Drug Combination Group Research Articles

Mining the Synergistic Core Allosteric Modules Variation and Sequencing Pharmacological Module Drivers in a Preclinical Model of Ischemia.

Identifying the variation of core modules and hubs seems to be critical for characterizing variable pharmacological mechanisms based on topological alteration of disease networks. We first identified a total of eight core modules by using an approach of multiple modular characteristic fusing (MMCF) from different targeted networks in ischemic mice. Interestingly, the value of module disturbance intensity (MDI) increased in drug combination group. Second, we redefined a weak allosteric module and a strong allosteric module. Then, we identified 15 pharmacological module drivers (PMDs) by leave‐one‐out screening with a cutoff of two folds, which were at least, in part, validated by expression and variation of topological contribution. Finally, we revealed the fusional and emergent variation of PMD in core modules contributing to multidimensional synergistic mechanism in ischemic mice and rats. Our findings provide a new set of drivers that might promote the pharmacological modular flexibility and offer a potential avenue for disease treatment.

Experimental study on oral sulfhydryl as an adjuvant for improving nitrate ester tolerance in an animal model.

As an initial step in exploring the feasibility of oral sulfhydryl as an adjuvant for improving nitrate ester tolerance, this study was designed to experimentally test the adjuvant therapy in a rabbit model of atherosclerosis (AS). New Zealand white rabbits with induced AS were randomly divided into four groups: AS group, AS + nitrate ester group, AS + nitrate ester tolerance group, and AS + drug combination group. Additionally, four equivalent groups with healthy New Zealand white rabbits without AS were also conformed. After feeding the animals for 5 days, the concentrations of superoxide anion (•O2-), superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), and endothelin-1 (ET-1) in blood and the relaxation response of the aortic ring were determined in each subject. The vascular plaques in different treatment groups were assessed by Hematoxylin and eosin (HE) staining to investigate the therapeutic value of sulfhydryl as coadjuvant for improving nitrate ester tolerance, and changes in blood vessels in different treatment groups were studied by immunohistochemical assays. Our results showed no significant differences through time in the concentrations of •O2-, SOD, MDA, NO, ET-1 between the healthy control and the nitrate ester groups (p > 0.05). The levels of SOD and MDA in the nitrate ester tolerance group increased with time, however, the levels of •O2-, NO and ET-1 decreased gradually (p < 0.05). The NO, •O2- and ET-1 levels in both the AS and AS + nitrate ester tolerance groups were significantly decreased, but SOD and MDA were significantly increased (p < 0.05). SOD and MDA in the AS + nitrate ester group decreased gradually with time, but •O2-, NO- and ET-1 levels increased (p < 0.05). The levels of SOD and MDA in the AS + drug combination and the drug combination group decreased significantly with time, in contrast, those of •O2-, NO- and ET-1 increased (p < 0.05). The results of HE staining proved that the atherosclerosis model was established successfully. We conclude the use of a sulfhydryl compound as an adjuvant significantly reduced nitrate ester tolerance, and this strategy was safe and looks promising for humans.

Plasma exchange combining with plasma bilirubin adsorption effectively removes toxic substances and improves liver functions of hepatic failure patients.

Hepatic failure (HF) is a kind of complex disease characterizing with liver dysfunction and a few clinical complications. Artificial liver support system (ALSS) has been applied to HF patients to improve dysfunctional liver in recent years. This study aims to evaluate therapeutic effects of ALSS approaches, including plasma exchange (PE), plasma diafiltration (PDF) and plasma bilirubin adsorption (PBA), on liver function of HF patients. This study is a retrospective analysis involving 516 patients diagnosed as HF between February 2014 and February 2015. Patients were randomly divided into PE, PDF, PE plus PBA, and PDF plus PBA group. Meanwhile, single-drug group and combined-drug group were also divided. The liver functions, capability of removing toxic substances and coagulation functions were evaluated both pre-treatment and post-treatment. The side effects and hospital improvement rate were also observed post-treatment. Hospital improvement rate achieves to 69.6%. TBIL levels and MELD scores were significantly decreased post-treatment compared to pre-treatment (p<0.05). PTA values were significantly increased post-treatment compared to pre-treatment (p<0.05). Reduction value in PE+PBA group was significantly higher compared to PE and PDF group (p=0.002, 0.002, respectively). MELD scores were significantly decreased post-treatment compared to pre-treatment in each group (p<0.05). Combined-drug treatment is superior to single-drug treatment for removing toxic substances and improving liver functions. PE treatment, PDF treatment and PE+PBA treatment induced more side effects compared to PDF+PBA treatment. PE combining with PBA plays a better role in removing toxic substances, improving liver functions of HF patients.

DRES-09. THERAPEUTIC TARGETING OF CLK2 AND PI3K/mTOR IN GLIOBLASTOMA STEM CELLS

AbstractKey signaling genes of the PI3K pathway are mutated in a large proportion of human glioblastoma (GBM), leading to persistent activation of the pathway. To determine the efficacy of inhibiting multiple components or the RTK/PI3K pathway, we examined GSK2126458 (PI3K/mTOR inhibitor) in a panel of glioma stem cell (GSC) lines. GSK2126458, a dual phophoinositide 3-kinase/mammalian target of rapamycin, had the lowest IC values in all of the GSCs with an IC below 0.5 μM. We confirmed the efficacy of these inhibitors in two orthotopic xenograft models of glioma. GSK2126458 prolonged survival in GSC272 implanted mice but not in the resistant line GSC7-2. Dual targeting of CLK2 and PI3K was pursued to overcome heterogeneity-related resistance from multiple signaling pathways. The survival of mice orthotopically implanted with CLK2 knockdown cells was significantly prolonged compared to mice implanted with GSC272 and GSC7-2-shRNA control cells. No significant prolongation in survival was observed with CLK2 knockdown combined with GSK212468 but tumor volumes were significantly decreased in the CLK2 knockdown plus drug combination groups with GSK212468 compared to drug treatment alone. Our findings suggest that the combination of CLK2 knockdown with PI3K/mTOR inhibitor may provide an effective therapeutic approach for glioblastoma.

In situ controlled release of stromal cell-derived factor-1α and antimiR-138 for on-demand cranial bone regeneration

Bone regeneration involves complex physiological processes, which is generally regulated and controlled by multiple bioactive molecules. In situ controlled release of combined bioactive factors in a spatiotemporal sequence for adapting the demand of bone regeneration is a desired strategy. In this study, nanoparticle/hydrogel composite system was constructed by incorporating stromal cell derived factor-1α (SDF-1α) and chitosan/tripolyphosphate/hyaluronic acid/antimiRNA-138 nanoparticles (CTH/antimiR-138 NPs) in chitosan/β-sodium glycerol phosphate (CS/GP) hydrogel for rat critical-size calvarial bone regeneration. The fast release of SDF-1α promoted the migration of mesenchymal stem cells (MSCs) for 6 d, while the sustained release of antimiR-138 from the nanoparticle/hydrogel compound enhanced the osteogenic differentiation of MSCs over 21 d. 8 weeks after surgery, calvarial specimens were evaluated by microcomputed tomography (μ-CT), histological analysis and immunohistochemistry. Comparing with blank group and hydrogel group, hydrogels incorporated with SDF-1α and/or CTH/antimiR-138 NPs significantly enhanced bone regeneration (p<0.05). In addition, the expression of collagen type-1 (COL-1), osteopontin (OPN) and osteocalcin (OCN) proteins were enhanced in the combined drug group (incorporated both SDF-1α and CTH/antimiR-138 NPs) in comparison to the hydrogel group. Our research indicated the in situ formation of NPs/hydrogel composite could provide temporal sequence-release of SDF-1α and CTH/antimiR-138 NPs for on-demand MSCs homing and cranial bone regeneration.

Puerarin increases the chemosensitivity of hepatocellular carcinoma cells.

The present study investigated the effect of puerarin (Pu) on the sensitivity of HepG2 human hepatocellular carcinoma (HCC) cells to chemotherapeutic drugs to determine the possible mechanism. HepG2 cells were treated with different concentrations of Pu and cisplatin (CDDP), alone or in combination. MTT assay was used to determine the inhibitory effects of the different drugs on HepG2 cells. Cell morphology was observed by inverted microscopy. The expression of B-cell lymphoma 2 (Bcl-2) and Bax protein was measured by western blot analysis. Pu and CDDP, alone or in combination, inhibited the proliferation of HepG2 cells. The inhibitory effect of CDDP combined with Pu on HepG2 cells was significantly higher than that of the single drug treatments (p<0.01). In addition, compared with the single drug groups, cellular morphology was significantly altered and the apoptotic rate of cells and the expression of Bax protein were significantly increased (p<0.01). However, the expression of Bcl-2 protein was significantly decreased (p<0.01) in the combined drug group. In conclusion, Pu can increase the sensitivity of HCC to chemotherapeutic drugs, enhance the inhibitory effect of chemotherapeutic drugs on cell proliferation and synergistically induce apoptosis of HepG2 cells. The mechanism is likely related to the upregulation of Bax protein and the downregulation of Bcl-2 protein.

Mechanism of Bushen Jianpi decoction in preventing and treating osteoporosis caused by aromatase inhibitors in breast cancer treatment.

Research on the mechanism of Bushen Jianpi decoction (BJD) for preventing and treating osteoporosis caused by aromatase inhibitors (AI) during treatment for breast cancer resection. An ovariectomized mouse model was established using random division into 6 groups: a sham ovariectomized group, a blank control group, a control group, an alendronate group, a BJD group, and a drug combination group. Mice breast cancer cell lines (4T1) were cultured and seeded into the armpits of 6 groups of BALB/c mice. The mouse breast cancer postoperative model was built when resecting the tumor after 3 weeks following seeding tumor. After 1 weeks, the 6 groups of mice were given different drugs. Then the following analyses were made: estradiol (E2) levels and alkaline phosphatase (ALP) levels in the serum; detection of in vitro bone density and calcium and bone phosphorus content; tumor pathology and immunohistochemistry detection. The results suggested that BJD decreased levels of ALP in ovariectomized mice, and there was a trend for improved bone loss. BJD strengthened the trend of alendronate to improve bone loss, improved bone density, bone calcium and phosphorous, and reduced ALP. BJD had a certain role on the promotion of the expression of estrogen receptors (ERs) in the relapse of the tumor tissue. Combined therapy with BJD and alendronate can act synergistically against osteoporosis, which was possibly related to a reduced bone conversion rate through inhibiting bone resorption. BJD may block the MAPK signal pathway in breast cancer cells, increasing the expression of ERs and making cancer cells sensitive to endocrine treatment.

Immunosuppressive Treatment Alters Secretion of Ileal Antimicrobial Peptides and Gut Microbiota, and Favors Subsequent Colonization by Uropathogenic Escherichia coli

Transplant recipients are treated with immunosuppressive (IS) therapies, which impact host-microbial interactions. We examined the impact of IS drugs on gut microbiota and on the expression of ileal antimicrobial peptides. Mice were treated for 14 days with prednisolone, mycophenolate mofetil, tacrolimus, a combination of these 3 drugs, everolimus, or water. Feces were collected before and after treatment initiation. Ileal samples were collected after sacrifice. Fecal and ileal microbiota were analyzed by pyrosequencing of 16S rRNA genes and enumeration of selected bacteria by culture, and C-type lectins were assessed in ileal tissues by reverse transcriptase-quantitative polymerase chain reaction. Prednisolone disrupted fecal microbiota community structure, decreased Bacteroidetes, and increased Firmicutes in the feces. Prednisolone, tacrolimus, and mycophenolate mofetil modified fecal microbiota at the family level in each experimental replicate, but changes were not consistent between the replicates. In ileal samples, the genus Clostridium sensu stricto was dramatically reduced in the prednisolone and combined IS drug groups. These modifications corresponded to an altered ileal expression of C-type lectins Reg3γ and Reg3β, and of interleukin 22. Interestingly, the combined IS treatment enabled a commensal Escherichia coli to flourish, and dramatically increased colonization by uropathogenic E. coli strain 536. IS treatment alters innate antimicrobial defenses and disrupts the gut microbiota, which leads to overgrowth of indigenous E. coli and facilitates colonization by opportunistic pathogens.

Tanshinone IIA combined with adriamycin inhibited malignant biological behaviors of NSCLC A549 cell line in a synergistic way.

BackgroundThe study was designed to develop a platform to verify whether the extract of herbs combined with chemotherapy drugs play a synergistic role in anti-tumor effects, and to provide experimental evidence and theoretical reference for finding new effective sensitizers.MethodsInhibition of tanshinone IIA and adriamycin on the proliferation of A549, PC9 and HLF cells were assessed by CCK8 assays. The combination index (CI) was calculated with the Chou-Talalay method, based on the median-effect principle. Migration and invasion ability of A549 cells were determined by wound healing assay and transwell assay. Flow cytometry was used to detect the cell apoptosis and the distribution of cell cycles. TUNEL staining was used to detect the apoptotic cells. Immunofluorescence staining was used to detect the expression of Cleaved Caspase-3. Western blotting was used to detect the proteins expression of relative apoptotic signal pathways. CDOCKER module in DS 2.5 was used to detect the binding modes of the drugs and the proteins.ResultsBoth tanshinone IIA and adriamycin could inhibit the growth of A549, PC9, and HLF cells in a dose- and time-dependent manner, while the proliferative inhibition effect of tanshinone IIA on cells was much weaker than that of adriamycin. Different from the cancer cells, HLF cells displayed a stronger sensitivity to adriamycin, and a weaker sensitivity to tanshinone IIA. When tanshinone IIA combined with adriamycin at a ratio of 20:1, they exhibited a synergistic anti-proliferation effect on A549 and PC9 cells, but not in HLF cells. Tanshinone IIA combined with adriamycin could synergistically inhibit migration, induce apoptosis and arrest cell cycle at the S and G2 phases in A549 cells. Both groups of the single drug treatment and the drug combination up-regulated the expressions of Cleaved Caspase-3 and Bax, but down-regulated the expressions of VEGF, VEGFR2, p-PI3K, p-Akt, Bcl-2, and Caspase-3 protein. Compared with the single drug treatment groups, the drug combination groups were more statistically significant. The molecular docking algorithms indicated that tanshinone IIA could be docked into the active sites of all the tested proteins with H-bond and aromatic interactions, compared with that of adriamycin.ConclusionsTanshinone IIA can be developed as a novel agent in the postoperative adjuvant therapy combined with other anti-tumor agents, and improve the sensibility of chemotherapeutics for non-small cell lung cancer with fewer side effects. In addition, this experiment can not only provide a reference for the development of more effective anti-tumor medicine ingredients, but also build a platform for evaluating the anti-tumor effects of Chinese herbal medicines in combination with chemotherapy drugs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2921-x) contains supplementary material, which is available to authorized users.

EFEK SINERGIS KLOROKUIN DAN N-ACETYL CYSTEINE TERHADAP PENURUNAN PARASITEMIA DAN PENURUNAN KADAR MALONDYALDEHYDE ERITROSIT MENCIT YANG DIINFEKSI PLASMODIUM BERGHEI

Malaria is still a global health problem around the world particularly in Indonesia. Chloroquine is one of the anti-malarial drugs which is still used in Indonesia. The aim of this research is to know the synergic effect of Chloroquine and N-Acetyl Cysteine towards parasitemia and erythrocyte malondyaldehyde levels in mice infected with Plasmodium berghei. This was an experimental research using Plasmodium berghei and BALB/c strain mice, 6−8 weeks old, with body weight 20−30 grams. After infected with Plasmodium berghei, the mice were divided into eight (8) groups: control, chloroquine, NAC 0.25 mg/gBb; NAC 0.5 mg/gBb; NAC 1 mg/gBb and combined drugs of chloroquine and NAC 0.25 mg/gBb; NAC 0.5 mg/gBb; NAC 1 mg/gBb. On the 3rd, 5th, and 7th day after treatment, the mice were killed and their parasitemia and Malondialdehyde (MDA) levels were measured. By using MANOVA and Tukey HSD test, a signifi cant difference in parasitemia level was found between the control and chloroquine group, as well as the control and combined drug groups. In the NAC groups, there was no decrease. The decrease of MDA level began on the 7th day. It can be concluded that there was a synergic effect between the chloroquine and N-Acetyl Cysteine in decreasing parasitemia and malondyaldehyde levels in mice BALB/c infected with Plasmodium berghei.

Role of high-fat diet on the effect of pioglitazone and melatonin in a rat model of breast cancer.

The risk of cancer may be modulated by drugs with pleiotropic effects and diet has been implicated in the efficacy of treatment. The oncopreventive effects of the antidiabetic drug pioglitazone (PIO) and the anti-insomnia drug melatonin (MT), in vivo, have been proven before, but using a standard-type diet. This study evaluated the impact of a high-fat diet on their efficacy in chemically induced mammary carcinogenesis in Sprague-Dawley rats. Mammary tumours were induced by N-methyl-N-nitrosourea (50 mg/kg, intraperitoneal, on the 41st postnatal day). PIO and MT administration was initiated 11 days before the carcinogen application and lasted until the termination of the experiment at 16 weeks. PIO was administered in a diet (10% fat) at a concentration of 100 ppm and MT was administered in tap water (20 mg/l). PIO, MT and the combination did not significantly alter the basic tumour growth parameters. However, histopathology showed a decrease in the high-grade/low-grade tumour ratio, particularly in animals that received combined treatment (P<0.01). Semiquantitative immunohistochemistry indicated the proapoptotic effect of chemoprevention, particularly in the drug combination group (P<0.01), but no changes in tumour cell proliferation and angiogenesis were recorded. Results were evaluated by one-way analysis of variance or the Mann-Whitney U-test, respectively. PIO and MT, alone or in combination, administered to rats fed a high-fat diet reduced the proportion of high-grade tumours and promoted apoptosis in an in-vivo breast cancer model, although it did not suppress tumour growth. The impact of high dietary fat content on the chemopreventive efficacy of these and other substances should be considered in human studies.

Effect of Fuke Qianjin tablets on pharmacokinetics of Azithromycin in rats with chronic pelvic inflammatory disease

To investigate the effect of Fuke Qianjin tablets on the pharmacokinetics of Azithromycin(AZM) in rats with chronic pelvic inflammation, and evaluate the rationality of the drug combination. The animal models of chronic pelvic inflammatory disease were established by intrauterine injection of mixed bacterial suspension plus mechanic injury in female Sprague-Dawley(SD) rats. The model rats were randomly divided into control group and drug combination group. The rats in control group were given with Azithromycin 10 mg•kg⁻¹•d⁻¹ by intragastric administration, while the rats in drug combination group were given with Fuke Qianjin tablets 1.66 g•kg⁻¹•d⁻¹ by intragastric administration 2 hours after the equivalent dose of azithromycin, once a day, consecutively for 7 days. Plasma samples were collected at different time points and the concentration of AZM in plasma was determined by high performance liquid chromatography coupled with mass spectrometry(HPLC-MS). ADAPT 5.1 software was used to calculate the pharmacokinetic parameters of each group by Inverse Gaussian Function model. SPSS software was used for the statistical analysis of data in each group. The results showed that Fuke Qianjin tablets had no significant effects on the main pharmacokinetic parameters of AZM, including CLt, CLd, MIT and F. The study showed that Fuke Qianjin tablets can be combined with azithromycin for treatment of chronic pelvic inflammation disease.

Combined Effects of Acamprosate and Escitalopram on Ethanol Consumption in Mice.

Major depression is one of the most prevalent psychiatry comorbidities of alcohol use disorders (AUD). As negative emotions can trigger craving and increase the risk of relapse, treatments that target both conditions simultaneously may augment treatment success. Previous studies showed a potential synergistic effect of Food and Drug Administration approved medication for AUD acamprosate and the antidepressant escitalopram. In this study, we investigated the effects of combining acamprosate and escitalopram on ethanol (EtOH) consumption in stress-induced depressed mice. Forty singly housed C57BL/6J male mice were subjected to chronic unpredictable stress. In parallel, 40 group-housed male mice were subjected to normal husbandry. After 3weeks, depressive- and anxiety-like behaviors and EtOH consumption were assessed. For the next 7days, mice were injected with saline, acamprosate (200mg/kg; twice/d), escitalopram (5mg/kg; twice/d), or their combination (n=9 to 11/drug group/stress group). Two-bottle choice limited-access drinking of 15% EtOH and tap water was performed 3hours into dark phase immediately after the daily dark phase injection. EtOH drinking was monitored for another 7days without drug administration. Mice subjected to the chronic unpredictable stress paradigm for 3weeks showed apparent depression- and anxiety-like behaviors compared to their nonstressed counterparts including longer immobility time in the forced swim test and lower sucrose preference. Stressed mice also displayed higher EtOH consumption and preference in a 2-bottle choice drinking test. During the drug administration period, the escitalopram-only and combined drug groups showed significant reduction in EtOH consumption in nonstressed mice, while only the combined drug group showed significantly reduced consumption in stressed mice. However, such reduction did not persist into the postdrug administration period. The combination of acamprosate and escitalopram suppressed EtOH intake in both nonstressed and stressed mice; hence, this combination is potentially helpful for AUD individuals with or without comorbid depression to reduce alcohol use.

Effects of NVP-BEZ235 on the proliferation, migration, apoptosis and autophagy in HT-29 human colorectal adenocarcinoma cells.

The phosphoinositide 3 kinase (PI3K)/Akt/mammalian target of the rapamycin (mTOR) pathway plays a significant role in colorectal adenocarcinoma. NVP-BEZ235 (dactolisib) is a novel dual inhibitor of PI3K/mTOR. The effects of NVP-BEZ235 in human colorectal adenocarcinoma are still unclear. In the present study, we aimed to explore the proliferation, migration, apoptosis and autophagy in HT-29 human colorectal adenocarcinoma cells. HT-29 human colorectal adenocarcinoma cells were treated with NVP-BEZ235 (0, 0.001, 0.01, 0.1, 1 and 3µM) for 24 and 48h, respectively. Cells were also treated with NVP-BEZ235 (0.1µM), DDP (100, 300 and 1,000µM), and NVP-BEZ235 (0.1µM) combined with DDP (100, 300 and 1,000µM) respectively, and cultured for 24h after treatment. MTT assay was utilized to evaluate the effects of NVP-BEZ235 alone or NVP-BEZ235 combined with cis-diamminedichloroplatinum (DDP) on proliferation of HT-29 cells. Cell wound-scratch assay was used detect cell migration. In addition, expression of microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B and LC3B) in HT-29 cells was detected by immunofluorescence at 48h after NVP-BEZ235 (1µM) treatment. Expression of proteins involved in cell cycle and proliferation (p-Akt, p-mTOR and cyclinD1), apoptosis (cleaved caspase-3), and autophagy (cleaved LC3B and Beclin-1) were detected by western blot analysis. NVP-BEZ235 inhibited the proliferation and migration of HT-29 human colorectal adenocarcinoma cells. NVP-BEZ235 decreased protein expression of p-Akt, p-mTOR and cyclinD1, and increased protein expression of cleaved caspase-3, cleaved LC3B and Beclin-1 as the concentrations and the incubation time of NVP-BEZ235 increased. In addition, NVP-BEZ235 and DDP had synergic effects in inhibiting cell proliferation and migration. The expression of protein involved in apoptosis (cleaved caspase-3) was higher in drug combination group compared to the NVP-BEZ235 single treatment group. NVP-BEZ235 inhibited the proliferation and migration, and induced apoptosis and autophagy of HT-29 human colorectal adenocarcinoma cells.

Mechanism of all-transretinoic acid increasing retinoblastoma sensitivity to vincristine.

To explore the mechanism of all-transretinoic acid (ATRA) increasing retinoblastoma (RB) sensitivity to vincristine, and the inhibiting effect of vincristine combined with ATRA treatment on the SO-RB50cell proliferation. SO-RB50cells were cultivated by routine culture method. Different concentrations of vincristine or ATRA were added into culture solution. After 48h, cell counting kit-8 was used to detect the median inhibitory concentration (IC50) of vincristine combined with ATRT treatment to SO-RB50cells. SO-RB50cells were divided into drug combination group, vincristine group, ATRA group and control group. Different drugs were added into the culture solution respectively for cell culture based on the IC50 value. Cell counting kit-8 was used to detect the cell proliferation every 24-h cultivation. After continuous determination for 6d, data was processed to draw the cell growth curve. After drug use for 72h, flow cytometry was used to detect the proportion of different cell cycles of SO-RB50cells in each group. After drug use for 48h, annexin V/propidium iodide method was used to detect the SO-RB50cell apoptosis in each group. The IC50 value of vincristine treatment on the SO-RB50cells was 0.11μmol/L, and ATRT was 12.84μmol/L. The cell growth curve in control group rose gradually along with the extended culture time, but after vincristine and ATRA treatment, the cell growth curve was smooth and steady. The cell increment was the least in drug combination group and its cell growth curve was the smoothest. There was significant difference in A450 48h and 72h after treatment (Fgrouping=77.316, P<0.001; Ftime=86.985, P<0.001). Compared with control group, A450 value in drug combination group, vincristine group, ATRA group was significant lower (P<0.001). Compared with control group, the G2/M phase cell proportion in vincristine group was significantly increased, while the G0/G1 phase cell proportion was significantly decreased; the G0/G1 phase cell proportion in ATRA group was significantly increased, while the S phase cell proportion was significantly decreased (FG0/G1=85.878, Fs=56.455, FG2/M=85.878, P<0.001). After 48h, there was significant difference in SO-RB50cell apoptosis rate among groups (F=11.312, P<0.05). The apoptosis rate in drug combination group was significantly higher than that of other groups (P<0.001). ATRA can increase the sensitivity of SO-RB50cells to vincristine. Vincristine combined with ATRA treatment can significantly increase the inhibiting effect on SO-RB50cells, which may be related with promoting cell apoptosis and involving in cell cycle control.

Inhibitory effect of a Chinese medicine Xiaoaiping combined with cisplatin on the proliferation, invasion and apoptosis in ovarian cancer HO-8910 PM cells in vitro and in vivo

To explore the effect of a Chinese medicine, Xiaoaiping, in combination with cisplatin on the proliferation, invasion and apoptosis in human ovarian cancer HO-8910PM cells in vitro and vivo. CCK-8 assay was used to detect the inhibitoty effect of Xiaoaiping alone or in combination with cisplatin on the proliferation of human ovarian cancer HO-8910PM cells. Flow cytometry was used to detect the apoptosis and cell cycle distribution. Transwell migration test was used to assay the effect of drugs on the cell invasive capability. Changes of the tumor volume in nude mice were observed to evaluate the antitumor effects in vivo. CCK-8 assay Xiaoaiping alone or combined with cisplatin could inhibit the proliferation of HO-8910PM cells with a dose-dependent manner. The inhibition rates of Xiaoaiping combined with cisplatin were (53.4±3.0)%, significantly increased than those with single drug (P<0.05). Flow cytometry showed that G0/G1 fraction was increased respectively from (64.2±1.6)% to (74.1±1.6)% and (68.6±1.6)%. The percentages of apoptotic cells were increased from (2.2±1.6)% to (16.1±1.6)%, (35.6±1.6)% after treated with Xiaoaiping, Cisplatin and combination drugs (P<0.05 for all). Transwell chamber with matrigel assay showed that number of cells penetrating through membrane in HO-8910PM cells was (89.2±20.7)/HPF in the drug combination group, significantly less than that in the control group(187.2±24.6)/HPF, Xiaoaiping(141.8±13.7 )/HPF or cisplatin group (155.8±19.4)/HPF (P<0.01 for all). The inhibition rate of drug combination group in the nude mouse transplanted tumors, compared with that of single Xiaoaiping and cisplatin group, was increased significantly (59.0% vs. 23.4% and 34.2%), (P<0.01 for both). The results of our in vitro and vivo experiments indicate that Xiaoaiping can inhibit cell proliferation, increase G0/G1 arrest, promote apoptosis, inhibit cell migration of human ovarian cancer HO-8910 PM cells, and can synergistically enhance the antitumor activity of cisplatine.

Atorvastatin and fenofibrate combination induces the predominance of the large HDL subclasses and increased apo AI fractional catabolic rates in New Zealand white rabbits with exogenous hypercholesterolemia.

The anti-atherogenic properties of high-density lipoproteins (HDLs) may be related to their structure and metabolism. The HDL physicochemical characteristics that determine their plasma clearance during treatment with statins and fibrates are not well understood. In this study, we analyzed HDL-apo AI fractional catabolic rates (FCRs), size distributions, and the lipid composition of the HDL subclasses in New Zealand white rabbits with exogenous dyslipidemia that received low doses of atorvastatin and fenofibrate. Hypercholesterolemia decreased only partially with the combination of both drugs. HDL size distribution shifted toward larger particles among the groups of rabbits that received atorvastatin, fenofibrate, or their combination, compared with both the control group and the dyslipidemic group. The HDL subclasses were significantly rich in cholesterol in each of the groups compared with controls. The structural changes noted in the HDL subclasses were not associated with impaired plasma paraoxonase-1 (PON1) activity. The groups receiving monotherapy and the drug combination group were all associated with a higher apo AI FCR value compared with both the dyslipidemic rabbits and the control group. In conclusion, the combination of atorvastatin and fenofibrate induced a more favorable HDL subclass profile than did the individual use of these drugs. Similarly, the apo AI FCR values were augmented in every group receiving drug treatment (either monotherapy or combination therapy) in the setting of hypercholesterolemia. The anti-atherogenic properties of HDLs, excluding their capacity to bind PON1, may be enhanced by the structural and metabolic modifications induced by the combination of atorvastatin and fenofibrate.

Antipsychotic Use Pattern in People with Psychotic Disorder Living in Board and Care Facilities.

The aim of this survey is to determine the pattern of antipsychotic drug use in patients with psychotic disorders, living in board and care facilities and to investigate the related factors. We evaluated the antipsychotic drug use pattern in outpatients with psychotic disorders according to DSM-IV, living in board and care facilities. Patients using polypharmacy at least one month were compared with patients using monotherapy in terms of clinical and demographic characteristics. Antipsychotic polypharmacy (with two: 34%, with more than two: 28%) was identified in 62% of the patients. The most frequently prescribed combination was olanzapine+quetiapine (13%), the rate of first and second generation combination was 50%, the rate of second generation antipsychotic combination was 44%, and the rate of first generation anytipsychotic combination was 4% in the two antipsychotic drug combination group. The rate of clozapine use was 3%. Use of polypharmacy was associated with the diagnosis of schizophrenia and schizoaffective disorder, young age, suicidal behavior, multiple hospitalizations, clinical severity, and the need of anticholinergic drug. The ratio of using more than two antipsychotic drug combination is high (28%) in psychotic patients living in board and care, and rate of clozapine use is low, which shows that clinical practice is inconsistent with the treatment guidelines recommendations. It appears that further education to rationale antipsychotic drug use in psychiatric practices is required.

In vitro and in vivo analysis of antimicrobial agents alone and in combination against multi-drug resistant Acinetobacter baumannii.

Objective: To investigate the in vitro and in vivo antibacterial activities of tigecycline and other 13 common antimicrobial agents, alone or in combination, against multi-drug resistant Acinetobacter baumannii.Methods: An in vitro susceptibility test of 101 A. baumannii was used to detect minimal inhibitory concentrations (MICs). A mouse lung infection model of multi-drug resistant A. baumannii, established by the ultrasonic atomization method, was used to define in vivo antimicrobial activities.Results: Multi-drug resistant A. baumannii showed high sensitivity to tigecycline (98% inhibition), polymyxin B (78.2% inhibition), and minocycline (74.2% inhibition). However, the use of these antimicrobial agents in combination with other antimicrobial agents produced synergistic or additive effects. In vivo data showed that white blood cell (WBC) counts in drug combination groups C (minocycline + amikacin) and D (minocycline + rifampicin) were significantly higher than in groups A (tigecycline) and B (polymyxin B) (P < 0.05), after administration of the drugs 24 h post-infection. Lung tissue inflammation gradually increased in the model group during the first 24 h after ultrasonic atomization infection; vasodilation, congestion with hemorrhage were observed 48 h post infection. After 3 days of anti-infective therapy in groups A, B, C, and D, lung tissue inflammation in each group gradually recovered with clear structures. The mortality rates in drug combination groups(groups C and D) were much lower than in groups A and B.Conclusion: The combination of minocycline with either rifampicin or amikacin is more effective against multi-drug resistant A. baumannii than single-agent tigecycline or polymyxin B. In addition, the mouse lung infection by ultrasonic atomization is a suitable model for drug screening and analysis of infection mechanism.

Inhibitory effect of all trans retinoic acid with vincristine on the proliferation of retinoblastoma cells

Background Drug resistance is the main cause of failure after chemotherapy of retinoblastoma(RB), and how to improve the sensitivity of RB cells to chemotherapic drug become an urgent issue.All trans retinoic acid (ATRA) can inhibit the growth of tumor cells.However, whether ATRA increases the sensitivity of RB cells to chemotherapic drug is unclear. Objective This study aimed to investigate the inhibitory effect of ATRA with vincristine on the proliferation of SO-RB50 cells. Methods SO-RB50 cells were cultured routinely.Different concentrations of ATRA or vincristine were added into the medium for 48 hours for the determination of IC50 by cell counting kit-8(CCK-8) method.Cultured cells were divided into normal control group, ATRA group, vincristine group and combined drug group.The cells were treated by ATRA or vincristine with the dose of IC50, and the proliferation of the cells was detected every day at the 24-hour interval for 6 consecutive days.The percentage of the cells in different cell cycles was analyzed 72 hours after treatment using flow cytometry.Cell apoptosis rate was detected and calculated 48 hours after treatment by annexin V/propidium iodide(PI) method. Results The IC50 of ATRA was approximately 12.84 μmol/L, and that of vincristine was 0.11 μg/ml.The growth curve of SO-RB50 cells was gradually raised as the lapse of time, but the curves were relatively low in the ATRA group, vincristine group and combined drug group, with the lowest curve in the combined drug group.The proliferation values of the cells (A450) were 1.078±0.022, 0.611±0.038, 0.596±0.031 and 0.483±0.030 in 48 hours after treatment, and those in 72 hours were 1.380±0.021, 0.799±0.016, 0.668±0.041 and 0.532±0.033 in normal control group, ATRA group, vincristine group and combined drug group, showing significant differences among the groups and various time points (Fgroup=1 115.207, P=0.000; Ftime =257.781, P=0.000). The A450 values of the ATRA group, vincristine group and combined drug group were significantly lower than those of normal control group (all at P<0.05). The percentage of the cells in different cell cycles was changed after 72 hours' treatment and the differences were statistically significant (FG0/G1=130.565, FS=57.435; FG2/M=114.290, P<0.05). Compared with the normal control group, the percentage of G0/G1 phase cells was increased and S phase cells was decreased significantly in ATRA group, the percentage of G0/G1 phase cells was decreased and G2/M phase cells was increased significantly in vincristine group(P<0.05). The apoptotic rate of the cells was (7.17±0.18) %, (27.34±1.36)%, (27.49±2.45)% and (34.50±1.84) % in normal control group, ATRA group, vincristine group and combined drug group, with a significant difference among the groups (F=147.555, P<0.05), and the apoptotic rate in the combined drug group was remarkedly lower than that of normal control group, ATRA group and vincristine group (all at P=0.000). Conclusions ATRA can improve the sensitivity of SO-RB50 cells to chemotherapeutic drug.The combined application of ATRA and vincristine enhance the inhibitory effect on the cells probably by regulating cell cycle and inducing apoptosis. Key words: All trans retinoic acid; Vincristine; Retinoblastoma; Proliferation; Cell cycle; Apoptosis