Intrapleural chemo- and hyperthermotherapies for malignant pleural effusion: a randomized prospective study.

OBJECTIVE: The current prospective randomized study was designed to evaluate the safety and efficacy of combined intrapleural cisplatin and OK-432 (picibanil) plus hyperthermotherapy in patients with malignant pleural effusion (MPE). METHODS: A total of 358 patients with MPE due to end-stage malignancies were enrolled and randomly divided into two groups, A and B: the intrapleural combination of cisplatin and OK-432 with hyperthermotherapy (n = 179) or without hyperthermotherapy (n = 179), respectively. Mild toxicities such as nausea, vomiting or anorexia, bone marrow depression, and pyrexia were similar in both groups. RESULT: Patients in Group A (with hyperthermotherapy) showed a significantly higher overall response (93.4%) compared to those in Group B (79.8%, χ(2) = 43.11, p < .05). The median survival time for patients in Group A and Group B were 8.9 and 6.2 months, respectively (p > .05). After treatment, the quality of life scores were significantly increased in both groups as compared to prior treatment (p < .05). CONCLUSION: In conclusion, our study suggests that combined intrapleural cisplatin and OK-432 followed by hyperthermotherapy are more effective in the control of MPE and improve patients' quality of life.

[Endostatin in different administration routes combined with adriamycin chemotherapy in the treatment of liver cancer xenograft in mice].

OBJECTIVE: To study the antiangiogenetic and tumor inhibitory effects of endostatin (Es) by intratumoral versus intravenous administration combined with adriamycin (Adm) for treatment of transplanted tumor in mice. METHODS: Forty mice were subjected to subcutaneous implantation of H22 cells and randomly divided into 4 groups by the body weight when the tumor diameter reached 1 cm, namely the control group (with intratumoral and intravenous injection of normal saline), Es intratumoral group (with intratumoral injection Es and intraperitoneal Adm injection), Es vein group (with intravenous Es injection and intraperitoneal Adm injection), and Adm group (with intratumoral saline injection and intraperitoneal Adm injection). The tumor volumes and tumor inhibition rates were calculated, and the expression of vascular endothelial growth factor (VEGF) and the microvessel density (MVD) of the tumors were examined, with the survival time of the mice also observed. RESULTS: The tumor volume was smaller in Es intratumoral group than in the other groups (P<0.05). The expression of VEGF and M VD in Es intratumoral group was significantly decreased as compared with that in the other groups (P<0.05). The survival time was significantly longer in Es intratumoral group and Es vein group than in the other groups (P<0.05), but showed no significant difference between Es intratumoral group and Es vein group (P>0.05). CONCLUSION: In combination with Adm regimen, Es given intratumoral injection produces better effect than intravenous Es injection against angiogenesis and tumor growth, no significant difference can be found in the survival time between them.

[Effect of monoclonal antibodies against LI-cadherin on the proliferation of human hepatocellular carcinoma cells].

OBJECTIVE: To obtain monoclonal antibodies (mAb) against LI-cadherin and investigate their effects on the proliferation of human hepatocellular carcinoma cells. METHODS: Balb/c mice were immunized with recombinant LI-cadherin, and hybridoma cell lines secreting monoclonal antibodies against LI-cadherin were established with routine cell fusion and subcloning approach. The specificity of these mAbs was determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The effect of the mAbs obtained on the growth of HepG2 cells was assessed using inverted microscope and MTT assay. RESULTS: Two hybridoma cell lines (F001 and F002) stably secreting specific mAbs were obtained. Western blot analysis showed that the two antibodies specifically recognized LI-cadherin antigen derived from human eucaryotic cells or tissue. Treatment of the HepG2 cells with the mAbs resulted in reduced viable cell number and changes in the cell morphologies, and the two mAbs inhibited the proliferation of HepG2 cells in a concentration-dependent manner (P<0.05). CONCLUSION: The two specific mAbs obtained can inhibit the proliferation of HepG2 cells in vitro, which facilitates further study of the relationship between LI-cadherin and tumors.

[Effect of KDR recombinant adenovirus containing double suicide gene on the proliferation of human colon adneocarcinoma SW620 cells].

OBJECTIVE: To study the effect of adenovirus (Ad)-mediated fusion gene system driven by the KDR promoter on the proliferation of human colon adenocarcinoma SW620 cells. METHODS: The KDR-expressing SW620 cells and LS174T cells not expressing KDR were both infected with AdEasy-KDR-CDglyTK followed by treatment with the prodrugs 5-FC and/or ganciclovir at different concentrations. The effect of the transfection on the cell proliferation was evaluated. RESULTS: The expression of green fluorescent protein (GFP) was observed in 95% of the infected SW620 and LS174T cells with a multiplicity of infection (MOI) of 100. Significant difference was not founded in the growth of SW620 and LS174T cells with or without the transfection. The infected SW620 cells exhibit high sensitivity to the prodrugs, but the infected LS174T cells did not (P<0.01). The CDglyTK fusion gene produced much stronger killing effect of on the target cells than either of the single suicide genes (P<0.01). CONCLUSION: CDglyTK fusion gene system driven by the KDR promoter selectively kills the KDR-CDglyTK SW620 cells and inhibits the cell proliferation.

[Synthesis and characterization of folic acid-conjugated chitosan nanoparticles as a tumor-targeted drug carrier].

OBJECTIVE: To synthesize and characterize paclitaxel (PTX)-loaded folate-conjugated chitosan (FA-CTS/PTX) nanoparticles and evaluate its cytotoxicity in vitro. METHODS: CTS/PTX and FA-CTS/PTX nanoparticles were prepared using reductive amidation and ionic gelation of chitosan with tripolyphosphate anions (TPP). The particle size was determined by laser scattering and the morphology observed using transmission electron microscopy, and the PTX content in the nanoparticles was determined using ultraviolet spectrophotometer at 227 nm. The in vitro cytotoxicity of the nanoparticles against HeLa cells was evaluated by MTT assay. Fluorescence microscopy was used to observe the HeLa cells incubated with FA-chitosan nanoparticles in the presence or absence of folic acid in the culture medium. RESULTS: PTX loading did not cause adhesion of the FA-CTS nanoparticles, which presented with uniform spherical morphology with an average diameter of 282.8 nm. The loading and encapsulation efficiencies of FA-CTS/PTX were 9.0% and 75.4%, respectively. The FA-CTS nanoparticles showed a greater extent of intracellular uptake in the absence of folic acid, indicating that the cellular uptake of the nanoparticles occurred through endocytosis mediated by the folate receptors. The PTX-loaded FA-CTS nanoparticles exhibited potent cytotoxicity against HeLa cells, an effect 2- to 3-fold stronger than that of PTX-loaded CTS nanoparticles. CONCLUSION: FA-CTS can be a promising drug carrier with high efficiency in condensing drug, good tumor-targeting ability and low cytotoxicity.

[Preparation of long-circulating nolatrexed dihydrochloride liposomes and its antitumor activity].

OBJECTIVE: To prepare long-circulating liposome (LCL) for sustained release of nolatrexed dihydrochloride and evaluate the effect of this preparation against the growth of hepatocarcinoma cells in mice. METHODS: The long-circulating nolatrexed dihydrochloride liposome was prepared by film dispersion-extrusion combined with ammonium sulphate gradient method. Amphipathic polyethylene glycol-distearoyl phosphatidylethanolamine (PEG-DSPE) was added to modify the property of the liposome membrane. The drug entrapment efficiency of the nolatrexed dihydrochloride-containing liposome was determined using UV detector with Sephadex G50. Electron microscopy and laser particle analyzer were employed to determine the size of the nolatrexed dihydrochloride liposome. For in vivo evaluation of the effect of the liposomal preparation, H22 mouse hepatoma carcinoma cells were transplanted subcutaneouly in mice in the axillary region of the right hind limb to induce growth of solid tumors, which were evaluated for tumor weight inhibition rate and tumor volume changes after administration of the LCL preparations. RESULTS: The mean diameter of the long-circulating nolatrexed dihydrochloride liposomes was 109 nm, with an entrapment efficiency of 68.5%. In vivo antitumor experiment showed that both the common liposomal and LCL preparations of nolatrexed dihydrochloride produced antitumor effect in vivo, and the latter had weaker antitumor effect than free and common liposomal preparation of nolatrexed dihydrochloride, but in the long term, the LCL preparation showed stronger antitumor effect with a tumor weight inhibition rate of 41.68%. CONCLUSION: LCL allows sustained release of nolatrexed dihydrochloride in vivo, and may effectively lengthen the relatively short half life of this drug after administration.

Nonvariational time-dependent multiconfiguration self-consistent field equations for electronic dynamics in laser-driven molecules.

A time-dependent multiconfiguration self-consistent field (TDMCSCF) scheme is developed to describe the time-resolved electron dynamics of a laser-driven many-electron atomic or molecular system, starting directly from the time-dependent Schrodinger equation for the system. This nonvariational formulation aims at the full exploitations of concepts, tools, and facilities of existing, well-developed quantum chemical MCSCF codes. The theory uses, in particular, a unitary representation of time-dependent configuration mixings and orbital transformations. Within a short-time, or adiabatic approximation, the TDMCSCF scheme amounts to a second-order split-operator algorithm involving generically the two noncommuting one-electron and two-electron parts of the time-dependent electronic Hamiltonian. We implement the scheme to calculate the laser-induced dynamics of the two-electron H2 molecule described within a minimal basis, and show how electron correlation is affected by the interaction of the molecule with a strong laser field.

[Expression of DNA transcription- and repair-related genes in cisplatin-resistant human ovarian carcinoma cell line COC1/DDP].

OBJECTIVE: To study the molecular mechanism underlying cisplatin resistance in ovarian carcinoma by detecting the expressions of DNA transcription- and repair-related genes in cisplatin-resistant human ovarian carcinoma COC1 cell line. METHODS: The differential expression of DNA transcription- and repair-related genes between the parental COC1 and cisplatin-resistant COC1/DDP cell line was determined using cDNA microarray. RESULTS AND CONCLUSION: Compared with COC1 cells, 143 genes in COC1/DDP cells showed significant differential expression, among which 20 were DNA transcription- and repair-related genes including 13 significantly up-regulated genes and 7 down-regulated ones. Abnormality of DNA transcription and repair might be involved in the development of cisplatin resistance in COC1/DDP cells.

[Expression of glucosylceramide synthase mRNA in vincristine-resistant KBV200 cell line in association with multidrug resistance].

OBJECTIVE: To investigate the relationship between the expression of glucosylceramide synthase (GCS) mRNA in vincristine-resistant KBV(200) human cancer cell line and multidrug resistance (MDR) of the cancer cells. METHODS: Reverse transcriptional polymerase chain reaction (RT-PCR) was employed to analyze the differential expression of GCS mRNA between KBV(200) and KB cell lines and the changes in the mRNA expressions of GCS and mdr1 gene in KBV(200) cells after reversion of MDR. The effects of de-phenyl-z-palmaitoylamino-3-morpholine-1-propanol (DL-PPMP) and verapamil in reversing MDR of the cells were evaluated by MTT assay. RESULTS: KBV(200) cells exhibited significantly increased expressions of GCS and mdr1 gene, whereas mdr1 gene failed to be detected in the parental KB cells. DL-PPMP within the concentrations ranging from 5 to 25 micromol/L could inhibit the expression of GCS gene, with the maximum inhibition achieved at 25 micromol/L. Verapamil at the concentration of 10 micromol/L was already sufficient to induce inhibition of GCS expression in KVB(200) cells, which was more manifest with the concentration of 15 micromol/L. DL-PPMP and verapamil were found to inhibit mdr1 gene expression in KBV(200) cells at the mRNA level, and complete inhibition occurred after a 48-hour DL-PPMP treatment at 25 micromol/L. CONCLUSION: The inhibition of GCS and mdr1 gene expressions is positively correlated with the concentrations of DL-PPMP and verapamil, which can reverse MDR by inhibiting synthesis of GCS and mdr1 gene, indicating the positive correlation between the expression of GCS gene and MDR in KBV(200) cells. GCS gene might play an important role in MDR during tumor progression.

[Morphological observation on NK/LAK cell-mediated lysis of human oral carcinoma cells].

OBJECTIVE: To understand the relation between cytotoxic activity of immunologic effector cells and multidrug resistance of the tumor cells. METHODS: Continuous observation of the morphological changes and MTT colorimetry were employed to evaluate the cytotoxic activity of lymphokine-activated killer (LAK) cells and natural killer (NK) cells against multidrug-resistant (MDR) human oral carcinoma cell line-KBV200 (before and after reversal of MDR) and parental drug-sensitive cell line KB. The morphologic changes of LAK cells and the 3 target cell lines were observed continuously under inverted microscope 3 h after co-culture of LAK cells with one of three target cell lines respectively. The lysis rates of three tumor cell lines in response to co-culture with LAK or NK cells were determined using MTT colorimetry. RESULTS: In comparison with the parental drug-sensitive cell line KB, both KBV200 and its reserved cell line by verapamil (KBVV) showed earlier adherence and greater number of cells lysed by LAK. In MTT colorimetry assay, the cytotoxicity of both LAK and NK cells against the 3 cell lines was associated with the effector-to-target (E/T) cell ratio; the lysis rates of KBV200 and reversed KBV200 cells by verapamil in response to LAK and NK cells were higher than that of KB cells (P<0.05), but KBV200 and KBVV did not significantly differ (P>0.05). At the same E/T ratio, LAK cells possessed stronger cytotoxicity than NK cells against all the tumor cell lines (P<0.05). CONCLUSIONS: Immunologic effector cells possess strong cytotoxic activity against multidrug-resistant cell line KBV200. Modulation of MDR does not decrease the cytotoxic activity of the immunologic effector cells. The results of this study suggest that adoptive cell immunotherapy with immunologic effector cells may be of value in controlling the progress of MDR tumors.

[Comparative study of three local ablation methods for transplanted hepatocellular carcinoma in mice].

OBJECTIVE: To observe the effects of three commonly used local ablation methods in the treatment of transplanted hepatocellular carcinoma in mice to provide experimental evidence for treating hepatocellular carcinoma that defies surgical removal. METHODS: Mouse models of hepatocellular carcinoma were established by means of subcutaneous transplantation, and treatment results of the three ablation methods, namely percutaneous ethanol injection therapy (PEIT), percutaneous acetic acid injection therapy (PAIT) and percutaneous local cryosurgery therapy (PLCT), were compared. RESULTS: The tumor inhibition rates of PLCT, PAIT and PEIT were 99.2%, 85.3%, and 72.8%, respectively. The rates for complete necrosis of the tumors were 100% (5/5), 60% (3/5), and 40% (2/5), with the survival time of 88.11+/-5.67, 86.67+/-7.26, and 72.89+/-12.86 days respectively. CONCLUSIONS: All of the three local ablation methods can inhibit the tumor growth to various degrees and prolong the survival time of the tumor-bearing mice. PLCT may yield relatively better result than the other two methods.

Synthesis of targeted drug delivery system for fluorouracil using sulfadiazine as the carrier.

OBJECTIVE: To synthesize a targeted drug delivery system for 5-fluorouracil (5Fu) using sulfadiazine (SF) as a carrier with reduced side-effects and strong antitumor activity. METHODS: SF-poly (ethylene glycol) (PEG) conjugate was initially synthesized. 5Fu was subjected to reaction with trichloromethyl chloroformate to prepare chloroformyl 5Fu, which was linked to a spacer hydroxyl group of PEG that served as a macromolecular linking arm between SF and 5Fu. The content of 5Fu in the conjugate was determined by ultraviolet spectrophotometry. Spectrum of ultraviolet and infrared along with differential scanning calorimetry were employed to identify the structure of the conjugate of SFPEG-end capped 5Fu. RESULTS: The drug loading content of the conjugate was 3.2 %, and structural analysis confirmed the linkage between 5Fu and SF via PEG. CONCLUSION: Targeted drug delivery system for 5Fu using SF as a carrier has been successfully synthesized by this means.

[Combination chemotherapy with Taxol and cisplatin for 57 patients with non-small cell lung cancer by intraartery and intravenous infusion].

BACKGROUND & OBJECTIVE: Although previous studies showed that intravenous infusion chemotherapy combined with Taxol and cisplatin (DDP) can improve the effects of the treatment of advanced non-small cell lung cancer (NSCLC), the results remained unsatisfactory because of the lower concentration of local chemotherapeutic agent. This study was designed to administrate the chemotherapeutic agents through the local artery and to evaluate the efficacy of the intraartery (IA) and the intravenous (i.v.) infusion chemotherapy combined Taxol with cisplatin (DDP) in the treatment of NSCLC. METHODS: Fifty-seven patients with advanced NSCLC were enrolled and randomized into two groups. Group A (27 patients) was given by IA chemotherapy consisting of Taxol 135 mg/m2 and DDP 80 mg/m2 within 3 hours in the first day. Group B (30 patients) was given by i.v. chemotherapy consisting of Taxol 135 mg/m2 and DDP 80 mg/m2 within 3 hours in the first day. The patients in two groups were repeatedly treated every 3-4 weeks for at least 2 cycles. RESULTS: The recent effect was evaluated by the CT or X-ray results. Of the 57 patients, 1 case acquired complete response, 29 partial response. The overall response rate was 52.63% with 66.67% (18/27) in group A and 40.00% (12/30) in group B. The median TTP was 5.5 months in group A and 4.0 months in group B. The median survival time was 13.5 months in group A and 10.5 months in group B. The 0.5, 1, 2, and 3-year survival rates were 85.18%, 66.67%, 48.15%, and 18.51% months in group A, respectively. The 0.5, 1, 2, and 3-year survival rates were 70.00%, 53.33%, 30.00%, and 6.67% months in group B, respectively. The main side effects were myelosuppression, gastrointestinal reactions and peripheral neuropathy. They could be effectively treated with large amount of vitamines, antacid, guastil, white blood cell stimulating drugs and liquid compensation. CONCLUSION: The artery infusion Taxol and DDP chemotherapy is one of the useful methods in the treatment of NSCLC patients. The local response rate in the patients treated by IA may be higher than those treated by i.v.