High-dose Combination Group Research Articles

Synergistic anti-tumorigenic effect of diosmetin in combination with 5-fluorouracil on human colon cancer xenografts in nude mice

5-Fluorouracil (5-FU) is frequently used to treat colorectal cancer (CRC), but its clinical application is limited by its toxicity. Natural compounds have been combined with chemotherapeutic drugs to reduce chemotherapy-related toxicity. Diosmetin, a natural flavonoid, has demonstrated anticancer effects against CRC. This study investigated diosmetin's potential in combination with 5-FU using a murine model of HCT-116 colon cancer xenografts in nu/nu nude mice. HCT-116 cells were injected into the right flanks of mice, and once tumors reached a size of 50 mm3, the mice were treated with diosmetin (100 mg/kg), 5-FU (30 mg/kg), or a combination of both at two dose levels (100 + 30 mg/kg and 50 + 15 mg/kg) for 4 weeks. Blood and tumors were collected on the final day for further analysis. Mice treated with the higher combination dose exhibited the smallest tumor volume (330.91 ± 88.49 mm3). Biochemistry and histology analysis showed no toxicity or abnormalities in the liver, kidney, and heart with the combination therapy. Immunohistochemistry results revealed a notable reduction in the proliferation marker (Ki67) and inflammation marker (TLR4) in tumors from high-dose combination-treated mice. Moreover, immunofluorescence data indicated increased levels of apoptotic markers (Bax, Caspase-3, p53, p21) and downregulation of anti-apoptotic protein (Bcl-2) in the high-dose combination group. The findings suggest that 100 mg/kg of diosmetin combined with 30 mg/kg 5-FU significantly reduced tumor volume and had a less toxic effect on the heart compared to 5-FU monotherapy.

Exploring the combined therapeutic efficacy of bexarotene and icariin in type 2 diabetic rats.

The aim of this study was to determine the single and combined antidiabetic activity and side effects of the retinoid X receptor agonist bexarotene and the thioredoxin-interacting protein inhibitor and peroxisome proliferator-activated receptor γ and AMP-activated protein kinase activator icariin. The rats were grouped as healthy (control), diabetes, diabetes + bexarotene (20mg/kg), diabetes + icariin (60mg/kg), diabetes + bexarotene (10mg/kg) + icariin (30mg/kg) low-dose combination and diabetes + bexarotene (20mg/kg) + icariin (60mg/kg) high-dose combination groups. Icariin treatment led to a significant reduction in glucose levels compared with the diabetes control group, a remarkable outcome observed 45 days after the initial application. HbA1c levels of the icariin and low-dose combination treatment groups were significantly lower than in the diabetes group. Notably, icariin treatment also significantly elevated HOMA-β levels, which is indicative of improved β-cell function. Icariin significantly decreased glucose levels at 30 and 120min in the oral glucose tolerance test. Moreover, it ameliorated hepatocyte degeneration, hepatic cord dissociation, congestion, mononuclear cell infiltration in the liver, and degeneration in the pancreas. Icariin treatment exhibited robust antidiabetic effects with fewer side effects than other treatment options in this study. In future studies, long-term and varying doses of icariin will contribute to the development of novel antidiabetic drugs.

Combination of shikonin and gefitinib reverses drug resistance in human non-small cell lung cancer and its mechanism

The occurrence and development of tumors are associated with the cell energy metabolism. Inhibiting energy metabolism of lung cancer cells is an important strategy to overcome drug resistance. Based on the cellular energy metabolism pathway, this study observed the effect of combination of shikonin(SKN) and gefitinib(GFB) on the drug resistance in non-small cell lung cancer and explored the underlying mechanism. The human non-small cell lung cancer line HCC827/GR resistant to gefitinib was used as the cell model in vitro. The CCK-8 assay and flow cytometry were employed to investigate the cell viability and apoptosis, respectively. The high performance liquid chromatography was employed to measure the intracellular accumulation of GFB. A Seahorse XFe96 Analyzer was used to detect the changes of cellular energy metabolism. Western blot was employed to determine the expression of the proteins involved in the drug resistance. The tumor-bearing nude mouse model was used to verify the efficacy of SKN+GFB in overcoming drug resistance in vivo. The results showed that SKN+GFB significantly reduced the IC_(50) of GFB on HCC827/GR cells, with the combination index of 0.628, indicating that the combination of the two drugs had a synergistic effect and promoted cell apoptosis. SKN increased the intracellular accumulation of GFB. SKN+GFB lowered the oxygen consumption rate(OCR) and glycolytic proton efflux rate(GlycoPER) in cell energy metabolism, and down-regulated the overexpression of PKM2, p-EGFR, P-gp, and HIF-1α in drug resistance. The results of reversing drug resistance test in vivo showed that GFB or SKN alone had no significant antitumor effect, while the combination at different doses induced the apoptosis of the tumor tissue and inhibited the expression of PKM2 and P-gp, demonstrating a significant antitumor effect. Moreover, the tumor inhibition rate in the high-dose combination group reached 64.01%. In summary, SKN+GFB may interfere with the energy metabolism to limit the function of HCC827/GR cells, thus reversing the GFB resistance in non-small cell lung cancer.

A serum metabolomics study based on LC-MS: Chemosensitization effects of Rauvolfia vomitoria Afzel. combined with 5- fluorouracil on colorectal cancer mice

Colorectal cancer (CRC) is one of the malignant tumors with high incidence, and is mainly treated by chemotherapy at present. However, during CRC treatment, long-term use of traditional chemotherapeutic drugs will reduce the sensitivity of chemotherapy. Our previous studies have shown that Rauvolfia vomitoria total alkaloids (RVA) played an important role in 5-fluorouracil (5-FU) chemosensitization in CRC therapy, but its intervention mechanism has not been clarified completely in the metabolic level. Therefore, in this study, LC-MS based metabolomics was employed to explore the mechanism of 5-FU chemosensitization in CRC induced by the combination of RVA and conventional chemotherapeutic with 5-FU. The results showed that the final tumor weight of the high-dose combined group was significantly different from that of the 5-FU alone group. To evaluate the chemosensitization effects of RVA, serum samples collected from six groups (six mice in each group) with different administration methods were analyzed by HPLC-Q-Exactive Orbitrap/MS. After multivariate statistical analysis and metabolites identification, 25 different metabolites were identified between the 5-FU treatment group and combined high-dose treatment group, among which lipid and fatty acid metabolism pathways were mostly affected. These results suggest that RVA may sensitize traditional chemotherapeutic drug 5-FU and exert anti-tumor activity through influencing lipid metabolism and cell energy metabolism. Metabolomics provided a new insight into estimate of the therapeutic effect and dissection of the potential mechanisms of traditional Chinese medicine in treating colorectal cancer.

Immunohistochemical and Histochemical Studies of Β-Cells Insulin Up-Regulation in Pancreatic Tissues of Streptozotocin-Induced Diabetic Albino Rats Treated with Melatonin and Magnesium

Background: Improve insulin secretion and cellular availability to reduced blood glucose levels in diabetic subjects by bioactive compound especially antioxidants are the new focus to ameliorate the complication with diabetes mellitus. Aims and objectives: The aim of this study was to evaluate the effects of administration of melatonin and magnesium on the cytoarchitecture of the pancreatic tissue and to access immunohistochemically insulin release in streptozotocin-induced diabetic Albino rats. Materials and methods: To achieve this aim six normoglycaemic rats and fourty eight Streptozotocin (STZ) induced diabetic rats was used in the study after two weeks acclimatization period. The animals were assigned into nine groups as follows, Normal control group (NC), Diabetic control (DC) group, Melatonin at 10 mg/kgb (MLD), magnesium dose group of 240 mg/kgbw (MgLD), melatonin and magnesium combined group of 10mg/kgbw+240mg/kgbw (MMgLD), melatonin group of 20mg/kgbw (MHD), melatonin and magnesium high dose combined group of 20mg/kgbw+480mg/kgbw (MMgHD) and insulin at 500mg/kgbw group (IN). Melatonin and insulin were administered through intraperitoneal injections (IP) while magnesium was by oral administration. The control groups were given placebo and all groups’ treatment was for twenty-one days. At the end of the study, the animals were aestheticized and euthanized to harvest pancreatic organ. The organs were fixed in neutral buffered formaldehyde (NBF). They were histologically prepared and stained using haematoxylin and eosin and immunohistochemically stained using insulin antibody to access insulin release. Results: Melatonin treatment at 10mg/kgbw and at 20 mg/kgbw showed histological improvement in histological tissues and insulin release while when combined with magnesium at dose of 10mg/kgbw and at 240 mg/kgbw showed better results. The administration of magnesium at 240 mg/kgbw, 480 mg/kgbw and when combined with melatonin at high doses does not show significant improvement in islet β-cell proliferation and insulin release. Conclusion: The administration of melatonin and magnesium at low doses regenerates pancreatic islet histoarchitecture and augments insulin release from treated diabetic albino rats.

Atractylenolide Ⅰ improves acetaminophen-induced acute liver injury in mice by inhibiting MAPK/NF-κB signaling pathway

This study explored the protective effect of atractylenolide Ⅰ(AO-Ⅰ) against acetaminophen(APAP)-induced acute liver injury(ALI) in mice and its underlying mechanism. C57 BL/6 J mice were randomly divided into a control group, an APAP group(500 mg·kg~(-1)), a low-dose combination group(500 mg·kg~(-1) APAP + 60 mg·kg~(-1) AO-Ⅰ), and a high-dose combination group(500 mg·kg~(-1) APAP + 120 mg·kg~(-1) AO-Ⅰ). ALI was induced by intraperitoneal injection of APAP(500 mg·kg~(-1)). AO-Ⅰ by intragastric administration was performed 2 hours before APAP treatment, and the control group received the same dose of solvent by intragastric administration or intraperitoneal injection. The protective effect of AO-Ⅰ against APAP-induced ALI was evaluated by detecting alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels in the plasma and H&E staining in liver tissues of mice. The malondialdehyde(MDA) and glutathione(GSH) content and catalase(CAT) activity in mouse liver tissues were detected to evaluate the effect of AO-Ⅰ on APAP-induced oxidative stress in the liver. The proteins in the liver p38 mitogen-activated protein kinase(p38 MAPK), c-jun N-terminal kinase(JNK), and nuclear factor kappa-B p65(NF-κB p65) signaling pathways were measured by Western blot, and the liver inflammatory cytokines interleukin-1β(IL-1β) and interleukin-6(IL-6) were detected by real-time PCR. Compared with the APAP group, the combination groups showed reduced APAP-induced ALT level and liver MDA content, potentiated liver CAT activity, and elevated GSH content. Mechanistically, AO-Ⅰ treatment significantly inhibited APAP-up-regulated MAPK phosphorylation and NF-κB p65, and significantly reduced the transcriptional activities of IL-1β and IL-6, downstream targets of NF-κB p65. AO-Ⅰ can improve APAP-induced ALI and the underlying mechanism is related to the inhibition of the MAPK/NF-κB p65 signaling pathway in APAP-challenged mice.

Magnesium and Melatonin Co-Administration Attenuates Blood Glucose Levels in Streptozotocin-Induced Diabetic Rats

Diabetes mellitus (DM) is a serious metabolic disorder characterized by elevated blood glucose. The raised blood glucose level if not properly managed can results in damage and failure of several vital organs such as the eyes, kidneys, nerves, heart, and blood vessels. The aim of this study was to evaluate the effects of co-administration of melatonin and magnesium on blood glucose level of streptozotocin-induced Type 1 diabetic Wistar rats. To achieve this aim sixty-four Wistar rats were used in the study. Streptozotocin (STZ) was used to induce chemical type 1 diabetes mellitus (T1DM) after two weeks acclimatization period in fifty-eight Wistar rats. Fifty-three rats were diabetic and fourty-eight diabetic rats were randomly distributed in eight groups and six normal normoglycaemic rats were used as control. The animals were assigned into nine groups as follows, Normal control group (NC), Diabetic control (DC) group, Melatonin Low dose group of 10 mg/kgb (MLD), magnesium low dose group of 240 mg/kgbw (MgLD), melatonin and magnesium combined low dose group of 10mg/kgbw+240mg/kgbw (MMgLD), melatonin high dose group of 20mg/kgbw (MHD), magnesium high dose group of 480mg/kgbw (MgHD), melatonin and magnesium high dose combined group of 20mg/kgbw+480mg/kgbw (MMgHD) and insulin at 500mg/kgbw group (IN). Melatonin and insulin were administered through intraperitoneal injections (IP) while magnesium was by oral administration. The control groups were given placebo and all group treatment was for twenty-one days. Blood glucose was measured weekly in all groups by day 8, day 15 and day 22. At the end of the evaluation Day 22, the blood glucose levels showed a highly significant decrease (p˂0.05) in melatonin treatment group at low dose (MLD), melatonin and magnesium combined group at low doses (MMgLD), melatonin treatment group at high doses (MHD) and insulin treated group (IN) when compared with normoglycaemic group (NC) while groups treated with Magnesium at low doses (MgLD), magnesium treated group at high doses and melatonin and magnesium treatment group at high doses (MMgHD) showed no significant decrease (p˂0.05) in blood glucose levels as compared with diabetic control (DC). Melatonin and magnesium co-administration has hypoglycaemic effects on blood glucose levels in STZ-induced type 1 diabetic Wistar rats.

The role of intermittent PTH administration in conjunction with allogenic stem cell treatment to stimulate fracture healing.

AimsA growing number of fractures progress to delayed or nonunion, causing significant morbidity and socioeconomic impact. Localized delivery of stem cells and subcutaneous parathyroid hormone (PTH) has been shown individually to accelerate bony regeneration. This study aimed to combine the therapies with the aim of upregulating fracture healing.MethodsA 1.5 mm femoral osteotomy (delayed union model) was created in 48 female juvenile Wistar rats, aged six to nine months, and stabilized using an external fixator. At day 0, animals were treated with intrafracture injections of 1 × 106 cells/kg bone marrow mesenchymal stem cells (MSCs) suspended in fibrin, daily subcutaneous injections of high (100 μg/kg) or low (25 μg/kg) dose PTH 1-34, or a combination of PTH and MSCs. A group with an empty gap served as a control. Five weeks post-surgery, the femur was excised for radiological, histomorphometric, micro-CT, and mechanical analysis.ResultsCombination therapy treatment led to increased callus formation compared to controls. In the high-dose combination group there was significantly greater mineralized tissue volume and trabecular parameters compared to controls (p = 0.039). This translated to significantly improved stiffness (and ultimate load to failure (p = 0.049). The high-dose combination therapy group had the most significant improvement in mean modified Radiographic Union Score for Tibia fractures (RUST) compared to controls (13.8 (SD 1.3) vs 5.8 (SD 0.5)). All groups demonstrated significant increases in the radiological scores – RUST and Allen score – histologically compared to controls.ConclusionWe demonstrate the beneficial effect of localized MSC injections on fracture healing combined with low- or high-dose teriparatide, with efficacy dependent on PTH dose.Cite this article: Bone Joint Res 2021;10(10):659–667.

Effects of molybdenum and cadmium on the oxidative damage and kidney apoptosis in Duck

Molybdenum (Mo) is an essential element for human beings and animals; however, high dietary intake of Mo can lead to adverse reactions. Cadmium (Cd) is one of the major transitional metals which has toxic effects in animals. To investigate the co-induced toxic effects of Mo and Cd on oxidative damage and kidney apoptosis in duck, 120 ducks were randomly divided into control group and 5 treatment groups which were treated with a commercial diet containing different dosages of Mo and Cd. Kidney samples were collected on the 60th and 120th days to determine the mRNA expression levels of ceruloplasmin (CP), metallothionein (MT), Bak-1, and Caspase-3 by quantitative RT-PCR. Additionally, we also determined the antioxidant activity indexes and contents of Mo, Cd, copper (Cu), iron (Fe), zinc (Zn), and selenium (Se) in serum. Meanwhile, ultrastructural changes of the kidney were observed. The results showed that glutathione reductase (GR) activity and CP level in serum were decreased in combination groups. In addition, the antioxidant indexes were decreased in co-treated groups compared with single treated groups. The mRNA expression levels of Bak-1 and Caspase-3 increased in co-treated groups. The mRNA expression level of CP in high-dose combination group was downregulated, while the mRNA expression of MT was upregulated except for low-dose Mo group. Additionally, in the later period the content of Cu in serum decreased in joint groups while the contents of Mo and Cd increased. In addition, ultrastructural changes showed mitochondrial crest fracture, swelling, deformed nuclei, and karyopyknosis in co-treated groups. Taken together, it was suggested that dietary Mo and Cd might lead to oxidative stress, kidney apoptosis and disturb homeostasis of trace elements in duck, and it showed a possible synergistic relationship between the two elements.

What is the Ideal Route of Administration of Tranexamic Acid in TKA? A Randomized Controlled Trial.

TKA commonly involves substantial blood loss and tranexamic acid has been used to reduce blood loss after TKA. Numerous clinical trials have documented the efficacy and safety of intravenous (IV) or intraarticular (IA) use of tranexamic acid. Combined administration of tranexamic acid also has been suggested; however, there is no consensus regarding the ideal route of tranexamic acid administration. (1) To compare the efficacy of tranexamic acid in terms of total blood loss and the allogeneic transfusion rate among three routes of administration: IV alone, IA alone, and combined IV and IA. (2) To compare these regimens in terms of venous thromboembolism (VTE) and the frequency of wound complications. In total, 376 patients undergoing TKA between March 2014 and March 2015 were randomized to four groups by the route of tranexamic acid administration: IV only, IA only, low-dose combined (IV + IA injection of 1 g), and high-dose combined (IV + IA injection of 2 g). The calculated total blood loss, allogeneic transfusion rate, decrease in hemoglobin, the frequency of symptomatic deep vein thrombosis and pulmonary embolism, wound complications, and periprosthetic joint infection were compared among the groups. Total blood loss was calculated using estimated total body blood volume and hemoglobin loss. The decision regarding when to transfuse was determined based on preset criteria. The high- and low-dose combined groups and the IA-only group had lower total blood loss (564 ± 242 mL, 642 ± 242 mL, and 633 ± 205 mL, respectively) than the IV-only group (764 ± 217 mL; mean differences = 199 mL [95% CI, 116-283 mL], p < 0.001; 121 mL [95% CI, 38-205 mL], p = 0.001; 131 mL [95% CI, 47-214 mL], p < 0.001); no differences were found among the other three groups. No patients in any study group received an allogeneic transfusion. One patient in the IV-only group had a symptomatic pulmonary embolism develop, but no other symptomatic VTE events occurred in any group. In addition, no differences were observed in wound complications, such as superficial wound necrosis (one patient in the IV-only and the high-dose combined group, respectively) and oozing (IV-only, IA-only, low-dose combined, high-dose combined = 3%, 4%, 4%, and 7%; p = 0.572) between the groups. No patients had a periprosthetic joint infection. IA tranexamic acid administration further reduces blood loss after TKA in comparison to IV use alone; no additional effect in further reducing blood loss was found in combination with IV tranexamic acid. Appropriately powered studies are needed to confirm the safety of this route of administration as the preferred route of administration in TKA. Level I, therapeutic study.

PHARMACODYNAMIC INTERACTION OF QUERCETIN AND SILYMARIN AGAINST CYCLOPHOSPHAMIDE INDUCED CARDIO-TOXICITY

The study was designed to investigate the cardioprotective effect of different combination of Silymarin and Quercetin against Cyclophosphamide induced cardiotoxicity in experimental rats . The Albino rats of either sex were divi ded into five groups of six animals . Group 1 and group 2 served as normal and control group respectively . Other groups treated with Silymarin ( 60 mg / kg, p . o . ) alone, high ( 30 mg / kg, p . o . ) and low ( 15 mg / kg, p . o . ) dose of Silymarin combined with Quercetin ( 10 mg / kg, p . o .). Myocardial damage was induced by the administration of Cyclophosphamide ( 200mg / kg, i . p . ) on the first day and the treatment was conducted for ten days . Twenty - four hours after the last administration, the blood was collected by the retro - o rbital puncture method and the serum biomarker level of Lactate Dehydrogenase ( LDH ) , Creatinine Kinase - NAC ( CK - NAC ) , Creatinine Kinase - MB ( CK - MB ) , Aspartate Transaminase ( AST ) , Alanine Transaminase ( ALT ) and Alkaline Phosphatase ( ALP ) in serum were evaluat ed . Electro - cardiograph parameters such as Heart rate, QRS interval, QT segment, PR interval and RR interval also carried out . Compared to the control group all treated group showed significant activity and high and low dose combination group showed profou nd car diac protection compare to the S ilymarin alone . Among the entire groups high dose combination group ( SILLY30 + QUE10 ) showed most effective Cardio - protection and was also responsible for the significant improvement in the ECG parameters .

Direct and protective effects of single or combined addition of vincristine and ε-viniferin on human HepG2 cellular oxidative stress markers in vitro.

The objective of this study is to examine the direct effects of low doses and high doses of ε-viniferin, a substance known to be an antioxidant, and vincristine sulphate, a chemotherapeutic agent, alone and in combination [ε-viniferin+vincristine] on HepG2 cell strain, as well as evaluate oxidative stress after incubation periods of 3, 6, and 24h. Direct effect was determined right after the incubation period; however, for protective effect, antioxidant protection response was determined after the treatment for 1h with 500μM H2O2, which is an oxidative stressor. For this purpose, superoxide dismutase was determined for enzyme activity, and lipid hydroperoxide (LPO) and reduced glutathione concentrations were studied as indicators of oxidative stress. Results show that low [3.63µM vincristine+3.75µM ε-viniferin] and high [11.25µM vincristine+15.8µM ε-viniferin] doses of combination groups showed similar direct antioxidant effect on LPO levels as protective when compared to the H2O2 control group (p<0.05). Superoxide dismutase enzyme showed a direct antioxidant effect in low and high dose combination groups. In addition, when the incubation period was increased to 24h, a protective effect was observed in both dose groups (p<0.05). Reduced glutathione activities showed a direct effect in the low dose combination group, and a protective effect in both the low and high doses in the 24h. These results show that combined usage of drugs in HepG2 cell strain possesses a protective effect against exogenically produced oxidative stress conditions.

Effect of initial combination therapy with sitagliptin and metformin on β‐cell function in patients with type 2 diabetes

To examine the effect of sitagliptin and metformin, alone and in combination, on modelled parameters of β-cell function in patients with type 2 diabetes. The data used in the present analyses are from a 104-week study, which included a 24-week, placebo- and active controlled phase followed by a 30-week, active controlled, continuation phase and an additional 50-week, active controlled extension phase. Patients were randomised to one of six blinded treatments: sitagliptin 50 mg + metformin 1000 mg b.i.d., sitagliptin 50 mg + metformin 500 mg b.i.d., metformin 1000 mg b.i.d., metformin 500 mg b.i.d., sitagliptin 100 mg q.d. or placebo. Patients on placebo were switched in a blinded manner to metformin 1000 mg b.i.d. at week 24. Subsets of patients volunteered to undergo frequently sampled meal tolerance tests at baseline and at weeks 24, 54 and 104. β-cell responsivity was assessed with the C-peptide minimal model. The static component (Φ(s)) estimates the rate of insulin secretion related to above-basal glucose concentration. The dynamic component (Φ(d)) is related to the rate of change in glucose. The total index (Φ(total)) represents the overall response to a glycaemic stimulus and is calculated as a function of Φ(s) and Φ(d). Insulin sensitivity was estimated with the Matsuda index (ISI). The disposition index, which assesses insulin secretion relative to the prevailing insulin sensitivity, was calculated based on the Φ(total) and ISI. At week 24, substantial reductions in postmeal glucose were observed with all active treatment groups relative to the placebo group. Φ(s), Φ(total) and the disposition index were significantly improved from baseline at week 24 with all active treatments relative to placebo. Generally larger effects were observed with the initial combination of sitagliptin and metformin relative to the monotherapy groups. When expressed as median percent change from baseline, Φ(s) increased from baseline by 137 and 177% in the low- and high-dose combination groups and by 85, 54, 73 and -9% in the high-dose metformin, low-dose metformin, sitagliptin monotherapy and placebo groups, respectively. At weeks 54 and 104, the combination treatment groups continued to demonstrate greater improvements in β-cell function relative to their respective monotherapy groups. After 24 weeks of therapy, relative to placebo, initial treatment with sitagliptin or metformin monotherapy improved β-cell function; moreover, initial combination therapy demonstrated larger improvements than the individual monotherapies. Improvements in β-cell function were found with treatments for up to 2 years.

Effect of Combining Extended‐Release Carvedilol and Lisinopril in Hypertension: Results of the COSMOS Study

Hypertension treatment commonly requires multiple agents to achieve target blood pressure (BP). β-blockers and angiotensin-converting enzyme inhibitors (ACEIs) are commonly co-prescribed in clinical practice although few data are available that test their additivity on BP lowering. The efficacy and safety of once-daily extended-release carvedilol (carvedilol CR) combined with the ACEI lisinopril in a double-blind, randomized, factorial design study were studied. Patients (N=656) with stage 1 or 2 hypertension were randomized evenly to 1 of 15 groups for 6 weeks: carvedilol CR monotherapy 20 mg, 40 mg, or 80 mg/d; lisinopril monotherapy 10 mg, 20 mg, or 40 mg/d; or 1 of 9 combinations of carvedilol CR plus lisinopril initiated simultaneously. Primary efficacy measures (assessed by ambulatory BP monitoring [ABPM]) were change from baseline in 24-hour mean diastolic BP (DBP) and in trough (20-24 hours) DBP. Continuous efficacy variables were assessed using analysis of covariance. Whether any combination dose was superior to its monotherapy components was assessed using the Hung AVE procedure. Despite the presence of additional BP lowering observed with most of the combinations compared with their monotherapy components, the Hung AVE test was not significant for either primary efficacy measures. Post hoc analyses of the high-dose combination groups (carvedilol CR/lisinopril regimens of 80/10 mg, 80/20 mg, 80/40 mg, 20/40 mg, and 40/40 mg) showed a significant treatment difference compared with both carvedilol CR 80 mg and lisinopril 40 mg for 24-hour mean DBP but not for trough DBP. With the exception of dizziness, individual adverse events did not increase with ascending doses or combinations. The superiority of initiating combination treatment with carvedilol CR and lisinopril compared with the monotherapy components was not demonstrated with the ABPM measurements. Nonetheless, the post hoc assessment combining all high-dose groups did produce significant 24-hour mean BP reduction when compared with the high-dose monotherapy groups. The tolerability profile of initiating combination therapy was generally comparable to the initiation of treatment with monotherapy.

Effects of astragalosides and Panax notoginseng saponins combination on oxidative stress of cerebral ischemic reperfusion injury in mice

To investigate the influence of astragaloside (AST) and Panax notoginseng saponins (PNS) combination on oxidative stress of brain tissues in C57BL/6 mice with cerebral ischemia-reperfusion injury. Eighty C57BL/6 mice were randomly divided into sham-operated group, untreated group, high-dose combination group (AST at a dose of 220 mg/kg plus PNS at a dose of 230 mg/kg), medium-dose combination group (AST at a dose of 110 mg/kg plus PNS at a dose of 115 mg/kg), low-dose combination group (AST at a dose of 55 mg/kg plus PNS at a dose of 57.5 mg/kg), AST (110 mg/kg) group, PNS (115 mg/kg) group and edaravone (4 mg/kg) group. AST and PNS were administered by gavage once daily for 4 days and edaravone was administered by intraperitoneal injection twice daily for 4 days. On the fourth day, bilateral common carotid arteries were ligated for 20 minutes to induce cerebral ischemia, followed by 60 minutes of reperfusion. Ischemic brain tissue was used to prepare tissue homogenate, then contents of malonaldehyde (MDA), glutathione (GSH) and nitric oxide (NO), and activities of superoxide dismutase (SOD) and nitric oxide synthase (NOS) in the homogenate were detected. Two x two analysis of variance of factorial design was used to analyze whether there was an interaction between AST at 110 mg/kg and PNS at 115 mg/kg. Compared with sham-operated group, contents of MDA and NO, and activity of NOS in the untreated group were remarkably increased (P<0.01), activity of SOD and content of GSH were decreased (P<0.01). Compared with the untreated group, content of MDA in the AST group was decreased (P<0.01) and activity of SOD was increased (P<0.01), however, contents of GSH and NO and activity of NOS had no obvious changes (P>0.05). Contents of MDA and NO in the PNS group was decreased as compared with the untreated group (P<0.01), but activities of SOD and NOS and content of GSH had no changes (P>0.05). Contents of MDA and NO and activity of NOS in brain tissues in the edaravone group were decreased (P<0.01, P<0.05), and activity of SOD was increased (P<0.05), while content of GSH had no changes (P>0.05). Contents of MDA and NO and activity of NOS in brain tissue in the AST and PNS combination groups were decreased (P<0.01, P<0.05), the activity of SOD increased (P<0.01, P<0.05), the content of GSH increased (P<0.01, P<0.05), and activity of SOD and content of GSH were increased (P<0.01, P<0.05). The results of analysis of variance of factorial design showed that there were interactions between AST (110 mg/kg) and PNS (115 mg/kg) (P<0.01). Combination of AST (110 mg/kg) and PNS (115 mg/kg) has a restraint effect on the early oxidative stress injury in the brain after ischemia-reperfusion, and the combination has a synergistic or additive effect.

Reduced transplant arteriosclerosis after treatment with MMF and MMF in combination with ganciclovir in a mouse aortic transplant model

Objectives: The aim of this study was to investigate whether Ganciclovir and Mycophenolate Mofetil (MMF), alone and in combination, have a beneficial influence on the development of transplant arteriosclerosis (TxA). Methods: Fully allogeneic C57BL/6 (H2b) donor aortas were transplanted into CBA (H2k) recipients. Recipient mice were treated with a single medication of MMF (100mg/kg or 300mg/kg) or a combination of MMF and Ganciclovir (10mg/kg Ganciclovir and 100mg/kg MMF or 72mg/kg Ganciclovir and 300mg/kg MMF) per day. Grafts were analysed by histology and morphometry on day 30 after transplantation. A potential effect of MMF or Ganciclovir on Tregs was investigated by FACS analysis. Results: Two doses of 150mg/kg MMF per day significantly reduced the development of TxA compared to untreated controls (intima proliferation of 23%±8% vs. 68%±7% [control]). 50mg/kg of MMF did not have any effect on the development of TxA. Strikingly, combination of Ganciclovir and low-dose MMF reduced the formation of TxA (intima proliferation of 27%±2% vs. 68%±7% [control]). The high-dose combination group (300mg MMF/72mg Ganciclovir) showed an intima proliferation of 24%±4%. FACS analysis of the spleen within this group revealed 12.73% CD4+cells with 14.70% CD4+CD25+FoxP3+cells. As compared to 17.5% CD4+cells with 11.1% CD4+CD25+FoxP3+cells in the control group. Single treatment with MMF (300mg) showed 10.55% CD4+ cells and 10.87% CD4+CD25+FoxP3+ cells within the analysed spleens. Conclusion: Our results demonstrate a dose-dependent positive effect of MMF on the development of TxA and a synergistic effect of Ganciclovir in combination with MMF.

Teratogenic effects in rabbits of simultaneous exposure to ochratoxin A and aflatoxin B 1 with special reference to microscopic effects

To elucidate the teratogenic effects, ochratoxin A (OTA) and aflatoxin B 1 (AFB 1) were dissolved in corn oil and administered in combination to New Zealand White rabbits during 6–18 days of gestation orally with the dose levels of OTA + AFB 1, 0.05 + 0.05 and 0.1 + 0.1 mg/kg body weight. To assess pathomorphological features of the anomalies, the fetal serial sections were histologically examined. There was no mortality in any of the treated groups. Body weights and body weight gains of dams in the combined treatment groups were comparable with those of controls and individual treatments. The mean crown to rump lengths in both the combination dose groups and mean fetal weights in high dose combination group were significantly decreased. In the high dose combination, there was increase in the percent of implants resorbed and significant increase in the incidence of visceral anomalies. The combination treatment resulted in various gross, skeletal and visceral anomalies such as wrist drop, scoliosis, bent metacarpals, rudimentary ribs, cardiac defects and microphthalmia. There was a dose-related increase in the percent of litters showing the histopathological changes in the fetal tissues. The incidence of histopathological changes in the tissue sections prepared from fetal liver, kidneys, brain, heart and eyes was found increased in the high dose combination group. The comparative evaluation of the results of combination versus individual treatments revealed that certain anomalies observed in the individual treatment of OTA such as knuckling of fetlock, rudimentary tail or agenesis of tail, wavy ribs, hydrocephalus and agenesis of kidney and AFB 1 as enlarged eye sockets and enlarged liver were absent in the combination treatment. However, some new manifestations such as cardiac defects and scoliosis were seen. The results of the present study indicated that in combination, OTA and AFB 1 have antagonistic interaction. The presence of subtle lesions histologically due to an interference with normal development suggested that microscopic examination of the fetal tissues could provide additional, useful information to a developmental toxicity study.

Glycoprotein IIb/IIIa antagonist, murine 7E3 F(ab') 2, and tissue plasminogen activator in focal ischemia: evaluation of efficacy and risk of hemorrhage with combination therapy.

Tissue hypoperfusion during cerebral ischemia results from occlusion of large and small vessels. Combination treatment strategies using fibrinolytics to thrombolyse an embolic clot and antiplatelet agents to prevent reocclusion and the formation of new platelet thrombi in the microcirculation may offer advantages over single-agent therapy. The authors report on the effects of tissue plasminogen activator (rt-PA), a glycoprotein (GP) IIb/IIIa receptor antagonist, 7E3 F(ab') 2, or a combination of the two agents in a focal embolic model of cerebral ischemia in Wistar rats. Focal ischemia was produced by introducing an autologous thrombus into the right side middle cerebral artery. Forty-six male Wistar rats were randomly divided into 6 groups: control (n = 8), 7E3 F(ab') 2 (n = 9, 6 mg/kg), rt-PA (n = 9, 10 mg/kg), rt-PA (n = 6, 20 mg/kg), and 7E3 F(ab') 2 with either 10 mg/kg (n = 10) (low-dose combination) or 20 mg/kg (n = 6) (high-dose combination) rt-PA. Evaluation of neurobehavioral scores, cerebral angiography, bleeding time, and measurement of brain infarction volume were used to determine efficacy. All actively treated groups showed a significant reduction in the infarct volume. Animals treated with 7E3 F(ab') 2 showed reduced infarction volumes (24.0 +/- 5.1%) compared with controls (42.43 +/- 5.6%, P < 0.02). Treatment with rt-PA significantly reduced infarction volume (20.7 +/- 3.3, = 0.01) at 10 mg/kg and at 20 mg/kg (19.5 +/- 8.2%, P < 0.05). Compared with vehicle-treated animals, the low-dose combination (16.4 +/- 5.5, P < 0.003) and high-dose combination (23.7 +/- 6.2%, P < 0.05) showed significant reduction in infarction volume. Cerebral angiography revealed significantly better recanalization in the combination group (5/6 animals in the high dose and 4/6 in low dose) compared with animals treated with 7E3 F(ab') 2 (3/10) or rt-PA alone (2/6). Bleeding time significantly increased from 11.25 +/- 1.9 minutes in the control group to 17 +/- 3.1 minutes in the rt-PA group, 24.5 +/- 2.6 minutes in the 7E3 F(ab') 2 group, 25.7 +/- 3.1 minutes in the low-dose combination group, and 32.5 +/- 4.7 minutes in the high-dose combination group. The incidence of intercerebral hemorrhage was highest in the high-dose combination group (6 of 6 animals) and lowest in the single treatment with 7E3 F(ab') 2 alone (1 of 10 animals) ( P < 0.05). Our data show that murine 7E3 F(ab') 2 alone has therapeutic effects when used after cerebral ischemia. Although this study suggests that higher doses of thrombolytic combined with anti-GPIIb/IIIa therapy may increases the risk of intracranial hemorrhage, the data also support the notion that anti-GPIIb/IIIa agents can safely be combined with low doses of thrombolytic agent to produce significant attenuation of neuronal damage with no increase in the incidence of cerebral hemorrhage.

Carcinogenicity of antioxidants BHA, caffeic acid, sesamol, 4-methoxyphenol and catechol at low doses, either alone or in combination, and modulation of their effects in a rat medium-term multi-organ carcinogenesis model.

The carcinogenicity of low dietary levels of the antioxidants butylated hydroxyanisole (BHA), caffeic acid, sesamol, 4-methoxyphenol (4-MP) and catechol, known to target the forestomach or glandular stomach, were examined alone or in combination in a 2-year long-term experiment and their modifying effects assessed in a medium-term multiorgan model. In the carcinogenicity study, groups of 30-31 male F344 rats were treated with 0.4% BHA, 0.4% caffeic acid, 0.4% sesamol, 0.4% 4-MP and 0.16% catechol either alone or in combination for up to 104 weeks and then killed. In the medium-term multi-organ model, groups of 10 to 15 male F344 rats were given diethylnitrosamine (DEN), N-methylnitrosourea (MNU), 1,2-dimethylhydrazine (DMH), N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) and 2,2'-dihydroxy-di-n-propylnitrosamine (DHPN) for a total multiple initiation period of 4 weeks (DMBDD treatment). BHA, caffeic acid, sesamol and 4-MP, each at doses of 0.4% or 0.08%, and catechol at doses of 0.16% or 0.032% were administered in the diet either alone or in combination after completion of the initiation regimen. All surviving animals were killed at the end of week 28, and major organs were examined histopathologically. In the carcinogenicity study, slightly increased incidences of forestomach papillomas were found in the sesamol- (15.8%), caffeic acid- (14.8%), catechol- (3%) and 4-MP- (11.5%) treated groups as compared with basal diet (0%), and a significant increase was observed with the five antioxidants in combination (42.9%, P < 0.001). In a medium-term multiorgan carcinogenesis model, incidences of forestomach papillomas and/or carcinomas were increased in each high dose group, but additive or synergistic effects were not found in the combination group. In the low dose case, the incidence of forestomach papillomas was significantly increased only in the combination group. With regard to other organs, the incidence of colon tumors was significantly decreased only in the high dose combination group. The results indicate that even at low dose levels phenolic compounds can exert additive/synergistic effect on carcinogenesis.