Anti-tumor Effect and Mechanism of Pratia Extract on H22 Tumor-bearing Mice

[Objectives]This study was conducted to investigate the inhibitory effect of pratia extract on H22 tumor-bearing mice and the effects on immune organs.[Methods]With the application of H22 liver tumor-bearing mice as an animal model,the animals were divided into such three Pratia extract groups as the high,medium and low dose groups(400,200 and 100 mg/kg)and cyclophosphamide CTX group(20 mg/kg).15 d after the administration,the animals were killed by cervical dislocation,and the tumors,thymuses and spleens were taken and weighed,followed by the calculation of the tumor inhibitory rate and the thymus and spleen index,and the serum tumor necrosis factor-α(TNF-α)and interleukin-2(IL-2)levels were determined by ELISA assay.[Results]The inhibitory rates were 54.1,32.6 and 8.2%,respectively,and there were significant differences from the model group(P<0.05);and the spleen index of the tumor-bearing mice was reduced,while the thymus index was improved.The serological results showed that the drug-administrated groups significantly improved the IL-2 levels in the tumor-bearing mice,but had no effects on TNF-α.[Conclusions]Pratia extract has an antitumor effect on H22 tumor-bearing mice,and show certain dose-effect relationship,and its mechanism may be related to enhancing the immune function in tumor-bearing mice by regulating IL-2.

Detraining after tumor-bearing accelerates tumor growth while continuous training decreases tumor growth in mice

Objective:Previous studies have shown that exercise suppresses tumor growth.However,the effects of exercise with different intensities and exercise detraining after tumor-bearing on tumor progression remain unclear.The purpose of this study was to investigate the effects of continuous and disrupted free and exhausted swimming training after tumor-bearing on tumor progression in melanoma B16-F10-bearing C57BL/6 mice.Methods:C57BL/6 mice were subjected to free or exhausted swimming exercise training for 4 weeks prior to the injection of melanoma B16-F10 cells.Subsequently,the B16-F10-bearing mice were maintained with training consisting of free or exhausted swimming or without exercise for 2 weeks during the tumor challenge.Results:The tumor weight was increased by 42%and 109%in mice with 4-week exhausted swimming prior to B16-F10 tumor cells inoculation followed by 2-week training cessation compared with the tumor-bearing control(P<.05)and continuous training groups(P<.01).Tumor weights in groups with exercise detraining after tumor cell inoculation tended to be increased,while the proliferation of splenic T lymphocytes tended to be decreased compared with the group that maintained exercise intensity.After 6-weeks continuous free or exhausted swimming training,the tumor weight of mice was decreased and the proliferation of splenic T lymphocytes was increased compared with the tumor-bearing control group.The frequency of natural killer cells in tumors was increased in all exercise training groups of mice.Conclusions:These results suggest that maintaining exercise intensity after tumor-bearing slows tumor growth in mice,possibly because of the enhanced proliferative activity of splenic lymphocytes rather than natural killer cell infiltration.However,detraining after tumor-bearing might accelerate tumor progression because of the reduced proliferation of splenic T lymphocytes.

Effect of moxibustion on survival status and nutritional metabolic factor in tumor-bearing rats with gastric cancer

Objective:To observe the effect of moxibustion on the survival status and nutritional metabolic factors of tumor-bearing rats with gastric cancer and explore the inhibitory effect of moxibustion on the tumorbearing rats.Methods:A total of 40 SD rats were randomly divided into a blank group,a sham-operated group,a model group and a moxibustion group,10 rats in each group.Gastric cancer models were established in the model group and the moxibustion group.The rats of all the groups underwent the same constraining procedure for 20 min every day.Additionally,moxibustion was applied at acupoints for 20 min in the rats of the moxibustion group.There were two groups of acupoints.One group included"Zusanli(足三里ST 36)""Zhongwan(中脘CV12)"and"Guanyuan(关元CV4)".The other group included bilateral"Pishu(脾俞BL 20)"and"Weishu(胃俞BL 21)".Moxibustion was applied for 20 min every day at the acupoints of the two groups alternatively and lasted for 14 days.The survival status of rats was observed and the score of survival status and body weight of rats were recorded every day.At the end of intervention,the orbital blood was collected and the rats were sacrificed for sample collection.The viscera-free body weight was recorded.Using biochemical analyzer,the blood glucose(GLU),albumin(ALB),total protein(TP)and triglyceride(TG)in serum were detected.Results:(1)Regarding the score of the survival status,after modeling,the scores in the model group and the moxibustion group were all higher than those of the sham-operation group(both P<0.05).After intervention,the score in the moxibustion group was lower than that of the model group(P<0.05).(2)After modeling,the body weight in the moxibustion group and the model group was lower than those of the blank group,indicating a statistical significance(both P<0.05).The body weight in the moxibustion group was higher than that of the model group after intervention,indicating the statistical significance(P<0.05).(3)Regarding the viscera-free body weight,it was lower in the model group compared with the sham-operation group,indicating the statistical significance(P<0.05).Compared with the model group,the viscera-free body weight was higher in the moxibustion group,indicating the statistical significance(P<0.04).(4)Regarding the nutritional metabolic factors,compared with the sham-operation group,the levels of GLU and ALB in the model group were lower and TP was higher,presenting the statistical significance(all P<0.05).Compared with the model group,the levels of GLU and ALB were higher in the moxibustion group and TP was lower,presenting the statistical significance(all P<0.05).Conclusion:Moxibustion improves the survival status of tumor-bearing rats with gastric cancer,increases the body weight,the viscera-free body weight and the levels of GLU and ALB in serum and reduces TP of the tumor-bearing rats with gastric cancer and inhibits the growth of gastric cancer.

Possible mechanisms associated with immune escape and apoptosis on anti-hepatocellular carcinoma effect of Mu Ji Fang granules

BACKGROUND Hepatocellular carcinoma(HCC)is one of the most common digestive system cancers with high mortality rates worldwide.The main ingredients in Mu Ji Fang Granules(MJF)are alkaloids,flavonoids,and polysaccharides.MJF has been used in the clinical treatment of hepatitis,cirrhosis and HCC for more than 30 years.Few previous studies have focused on the mechanism of MJF on tumor immunology in the treatment of HCC.AIM To explore the mechanism of action of MJF on tumor immunology in the treatment of HCC.METHODS The absorbable ingredients of MJF were identified using Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight-Mass Spectrometry,and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis.Forty male mice were randomly divided into the Blank,Model,and MJF groups(1.8,5.4,and 10.8 g/kg/d)following 7 d of oral administration.Average body weight gain,spleen and thymus indices were calculated,tumor tissues were stained with hematoxylin and eosin,and Interferon gamma(IFN-γ),Tumor necrosis factorα(TNF-α),Interleukin-2,aspartate aminotransferase,alanine aminotransferase,alpha-fetoprotein(AFP),Fas,and FasL were measured by Enzyme-linked Immunosorbent Assay.Relevant mRNA expression of Bax and Bcl2 was evaluated by Real Time Quantitative PCR(RTqPCR)and protein expression of Transforming growth factorβ1(TGF-β1)and Mothers against decapentaplegic homolog(SMAD)4 was assessed by Western blotting.The HepG2 cell line was treated with 10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL of MJF,and another 3 groups were treated with TGF-β1 inhibitor(LY364947)and different doses of MJF.Relevant mRNA expression of TNF-α,IFN-γ,Bax and Bcl2 was evaluated by RT-qPCR and protein expression of TGF-β1,SMAD2,p-SMAD2,SMAD4,and SMAD7 was assessed by Western blotting.RESULTS It was shown that MJF improved body weight gain and tumor inhibition rate in H22 tumorbearing mice,protected immune organs and liver function,reduced the HCC indicator AFP,affected immunity and apoptosis,and up-regulated the TGF-β1/SMAD signaling pathway,by increasing the relative expression of TGF-β1,SMAD2,p-SMAD2 and SMAD4 and decreasing SMAD7,reducing immune factors TNF-αand IFN-γ,decreasing apoptosis cytokines Fas,FasL and Bcl2/Bax,and inhibiting the effect of LY364947 in HepG2 cells.CONCLUSION MJF inhibits HCC by activating the TGF-β1/SMAD signaling pathway,and affecting immune and apoptotic cytokines,which may be due to MJF adjusting immune escape and apoptosis.

Anti-breast cancer properties and toxicity of Dillenia suffruticosa root aqueous extract in BALB/c mice

Objective:To determine the anti-breast cancer activities and the safety oral consumption of Dillenia suffruticosa root aqueous extract(DRAE)in BALB/c mice.Methods:In the anti-breast cancer study,female BALB/c mice were divided into five groups(n=12),which were(1)positive control(with breast cancer,untreated),(2)negative control(without breast cancer,untreated)and other three groups of mice with breast cancer treated with 1 000,500 and 250 mg/kg of DRAE,respectively,by oral gavage for 28 days.All mice except from the negative control group were injected into the mammary fat pad with 4T1 cells(1×1054T1 cells/0.1 m L of phosphate buffer solution).DRAE was administered orally on Day 11 after the tumor has developed.Results:The tumor volume of the 1 000 mg/kg of DRAE group reduced significantly compared to the positive control while treatment with 500 mg/kg of DRAE had significantly inhibited metastasis to the heart.In the acute toxicity study,treatment with up to5 000 mg/kg of DRAE was not toxic to the animals,indicating its safety when a large amount of this plant extract was ingested.Based on the sub-acute toxicity study,treatment of the highest dose of DRAE(1 000 mg/kg)had mild liver toxicity indicated by mild focal hemorrhage.Conclusions:DRAE possesses anti-breast cancer properties but at the same time it shows mild toxicity to the liver.The non observable adverse effect dose for DRAE is500 mg/kg.

Usefulness of Magnetic Particle Imaging for Monitoring the Effect of Magnetic Targeting

Purpose: Magnetic targeting refers to the attachment of therapeutic agents to magnetizable particles such as magnetic nanoparticles (MNPs) and then applying magnetic fields to concentrate them to the targeted region such as solid tumors. The purpose of this study was to investigate the usefulness of magnetic particle imaging (MPI) for monitoring the effect of magnetic targeting using tumor-bearing mice. Materials and Methods: Colon-26 cells (1 × 106 cells) were implanted into the backs of eight-week-old male BALB/c mice. When the tumor volume reached approximately 100 mm3, the mice were divided into treated (n = 8) and untreated groups (n = 8). The tumors in the treated group were directly injected with MNPs (Resovist?, 250 mM) and a neodymium magnet was attached to the tumor surface, whereas the magnet was not attached to the tumor in the untreated group. The mice were imaged using our MPI scanner and the average and maximum MPI values were obtained by drawing a region of interest (ROI) on the tumor, with the threshold value for extracting the contour of the tumor being taken as 40% of the maximum MPI value in the ROI. The relative tumor volume growth (RTVG) was calculated from (V ? V0)/V0, where V0 and V represented the tumor volume immediately before and after the injection of MNPs, respectively. Results: The average and maximum MPI values in the treated group were significantly higher than those in the untreated group 3 days after the injection of MNPs, suggesting the effectiveness of magnetic targeting. There were no significant differences in RTVG between the two groups. Conclusion: Our preliminary results suggest that MPI is useful for monitoring the effect of magnetic targeting.

Rapid and Simple Analysis of <i>N</i>-Aspartylchlor in E6 (Talaporfin) Using Fluorescence Microtiterplate and Its Application for Determination in Cells, Tissues and Blood

N-aspartylchlorin e6 (talaporfin) concentrations in cancer cells, mouse liver tissues, and human plasma specimens were determined with fluorescence microtiter plate analysis. Talaporfin standard curves were obtained in each of the sample specimens of cells, mouse tissues, or human plasma including serial concentration of talaporfin. The correlation co-efficiencies (r2) of talaporfin standard curves were 0.99-1.00, and the CV less than 5%. Talaporfin incorporation into cells of human breast cancer cell line MCF-7 after incubating with 25 μg/mL talaporfin for up to 24 h revealed that the t-max of the drug incorporation was approximately 5 h, and the maximum drug concentration incorporated was 25 μg/107 cells. Talaporfin incorporation into MCF-7 cells was significantly decreased in the presence of 3 μg/mL cyc-losporine (p < 0.05). Balb/c nu/nu mice implanted human cholangiocarcinoma NOZ cells in liver were administered intravenously 5mg/mouse of talaporfin, and the tissues of normal liver and tumor, as well as plasma specimens, were analyzed for talaporfin concentrations. The mean (SD) of talaporfin concentration in plasma after 30 min of administration was 41.6 (2.3) μg/mL, while the level decreased to undetectable concentrations 2 h after administration. In contrast, the talaporfin concentrations in normal and tumor tissues after 30 min of administration were 1.1-7.8 μg/g tissue, and the level slightly increased or was almost maintained for up to 2-4 h after administration. The heparinized blood of the healthy subjects was incubated with 25 μg/mL talaporfin for up to 24 h. The plasma talaporfin concentration did not significantly change during the incubation, and thus talaporfin appears not to be incorporated into the blood cells. We established rapid and simple analysis procedures of talaporfin in biological specimens using a fluorescence microtiter plate assay. Using this assay procedure, the unique pattern of talaporfin disposition and pharmacokinetics were revealed in human cancer cells, liver tissues of tumor bearing mice, and human blood.

Regulatory effects of evodiamine on glucose metabolism-related factors in CT26 colorectal carcinoma-bearing mice

Objective:To investigate the therapeutic effect of evodiamine(EVO)on the expression of hexokinase(HK),lactate dehydrogenase A(LDHA),and pyruvate kinase M(PKM),key enzymes of glycolysis,in the tumor-bearing mice after modeling mouse colon cancer cells(CT26).Methods:A tumor-bearing mouse model was generated by administering axillary injection of CT26 and intraperitoneally injecting different doses of EVO.The therapeutic effects of EVO on CT26 tumor-bearing mice were evaluated by measuring the thymus and spleen indices,tumor volume,tumor suppression rate,and other related indicators in the tumor tissues of mice in each group after the administration of EVO,in addition,histopathological changes in the tumor tissues of the mice in the groups were studied by hematoxylin and eosin staining.The expression levels of HK,LDHA,and PKM in the tumor tissues of each group of mice were measured by performing Western blot to investigate the mechanism of EVO treatment in CT26 tumor-bearing mice.Results:EVO inhibited the growth of tumors in CT26-bearing mice and enhanced their splenic and thymic indices.Western blot results showed that EVO reduced the expression levels of HK,LDHA,and PKM proteins in the tumor tissues of CT26 tumor-bearing mice.Conclusion:EVO has a therapeutic effect on CT26 tumor-bearing mice,and its mechanism of action may be related to the low expression of key enzymes HK,LDHA and PKM of glycolysis in tumor tissues.

Anti-tumor immunity of BAM-SiPc-mediated vascular photodynamic therapy in a BALB/c mouse model

In recent decades, accumulating evidence from both animal and clinical studies has suggested that a sufficiently activated immune system may strongly augment various types of cancer treatment, including photodynamic therapy （PDT）. Through the generation of reactive oxygen species, PDT eradicates tumors by triggering localized tumor damage and inducing anti-tumor immunity. As the major component of anti-tumor immunity, the involvement of a cell-mediated immune response in PDT has been well investigated in the past decade, whereas the role of humoral immunity has remained relatively unexplored. In the present investigation, using the photosensitizer BAM-SiPc and the CT26 tumor-bearing BALB/c mouse model, it was demonstrated that both cell-mediated and humoral adaptive immune components could be involved in PDT. With a vascular PDT （VPDT） regimen, BAM-SiPc could eradicate the tumors of -70% of tumor-bearing mice and trigger an anti-tumor immune response that could last for more than 1 year. An elevation of Th2 cytokines was detected ex vivoafter VPDT, indicating the potential involvement of a humoral response. An analysis of serum from the VPDT-cured mice also revealed elevated levels of tumor-specific antibodies. Moreover, this serum could effectively hinder tumor growth and protect the mice against further re-challenge in a T-cell-dependent manner. Taken together, these results show that the humoral components induced after BAM-SiPc-VPDT could assist the development of anti-tumor immunity.

Development and progression of cancer cachexia:Perspectives from bench to bedside

Cancer cachexia(CC)is a devastating syndrome characterized by weight loss,reduced fat mass and muscle mass that affects approximately 80%of cancer patients and is responsible for 22%-30%of cancer-associated deaths.Understanding underlying mechanisms for the development of CC are crucial to advance therapies to treat CC and improve cancer outcomes.CC is a multi-organ syndrome that results in extensive skeletal muscle and adipose tissue wasting;however,CC can impair other organs such as the liver,heart,brain,and bone as well.A considerable amount of CC research focuses on changes that occur within the muscle,but cancer-related impairments in other organ systems are understudied.Furthermore,metabolic changes in organ systems other than muscle may contribute to CC.Therefore,the purpose of this review is to address degenerative mechanisms which occur during CC from a whole-body perspective.Outlining the information known about metabolic changes that occur in response to cancer is necessary to develop and enhance therapies to treat CC.As much of the current evidences in CC are from pre-clinical models we should note the majority of the data reviewed here are from preclinical models.