Adolescent ; Adult ; Antigens, Nuclear ; metabolism ; Brain Diseases ; complications ; diagnosis ; pathology ; surgery ; Brain Neoplasms ; pathology ; Child ; Child, Preschool ; Diagnosis, Differential ; Epilepsy ; etiology ; Female ; Ganglioglioma ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Malformations of Cortical Development ; classification ; complications ; diagnosis ; pathology ; surgery ; Malformations of Cortical Development, Group I ; Microtubule-Associated Proteins ; metabolism ; Neoplasms, Neuroepithelial ; pathology ; Nerve Tissue Proteins ; metabolism ; Neurofilament Proteins ; metabolism ; Retrospective Studies ; Vimentin ; metabolism ; Young Adult

OBJECTIVETo construct an eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and enhanced green fluorescent protein (EGFP) controlled by cytomegalovirus promoter using pIRES-EGFP vector.

METHODSThe mtHSP70 gene fragment was amplified by PCR from pVAX-mtHSP70-HSV2gD using specific primers. The PCR product was cloned into the vector pMD 18-T vector, and the correct clone was selected according to DNA sequence analysis. The interested mtHSP70 gene fragment was subcloned into pCMV-IRES-EGFP vector with XhoI and EcoR I digestion. The recombinant plasmid was transfected into mouse melanoma B16 cell line, and the green fluorescent cells were detected by fluorescence microscopy and mtHSP70 expression was detected by Western blotting.

RESULTSThe recombinant plasmid obtained was confirmed by enzyme digestion. The transfected mouse melanoma B16 cells exhibited green fluorescence under fluorescence microscopy and expressed mtHSP70 protein as demonstrated by Western blotting.

CONCLUSIONThe eukaryotic coexpression vector PCMV-mtHSP70-IRES-EGFP has been established to allow further investigation of the role of mtHSP70 gene in tumor immunotherapy.

Animals ; Base Sequence ; Cancer Vaccines ; Cell Line, Tumor ; Cytomegalovirus ; genetics ; metabolism ; Genetic Vectors ; biosynthesis ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Mice ; Molecular Sequence Data ; Mycobacterium tuberculosis ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA

Aged ; Coronary Angiography ; Coronary Restenosis ; diagnostic imaging ; etiology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Sirolimus ; administration & dosage ; Stents

OBJECTIVETo study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs.

METHODSHUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry.

RESULTSHUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%.

CONCLUSIONHighly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.

Cells, Cultured ; EGF Family of Proteins ; Endothelial Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Immunoglobulins ; TYK2 Kinase ; Umbilical Veins

OBJECTIVETo evaluate the effectiveness and safety of sirolimus-eluting stents (SESs) for treatment of in-stent restenosis (ISR).

METHODSAll 27 patients with ISR and clinical evidence of ischemia had been treated with SESs. Among them, 23 patients had diffuse and complex lesions, and 5 of them received 2 SESs. Clinical and angiographic follow-up were performed for all patients and the results were analyzed.

RESULTSAll stents were implanted successfully. There were no remained stenosis and major in-hospital complications. Average follow-up time was 8.9 +/- 2.1 (5-14) months, with a clinical follow-up rate of 96.3% and angiographic follow-up rate of 92.6%. During the follow-up, there was none of death. One patient had recurrent angina with an angiographic evidence of the proximal edge restenosis of the stent. Mild neointimal hyperplasia in the proximal edge was found in 2 patients, but the stenosis was less than 25%. No late lumen loss was found in other 24 patients. The late lumen loss of the in-stent averaged 0.09 +/- 0.02 mm, and of the distal edge vessel averaged 0.10 +/- 0.03 mm, and of the proximal edge vessel averaged 0.20 +/- 0.06 mm. The rate of target vessel revascularization was 3.8%.

CONCLUSIONThe SES implantation is safe and feasible for the treatment of in-stent restenosis, which could effectively prevent neointimal hyperplasia and recurrent restenosis of the lesion.

Adult ; Aged ; Coronary Angiography ; Coronary Restenosis ; diagnostic imaging ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Sirolimus ; administration & dosage ; Stents

OBJECTIVETo clone novel gene from suppression subtraction library established for screening down-regulated genes in gastric carcinoma, and the effects of novel gene on gastric tumorigenicity were analyzed.

METHODSSequencing results of 860 positive colonies chosen randomly were compared by Blast program in GenBank. Novel gene fragment was amplified by rapid amplification of cDNA ends (RACE). The mRNA expression of novel gene was detected by Northern blot and semi-quantitative PCR in 25 cases of gastric carcinoma tissue and counterpart normal gastric mucosa. The structure and chromosomal location of novel gene were investigated by Bio-message technique.

RESULTSA 233 bp novel gene fragment was screened out from 860 clones and a 802 bp novel gene was obtained by RACE. The novel gene was named as GDDM, registered in the number of AF494508 by GenBank. The mRNA expression of GDDM in gastric carcinoma tissue (4.496+/-0.637) was significantly lower than that in the counterpart normal gastric mucosa (36.919+/-6.290)(P<0.01). Chromosomal location of GDDM gene was at 4q31.

CONCLUSIONThe cloned novel gene, GDDM, is down-regulated in gastric carcinoma, and it is likely to be involved in gastric tumorigenicity.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Down-Regulation ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Gene Library ; Genes, Neoplasm ; Humans ; Molecular Sequence Data ; Stomach Neoplasms ; genetics ; metabolism

BACKGROUNDBlood transfusion has been found to be a devastating factor for outcomes of hepatectomy. This study was to assess the value of major hepatectomy without blood transfusion.

METHODSWe retrospectively studied 51 patients who had undergone major hepatectomy without blood transfusion, including 29 patients with primary liver cancer, from August 1997 to December 2000. Sixty patients undergoing major hepatectomy with blood transfusion including 48 patients with primary liver cancer served as controls. Hepatectomy was performed through normothermic interruption of the porta hepatis. Intraoperative ultrasonography was performed to define tumor margins, and an ultrasound dissector was used to dissect liver parenchyma.

RESULTSIn the study group, the operative mortality and morbidity and 1-, 2-, and 3-year recurrence rates were 0%, 9.8%, 24.1%, 27.6% and 31.0%, respectively. In the control group, they were 3.3%, 28.3%, 43.5%, 54.3% and 58.7%, respectively. Significant differences were seen in morbidity and recurrence rates of patients with liver cancer between the two groups (P < 0.05).

CONCLUSIONMajor hepatectomy without blood transfusion can reduce postoperative morbidity and recurrence rate of patients with liver cancer.

Adult ; Aged ; Blood Transfusion ; Female ; Hepatectomy ; methods ; mortality ; Humans ; Liver Neoplasms ; mortality ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Postoperative Complications ; prevention & control

A boy, aged 2 years and 5 months, had recurrent epistaxis, and the coagulation function examination showed that activated partial thromboplastin time (APTT) was significantly prolonged. Further laboratory examinations showed that the prolonged APTT was not immediately corrected in the APTT correction test, with positive lupus anticoagulant and low prothrombin activity. The boy was diagnosed with hypoprothrombinemia-lupus anticoagulant syndrome. The condition was improved after treatment with glucocorticoid, immunoglobulin, and vitamin K₁. The boy has been followed up for 6 months, and no epistaxis was observed. Prothrombin activity returned to normal, and lupus anticoagulant remained positive. This is a relatively rare disease, and for patients with bleeding symptoms and coagulation disorders, it is recommended to perform the tests such as APTT correction test, lupus anticoagulant testing, and coagulation factor dilution test, which can improve the detection rate of this disease, so as to achieve early diagnosis, provide rational treatment in the early stage, and improve the prognosis.
Antiphospholipid Syndrome/diagnosis* ; Blood Coagulation Disorders ; Child, Preschool ; Epistaxis/etiology* ; Humans ; Hypoprothrombinemias/diagnosis* ; Lupus Coagulation Inhibitor ; Male ; Partial Thromboplastin Time ; Prothrombin

OBJECTIVETo evaluate the protective effect of S-isopentenyl-L-cysteine,a new cysteine derivative,on DNA damage induced by radiation by using acute radiation injury animal models.

METHODSForty ICR mice were randomly divided into five groups:the control group,1.0Gy gamma irradiation group,1.0Gy gamma irradiation combined with S-isopentenyl-L-cysteine group,7.2Gy gamma irradiation group,and 7.2Gy gamma irradiation combined with S-isopentenyl-L-cysteine group,with 8 mice in each group.The comet assay and bone marrow polychromatic micronucleus experiments were performed to evaluate the double-strand DNA breaks in ICR mice exposed to 1.0 and 7.2Gy gamma-ray, respectively.

RESULTSThe tail DNA percentage,tail length,tail moment,and olive tail moment of peripheral blood lymphocytes in 7.2Gy gamma irradiation group were significantly higher than that of the control group (P<0.01).And it was also observed that above experimental indexes of 7.2Gy gamma irradiation combined with S-isopentenyl-L-cysteine group was significantly less than that of 7.2Gy gamma irradiation group (P<0.05). In addition,the micronucleus rate of 1.0Gy gamma irradiation group and 7.2Gy gamma irradiation group were both significantly higher than in the control group (P<0.01). In addition,in mice given S-isopentenyl-L-cysteine before irradiation,the micronucleus rate of ICR mice exposed to 1.0 and 7.2Gy gamma-ray decreased from (39.5000 ± 3.3141)‰ to (28.1667±4.1345)‰ (P=0.033) and from (76.5000 ± 4.6242)‰ to (22.8333 ± 3.6553)‰(P=0.000),respectively. The bone marrow polychromatic micronucleus experiment indicated that the value of polychromatic erythrocyte (PCE)/normochromatic erythrocyte(NCE) of ICR mice exposed to 1.0 and 7.2Gy gamma-ray was less than the control group(P<0.05). Meanwhile,after irradiating by certain dose,the value of PCE/NCE in mice given S-isopentenyl-L-cysteine before irradiation was significantly higher than the corresponding groups (P<0.05).

CONCLUSIONS-isopentenyl-L-cysteine has a good protective effect against DNA damage induced by radiation.

Animals ; Bone Marrow ; Cysteine ; DNA Damage ; Disease Models, Animal ; Gamma Rays ; Mice ; Mice, Inbred ICR ; Radiation Injuries ; Radiation-Protective Agents

To evaluate the safety and effectiveness of Shenbei Guchang capsules in treatment of diarrhea type irritable bowel syndrome (yang deficiency of spleen and kidney) under widely used conditions, an open, multicenter, controlled, phase Ⅳ clinical trial was conducted in the drug clinical trial centers of 16 domestic hospitals. 2 123 patients from June 10, 2011 to November 29, 2012 were enrolled in the trial. Drug clinical trial was approved by Sichuan Academy of Medical Sciences, Sichuan Provincial People's Hospital Ethics Committee before implementation. Before the start of trial, subjects were selected according to the research scheme and inclusion criteria, then they would step into the 14 d study after signing Informed Consent Form. All subjects were treated according to the research scheme, evaluated the conditions and filled in CFR sheet, to provide the evaluation data and information on safety and efficacy of Shenbei Guchang capsules. Shenbei Guchang capsules were used to treat diarrhea type irritable bowel syndrome in widely used conditions (2 123 cases), and 2 029 cases of them entered FAS set, cure+markedly effective in 1 921 cases, with a comprehensive curative effect rate of 94.68%; 2 010 cases of them entered PPS set, cure+markedly effective in 1 906 cases, with a comprehensive curative effect rate of 94.83%. The primary symptoms of IBS were abdominal pain and diarrhea. After treatment, both abdominal pain and diarrhea were improved, with significant differences (P<0.000 1). There were significant differences in traditional Chinese medicine symptom scores on both post-treatment day 7 and day 14 as compared with the conditions before treatment (P<0.000 1). 35 cases of adverse events occurred during the trial with an incidence of 1.65%, including 12 cases of drug-related adverse events (adverse reaction) with an incidence of 0.57%, mainly manifested as nausea, abdominal distension and dry mouth, most of which would be spontaneously relieved without any measures. No serious adverse events occurred. The commercially available Shenbei Guchang capsules are proved safe and effective for the treatment of diarrhea type irritable bowel syndrome (yang deficiency of spleen and kidney) under widely used conditions (2 123 cases), and can be continued for clinical promotion and application.