Biomedical Engineering ; Reconstructive Surgical Procedures ; Surgery, Plastic

Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

Objective To discuss the curative effect of transplantation using donation after cardiac death (DCD) according with Chinese standard Ⅲ (C-Ⅲ).Methods The organs were obtained from 4 DCDs from 2011 to 2012,and the clinical data of DCD transplantation were retrospectively analyzed.Withdrawal of life support occurred in the operating room.Donor warm ischemia time was 7-15 min.Results The biopsy of liver was performed on the 3rd DCD.Eight cases were subjected to renal transplantations,and 3 to liver transplantations.One patient exhibited delayed graft function of the kidney from the 4th DCD.All patients made an uneventful recovery and were discharged from the hospital without rejection or surgical complication.They were followed up in outpatient department.Conclusion The use of DCD according with C-Ⅲ is an effective way to increase the number of organs available for transplantation,and can obtain satisfactory effects.

OBJECTIVETo evaluate the role of p53 gene mutation in colorectal carcinoma and assess the value of peripheral blood p53 single nucleotide polymorphism (SNP) in early diagnosis of colorectal carcinoma.

METHODSNSP in axons 5-8 of p53 gene was detected using ligase detection reaction-polymerase chain reaction (LDR-PCR) in the peripheral blood of 100 patients with colorectal cancer and 100 healthy subjects.

RESULTSThe mutation rate of p53 gene was 24% (24/120) in colorectal carcinoma patients and 0% (0/120) in the healthy subjects (P<0.05).

CONCLUSIONp53 plays an important role in the carcinogenesis of colorectal carcinoma, and SNP analysis for p53 gene can be helpful in early diagnosis of colorectal carcinoma.

Adult ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; genetics ; DNA Mutational Analysis ; DNA, Neoplasm ; genetics ; Early Detection of Cancer ; Female ; Humans ; Male ; Mutation ; Polymorphism, Single Nucleotide ; Tumor Suppressor Protein p53 ; genetics

Objective To provide valid data and useful genetic counseling in the clinical application of non‐invasive prenatal test (NIPT) ,fetal chromosomal disorder were screened by massive parallel sequencing and made a follow‐up study .Methods Preg‐nant women with Down screening in high‐risk were screened by NIPT ;NIPT verified high‐risk individuals were suggested for kary‐otyping ;and we follow up on whoever showed low risk by NIPT before and after their deliveries .Results (1)Totally 1 676 cases of pregnant women were tested by NIPT ,25 cases prompted to be abnormal ,with an abnormal rate of 1 .49% ,karyotype analysis re‐sults in 12 cases of abnormalit ,the accuracies of NIPT for T21 ,T18 ,XO ,XXY ,and XYY were 99 .93% ,100 .00% ,99 .66% , 100 .00% ,100 .00% respectively ;the accuracy of NIPT for women with advanced paternal age and twins were both 100 .00% ;kary‐otyping positive individuals underwent abortion ,which gives a prenatal intervention rate of 100 .00% .(2)Out of 1 651 cases of NIPT low risk testers ,1 468 cases were successfully followed up ,with a 88 .91% success rate .We found chromosome abnormality with one case of inversion of chromosome 9 (maternal) .(3)Ultrasound‐detection possessed 98 .17% accuracy and 7 .69% in detec‐tion rate;in high‐risk pregnant woman ,Down screening had an accuracy of 0 .88% and false positive rate of 99 .12% ;98 .71%women were avoided prenatal diagnosis via NIPT .Conclusion Compare to ultrasound and maternal plasma screening ,NIPT is a far more accurate prenatal screening approach .To build effective follow‐up and service systems of NIPT is necessary to reduce birth de‐fects in medical institutions .

OBJECTIVETo study the potential aging effect on workers exposed to acrylonitrile (ACN).

METHODSThe deletion rates of mitochondrial DNA (mtDNA) in peripheral blood nucleate cells of 47 exposed workers and 47 non-exposed workers (as control), as well as 12 old people and 12 young people were measured with polymerase chain reaction (PCR).

RESULTSThe positive rates of mtDNA deletion in peripheral blood nucleate cells were 17.02% in the workers exposed to ACN and 25.00% in group of old people. However, the mtDNA deletion was not detected in the control group and young people.

CONCLUSIONSACN could induce mtDNA deletion in peripheral blood nucleate cells of the exposed workers. There may be a potential molecular effect of occupational ACN exposure on workers' aging.

Acrylonitrile ; toxicity ; Adolescent ; Aged ; Aged, 80 and over ; Aging ; drug effects ; Blood Cells ; drug effects ; ultrastructure ; DNA Damage ; DNA, Mitochondrial ; drug effects ; Humans ; Occupational Exposure

OBJECTIVETo further understanding of lymphoma of salivary gland through clinicopathologic analysis.

METHODSClinical findings, pathologic features, clinical staging, therapy and prognosis of 4 cases were reviewed and clinically analysed.

RESULTSDifferent treatment were received by the 4 patients, one had stage IIIA disease and three had stage IE disease. All patients got their illness completely remitted.

CONCLUSIONMucosa-associated lymphoid tissue lymphoma of the salivary glands is an indolent disease. Different treatments can all result in prolonged remission, and it has better outcome than other NHL.

Aged ; Female ; Humans ; Lymphoma, B-Cell, Marginal Zone ; diagnosis ; pathology ; therapy ; Middle Aged ; Parotid Neoplasms ; diagnosis ; pathology ; therapy ; Prognosis ; Submandibular Gland Neoplasms ; diagnosis ; pathology ; therapy

OBJECTIVETo explore apoptosis-inducing effects of realgar nanoparticle (nano-realgar) on drug-sensitive leukemia cells.

METHODPreparation of nano-realgar was mechanical milled using a high-energy planetary ball mill. Using drug-sensitive leukemia cells (K562) as target cells, MTT assay was used to detect the proliferating activity of K562 cells, and the cellular apoptosis was investigated with double staining of FITC-Annexin V and propidium iodide (PI) by flow cytometry. Flow cytometry (FCM) was employed to detect expression of intracellular Bax, Bcl-2, P-53 protein and the activity of Caspase-3.

RESULTThe raw realgar was made to ultra-fine powder by ball milling, and the average diameter of the nanoparticle was (72.72 +/- 22.18) nm measured with electron microscopes. Nano-realgar significantly inhibited the proliferation of K562 cells, Treated for 24, 48 and 72 hours, the 50% inhibitory concentration (IC50) was 43.48, 20.52, 16.07 mg x L(-1). After exposure to 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 hours, the apoptosis of K562 cells detected by Annexin V/PI staining was increased, the apoptotic rate of K562 cells was 10. 52% and 73.25%. After the target cells were treated with 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 h, the expression of P-53, Bax, Bcl-2 markedly increased in a time and dose-dependent manner. After administration of 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 h, the percentage of BCRP+, P-gp+ and co-expressing P-gp and BCRP cell population in K562 cells incrased dramatically.

CONCLUSIONNano-Realgar significantly induced apoptosis of drug-sensitive leukemia cells.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Leukemia ; drug therapy ; pathology ; Nanotechnology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Sulfides ; pharmacology ; Tumor Suppressor Protein p53 ; analysis

OBJECTIVE: To explore the correlation of genetic mutations and clinical features of myelodysplastic syndromes (MDS) with scores of Revised International Prognostic Scoring System (IPSS-R). METHODS: Eighty-seven patients with de novo MDS were enrolled. Mutations of MDS-related genes and clinical features were used to determine the incidence and subtype of mutations. Clinical features and IPSS-R scores of the patients with high frequency mutations involving TET2, TP53, ASXL1, RUNX1 and SF3B1 genes were compared. RESULTS: Fifty-four patients (62.1%) harbored at least one point mutation. The incidences of various mutations were significantly different, with the incidence of MDS-EB-2 being 100% and MDS-SLD being only 38.9%. Compared with the wild types, patients harboring mutations had higher lactate dehydrogenase, higher β2 microglobulin, higher percentage of bone marrow blast cells and lower hemoglobin levels (P=0.027, <0.01, <0.01, 0.046, respectively). The IPSS-R scores of MDS patients with mutations were significantly higher than the wild types (P<0.01). The IPSS-R scores of the TP53 mutation groups were 7.82±1.83, which was significantly higher than the control group (3.77±1.66, P<0.01). No difference was found between the IPSS-R between patients carrying TET2, ASXL1, RUNX1, and SF3B1 mutations or the wild types (P>0.05). CONCLUSION Genetic mutations are commonly found in MDS. MDS patients with mutations have unique clinical laboratory characteristics. Although the prognostic value of most genes is controversial, TP53 is an definite indicator of poor prognosis.
DNA Mutational Analysis ; Humans ; Incidence ; Mutation ; Myelodysplastic Syndromes ; genetics ; Prognosis ; Tumor Suppressor Protein p53 ; genetics