A simple and sensitive high performance liquid chromatography with fluorescence detection （HPLC-FD） has been developed for simultaneous quantification of doxorubicin （DOX） and its dipeptide conjugate prodrug （PDO...A simple and sensitive high performance liquid chromatography with fluorescence detection （HPLC-FD） has been developed for simultaneous quantification of doxorubicin （DOX） and its dipeptide conjugate prodrug （PDOX） in mice plasma. The chromatographic separation was carried out on an Amethyst C18-H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity （R2 〉 0.999） over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC-FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.展开更多
The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice.Two 6-mm wounds along the both sides of the spine were created on the back of each mouse（n=80） by using a punch biops...The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice.Two 6-mm wounds along the both sides of the spine were created on the back of each mouse（n=80） by using a punch biopsy.The mice were assigned randomly into two groups,with 40 animals in each group：a non-thermal plasma group in which the mice were treated with the non-thermal plasma;a control group in which the mice were left to heal naturally.Wound healing was evaluated on postoperative days（POD） 4,7,10 and 14（n=5 per group in each POD） by percentage of wound closure.The mice was euthanized on POD 1,4,7,10,14,21,28 and 35（n=1 in each POD）.The wounds were removed,routinely fixed,paraffin-embedded,sectioned and HE-stained.A modified scoring system was used to evaluate the wounds.The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group,earlier than in control group in which acute inflammation reached a peak on POD 7,and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7（P0.05）.The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group（P0.05）.The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10（P0.05 for all）.The count of bacterial colonies was 103 CFU/mL on POD 4 and 20 CFU/mL on POD 7,significantly lower than that in control group（109 CFU/mL on POD 4 and 1012 CFU/mL on the POD 7）（P0.05）.It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colo-nization.展开更多
Cold atmospheric-pressure plasma is a new technology, widely used in many fields of biomedicine,especially in cancer treatment. Cold plasma can selectively kill a variety of tumor cells, and its biological safety in c...Cold atmospheric-pressure plasma is a new technology, widely used in many fields of biomedicine,especially in cancer treatment. Cold plasma can selectively kill a variety of tumor cells, and its biological safety in clinical trials is also very important. In many cases, the patient’s immune level is relatively low, so we first studied the safety assessment of plasma treatment in an immunocompromised animal model. In this study, we examined the safety of immuno-deficient nude mice by oral lavage treatment of plasma-activated water, and studied the growth status, main organs and blood biochemical indexes. Acute toxicity test results showed that the maximum dose of plasma treatment for 15 min had no lethal effect and other acute toxicity. There were no significant changes in body weight and survival status of mice after 2 min and 4 min of plasma-activated water(PAW)treatment for 2 weeks. After treatment, the major organs, including heart, liver, spleen, lung and kidney, were not significantly changed in organ coefficient and tissue structure. Blood biochemical markers showed that blood neutrophils and mononuclear cells were slightly increased, and the others remained unchanged. Liver function, renal function, electrolytes, glucose metabolism and lipid metabolism were not affected by different doses of PAW treatment. The above results indicate that PAW treatment can be used to treat immuno-deficient nude mice without significant safety problems.展开更多
This study was performed to investigate the possible involvement of polyamincs in the development of cervical cancer. The objective of the present study was therefore to lind the specilic polyamine indicators, which c...This study was performed to investigate the possible involvement of polyamincs in the development of cervical cancer. The objective of the present study was therefore to lind the specilic polyamine indicators, which could be used as useful markers for the early determination of cervical cancer. A simple method for the simultaneous determination of plasma concentrations ol live polyamines in normal and UI4 model mice was developed by using HPLC-MS. The samples were derivatized by benzoyl chloride. The derived polyamines were separated on a C18 colunm by a gradient elution with methanol water, and then detected with HPLC-MS. The results showed tha! all polyamine levels in the U 14 model mice were higher than those in normal ones. Thc cadaverine. putrescine and 1,3-diaminopropane levels were significantly higher in U 14 model mice plasma than those in normal mice plasma, especially the putrescine and 1, 3-diaminopropane （P〈0.01）. The cadaverine, putrescine and 1, 3-diaminopropane levels were significantly higher in U 14 mice model urine than those in normal mice urine, especially the cadaverine and 1, 3-diaminopropane （P〈0.01）. Putrescine, cadaverine and I, 3-diaminopropane might be the indicators of the cervical cancer.展开更多
Immunologically mediated aplastic anemia in mice were used as animal models to study the the curative effect of Zaizhang-Ⅰin term of the changes of two pathogenetic aspects in aplastic mice,namely the defciency of he...Immunologically mediated aplastic anemia in mice were used as animal models to study the the curative effect of Zaizhang-Ⅰin term of the changes of two pathogenetic aspects in aplastic mice,namely the defciency of hematopoietic stem cells and the disturbance of immunology. Our results demonstrated that in aplastic mice, after treatment by Zaizhang-Ⅰ,the loss of mature hematopoietice cells (WBC, RBC, Plt) were reduced, and marrow cellular cytosis,and their clinical findings were improved, indicating a partial remission. The present data show that its curative mechanism lies in the action of promoting the recovery of colony forming unit-spleen (CFU-S) and reversing immunologically-induced plasma colony forming unit granulocyte/macrophage (CFU-GM ) inhibitory activity. Natural killer cells activity (Nka) and interleukin-2 tumor necrosis factors (TNF) were also examined to further understand the mechanism by which Zaizhang-Ⅰreverse plasma hematopoietic activity.展开更多
Melatonin (MEL) was investigated for protection against the anthracycline antibiotic doxorubicin (Dox) that is well known for its oxidative damage to various body organs. It was aimed to have a comparison of this prot...Melatonin (MEL) was investigated for protection against the anthracycline antibiotic doxorubicin (Dox) that is well known for its oxidative damage to various body organs. It was aimed to have a comparison of this protection to heart, liver and kidney in the treated subjects. In this study, groups of mice were treated with Dox and melatonin and their individual or combined effects were evaluated by assessing lipidperoxidation, non-protein sulfhydryls (NP-SH) and nitrate/nitrite (NO) contents in these tissues. Plasma aminotransferases, LDH and CK-MB enzyme activities were measured. Moreover, these tissues were subject to histopathological assessment. MEL co-treatment significantly prevented any rise in lipidperoxides more significantly in heart and liver as compared to kidney. In tandem, MEL prevented a decline in GSH that was observed by Dox alone in liver and kidney. Dox significantly increased total NO levels in all the tissues. Melatonin at both dose levels could not afford protection against nitrosative stress. MEL in combination treatment provided significant展开更多
基金supported by the National Natural Science Foundation of China (Grant nos. 21375101, 81573384 and 91417301)Natural Science Foundation of Hubei Province, China (No. 2014CFA077)Innovation Seed Fund and Translational Medical Research Fund of Wuhan University School of Medicine, China
文摘A simple and sensitive high performance liquid chromatography with fluorescence detection （HPLC-FD） has been developed for simultaneous quantification of doxorubicin （DOX） and its dipeptide conjugate prodrug （PDOX） in mice plasma. The chromatographic separation was carried out on an Amethyst C18-H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity （R2 〉 0.999） over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC-FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.
基金supported by grants from the National Natural Sciences Foundation of China(Nos.10875048,30700717)
文摘The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice.Two 6-mm wounds along the both sides of the spine were created on the back of each mouse（n=80） by using a punch biopsy.The mice were assigned randomly into two groups,with 40 animals in each group：a non-thermal plasma group in which the mice were treated with the non-thermal plasma;a control group in which the mice were left to heal naturally.Wound healing was evaluated on postoperative days（POD） 4,7,10 and 14（n=5 per group in each POD） by percentage of wound closure.The mice was euthanized on POD 1,4,7,10,14,21,28 and 35（n=1 in each POD）.The wounds were removed,routinely fixed,paraffin-embedded,sectioned and HE-stained.A modified scoring system was used to evaluate the wounds.The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group,earlier than in control group in which acute inflammation reached a peak on POD 7,and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7（P0.05）.The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group（P0.05）.The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10（P0.05 for all）.The count of bacterial colonies was 103 CFU/mL on POD 4 and 20 CFU/mL on POD 7,significantly lower than that in control group（109 CFU/mL on POD 4 and 1012 CFU/mL on the POD 7）（P0.05）.It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colo-nization.
基金supported by National Natural Science Foundation of China (Grant No. 51307135)China Postdoctoral Science Foundation (Grant No. 2017M610639)the Fundamental Research Funds for Central Universities
文摘Cold atmospheric-pressure plasma is a new technology, widely used in many fields of biomedicine,especially in cancer treatment. Cold plasma can selectively kill a variety of tumor cells, and its biological safety in clinical trials is also very important. In many cases, the patient’s immune level is relatively low, so we first studied the safety assessment of plasma treatment in an immunocompromised animal model. In this study, we examined the safety of immuno-deficient nude mice by oral lavage treatment of plasma-activated water, and studied the growth status, main organs and blood biochemical indexes. Acute toxicity test results showed that the maximum dose of plasma treatment for 15 min had no lethal effect and other acute toxicity. There were no significant changes in body weight and survival status of mice after 2 min and 4 min of plasma-activated water(PAW)treatment for 2 weeks. After treatment, the major organs, including heart, liver, spleen, lung and kidney, were not significantly changed in organ coefficient and tissue structure. Blood biochemical markers showed that blood neutrophils and mononuclear cells were slightly increased, and the others remained unchanged. Liver function, renal function, electrolytes, glucose metabolism and lipid metabolism were not affected by different doses of PAW treatment. The above results indicate that PAW treatment can be used to treat immuno-deficient nude mice without significant safety problems.
基金the National Science Foundation (81102784/H2803)the Liaoning Science Foundation (20102211)
文摘This study was performed to investigate the possible involvement of polyamincs in the development of cervical cancer. The objective of the present study was therefore to lind the specilic polyamine indicators, which could be used as useful markers for the early determination of cervical cancer. A simple method for the simultaneous determination of plasma concentrations ol live polyamines in normal and UI4 model mice was developed by using HPLC-MS. The samples were derivatized by benzoyl chloride. The derived polyamines were separated on a C18 colunm by a gradient elution with methanol water, and then detected with HPLC-MS. The results showed tha! all polyamine levels in the U 14 model mice were higher than those in normal ones. Thc cadaverine. putrescine and 1,3-diaminopropane levels were significantly higher in U 14 model mice plasma than those in normal mice plasma, especially the putrescine and 1, 3-diaminopropane （P〈0.01）. The cadaverine, putrescine and 1, 3-diaminopropane levels were significantly higher in U 14 mice model urine than those in normal mice urine, especially the cadaverine and 1, 3-diaminopropane （P〈0.01）. Putrescine, cadaverine and I, 3-diaminopropane might be the indicators of the cervical cancer.
文摘Immunologically mediated aplastic anemia in mice were used as animal models to study the the curative effect of Zaizhang-Ⅰin term of the changes of two pathogenetic aspects in aplastic mice,namely the defciency of hematopoietic stem cells and the disturbance of immunology. Our results demonstrated that in aplastic mice, after treatment by Zaizhang-Ⅰ,the loss of mature hematopoietice cells (WBC, RBC, Plt) were reduced, and marrow cellular cytosis,and their clinical findings were improved, indicating a partial remission. The present data show that its curative mechanism lies in the action of promoting the recovery of colony forming unit-spleen (CFU-S) and reversing immunologically-induced plasma colony forming unit granulocyte/macrophage (CFU-GM ) inhibitory activity. Natural killer cells activity (Nka) and interleukin-2 tumor necrosis factors (TNF) were also examined to further understand the mechanism by which Zaizhang-Ⅰreverse plasma hematopoietic activity.
文摘Melatonin (MEL) was investigated for protection against the anthracycline antibiotic doxorubicin (Dox) that is well known for its oxidative damage to various body organs. It was aimed to have a comparison of this protection to heart, liver and kidney in the treated subjects. In this study, groups of mice were treated with Dox and melatonin and their individual or combined effects were evaluated by assessing lipidperoxidation, non-protein sulfhydryls (NP-SH) and nitrate/nitrite (NO) contents in these tissues. Plasma aminotransferases, LDH and CK-MB enzyme activities were measured. Moreover, these tissues were subject to histopathological assessment. MEL co-treatment significantly prevented any rise in lipidperoxides more significantly in heart and liver as compared to kidney. In tandem, MEL prevented a decline in GSH that was observed by Dox alone in liver and kidney. Dox significantly increased total NO levels in all the tissues. Melatonin at both dose levels could not afford protection against nitrosative stress. MEL in combination treatment provided significant
