Objective: This study was to evaluate the effects of hepaCAM (hepatocyte cell adhesion molecule) on invasiveness of human renal cell carcinoma (RCC) cell line 786-0 and explore the underlying mechanism. Methods: A recombinant plasmid containing hepaCAM cDNA (hepaCAM-pEGFPN2) was transfected into 786-0 cells, then the positive cell clones were selected with G418 and amplified. The 786-0 cells transfected with pEGFPN2 and untreated 786-0 cells were as controls. The mRNA expressions of hepaCAM and matrix metalloproteinase (MMP)-2 and MMP-9 before and after stable transfection are detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time fluorescence PCR. The activities of MMP-2 and MMP-9 were analyzed by gelatin zymography. The invasiveness of 786-0 cells was determined by Transwell migration assay in vitro. Results: The mRNA transcription of hepaCAM was significantly increased in 786-0 cells after stable transfection of hepaCAM (P <0.01). The mRNA expressions and activities of MMP-2 and MMP-9 in 786-0 cells were markedly inhibited by hepaCAM (P <0.05). The transmembrane migration capacity of cells was significantly reduced after stable transfection of hepaCAM (P <0.01). Conclusion: HepaCAM inhibits the activities of gelatinase by down-regulating the expressions of MMP-2 and MMP-9, thereby preventing the invasiveness of renal cell carcinoma 786-0 cells.

OBJECTIVETo evaluate the advantages of combination therapy with interferon-alpha plus nucleoside analogue-lamivudine or HBV vaccine in children with HBeAg positive chronic hepatitis B.

METHODSA total of 120 patients with HBeAg positive chronic hepatitis B were divided into three groups, 40 patients per group. Each group was treated with one of the following therapies respectively: Group A IFN-alpha 1b 10 MU/m2 three times per week (Tiw); Group B IFN-alpha 1b 10MU/m2 three times per week (Tiw) plus lamivudine 3 mg/kg for 6 months. Group C IFN-alpha 1b 10 MU/m2 three times per week (Tiw) plus HBV vaccine 30 microg one a month.

RESULTSThere was no significant difference in normalizing rate of ALT among the three groups at end of treatment. There was more significant difference in negative rate (seroconversion) of serum HBV DNA and HBeAg in group B than group A and group C (P less than 0.05).

CONCLUSIONThe combination therapy of IFN-alpha 1b plus lamivudine seemed to be more effective than the therapy with IFN-alpha alone and the combination of IFN-alpha and HBV vaccine.

Anti-HIV Agents ; therapeutic use ; Child ; Child, Preschool ; DNA, Viral ; blood ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Hepatitis B Vaccines ; therapeutic use ; Hepatitis B e Antigens ; blood ; genetics ; immunology ; Hepatitis B virus ; drug effects ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Lamivudine ; therapeutic use ; Male ; Treatment Outcome

Humans ; Stroke ; epidemiology

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; ethnology ; Evoked Potentials, Somatosensory ; Evolution, Molecular ; Female ; Foot Deformities, Congenital ; genetics ; GTP Phosphohydrolases ; genetics ; GTP-Binding Proteins ; Genes, Dominant ; Humans ; Male ; Membrane Proteins ; Middle Aged ; Mutation ; Spastic Paraplegia, Hereditary ; genetics

To compare the efficacy and safety of Lamivudine (LAM) plus Adefovir dipivoxil (ADV) combination therapy and Entecavir (ETV) monotherapy for chronic hepatitis B patients. 120 patients with chronic hepatitis B managed in a single-centre clinical practice (median 96 weeks) were split into 2 cohorts, one was treated with de-novo combination Lamivudine (100 mg/day) plus Adefovir (10 mg/day) (LAM+ADV), the other with Entecavir (0.5 mg/day) monotherapy. Serum levels of ALT, creatinine, HBsAg, HBeAg and HBV viral load, together with genotypic resistence were analyzed at 0, 12, 24, 48, 96 weeks, respectively. HBV DNA was determined by real-time PCR. HBsAg and HBeAg were assessed by chemiluminescence. Serum levels of ALT and creatinine were detected by automatic biochemical analyzer. HBV genotypic resistence was tested by direct sequencing. (1) At the time point of 96 weeks, a total of 99 patients (51 cases in combination therapy cohort and 48 case in monotherapy cohort) were compared. The baseline characteristics as for HBV viral load, median age, serum levels of ALT and creatinine were compatible between combination therapy cohort and monotherapy cohort. (2) The rates of HBV DNA values is less than 300 copies/ml and HBV DNA values is less than 1000 copies/ml had no significant difference between LAM + ADV and ETV cohorts by the 12 and 24 weeks (P more than 0.05). (3) At the time point of 48 weeks, the rates of HBV DNA is less than 1000 copies/ml, HBeAg seroconversion, and ALT normalization were similar in both cohorts, though the rate of HBV DNA values is less than 300 copies/ml was obviously higher in combination therapy cohort than that of monotherapy cohort (90.7% vs 76%, P values is less than 0.05). (4) At the time point of 96 weeks, the rates of HBV DNA values is less than 300 copies/ml (96.1% vs 79.2%), HBV DNA values is less than 1000 copies/ml (98% vs 87.5%) and the HBeAg seroconversion (41.7% vs 16.7%) were markedly higher in combination therapy cohort than those of monotherapy cohort statistically (P values is less than 0.05 for all). The mean values of decreases for HBV viral loads and HBsAg levels were smilar in both cohorts at 48 and 96 weeks. (5) Elevated serum creatinine not be found in both cohorts at the end of treatment. (6) No virological breakthrough occurred in combination therapy cohort at the end of treatment. Four patients in monotherapy cohort were found with virological breakthrough at 96 weeks and three cases among were confirmed to be of variants associated with ETV resistance (rtL180M + T184L + M204V). Present study suggests that Lamivudine plus Adefovir dipivoxil de-novo combination therapy was more efficacious than Entecavir monotherapy for CHB patients and the tolerance is compatible.

Objective To investigate the risk factors related to outcome of chronic severe hepatitis B. Methods A total of 336 consecutive patients with chronic severe hepatitis B (CSHB) were analysed retrospectively. According to the outcome, objects were divided into survival group(n =137) and death group(n = 199), then to observe the differences between them in respect to age, sex, family history,prothrombin activity ( PTA ) , complications including ascites, infection, electrolyte disturbance, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal sydrome and the corresponding quantity of complications in each individual, antivirus therapy, artificial liver support system (ALSS) therapy, and alprostadil therapy. Finally, risk factors related to prognosis were selected by stepwise Logistic regression analyse. Results In univariate analyse, significant differences between the two groups were found related to age, PTA, complications and its quantity( P ＜ 0.01 for all), and antivirus therapy ( P ＜ 0. 05 ) rather than sex, family history and treatment of ALSS or alprostadil. Logistic regression revealed that risk factors comprised of PTA and quantity of complications, antivirus therapy was the only protective factor. Conclusion A numbers of factors including age,PTA,complications and its quantity, and antivirus therapy affect the prognosis of CSHB, among which, antivirus therapy can reduce the death rate.

OBJECTIVETo establish rhesus haploidentical hematopoietic stem cell transplantation model by nonmyeloablative conditioning, and examine the effects of mesenchymal stem cells (MSC) in haploidentical transplantation.

METHODSThe recipient haploidentical rhesus monkeys were conditioned with a nonmyeloablative regimen consisted of fludarabine, cyclophosphamide, 200 cGy total body irradiation, and rabbit anti-human thymocyte globulin. Cyclosporine A, mycophenolate mofetil and anti CD25 antibody were used for graft versus host disease (GVHD) prophylaxis. Rhesus monkeys in one group were given hematopoietic stem cell (HSC) only, while in the other group HSC combined with MSC. The differences in hematopoiesis recovery, chimerism level, and GVHD between the two groups were evaluated.

RESULTSStable chimerism could be achieved in recipient monkeys. Hematopoiesis recovery was mainly related with chimerism level. MSC seemed capable of facilitating HSC engraftment, as there were more mixed chimerism and less GVHD occurrence in the HSC combined with MSC recipient group.

CONCLUSIONA rhesus haploidentical hematopoietic stem cell transplantation model is successfully established by nonmyeloablative conditioning. MSC was of great benefit to haploidentical transplantation.

Animals ; Chimerism ; Graft vs Host Disease ; prevention & control ; Haploidy ; Hematopoietic Stem Cell Transplantation ; Macaca mulatta ; genetics ; surgery ; Mesenchymal Stem Cell Transplantation ; Models, Animal ; Transplantation Conditioning

BACKGROUNDThere are no reports on exnografting cultured human fetal neocortical cells in this infracted cavities of adult rat brains. This study was undertaken to observe whether cultured human cortical neurons and astrocytes can survive and grow in the infarcted cavities of adult rat brains and whether they interconnect with host brains.

METHODSThe right middle cerebral artery was ligated distal to the striatal branches in 16 adult stroke-prone renovascular hypertensive rats. One week later, cultured cells from human embryonic cerebral cortexes were stereotaxically transferred to the infarcted cavity of 11 rats. The other 5 rats receiving sham transplants served as controls. For immunosuppression, all transplanted rats received intraperitoneal injection of cyclosporine A daily starting on the day of grafting. Immunohistochemistry for glial fibrillary acidic protein (GFAP), synaptophysin, neurofilament, and microtubule associated protein-2 (MAP-2) was performed on brain sections perfused in situ 8 weeks after transplantation.

RESULTSGrafts in the infarcted cavities of 6 of 10 surviving rats consisted of bands of neurons with an immature appearance, bundles of fibers, and GFAP-immunopositive astrocytes, which were unevenly distributed. The grafts were rich in synaptophysin, neurofilament, and MAP2-positive neurons with long processes. The graft/host border was diffuse with dendrites apparently bridging over to the host brain, into which neurofilament immunopositive fibers protruded.

CONCLUSIONCultured human fetal brain cells can survive and grow in the infarcted cavities of immunodepressed rats and integrate with the host brain.

Animals ; Astrocytes ; transplantation ; Brain ; pathology ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cerebral Infarction ; metabolism ; pathology ; therapy ; Fetal Tissue Transplantation ; Glial Fibrillary Acidic Protein ; analysis ; Humans ; Microtubule-Associated Proteins ; analysis ; Neocortex ; cytology ; Neurons ; transplantation ; Rats ; Synaptophysin ; analysis

OBJECTIVETo elucidate the reconstruction of neovascularization that occurred in the superiorly and inferiorly based posterior pharyngeal flaps in different time postoperatively.

METHODSTen mongrel dogs were randomly divided into two experimental groups, which were performed superiorly or inferiorly based posterior pharyngeal flap surgery respectively. Each group was then subdivided into five subgroups, and were sacrificed immediately after operation or on 3, 7, 30, 90 day postoperative respectively. Microangiography was used to exhibite the vessel.

RESULTS1. The blood vessel reconstruction of the superiorly based posterior pharyngeal flap was more rapid compared with the inferiorly based flap. The 3-day flap has established an axial vascular network, which was mature on the 30-day flap. The superiorly based posterior pharyngeal flap was mainly supplied by the pedicle. 2. The blood vessels reconstruction of the inferiorly based posterior pharyngeal flap was firstly occurred in the pedicle and apex of the flap, which grew slowly to the middle of the flap. The inferiorly based posterior pharyngeal flap was supplied by the pedicle and the soft palate. A mature axial vascular network was exhibited on the 90-day flap, which was not mature on the 30-day flap.

CONCLUSIONBoth superiorly and inferiorly based posterior pharyngeal flap can establish an axial vascular network and gain ample blood supply.

Animals ; Dogs ; Neovascularization, Physiologic ; Palate, Soft ; blood supply ; Pharynx ; blood supply ; Random Allocation ; Reconstructive Surgical Procedures ; Surgical Flaps ; blood supply

OBJECTIVETo deduce the protocol, scoring criteria and interpretive guidelines for assessment of HER2 gene expression status by fluorescence in-situ hybridization (FISH) and to compare the results with those obtained by immunohistochemistry.

METHODSThe HercepTest kit from Dako Cytomation was employed for immunohistochemistry. FISH for HER2 gene expression status was performed using PathVysion DNA probe kit on the archival paraffin-embedded sections of breast cancer tissues from 28 Chinese female patients with immunohistochemical staining scores of (3 +), (2 +), (1 +) and 0.

RESULTSTen of the 12 patients with score (3 +) by immunohistochemistry were positive for HER2 by FISH, with 2 cases being polysomy. Two other cases with FISH-negative were also shown to be polysomy. Seven of the 10 patients with score (2 +) by immunohistochemistry showed HER2 gene amplification, with 1 case being polysomy. Two of the remaining 3 cases, which were FISH-negative, were shown to be polysomy. All the patients with scores (1 +, number = 3 ) or 0 ( number = 3) by immunohistochemistry failed to show amplification. One case of polysomy was noted in either group.

CONCLUSIONSImmunohistochemistry is useful as an initial screening tool for HER2 expression status. Because of the obvious discrepancies between protein expression and gene amplification, patients with score (2 +) by immunohistochemistry should undergo FISH testing as well. FISH is also required in selected examples with score (3 +) immunohistochemical results, especially in those with false-positive immunohistochemistry due to chromosome 17 aneuploidy.

Breast Neoplasms ; genetics ; metabolism ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; Chromosomes, Human, Pair 17 ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Genes, erbB-2 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Polyploidy ; Receptor, ErbB-2 ; metabolism