirmed in 9 patients (9.8%) by neuroimage, one of whom died. Transient symptomatic radiation edema occurred in 7 cases (7.6%) within 6-12 months after radiosurgery, and one patient underwent open surgery for cerebral decompression. The main pathological changes of cavernoma were coagulation necrosis and the vessels obliterated gradually after radiosurgery.</p><p><b>CONCLUSIONS</b>It is feasible to treat small and surgically high risk CMs by radiosurgery. The treatment has to be prudent in an acute bleeding and symptomatic progression. Optimal treatment timing and dose planning are prerequisites to reduce radiation-related complications. GKS is safe and effective to control the epilepsy caused by CMs, and also to bring down the rebleeding rate after a latency interval of several years.</p>
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Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Brain Neoplasms , Diagnosis , Pathology , General Surgery , Cerebral Hemorrhage , Diagnosis , General Surgery , Follow-Up Studies , Hemangioma, Cavernous, Central Nervous System , Diagnosis , Pathology , General Surgery , Magnetic Resonance Imaging , Radiosurgery , Retrospective StudiesABSTRACT
Objective To summary the clinical applied value of interventional therapy in hepatic venous outflow obstruction after orthotopic liver transplantation(OLT).Methods The clinical data of 27 patients suspect with hepatic outflow obstruction out of OLT patients were analyzed retrospec- tively.Most of them presented with liver dysfunction,like ascites,jaundice,or hydropsia of lower ex- tremity as BCS.These patients accepted venography and endovascular treatment if venous outflow ob- struction was found.Results By venography,one case of thrombus in inferior vena cava,one case of hepatic vein stenosis,13 cases of inferior vena cava stenosis(3 cases were associated with hepatic vein stenoses)were identified.Stent implantation was successfully performed on 10 patients,and balloon angioplasty on 4 patients.Rapid,dramatic resolution of symptoms was achieved in those patients. Hepatic vein restenosis occurred in one case 8 months after balloon dilatation,and treated with stent implantation.Inferior vena cava restenosis occurred in one case 2 years after balloon dilation,and trea- ted with another balloon expanding.Patients remained completely asymptomatic at 4 months to 5 years of follow-up.Conclusion The venacavographic balloon angioplasty and metallic stent replacement are safe and useful for post-OLT with venous outflow obstruction.