Objective To study whether peripheral blood (PB)CD34+ cell absolute count can reliably predict peripheral blood progenitor cell (PBPC) autograft CD34+ cell contents. Methods We examined the PB CD34+ cell count and PBPC CD34+ cell count with ProCOUNT by flow cytometry. Twenty-five collections were performed.The PBPC autograft parameters measured were the CD34+ cell?CFU-GM? CFU-E?BFU-E?CFU-MK and mononuclear cell (MNC) content. We analyzed the correlation between the PB CD34+ cell absolute count?CD34+%?WBC?NEU?MNC?PLT? RBC and Various parameters in the PBPC autograft with Spearman correlations analysis or Regression analysis. Results (1) There is a strong correlation between PB CD34+ cell/L and autograft CD34+ cell/kg (r=0.790,P

Objective:To investigate the treatment effect of using posterolateral approach under arthroscopic for senile ankle fractures.Methods: The clinical data from 30 cases of ankle fractures of the elder patients were received arthroscopic therapy (observation group) and 30 cases underwent surgical treatment(control group), both of the two groups were analyzed by retrospectively. The operation time, amount of bleeding and recovery time were compared between two groups. McGuire scoring standard was used to measure and compare the ankle function before and after surgery.Results: There were difference between two groups in the operation time, amount of bleeding and recovery time; and the observation group were less than control group in these indexes (t=26.82,t=23.54,t=31.21;P<0.05). Before surgery, there was no difference in term of McGuire scores between two groups. McGuire scores of every group after surgery was higher than those before surgery (t=12.34, P<0.05); and ankle function in observation group was improved more significantly than those in control group.Conclusion: Arthroscopic therapy could clearly find the joint injury, treatment in time, have little tissue damage and safety during operation, and it can improve the treatment effect.

OBJECTIVE:To apply the kinetic turbidimetric Limulus test to the endotoxin assay of Levocarnitine sodium chloride injection.METHODS:The16-fold dilution of Levocarnitine sodium chloride injection was prepared.The content of bacterial endotoxin in Levocarnitine sodium chloride injection was determined with kinetic turbidimetric Limulus test after screen test and validation test.RESULTS:The16-fold dilution of Levocarnitine sodium chloride injection was effective to e?liminate the interference in Limulus test.The average recovery was in the range of50%～200%.CONCLUSION:The kinetic turbidimetric Limulus test provides a new and quick method for the quantitative determination of bacterial endotoxin in Levo?carnitine sodium chloride injection.

Objective: To investigate hyperthermia enhanced expression of heat shock protein70 (HSP70) in breast cancer drug sensitive cell line MCF7 and multi-drug resistant (MDR) cell line MCF7/ADR, so as to increase cytotoxic activity of immunologic effector cells against the target cells, and to provide an experimental basis for cell immunotherapy based on hyperthermia. Methods: The immunological effector cells were induced and expanded by cytokines. Breast cancer cell lines MCF7 and MCF7/ADR were treated at 42 ℃ for an hour. Then after being incubated for additional 4 hours, 24 hours and 48 hours, the cells were digested. Flow cytometry (FCM) was used to detect the expression of HSP70 on the target cells. MTT assay was employed to evaluate cell proliferation and the cytotoxic activity of immunologic effector cells against target cells. Results: The expressions of HSP70 in both the target cells had significant difference. The expression of HSP70 in MCF7/ADR cells was higher than in MCF7 cells before hyperthermia. After hyperthermia the expression of HSP70 increased by 6 folds in MCF7/ADR cells, and 22 folds in MCF7 cells and was higher than in MCF7/ADR cells at hour 4. The proliferation inhibition fraction of hyperthermia against MCF7 was significantly lower than that of MCF7/ADR. The cytotoxic activity of immunologic effector cells against MCF7 cells was lower than against MCF7/ADR cells before hyperthermia. After hyperthermia the cytotoxic activity of immunologic effector cells against MCF7 cells rose by 86.23%, and was higher than against MCF7/ADR cells (rose by 30.32%). Conclusion: The expression of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR were enhanced significantly by hyperthermia. Cytotoxic activity of immunologic effector cells against both the target cells were increased by hyperthermia. The differential expressions of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR affect the cytotoxic activity of immunologic effector cells.

Objective To explore the efficacy of calmodulin antagonist berbamine(BBM)on multidrug resistance(MDR)reversal and its mechanism. Methods Human breast cancer cell line MCF7 and its adriamycin-resistant counterpart MCF7/ADR were used in the study.The cells were cultured with ADR and different concentration of BBM. MTT assay was used to analyze the effect of BBM on cell growth inhibition.According to the MTT assay,the 50% inhibitory concentration(IC 50 ),the multiples of drug resistance and increased sensitivity of ADR were calculated.The concentration of intracellular ADR and expression level of P-glycoprotein(P-gp)were detected by flowcytometry(FCM).The mRNA expression level of mdr1 gene was detected by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR)with ?-actin as internal reference. Results The IC 50 of ADR in MCF7 and MCF7/ADR cells were(0.98?0.06)?mol/L and(101.20?5.72)?mol/L,respectively.The resistant multiple of MCF/ADR cells to ADR was 103 folds higher than that of MCF7 cells.BBM increased the chemo-sensitivity of ADR in MCF7/ADR cells with dose-dependent relationship,i.e.when 5*!?mol/L ,10*!?mol/L and 20*!?mol/L BBM was added into the culture the chemo-sensitivity of ADR was increased to 2.76,5.88,and 28.26 folds(P

Objective:To investigate if the combined use of CDAⅡ and sodium butyrate can induce demethylation and re-expression of retinoic acid receptor?2(RAR?2)gene in cultured human breast cancer cells MCF7.To explore if the two drugs can inhibit cell growth and induce cell apoptosis synergetically.Methods:MCF7 cell line was treated with CDAⅡ,sodium butyrate,combination of the two drugs respectively.Methylation was assessed by methylation-specific polymerase chain reaction(MSP)for RAR?2 gene.Gene expression was evaluated by reverse transcription polymerase chain reaction(RT-PCR).Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL)and Hoechst33342/propidiumiodide(PI)staining.Cell growth inhibition was measured by MTT assay.Results:Neither CDAⅡ nor sodium butyrate induced demethylation and re-expression of RAR?2 gene,Combination of the two drugs partially demethylated gene promoter accompanied by re-expression of RAR?2.The apoptotic cells in the double-drug group were obvious following Hoechst33342/PI staining.The percentage of apoptotic cells in the double-drug group was significantly higher than that of the two single-drug group(39.5% vs 5.2%,8.1%)(P

The distribution of receptor sites for concanavalin A (Con A) on the cellsurface of human gastric cancer cell line (MGC 80-3) and fibroblast cell cultures wasstudied with electron and light microscope after application of Con A-peroxidasemethod.The cytochemical reaction of the majority of non-prefixed MGC 80-3 cellsshows a striking tendency to be more uneven distribution than on fibroblasts.Clustering,patching and capping of Con A binding sites on the cancer cells anduniform distribution on the fibroblasts were observed.The effects of incubationtemperature,prefix treatment,different phase of cell cycles and cell growth condi-tions on the distribution of Con A receptor complexes were observed.The possiblereasons for the more irregular distribution of the cytochemical reaction product onthe MGC 80-3 cells than the fibroblasts are discussed.

A rat monoclonal antibody of the IgG1 class, McAb B5A8, specific for the cAMP-dependent protein kinase(PKA) was produced. Western blot analysis revealed specific binding of the antibody to protein of 52-56 kd.The affinity-purified McAb BSA8 was labeled with FITC. Immunofluorescent localization of PKA was examined in human fibroblasts, gastric cancer cell line (MGc 80-3)and EAC cells. The distribution of PKA on the cytoplasmic microtubule network, Golgi region and nucleoli were observed. PKA was localized in the nuclear region in G2 phase of synchronized MGc 80-3 cells, and it was only found around the nuclei of MGc 80-3 cells which were incubated with DBcAMP. The changes in the distribution of PKA in cells are discussed.

Synchronous cells in different phase (G_1, S, G_2, M) of cell cycle were obtained from human gastric low-differentiated mucous adenocarcinoma cells (MGC 80-3), which werec ultured with nitrous oxide under high pressure and blocked with overdosage of TdR. Dynamic changes of the mitochondria stained with Rhodamine-123 and succinate dehydrogenase demonstrated by cytochemical method were observed in various phase of the cells at 0, 24, 36, 60 hours after treatment with HPD plus red light. The results showed that mitochonodria of all four phases are of impairment immediately after photoradiation, SDH reactivity is decreased slightly at 24 hours, the activities of SDH is the weakest. As time goes on, we observed that mitochondria gradually recovered to its original structure and then SDH returned to its normal level. The recovery rate of mitochondria and SDH was in the following order, i. e. S, G_1, G_2, andM. The relationship between these changes and cell killing is briefly discussed.

OBJECTIVETo investigate apoptosis in vivo in patients with leukemia at different stages of the first cycle of chemotherapy.

METHODSWe detected apoptosis of HL-60 cells and peripheral blood leukemia cells in 17 patients at different stages, using in situ terminal deoxynucleotidyl transferase (TdT) fluorescence measurement and DNA electrophoresis.

RESULTSWhen HL-60 cells were incubated with 0.02 mg/L harringtonine for 0 to 48 hours, agarose gel electrophoresis showed that DNA ladder patterns became evident only at 12 hour into the treatment. In situ TdT assay showed that apoptotic cells occurred after one hour of the treatment. Apoptotic cells were few (0 - 3.3%) before chemotherapy, but increased substantially (11.4% - 87.5%) during chemotherapy in patients with complete remission (CR) or partial remission (PR). Apoptotic cells were few (0 - 6.1%) during chemotherapy in ten patients with no remission (NR). DNA ladder cannot be detected by agarose gel electrophoresis either before, during or after chemotherapy. Wilcoxon signed rank test shows: P = 0.0012 < 0.01, apoptotic cells during chemotherapy were present in greater quantity than prior to chemotherapy. Wilcoxon rank sum test shows: P = 0.0011 < 0.01, with the median of apoptotic cells during chemotherapy in patients with CR or PR more than with NR.

CONCLUSIONSTdT assay can be used to detect apoptotic cells earlier and more sensitively than DNA agarose gel electrophoresis. In situ TdT assay is useful to detect apoptosis in vivo in the initial phase of chemotherapy for immediate modification of the chemotherapy regimen, whereas electrophoretic analysis is not sensitive enough to detect apoptotic cell in vivo. Where the median of apoptotic cells during chemotherapy in patients with CR or PR were greater than with NR, only effective drug therapy could induce apoptosis.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; DNA Damage ; Female ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; pathology ; Male ; Middle Aged