Objective To investigate the in vitro effects of levofloxacin combined with fosfomycin upon 30 strains of clinical isolates of Staphylo-coccus aureus (15 strains methicillin resistant and 15 strains methicillin sensitive Staphylococcus aureus ) . Methods A checkerboard method that adhered to the recommendations of the National Committee for Clinical Laboratory Standards was applied to assess the synergism effect between levofloxacin and fosfomycin. The FIC index was calculated according to the results. Results The MIC_(50) was reduced significantly for the combination of levofloxacin plus fosfomycin against Staphylococcus aureus. The FIC indexes of methicillin sensitive Staphylococcus aureus (MSSA ) less than 0. 5, from 0. 5 to 1, from 1 to 2, more than 2 were 36. 4% ,63. 6% ,0,0 respectively. The FIC indexes of methicillin resistant Staphylococcus aureus ( MRSA) less than 0. 5 ,from 0. 5 to 1 ,from 1 to 2,more than 2 were 81. 8% , 18. 2% ,0,0, respectively. Conclusion In vitro the combination of subinhibitory concentration of levofloxacin and fosfomycin presented synergistic and additive effect There were no antagonism.

coefficient of variation, corneal endothelium hexagonal cell ratio, anterior corneal surface curvature ratio of horizontal(HK) and vertical curvature(VK) were not statistically significant before and after wearing for 6mo, 1, and 2a (P > 0. 05). The uncorrected visual acuity increased significantly, and the diopter decreased significantly after their wearing (P<0.05). There was no significant difference in axial length after wearing OK lens for 6mo,1,and 2a (P>0.05). CONCLUSION: The orthokeratology lenses can significantly increase uncorrected visual acuity and improve refractive power for juvenile myopia without severe corneal or conjunctival complications occurred, which has little influence on corneal endothelial cells and corneal thickness with a certain degree of safety.

OBJECTIVETo investigate the expressions of TopI gene in small cell lung cancer cell line H446, and explore the influence of TopI on the chemosensitivity of the cell line to topotecan (TPT).

METHODSWestern blot was performed to detect the TopI expression in H446 cells. Lipofectamine 2000 was used for the transient transfection of H446 cells by siRNA, and the transfection efficacy was detected. TopI mRNA was analyzed by quantitative RT-PCR and TopI protein was detected by Western blot to selected effective siRNA. The drug-sensitivity to topotecan (TPT) was evaluated by MTT assay.

RESULTSTopI gene was expressed in H446 cells. Lipofectamine 2000 mediated the siRNA effectively (88.67%). Compared with its parental cells, RT-PCR results showed that TopI mRNAs in transfected cells were reduced by (95.7 +/- 1.6)%, (90.8 +/- 1.6)%, (96.1 +/- 2.7)% and (96.3 +/- 1.8)%, respectively, and decreased significantly at protein level. By MTT assay, the inhibition rate of TPT to H446 cells transfected by siRNA was lower than that of control group at same concentrations (P < 0.01).

CONCLUSIONsiRNAs can silence the expression of TopI and decrease the drug-sensitivity of H446 cells to TPT.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type I ; genetics ; metabolism ; Down-Regulation ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Topotecan ; pharmacology ; Transfection

OBJECTIVETo investigate the prognostic factors of small cell lung cancer (SCLC) and establish a reliable model of clinical prognostic index.

METHODSKaplan-Meier and Cox regression were used to analyze the relationship between survival time and prognostic factors in 60 cases of SCLC. The prognostic factors included clinical and laboratory parameters, serum cytokeratin fragment 19 (CYFRA21-1), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).

RESULTSKaplan-Meier analysis showed that poor prognosis was in patients with KPS < 80 or extensive disease and unrelated to other clinical parameters such as age, sex and smoking index, and in patients with serum NSE > 30 micro g/L, CEA > 5.0 micro g/L, CA125 > 37 KU/L and sIL-2R > 500 KU/L. Serum IL-2 and CYFRA21-1 were also elevated, but had no significant prognostic value. Multivariate analysis indicated that serum NSE, stage and treatment of disease were independent prognostic factors. The three prognostic factors enabled establishment of a prognostic index (PI) based on a simple algorithm: PI = NSE (0 if < or = 30 micro g/L, 1 if > 30 microg/L) + stage (0 = LD, 1 = ED) + CEA (0 if < or = 5.0 microg/L, 1 if > 5.0 microg/L).

CONCLUSIONThe stage of disease, systemic treatment and the level of serum NSE are independent prognostic factors. Without considering the influence of treatment-related factors on survival, the levels of serum CEA, NSE and stage of disease before treatment are significant independent prognostic factors. PI calculated on the basis of CEA, NSE and stage is recommended to predict the survival of SCLC.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Brain Neoplasms ; secondary ; Carcinoma, Small Cell ; mortality ; secondary ; therapy ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; secondary ; Lung Neoplasms ; mortality ; pathology ; therapy ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Survival Rate

OBJECTIVETo investigate the prognostic factors in non-small cell lung cancer (NSCLC) at stage III and IV and establish a reliable model of clinical prognostic index.

METHODSKaplan-Meier and Cox regression were used to analyze the relationship between the prognostic factors and survival time in 114 cases of NSCLC. The prognostic factors included clinical-pathological features and serum levels of cytokeratin fragment 19 (Cyfra21-1), CEA, neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).

RESULTSKaplan-Meier analysis showed that KPS, sex, disease stage, treatment, Cyfra21-1, sIL-2R and CA125 were related to prognosis. Multivariate analysis indicated that Cyfra21-1, stage and treatment were independent prognostic factors. When Cyfra21-1 > 3.5 mg/L, stage IV and chemotherapy < 3 cycles, the relative risk (RR) was 1.691, 2.229 and 3.035, respectively. In patients given 3 or more cycles of chemotherapy, serum Cyfra21-1, sIL-2R and stage at diagnosis were significantly independent prognostic factors. Three of these prognostic factors were used to establish a prognostic index (PI) model based on a simple algorithm: PI = Cyfra21-1 + sIL-2R + stage. The median survival period of patients with 3 or more cycles of chemotherapy were 18 months if PI = 0, 8 months if PI = 1 or 2, and 5 months if PI = 3.

CONCLUSIONThe serum Cyfra21-1, sIL-2R and disease stage in unresectable NSCLC were independent prognostic factors. PI calculated on the basis of Cyfra21-1, sIL-2R and stage is recommended to predict the survival period of NSCLC.

Antigens, Neoplasm ; blood ; Biomarkers, Tumor ; blood ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; mortality ; pathology ; Female ; Follow-Up Studies ; Humans ; Keratin-19 ; Keratins ; Lung Neoplasms ; drug therapy ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Receptors, Interleukin-2 ; blood ; Survival Rate

OBJECTIVEThis article was to explore the impact of temperature on hepatitis B virus infectivity.

METHODSHBV positive serum with a HBV DNA titer of 1.33 × 10(8) copies/ml was aliquots into 23 Ep tubes with 1.5 ml, 100 µl in one tube.15 tubes were incubated at 37, 56 and 65°C for 0, 30, 60, 120 and 600 minutes, respectively. The other 8 tubes were incubated at 98°C for 0, 5, 10 and 30 minutes, respectively. Post-treated serum at all time points were selected to infect HepG-2 cell. When 18 hours after infection, these cells were extensively washed with phosphate buffered saline. Cells were harvested after the addition of fresh culture medium to culture cells for 48 hours. HBV DNA was detected by FQ-PCR.

RESULTSHBV DNA was detected in cells that were infected by serum at 37°C and 56°C for 30, 60, 120 and 600 minutes, respectively. The titers for the cells incubated at 37°C were (4.85 ± 1.71) × 10(5), (3.85 ± 1.76) × 10(5), (1.67 ± 0.67) × 10(5), (7.86 ± 1.03) × 10(4) copies/ml, and those for the cells incubated at 56°C were (4.01 ± 0.16) × 10(5), (9.77 ± 0.97) × 10(4), (6.36 ± 0.65) × 10(4), (5.05 ± 0.24) × 10(3) copies/ml at different incubation time points. For the cells incubated at 65°C for 60 and 120 minutes, HBV DNAs were (5.15 ± 7.28) × 10(3) and (7.56 ± 10.60) × 10(2) copies/ml, respectively, which were much lower than those in the controls cells ((6.79 ± 1.48) × 10(5) copies/ml). The results of HBV DNA were different (F = 104.4, P < 0.001) in groups treated with different temperature, and results of HBV DNA were also different (F = 144.0, P < 0.001) in groups processed for different period of time. Temperature and processing time had interaction (F = 23.6, P < 0.001). After heating at 98°C for 10 minutes and boiling for 5 minutes, the HBV DNA copy number ((3.02 ± 4.26) × 10(2), (4.31 ± 6.09) × 10(2) copies/ml) in infected cells decreased by about 10 folds than that in the control group ((6.79 ± 1.48) × 10(5) copies/ml). HBV DNAs were not detected in cells that were infected by serum which was heated at 98°C for 30 minutes and boiled for 10 minutes.

CONCLUSIONThe infectivity of HBV serum in vitro was relatively stable at low temperature, and it would lose its infectivity in short period of time at high temperature.

Hep G2 Cells ; Hepatitis B virus ; pathogenicity ; physiology ; Hot Temperature ; Humans ; Serum ; virology

This study was aimed to investigate the effect of MK886, a mPGES-1 inhibitor, on apoptosis and drug resistance of leukemia HL-60/A cell line. Expression of mPGES-1 was assayed by QT-PCR and Western blot. The effect of MK886 on HL-60/A cell proliferation was assayed by CCK-8 method, and flow cytometry was used to detect cell apoptosis. The expression of Akt and P-Akt was detected by Western blot. PGE2 was measured by ELISA. Effect of MK886 (10 µmol/L) on the chemotherapeutic sensitivity of HL-60/A cells and expression of mdr-1 mRNA and P170 protein were investigated too. The results indicated the expression of mPGES-1 was higher in HL-60/A cells. MK886 inhibited HL-60/A cell proliferation and induced apoptosis in a time- and concentration-dependent manner. Expression of mPGES-1 and P-Akt and synthesis of PGE2 decreased significantly. MK886 reduced expression of mdr-1 and P170 protein and enhanced the sensitivity of HL-60/A cells to chemotherapeutic drugs. It is concluded that MK886 can inhibit HL-60/A cell proliferation, induce apoptosis and enhance sensitivity to chemotherapeutic drugs, the mechanism of which possibly associates to down-regulation of mPGES-1/PGE2 synthesis, reduction P-Akt expression and decreasing mdr-1 and P170 protein expression.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Indoles ; pharmacology

OBJECTIVETo explore the effects of chloride channels on the regulation of platelet cytoplasmic free calcium concentration ([Ca2+]i) and platelet aggregation (PAG).

METHODSFreshly separated platelets were activated by thrombin. Chloride channel blockers DIDS or NFA and calcium channel blockers SK&F96365 or nifedipine were added to study the effects on platelet [Ca2+]i and PAG by a single reagent or the combination of reagents and find out the interactions among DIDS, NFA, SK&F96365 and nifedipine.

RESULTSBoth DIDS and NFA could inhibit the thrombin (1 U/ml) induced PAG in a dose-dependent manner, whereas had little effect on resting [Ca2+]i. As compared with the control group, DIDS, SK&F96365 and Nifedipine could significantly reduce the PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05). The combination of DIDS and SK&F96365 had greater effects in reducing the PAG, Ca2+ release and Ca2+ influx than either reagent alone (P < 0.05). The combination of DIDS and nifedipine also had greater effect than each alone in reducing Ca2+ release (P < 0.05). The combination of NFA and SK&F96365 weakened each other's effect on Ca2+ release (P < 0.05), while NFA and nifedipine weakened each other's effects on PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05).

CONCLUSIONDIDS and NFA have no effect on the resting [Ca2+]i and the leak calcium influx of platelet. DIDS can inhibit the Ca2+ release, Ca2+ influx and PAG of platelet induced by thrombin, while NFA can only inhibit the Ca2+ release. The chloride channel and calcium channel blockers have interactions in affecting resting [Ca2+]i and PAG of platelet. The opening of chloride channel can influence the cellular calcium movement of platelet.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid ; pharmacology ; Adult ; Blood Platelets ; cytology ; drug effects ; metabolism ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; physiology ; Cytoplasm ; drug effects ; metabolism ; Drug Interactions ; Humans ; Imidazoles ; pharmacology ; Nifedipine ; pharmacology ; Niflumic Acid ; pharmacology ; Platelet Aggregation ; drug effects ; Thrombin ; pharmacology

OBJECTIVETo investigate the effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms.

METHODSHL-60/HT cells were derived from HL-60 cells induced by harringtonine (HT) in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27(Kip1) expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry.

RESULTSThe multidrug-resistant HL-60/HT was acquired, which showed a stable drug-resistant index with increased IC(50) of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27(Kip1) in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells (P<0.05). The expressions of P27(Kip1) and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that of valproic acid or Ara-C alone in HL-60/HT cells and HL-60 cells (q=1.37 and 1.51, respectively). HL-60/HT and HL-60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27(Kip1) and the G(1)-phase cells (P<0.05), but the expression of P170 underwent no significant changes (P>0.05).

CONCLUSIONHL-60/HT cells have lower P27(Kip1) expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27(Kip1) expression and causing cell cycle arrest in G(1) phase.

Antineoplastic Agents ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Cytarabine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Glycoproteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Valproic Acid ; pharmacology

OBJECTIVETo study the anatomy of lateral crural skin flap nourished by cutaneous branches of peroneal artery and its clinical application as vascularized skin flap transfer.

METHODSIn 20 cadavers specimen with 40 lower limbs, the cutaneous branches of the peroneal artery were dissected and their measurements were recorded. In the other 30 adult legs, their perforating points of the cutaneous arteries of peroneal artery were detected with supersonic Doppler flow meter. With the aid of anatomic and supersonic Doppler flow meter study, vascularized transfer of lateral crural skin flap pedicled by cutaneous branches of peroneal artery were successfully performed in 21 clinical cases.

RESULTSIn altogether 40 legs studied, 140 cutaneous branches were found. One to seven branches were found on one specimen, the average was 3.5 branches, in one leg was a high perforating skin branch. The perforating points of the cutaneous branches were mostly (76% cases) appeared within 7 - 21 cm length below the protruding point of head of fibula. The external diameter of the thickest cutaneous branch of each leg was (1 .4 - 2.9) mm, (1.8 +/- 0.4) mm, while the external diameters of two vena concomitants were (3.0 +/- 0.5) mm and (2.4 +/- 0.4) mm. 145 artery perforating points in 30 legs were detected by Doppler, with an average points of 4.8. The skin flaps taken in the 21 clinical cases were 5.0 cm x 3.5 cm - 28 cm x 11 cm in size. All the transferred free flaps survived uneventfully.

CONCLUSIONSThe lateral crural skin flap is nourished by a variable number of cutaneous branches of peroneal artery. The main branch can meet the demand of microvascular anastomosis. The free transfer of lateral crural flap by anastomosis of cutaneous branch of peroneal artery is superior to lateral skin flap transfer by anastomosis of main trunk of peroneal artery with the merit of simple procedure, minimal trauma and more physiological circulation established.

Adult ; Aged ; Arteries ; anatomy & histology ; surgery ; Humans ; Leg ; blood supply ; Middle Aged ; Skin ; anatomy & histology ; blood supply ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply ; Young Adult