Aged ; Anastomosis, Surgical ; Duodenum ; blood supply ; surgery ; Female ; Gastrointestinal Hemorrhage ; etiology ; surgery ; Humans ; Hypertension, Portal ; complications ; Jejunum ; surgery ; Rupture, Spontaneous ; Varicose Veins ; complications

The purpose of this study is to investigate the Sal I, Nru I and Mse I restriction fragment length polymorphisms (RFLPs) of factor IX gene in Chinese Han people. The frequencies of FIX-192 and FIX-793 for A and G, and FIX-698 for T and C were analyzed by polymerase chain reaction (PCR) in unrelated normal Chinese Han people. A sample of 214, 210 and 206 unrelated X chromosomes were analyzed for FIX-192 and FIX-793 and FIX-698, respectively. The results showed that the frequencies for FIX-192 were 0.878 for A and 0.122 for G, with a heterozygosity rate of 0.213, and the frequencies for FIX-793 were 0.552 for A and 0.448 for G, with a heterozygosity rate of 0.494, the frequencies for FIX-698 were 0.311 for T and 0.689 for C, with a heterozygosity rate of 0.429. It was concluded that the SalIand NruI and MseI RFLPs of FIX gene may be useful markers for carrier detection and prenatal diagnosis in Chinese families with hemophilia B patients.
China ; DNA ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Factor IX ; genetics ; Female ; Gene Frequency ; Heterozygote ; Humans ; Male ; Polymorphism, Restriction Fragment Length

OBJECTIVERecombinant adeno-associated virus type 8 (rAAV8) mediating transgene expression in mice was investigated using co-expressed report gene of secreted Gaussia princeps luciferase (Gluc) and non-secreted firefly luciferase(Fluc).

METHODSrAAV8-Gluc/Fluc was prepared and infected HEK293 cells to test its performance in vitro. BALB/c mice were received rAAV8-Gluc/Fluc at different doses by intravenous injection (iv) or intramuscular injection (im). Then Glue activities in blood were measured,the whole-body images for Flue activities were performed and Flue activities of tissue lysate were also detected.

RESULTSrAAV8-Gluc/Fluc was successfully prepared and could infected HEK293 cells. The Glue was mainly detected in the culture media while the Flue was mainly detected within cells. The blood Glue activities of mice received rAAV8-Gluc/Fluc by iv or im peaked at 10-20 d post injection and persisted for at least 120d. The blood Gluc activities increased at the rAAV8-Gluc/Fluc dose-dependent manner. For mice received rAAV8 by iv, Fluc mainly expressed in liver and minor Fluc expression was also found in cardiac muscle and skeletal muscle. For mice received rAAV8-Gluc/Fluc by im, Fluc mainly expressed in local skeletal muscle and secondly in liver.

CONCLUSIONrAAV8-Gluc/Fluc has been prepared successfully and its mediating transgene expression in mice has been investigated. This research will facilitate preclinical studies for rAAV8-mediated gene therapy.

Animals ; Dependovirus ; genetics ; metabolism ; Gene Expression ; Genes, Reporter ; Genetic Therapy ; instrumentation ; Genetic Vectors ; genetics ; metabolism ; HEK293 Cells ; Humans ; Liver ; enzymology ; Luciferases ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Muscle, Skeletal ; enzymology

OBJECTIVETo investigate the apoptosis effect of isothiocyanates (ITCS) on human liver cancer cells HepG-2, and its mechanism.

METHODHepG-2 cells were treated with different concentrations of ITCS. MTT assay was used to evaluate the influence of ITCS on cell proliferation. Flow cytometry was used to test ROS levels, intracellular mitochondrial transmembrane potential (deltapsim) , and hypodiploid apoptosis peak in HepG-2 cells.

RESULTITCS obviously inhibited proliferation of HepG-2 cells. When treated with 15, 30, 60, 120, 240 microg x mL(-1) of ITCS for 24 h, ROS levels were (23.1+/-1. 8)%, (53.3+/-3.3)%, (57.9+/-2.0)%, (79.9+/-3.4)%, (93.4+/-2. 6)% respectively; and deltapsim were (94.8+/-5.5)%, (91.8+/-5.4)%, (66.0+/-5.6)%, (65. 5+/-6.6)% and (44.3+/-2.7)% respectively; when treated with 60, 120, 240 microg x mL(-1) of ITCS for 48 h, cell apoptotic rates were (16.6+/-2.8)%, (21.9+/-4.4) % and (70.2+/-5.3) % respectively.

CONCLUSIONITCS generates ROS in gastric cancer HepG-2 cells, which causes mitochondrial membrane permeabilization and deltapsim decrease, therefore, leads to apoptosis of HepG-2 cells.

Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Brassica ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Isothiocyanates ; isolation & purification ; pharmacology ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria, Liver ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species ; metabolism

Alzheimer's disease is the most common disease which causes dementia in senior citizens. It is a neurodegenerative disease which is characterized by the degradation of the neurons and synapses in the brain. Usually,the irreversible and progressive neuron loss particular in the regions of cortex and hippocampus,the plaques of the accumulation of amyloidosis outside the cells,as well as the neurofibrillary tangles which is made by the hyperphosphorylation of Tau microtubule proteins can be seen in the lesion loca-tion of the patients. The Insulin malfunction in the central nervous system is now regarded as an important pathogenesis of Alzheimer's disease. This review focuses on the development of the knowledge of the mechanism of the malfunction of insulin in the central nervous system leading to the sporadic type of Alzheimer's disease and its applications,aiming to provide a reference in the study of Alzheimer 's disease.

We recently developed a mouse model of hepatitis B virus (HBV) chronic infection by intravenous (i.v.) injection with rAAV8-1. 3HBV to C57BL/6 mice. To define the responses of different mouse strains after injection with rAAV8-1. 3HBV, we intravenously injected rAAV8-1. 3HBV at doses of 4 x10(9) (Viral genome,vg), 4 x 10(10) vg and 4 x 10(11) vg to C57BL/6 and BALB/c mice,respectively, and determined the levels of serum HBV antigen and antibody by ELISA,serum viral DNA by real-time PCR,and HBcAg expression in liver by immunohistochemical staining. For C57BL/6 mouse strain with injection of rAAV8-1. 3HBV at three doses, 100% of the mice carried HBV for more than 8 months. The levels of serum HBsAg and HBeAg, serum viral DNA and HBcAg-positive hepatocytes increased in a rAAV8-1. 3HBV dose-dependent manner. For C57BL/6 mice injected with rAAV8-1. 3HBV at the dose of 4 x 10(11) vg,over 40% of hepatocytes expressed HBcAg and serum viral DNA reached over 10(5) IU/mL. No HBV antibody was detected in sera of C57BL/6 mice. For BALB/c mice with injection of rAAV8-1. 3HBV at three doses, serum HBeAg, serum viral DNA and HBcAg-positive hepatocytes persisted for more than 8 months, but serum HBsAg declined remarkably at 2 weeks after injection. The levels of serum HBeAg and HBcAg-positive hepatocytes in BALB/c mice increased in a rAAV8-1. 3HBV dose-dependent manner. Injection with rAAV8-1. 3HBV at the dose of 4 x 10(11) vg resulted in over 50% of BALB/c mice hepatocytes expressing HBcAg. Serum anti-HBsAg were detected in BALB/c mice with rAAV8-1. 3HBV injection at the dose of 4 x10 (10) vg. In conclusion, both C57BL/6 and BALB/c strains can be developed to chronic HBV infection mouse models by i. v. injection with rAAV8-1. 3HBV at doses of 4 x10(9) - 4 x 10(11) vg and the levels of HBV replication increase in a rAAV8-1. 3HBV dose-dependent manner. In contrast to C57BL/6 strain, the BALB/c mice carry out humoral immunity to HBsAg, but fail to mediate HBV clearance.
Animals ; Dependovirus ; genetics ; metabolism ; Disease Models, Animal ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; genetics ; immunology ; physiology ; Hepatocytes ; immunology ; virology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transduction, Genetic ; Virus Replication

OBJECTIVETo investigate the clinicopathological characteristics, and treatment of malignant gastrointestinal stromal tumors (MGIST).

METHODSImmunohistochemistry was used to detect CD117, CD34, S100, vimentin and SMA expressions. The postoperative curative effect was compared between the patients with or without imatinib treatment.

RESULTSRadical resection was performed in 60 cases. Twenty-two tumors with a mean diameter of 5.3 cm were potentially malignant, and 38 tumor with a mean diameter of 9.2 cm were malignant. Microscopical examination revealed haemorrhagia or necrosis, abundant tumor cells, heteromorphism and caryocinesia of the tumors. 54 Cases were CD117 positive, 53 cases CD34 positive, 48 cases vimentin positive, 27 cases S100 positiveì16 cases SMA positive. The two-year recurrence rate was 80.5% in the patients without postoperative imatinib treatment, significantly higher than 21.1% in the patients with postoperative imatinib treatment(P< 0.05).

CONCLUSIONSCD117 and CD34 markers are most valuable diagnostic indexes of MGIST, but its final diagnosis depends on pathology. Postoperative imatinib treatment is most effective to control recurrence and metastasis.

Adult ; Aged ; Antigens, CD34 ; metabolism ; Antineoplastic Agents ; therapeutic use ; Benzamides ; Female ; Gastrointestinal Stromal Tumors ; diagnosis ; therapy ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Proto-Oncogene Proteins c-kit ; metabolism ; Pyrimidines ; therapeutic use ; Retrospective Studies

In this report, we developed a HBV infection model in C57BL/6 mouse line by in vivo injection of a recombinant adeno-associated virus 8 vector carrying 1. 3 copies of HBV genome (ayw subtype) (rAAV8-1. 3HBV). We firstly prepared and purified the rAAV8-1. 3HBV and then injected it into three C57BL/6 mice with the dose of 2 x 10e11vg, respectively. HBsAg and HBeAg were assayed in sera collected at different time points post injection. Ten weeks post injection, the three mice were sacrificed and blood and liver tissue were taken for assay. Copies of HBV DNA were detected by real time PCR and the way of HBV DNA replication was identified by PCR. Subsequently, detection of HBV antigen by immunohistochemistry and pathology analysis of liver tissue of mice were performed. The results suggested that expression of HBsAg and HBeAg lasted for at least 10 weeks in mice sera. Among mice injected with rAAV8-1. 3HBV, HBsAg levels were showed an 'increasing-decreasing-increasing' pattern (the lowest level at the 4th week post injection), while HBeAg levels were kept high and relatively stable. HBV DNA copies were 4.2 x 10(3), 3.6 x 10(3), 2.5 x 10(3) copies/mL in sera and 8.0 x 10(6), 5.7 x 10(6), 2.6 x 10(6) copies/g in hepatic tissues of three mice, respectively. We found that the linear 1. 3HBV DNA in the rAAV8-1. 3HBV could self form into circular HBV genome and replicate in livers of HBV transfected mice. HBsAg and HBcAg were both positive in liver tissue of mice injected with rAAV8-1. 3HBV and no obvious pathological characters were found in liver of mice injected with rAAV8-1. 3HBV. In conclusion, we successfully developed a HBV chronic infection model in C57BL/6 mouse line by in vivo transduction with the recombinant virus rAAV8-1. 3HBV, in which HBV genes could be continuously expressed and replicated over 10 weeks, and paved a way for further characterization of the human chronic hepatitis B virus infection and evaluation of vaccine and anti-HBV agents.
Animals ; Dependovirus ; genetics ; metabolism ; Disease Models, Animal ; Gene Dosage ; Genetic Vectors ; genetics ; metabolism ; Genome, Viral ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; virology ; Humans ; Mice ; Mice, Inbred C57BL ; Transduction, Genetic ; Virus Replication

OBJECTIVETo discuss the reliability of SARS-CoV antibody detection for SARS diagnosis.

METHODSUsing SARS-CoV ELISA kit to detect relevant antibody in fresh serum of healthy, fever, probable, and suspect cases.

RESULTSThe positive rate is 0%, 40%, and 95% respectively in healthy, probable, and suspect cases.

CONCLUSIONSIt is reliable to detect SARS-CoV antibody in late suspect patients, but there will be high false-positive result in ordinary fever cases.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; blood ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Female ; Fever ; diagnosis ; Humans ; Male ; Middle Aged ; SARS Virus ; immunology ; isolation & purification ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology ; Time Factors

This study was purposed to investigate the impact of human umbilical cord-derived mesenchymal stem cells (hUC-MSC) on the sensitivity of HL-60 cells to therapeutic drugs so as to provide more information for exploring the regulatory effect of hUC-MSC on leukemia cells. Transwell and direct co-culture systems of HL-60 and hUC-MSC were established. The apoptosis and cell cycle of HL-60 cells were detected by flow cytometry. RT-PCR and Western blot were used to detect the mRNA and protein levels of Caspase 3, respectively. The results showed that the apoptosis of HL-60 induced by cytarabine (Ara-C) decreased significantly after direct co-cultured with hUC-MSC cycle mRNA (P < 0.05). The similar phenomenon was observed in transwell co-culture system. Cell cycle of HL-60 cells were arrested at G0/G1 phase and did not enter into S phase (P < 0.05) and the expression of Caspase-3 mRNA and protein in HL-60 cells were reduced (P < 0.05). It is concluded that hUC-MSC protected HL-60 from Arc-C induced apoptosis through regulating the cell cycle and down-regulating expression of Caspase 3 in HL-60 cells. In addition, this effect is caused by the soluble factors from hUC-MSC.
Apoptosis ; Caspase 3 ; metabolism ; Coculture Techniques ; Cytarabine ; pharmacology ; HL-60 Cells ; Humans ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology