KM Mice Research Articles

White tea (Camellia sinenesis (L.)) ethanol extracts attenuate reserpine-induced gastric ulcers in mice

The gastroprotective effect of white tea extract ethanol (WTEE) on reserpine induced gastric ulcers in KM mice was investigated. Gastric juice secretion, the pH of gastric juice, serum neuropeptides, including motilin (MOT), substance P (SP), vasoactive intestinal peptide (VIP), and somatostatin (SS) levels, and serum levels of the inflammatory cytokines TNF-α, IL-6, IL-12, and IFN-γ were measured. WTEE dose-dependently reduced reserpine-induced gastric juice secretion and increased the pH of gastric juice. WTEE increased serum levels of VIP and SS and reduced levels of MOT, SP, and inflammatory cytokines in the serum. WTEE modulated gastric expression of occludin and p38 phosphorylation in ulcer-bearing mice. WTEE has a gastroprotective effect against reserpine-induced gastric ulcers in KM mice via reduction of gastric juice secretion, modulation of serum neuropeptide levels, attenuation of serum inflammatory cytokines, and regulation of gastric levels of occludin and p38 phosphorylation.

Licochalcone A-induced human gastric cancer BGC-823 cells apoptosis by regulating ROS-mediated MAPKs and PI3K/AKT signaling pathways.

Both phosphatidylinositol 3-kinase (PI3K)/AKT and mitogen activated protein kinase (MAPK) signaling cascades play an important role in cell proliferation, survival, angiogenesis, and metastasis of tumor cells. In the present report, we investigated the effects of licochalcone A (LA), a flavonoid extracted from licorice root, on the PI3K/AKT/mTOR and MAPK activation pathways in human gastric cancer BGC-823 cells. LA increased reactive oxygen species (ROS) levels, which is associated with the induction of apoptosis as characterized by positive Annexin V binding and activation of caspase-3, and cleavage of poly-ADP-ribose polymerase (PARP). Inhibition of ROS generation by N-acetylcysteine (NAC) significantly prevented LA-induced apoptosis. Interestingly, we also observed that LA caused the activation of ERK, JNK, and p38 MAPK in BGC-823 cells. The antitumour activity of LA-treated BGC-823 cells was significantly distinct in KM mice in vivo. All the findings from our study suggest that LA can interfere with MAPK signaling cascades, initiate ROS generation, induce oxidative stress and consequently cause BGC cell apoptosis.

Energy intake, oxidative stress and antioxidant in mice during lactation.

Reproduction is the highest energy demand period for small mammals, during which both energy intake and expenditure are increased to cope with elevated energy requirements of offspring growth and somatic protection. Oxidative stress life history theory proposed that reactive oxygen species (ROS) were produced in direct proportion to metabolic rate, resulting in oxidative stress and damage to macromolecules. In the present study, several markers of oxidative stress and antioxidants activities were examined in brain, liver, kidneys, skeletal muscle and small intestine in non-lactating (Non-Lac) and lactating (Lac) KM mice. Uncoupling protein (ucps) gene expression was examined in brain, liver and muscle. During peak lactation, gross energy intake was 254% higher in Lac mice than in Non-Lac mice. Levels of H2O2 of Lac mice were 17.7% higher in brain (P<0.05), but 21.1% (P<0.01) and 14.5% (P<0.05) lower in liver and small intestine than that of Non-Lac mice. Malonadialdehyde (MDA) levels of Lac mice were significantly higher in brain, but lower in liver, kidneys, muscle and small intestine than that of Non-Lac mice. Activity of glutathione peroxidase (GSH-PX) was significantly decreased in brain and liver in the Lac group compared with that in the Non-Lac group. Total antioxidant capacity (T-AOC) activity of Lac mice was significantly higher in muscle, but lower in kidneys than Non-Lac mice. Ucp4 and ucp5 gene expression of brain was 394% and 577% higher in Lac mice than in Non-Lac mice. These findings suggest that KM mice show tissue-dependent changes in both oxidative stress and antioxidants. Activities of antioxidants may be regulated physiologically in response to the elevated ROS production in several tissues during peak lactation. Regulations of brain ucp4 and ucp5 gene expression may be involved in the prevention of oxidative damage to the tissue.

Fabrication of poly(γ-glutamic acid)-based biopolymer as the targeted drug delivery system with enhanced cytotoxicity to APN/CD13 over-expressed cells

Poly(γ-glutamic acid)-based targeted drug delivery system (PAMCN) targeting transmembrane metalloprotease aminopeptidase-N (APN/CD13) was fabricated and evaluated for the enhancement of targeting efficiency and cytotoxicity. The cisplatin (CDDP) loading content of PAMCN was about 36 ± 5% and PAMCN showed a sustainable release profile with a half-maximal release time (t1/2) of 23 h. The average size of PAMCN was 132 ± 18 nm determined by light scattering (LS) and 158 ± 67 nm by atomic force microscopy (AFM). Flow cytometry and fluorescence microscope analysis showed that the drug carrier (PAMN) could specifically bind to human umbilical vein endothelial cells (HUVEC). PAMCN enhanced the efficacy of CDDP to HUVEC cells with the half maximal inhibitory concentration (IC50) value decreased to 90.83 ± 33.00 μg/ml comparing with free CDDP treatment and showed less tube formation amounts (p < 0.01) than free CDDP in matrigel angiogenesis inhibition assay in vitro. In vivo toxicity experiment indicated that the survival rate of KM mice in PAMCN group was 100% and PAMCN reduced the hepatic and renal toxicity significantly compared to free CDDP group. Therefore, this novel drug delivery system presents a promising potential for antiangiogenic chemotherapy.

Cloprostenol and eCG influence oestrus synchronisation and uterine development in mice

The aim of this study was to investigate the efficacy of equine chorionic gonadotropin (eCG) and cloprostenol (CLO) administration on oestrus synchronisation, uterine development as well as serum LH and FSH concentrations in mice. One hundred and five KM mice were assigned into eCG-1, eCG-2, eCG-3, CLO-1, CLO-2, CLO-3 and control groups (CG, n = 15). The eCG-1, eCG-2 and eCG-3 groups were intramuscularly injected with 10, 20 and 40 IU eCG. CLO-1, CLO-2 and CLO-3 were intramuscularly injected with 10, 15 and 20 μg cloprostenol acetate. The results showed that 93.33% and 66.67% of synchronised mice displayed oestrus within 18.68-37.59 h. Oestrus numbers, oestrus onset time (EOT) and oestrus rate in CLO and eCG groups were greater than in CG ( P < 0.05). EOT in CLO and eCG groups were 19.88 ± 2.91 h and 34.84 ± 5.05 h. Uterine weights of treatment groups were larger than CG. Uterine weights of the eCG group were higher than those in the CLO group. Uterine horn longitudial diameters (ULD) in treatment groups were larger than CG during the experiment. ULD in eCG-2 and eCG-3 were significantly greater when compared to CG (P < 0.05 or P < 0.01). On days 14 and 21, uterine horn transverse diameters (UTD) in CLO-1, eCG-1, eCG-2 and eCG-3 subgroups were significantly larger than that of CG ( P < 0.05 or P < 0.01). Serum LH concentrations in eCG and CLO increased. Increments in eCG and CLO groups were greater than that of CG. FSH concentrations in eCG mice were higher than those in CLO and CG mice ( P < 0.05) on day 21. Thus, eCG and cloprostenol treatments in mice can improve uterine development and promote the secretion of LH and FSH.

Anti-Inflammatory Effects of Moxibustion on Mice with Adjuvant Arthritis: Role of TRPV1

Aim and Objective: This research was done to observe the anti-inflammatory effect of moxibustion and explore the role of Transient Receptor Potential Vanilloid sub type 1 (TRPV1) on Mice with adjuvant arthritis. Method: Adjuvant arthritis (AA) was established by using the amount of 20 μL of Freund’s Adjuvant Complete (FCA) intradermal injected into right hindfoot plantar. 50 KM mice were randomized into 6 groups by random number table method: control group (n=10), model group (n=8), capsaicin group (n=8), capsazepine group (n=8), moxibustion group (n=8) and Moxibustion+Capsazepine (MC) group (n=8). 16 C57BL/6 wild-type mice are randomized into 2 groups by random number table method: Wild-Type (WT) model group and WT moxibustion group with 8 in each. 14 TRPV1 knockout mice are randomized into 2 groups by random number table method: knockout (KO) model group and KO moxibustion group with 7 in each. Each mouse in the capsaicin group is subcutaneously injected with the amount of 0.1 ml/10 g into the L5, L6 spinal cords. Each mouse in the capsazepine group was intraperitoneally injected with the amount of 0.1 ml/10 g. Similarly, each mouse in the moxibustion group, WT moxibustion group and KO moxibustion group was given a suspended moxibustion with 5 mm × 200 mm specially-made moxa stick for 20 minutes on L5 and L6 spinal cords. Kept the distance between the moxa stick and the skin where L5 and L6 spinal cords located was 10 ± 2 mm that the temperature was stabilized at 46 ± 1°C. Each mouse in MC group was intraperitoneally injected with the amount of 0.1 ml/10 g first, then after 15 minutes was given a suspended moxibustion for 20 minutes on L5 and L6 spinal cords. All the treatments above were performed at the same day when models were successfully establishedand performed once per day for 7 days in total. During the performance, mice in the control group, model group, WT model group and KO model group were of no treatment in any way. The right hindfoot paw volumes of mice in each group were measured and recorded on Day 1, Day 2, Day 4, Day 6 and Day 8 of the experiment. After all treatments, the serum IL-1β and TNF-α level was determined by ELISA. Results: Compared with the model group, the paw volumes in the moxibustion group was significantly decreased (P 0.05). Conclusions: TRPV1 plays a central role in the anti-inflammatory effect of moxibustion, while the effect may be lost if lacked of TRPV1-mediated.

The Relationship between Liver Dysfunction and Arsenic Methylation in Mice

The different methyl metabolic products of inorganic arsenic lead to various toxicities. Arsenic has been demonstrated to induce hepatotoxicity by oxidative stress. The relationship between hepatic injury and inorganic methylation is not yet known. This study was designed to explore the relationship between arsenic methylation and liver oxidative stress induced by arsenic trioxide (ATO). Forty healthy KM mice were randomly divided into control group (0.9% saline) and As2O3 (1.0 mg/Kg/day, 2.0 mg/Kg/day, 4.0 mg/Kg/day) groups with gastric perfusion for five weeks using high-efficiency liquid chromatography and hydride genesis atomic fluorescence spectroscopy (HPLC-HGAFS). The products of arsenic trioxiode methylating, including trivalent inorganic arsenic (iAs 3+ ), pentavalent inorganic arsenic (iAs 5+ ), mono methyl arsenic (MMA), and dimethyl arsenic (DMA) in the liver were determined. The indexes of arsenic methylation, including primary methyl index (PMI) and second methyl index (SMI) were calculated. The level of hepatic function and activity of MDA, GSH, SOD, and TAOC were detected with kits. We found that the remaining arsenic metabolic products in liver significantly increased with the increasing doses of arsenic trioxide and the liver function and oxidative stress deteriorated. Negative correlations were found between MMA%, PMI and GSH, SOD, and TAOC, while DMA% and SMI positively correlated with the levels of ALT and AST. PMI and SMI negatively correlated with TAOC, GSH, SOD, ALT, and AST, positively linked with the level of MDA. The present study demonstrates that the hepatotoxicity induced by the arsenic accounts for deteriorating oxidative injury activized by arsenic methylation metabolism, providing additional evidence to suggest a mechanism of arsenic poisoning. Therefore, reducing the process of arsenic methylation may be potentially benefical in treating and – more importantly – preventing arseniasis.

FGF21 treatment ameliorates alcoholic fatty liver through activation of AMPK-SIRT1 pathway.

Fibroblast growth factor 21 (FGF21), a recently identified member of the FGF superfamily, is mainly secreted from the liver and adipose tissues and plays an important role in improving metabolic syndrome and homeostasis. The aim of this study is to evaluate the role of FGF21 in alcoholic fatty liver disease (AFLD) and to determine if it has a therapeutic effect on AFLD. In this paper, we tested the effect of FGF21 on alcohol-induced liver injury in a murine model of chronic ethanol gavage and alcohol-treated HepG2 cells. Male KM mice received single dose of 5 g/kg ethanol gavage every day for 6 weeks, which induced significant fatty liver and liver injury. The alcohol-induced fatty liver cell model was achieved by adding ethanol into the medium of HepG2 cell cultures at a final concentration of 75 mM for 9 days. Results showed that treatment with recombinant FGF21 ameliorated alcoholic fatty liver and liver injury both in a murine model of chronic ethanol gavage and alcohol-treated HepG2 cells. In addition, FGF21 treatment down-regulated the hepatic expression of fatty acid synthetic key enzyme, activated hepatic AMPK-SIRT1 pathway and significantly down-regulated hepatic oxidative stress protein. Taken together, FGF21 corrects multiple metabolic parameters of AFLD in vitro and in vivo by activation of the AMPK-SIRT1 pathway.

Boehmerine, a New Amide Alkaloid from Piper boehmeriaefolium var. tonkinense, and its Antidepressant Activity

The antidepressant potential of boehmerine, a new amide alkaloid first isolated from Piper boehmeriaefolium (Miq.) C. DC. var. tonkinense, was investigated. The forced swimming test, tail suspension test, and open field test in KM mice were used to reveal the antidepressant-like effect of boehmerine. The results indicated that a significant and dose-dependent decrease in the immobility time of the tested mice was observed after boehmerine administration in both the forced swimming test and tail suspension test, without any accompanying changes in the number of crossings and rearings compared with the control group in the open field test. The results suggest that boehmerine possessed obvious antidepressant-like effects.

Wild-type KRAS inhibits oncogenic KRAS-induced T-ALL in mice.

The role of hyperactive RAS signaling is well established in myeloid malignancies but less clear in T-cell malignancies. The Kras2(LSL)Mx1-Cre (KM) mouse model expresses endogenous KRAS(G12D) in hematopoietic cells and is widely used to study mechanisms and treatment of myeloproliferative neoplasms (MPN). The model displays an intriguing shift from MPN to acute T-cell leukemia (T-ALL) after transplantation to wild-type mice, but the mechanisms underlying this lineage shift is unknown. Here, we show that KRAS(G12D) increases proliferation of both myeloid and T-cell progenitors, but whereas myeloid cells differentiate, T-cell differentiation is inhibited at early stages. Secondary mutations in the expanded pool of T-cell progenitors accompany T-ALL development, and our results indicate that the shift from myeloid to T-lymphoid malignancy after transplantation is explained by the increased likelihood for secondary mutations when the tumor lifespan is increased. We demonstrate that tumor lifespan increases after transplantation because primary KM mice die rapidly, not from MPN, but from KRAS(G12D) expression in nonhematopoietic cells, which causes intestinal bleeding and severe anemia. We also identify loss of the wild-type KRAS allele as a secondary mutation in all T-ALL cells and provide evidence that wild-type KRAS acts as a tumor suppressor in the T-cell lineage in mice.

Behavioral impairments and changes of nitric oxide and inducible nitric oxide synthase in the brains of molarless KM mice

More studies showed that as a common disorder in senior population, loss of teeth could adversely affect human cognitive function, and nitric oxide (NO) might play an important role in the cognitive function. However, the underlying mechanism has not yet been well-established. The objectives of this study are to evaluate behavior changes of KM mice after loss of molars, and levels of NO and inducible nitric oxide synthase (iNOS) in the brain in molarless condition. It is hypothesized that loss of molars of the mice tested results in the cognitive impairments and that the process is mediated by NO in the brain through the signaling pathways. Morris water maze is used to test the behavioral changes after 8 weeks of the surgery. The changes of NO and iNOS are evaluated by using Griess assay, western blot, and immunohistochemistry method. The results show that 8 weeks after loss of molars, the spatial learning and memory of KM mice impair and the levels of NO and iNOS in mice hippocampus increase. These findings suggest that molar extraction is associated with the behavioral impairment, and that the changes of NO and iNOS in the hippocampus may be involved in the behavioral changes in the molarless condition.

Effects of Nano-Selenium on Kindey Selenium Contents, Glutathione Peroxidase Activities and GPx-1 mRNA Expression in Mice

The experiment was conducted to evaluate kindey selenium status, glutathione peroxidase (GPx) activity and GPx-1 expression in mice fed with nanoselenium. Sixty KM mice, female and male in half, were randomly divided into control, sodium selenite and nanoselenium groups. 0.5 milliliter of water, sodium selenite (2 μg Se/mL) and nanoselenium (2 μg Se/mL) were respectively supplemented to the three groups in oral (ig) every day. Whole experiment lasted for 28 days. Kindey selenium contents, GPx activities and GPx-1 mRNA expression were analyzed at experiment trrmination. The results showed that kindey selenium contents and GPx activities in nanoselenium group and sodium selenite group were very significantly higher than those in control group (P&lt;0.01); kindey GPx activities in nanoselenium group were significantly higher than that in sodium selenite group (P&lt;0.05). Kindey mRNA expression of GPx-1 was approx 166% higher in nanoselenium group and approx 157% higher in sodium selenite group than that in control group. Kindey mRNA expression of GPx-1 was approx 3.50 % higher in nanoselenium group than that in sodium selenite group. The results indicated that nanoselenium supplementation could significantly enhance kindey selenium contents, GPx activities and GPx-1 mRNA expression in mice, nanoselenium was more available than sodium selenite in increasing kindey selenium contents, GPx activities and GPx-1 mRNA expression in mice.

Coencapsulation of epirubicin and metformin in PEGylated liposomes inhibits the recurrence of murine sarcoma S180 existing CD133+ cancer stem-like cells

Cancer stem cells (CSCs), also known as tumor-initiating cells, which constitute a subpopulation of tumor cells, are key drivers of tumorigenesis and potential recurrence of cancer. The CSC theory has brought new opportunities as well as challenges to the development of sophisticated drug delivery systems for treating cancer. In the present study, CD133+ cells were sorted from S180 cell lines by magnetic activated cell sorting and a fraction (approximately 1.01%) of CD133+ cells with higher proliferative potential and stronger tumorigenicity in vivo compared with CD133− cells was identified. Furthermore, a procedure for the coencapsulation of epirubicin (EPI) and metformin (MET) was developed with the primary goal of eradicating the bulk population of CD133− cells and the rare population of CD133+ cancer stem-like cells, thus ultimately preventing tumor relapse. The inhibitory effect of free MET was more potent in CD133+ cells than in CD133− cells; in addition, EPI- and MET-coencapsulated liposomes exhibited increased cytotoxicity against CD133+ cells compared with liposomal EPI alone. Meanwhile, tumors in KM mice were completely eliminated upon multiple intravenous injections of liposomal EPI and MET, and tumors virtually eliminated in the experimental period, which could be attributed to the arrest of CD133+ cells in the G0/G1 phase. The coencapsulation of an anti-CSC agent with conventional chemotherapy drugs in liposomes may be a promising drug delivery strategy for fighting cancer and eradicating tumor stem cells.

Effects of long-term exposure to low concentration sevoflurane on reproductive hormones in female mice

Objective To investigate the effects of long-term exposure to low concentration sevoflurane on reproductive hormones in female mice.Methods Thirty-two mature female KM mice were randomly divided into four groups,control group and sevoflurane groups (n=8).Three experimental groups were treated with 30,100,300 ppm (parts permillion1 × 10 6) sevoflurane accordingly 6 h inhalation qd for 4 weeks,while the control group breathed the fresh air only.The mice venous blood was taken before the experiment and after 4 weeks respectively by euzymelinked immunosorbent assay detection of serum luteinizing hormone (LH),follicle stimulating hormone (FSH) and estradiol (E2) concentration optical density (OD),the morphology of cells was observed in mice oocytes.Results Histological examination revealed a complete set of oocytes and cytoplasm uniform in the control group,30 ppm group and 100 ppm group.While 300 ppm group revealed oocytes complete,but no cytoplasmic vacuolization anomaly.Serum concentration of LH,FSH and E2 in 30 ppm and 100 ppm groups showed no significant difference compared to the level before the experiment and control group(P＞0.05).While serum concentrations of LH (3.15±0.52) U/L and FSH (4.9±1.6) U/L were significantly higher and concentrations of E2 (4.9±2.9) pmol/L were significantly lower in 300 ppm group than that of before the experiment,control group,30 ppm group and 100 ppm group (P＜0.05).Conclusions long-term exposure to lower 100 ppm sevoflurance had no effect on reproductive hormones in female mice,but higher than that of 300ppm sevoflurance might have a reproductive damage in mice. Key words: Sevoflurane; Mice; Reproduction

Study on effect of four traditional Chinese medicines distributed along lung meridian on TLR2 and NF-κB expressions in mice with lung heat syndrome

To investigate the effect of four traditional Chinese medicines distributed along lung meridian, namely Ephedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex, on TLR2 and NF-κB expressions in mice with lung heat syndrome, in order to study the intervention effect of the four traditional Chinese medicines (TCMs) on the lung heat syndrome. One hundred KM mice were randomly divided into the normal control group, the model group, the Ephedrae Hebra group, the Zingiberis Rhizoma group, the Scutellariae Radix group and the Mori Cortex group (20, 10 g x kg(-1)), nasally dripped with streptococcus pneumoniae to establish the mouse lung heat syndrome model, and then administered with different TCMs. The expressions of TLR2, NF-κB p65 proteins in lung tissues were analyzed by the immunohistochemical method. The expressions of TLR2, NF-κB p65 mRNA were measured by real time PCR. Compared with the normal control group, the expressions of TLR2 and NF-κB p65 proteins in lung tissues in the model group were higher (P < 0.01), and the expressions of TLR2 and NF-κB p65 mRNA in lung tissues were up-regulated (P < 0.01 or P < 0.05). Compared with the model group, Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group and the Scutellariae Radix high dose group showed decreased expression of TLR2 protein (P < 0.05 or P < 0.01); Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group, Scutellariae Radix high and low dose groups and Mori Cortex high and low dose groups showed reduced expression of NF-κB p65 protein (P < 0.05 or P < 0.01). Ephedrae Hebra high and low dose groups, Zingiberis Rhizoma high and low dose groups, Scutellariae Radix high dose group and Mori Cortex high dose group showed down-regulated expression of TLR2 mRNA (P < 0.01 or P < 0.05). Ephedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex can induce the TLR2/NF-κB inflammatory signal pathways by down-regulating the expressions of TLR2 and NF-κB p65 in protein and mRNA, so as to alleviate the lung tissue injury in mice with lung heat syndrome.

The Effects of Soybean Trypsin Inhibitor on Free Radicals Levels in Pancreatic Mitochondria of Mice

This paper aims to explore the effect of soybean trypsin inhibitor（STI） on oxygen free radicals levels in pancreatic mitochondria of mice. 36 male KM mice were randomly divided into three groups and fed with different diets as follows: control group was fed on control diet, STI group was fed on control diet containing STI, and VC (vitamin C) group was fed on STI diet supplemented with vitamin C. Mice were sacrificed by decapitation after being fed for 3 weeks. Pancreas was quickly removed and mitochondria were prepared for determining the oxidative and antioxidative index. The results showed that, compared with mice fed with control diet, malondialdehyde (MDA) content and activity of nitric oxide synthase (NOS) in pancreatic mitochondria of mice fed with STI diet were significantly increased (<i>P</i><i> </i>< 0.05), the content of glutathione (GSH), total antioxidant capacity (T-AOC), the activities of glutathione peroxidase (GSH-Px), catalase(CAT) and superoxide dismutase (SOD) were significantly dropped. After STI diet was supplemented with antioxidant, the contents of oxidation parameters were significantly decreased (<i>P</i><i> </i>< 0.05), but their values were still higher than those of control group; the activities of antioxidant parameters in pancreatic mitochondria were significantly increased (<i>P </i>< 0.05), but still lower than those of control group. These results supported that STI could cause oxidative stress in pancreatic mitochondria. In addition, vitamin C could interfere with the role of STI on the organism and effectively improve the status of oxidative stress.

Study on safety of Tibetan medicine Zuotai and preliminary study on clinical safety of its compound Dangzuo

Zuotai (gTso thal) is a typical representative of Tibetan medicines containing heavy metals, but there is still lack of modem safety evaluation data so far. In this study, acute toxicity test, sub-acute toxicity test, one-time administration mercury distribution experiment, long-term mercury accumulative toxicity experiment and preliminary study on clinical safety of Compound Dangzuo were conducted in the hope of obtain the medicinal safety data of Zuotai. In the acute toxicity test, half of KM mice given the lethal dose of Zuotai were not died or poisoned, and LD50 was not found. The maximum tolerated dose of Zuotai was 80 g x kg(-1). In the subacute toxicity test, Zuotai could reduce ALT, AST, Crea levels in serums under low dose (13.34 mg x kg(-1) x d(-1)) and medium dose (53.36 mg x kg(-1) x d(-1)), with significant difference under low dose, and increase the levels of ALT, AST, MDA, Crea in serums under high dose (2 000 mg x kg(-1) x d(-1)); besides, the levels of BUN and GSH in serums reduced with the increase in dose of Zuotai, indicating a significant dose-effect relationship. In the one-time administration distribution experiment, the content of mercury in rat kidney, liver and lung increased after the one-time administration with Zuotai, with a significant dose-dependent relationship in kidney. In the long-term mercury accumulative toxicity experiment, KM mice were administered with equivalent doses of Zuotai for 4.5 months and then stopped drug administration for 1.5 months. Since the 2.5th month, they showed significant mercury accumulation in kidney, which gradually reduced after drug withdrawal, without significant change in mercury content in liver, spleen and brain and ALT, AST, TBIL, BUN and Crea in serum. At the 4.5th month after drug administration, KM mice showed slight structural changes in kidney, liver and spleen tissues, and gradually recovered to normal after drug withdrawal. Besides, no significant difference in weight gain was found between the Zuotai group and the control group. According to the findings of the clinical safety study of Dangzuo, after subjects administered Dangzuo under clinical dose for one month, their serum biochemical indicators, blood routine indicators and urine routine indicators showed no significant adverse change. This study proved that traditional Tibetan medicine Zuotai was slightly toxic, with a better safety in clinical combined administration and no adverse effects on bodies under the clinical dose and clinical medication cycle. However, long-term high-dose administration of Zuotai may have a certain effect on kidney.

Tumor inhibition effects and mechanisms of Angelica sinensis and Sophorae flavescentis ait decoction combined with cisplatin in xenograft mice.

To investigate tumor inhibition effects and mechanisms of Angelica sinensis and Sophorae flavescentis ait decoction (ASSF) combined with diamine-dichloroplatinum (DDP). Bodyweight, tumor inhibition rate and q value were calculated for single ASSF or ASSF combined with DDP on H22 carcinoma xenograft KM mice. Biochemical methods for serum LDH, AST, ALT, and AKP, ELISA method for serum HIF-1α, pathological assessemnt of thymus, immunohistochemistry detection of tumor tissue caspase3 and mutant p53 protein, and qRT-PCR detection of bax/ bcl-2 mRNA were applied. Compared with DDP control group, the bodyweight increased in ASSF-DDP group (p<0.01). Tumor inhibition rates for DDP, ASSF, ASSF-DDP were 62.7%. 43.7% and 71.0% respectively, with a q value of 0.90. Compared with other groups, thymus of DDP control group had obvious pathological injury (p<0.01), serum LDH, AST, ALT, AKP increased significantly in DDP control group (p<0.01), while serum HIF-1α was increased in the model control group. Compared with this latter, the expression of mutant p53 protein and bcl-2 mRNA were decreased in all treatment groups (p<0.01), but there were no statistical difference between DDP control p and ASSF-DDP groups. The expression of caspase3 protein and bax mRNA was increased in all treatment groups, with statistical differences between the DDP and ASSF-DDP groups (p<0.01). ASSF can inhibit bodyweight decrease caused by DDP, can inhibit tumor growth synergistically with DDP mainly through increasing serum HIF-1α and pro-apoptotic molecules such as caspase 3 and bax, rather than through decreasing anti-apoptotic mutant p53 and bcl-2. ASSF can reduce DDP toxicity due to decreasing the release of LDH, AST, ALT, AKP into blood and enhancing thymus protection.

Kit formulated asialoglycoprotein receptor targeting tracer based on copolymer for liver SPECT imaging.

Specific targeting of galactose-carrying molecule to ASGP-R in normal hepatocytes has been demonstrated before. In this study, galactosyl polystyrene was synthesized from controllable ratio of functional monomers and radio-labelled with (99m)Tc by formulated kit for SPECT imaging of hepatic function. p(VLA-co-VNI)(46:54) was synthesized by free-radical copolymerization initiated by AIBN, purified by dialysis, lyophilized to kit with Tricine and TPPTS as co-ligands for (99m)Tc labeling. Radiotracer (99m)Tc-p(VLA-co-VNI)(46:54)(Tricine)(TPPTS) was prepared and evaluated by in vitro stability, in vivo metabolism, ex vivo biodistribution and microSPECT/CT imaging in normal KM mice. MicroSPECT/CT and microMRI imaging were also performed in C57BL/b6 mice with xenograft hepatic carcinoma for hepatic function evaluation. (99m)Tc-p(VLA-co-VNI)(46:54)(Tricine)(TPPTS) was obtained in high radio chemical purity (RCP) (>99%) by using instant kit without further purification and excellent in vitro and in vivo stability. The result of biodistribution showed that liver had high uptake (90.49±10.68 ID%/g) at 30 min after injection and was blocked significantly by cold copolymer. MicroSPECT imaging in normal KM mice at 1h and 4h after injection showed good liver retention and targeting properties. Significant defect of activity was observed in the tumor site which was confirmed by MRI imaging. (99m)Tc-p(VLA-co-VNI)(46:54)(Tricine)(TPPTS) with lower ratio of targeting moiety has no observable effect on the specific binding affinity and liver uptake. This makes it possible to introduce more imaging units for multi-modality imaging. Furthermore, the instant kit preparation of (99m)Tc-labeling provides great potential for the evaluation of hepatocyte function in clinical application.

Chronic Exposure to Tributyltin Chloride Induces Pancreatic Islet Cell Apoptosis and Disrupts Glucose Homeostasis in Male Mice

It has been reported that organotin compounds such as triphenyltin or tributyltin (TBT) induce diabetes and insulin resistance. However, histopathological effects of organotin compounds on the Islets of Langerhans and exocrine pancreas are still unclear. In the present study, male KM mice were orally administered with TBT (0.5, 5, and 50 μg/kg) once every 3 days. The fasting plasma glucose levels significantly elevated, and the levels of serum insulin or glucagon decreased in the animals treated with TBT for 60 days. In animals treated for 45 days, the number of apoptotic cells in the islets and exocrine pancreas was elevated in a dose-dependent manner. The percentage of proliferating (PCNA-positive) cells was decreased in the islets, while it was increased in exocrine acinar cells. Immunohistochemistry analysis showed that estrogen receptor (ER) and androgen receptor (AR) were present in vascular endothelium, ductal cells, and islet cells, but absent from pancreatic exocrine cells. TBT exposure decreased the production of estradiol and triiodothyronine and elevated the concentration of testosterone, and resulted in a decrease of ERα expression and an elevation of AR in the pancreas measured by Western boltting. The results suggested that TBT inhibited the proliferation and induced the apoptosis of islet cells via multipathways, causing a decrease of relative islet area in the animals treated for 60 days, which could result in a disruption of glucose homeostasis. The different presence of ERs and AR between the islets and exocrine pancreas might be one of reasons causing different effects on cell proliferation.