Effects of invigorating-spleen and anticancer prescription on extracellular signal-regulated kinase/mitogen-activated protein kinase signaling pathway in colon cancer mice model

BACKGROUND Colon cancer(CC)is one of the most common malignant tumors in the gastrointestinal system.Overall,CC had the third highest incidence but the second highest mortality rate globally in 2020.Nowadays,CC is mainly treated with capecitabine chemotherapy regimen,supplemented by radiotherapy,immunotherapy and targeted therapy,but there are still limitations,so Chinese medicine plays an important role.AIM To investigate the effects of invigorating-spleen and anticancer prescription(ISAP)on body weight,tumor inhibition rate and expression levels of proteins in extracellular-signal-regulated kinase(ERK)/mitogen-activated protein kinase(MAPK)signaling pathway in CC mice model.METHODS The CC mice model were established and the mice were randomly divided into 5 groups,including the control group,capecitabine group,the low-dose,mediumdose and high-dose groups of ISAP,with 8 mice in each group,respectively.After 2 weeks of intervention,the body weight and tumor inhibition rate of mice were observed,and the expression of RAS,ERK,phosphorylated ERK(p-ERK),C-MYC and matrix metalloproteinase 2(MMP2)proteins in the tissues of tumors were detected.RESULTS Compared with the control group,the differences of body weight before and after treatment was much smaller in the groups of ISAP,with the smallest difference in the high-dose group of ISAP,while the capecitabine group had the greatest difference,indicating ISAP had a significant inhibiting effect on the growth of transplanted tumor in mice.The expression of RAS protein was decreased in the low-and medium-dose groups of ISAP,and the change of p-ERK was significant in the medium-and high-dose groups of ISAP.MMP2 protein expression was significantly decreased in both the low-dose and medium-dose groups of ISAP.There were no significant changes in ERK in the ISAP group compared to the capecitabine group,while RAS,MMP2,and C-MYC protein expression were reduced in the ISAP group.The expression level of C-MYC protein decreased after treated with ISAP,and the decrease was the most significant in the medium-dose group of ISAP.CONCLUSION ISAP has a potential inhibiting effect on transplanted tumor in mice,and could maintain the general conditions,physical strength and body weight of mice.The expression levels of RAS,p-ERK,MMP2 and c-myc were also decreased to a certain extent.By inhibiting the expression of upstream proteins,the expression levels of downstream proteins in ERK/MAPK signaling pathway were significantly decreased.Therefore,it can be concluded that ISAP may exert an anti-tumor effect by blocking the ERK/MAPK signaling pathway and inhibiting the expression of MMP2 and c-myc proteins.

Insights into sensitizing and eliciting capacity of gastric and gastrointestinal digestion products of shrimp(Penaeus vannamei)proteins in BALB/c mice

Shrimp(Penaeus vannamei)proteins have been shown an allergenic potential;however,little information is available on the sensitizing and eliciting capacity of shrimp protein digestion products.In this study,a BALB/c mice model was used to explore the allergenicity of shrimp protein sample(SPS)and their gastric and gastrointestinal digestion products(GDS/GIDS).As compared with the SPS groups,the GDS/GIDS groups caused lower specific immunoglobulins(Ig E/Ig G1)levels(P<0.05),but higher than the control groups,indicating that the digestion products sensitized the mice.Meanwhile,spleen index,mouse mast cell protease-1(m MCP-1)concentration and proportion of degranulated mast cells were significantly reduced in the GDS/GIDS groups(P<0.05);simultaneously,allergic symptoms,vascular permeability and histopathological changes of tissues were alleviated.Nevertheless,the allergenicity of digestion products cannot be eliminated and still cause systemic allergic reactions in mice.The study showed that the digestion products of shrimp still had high sensitizing and eliciting capacity.

Targeting Effect Study of ￣(3) H-Mitoxantrone Nanosphereson Hepatocellular Carcinoma(HCC) Model in Nude Mice

The distribution of ￣(3)H-mitoxantrone polybutyl cyanoacrylate nanospheres(￣(3)H-DHAQ-PBCA-NS)in the viscera,muscle and tumors of human hepatocellular carcinoma (HCC)model in nude mice was studied with liquid scintillation counting techniique. The results showed that the ￣(3)H-DHAQ-PBCA-NS had remarkable liver targeting effect. The content of ￣(3)H-DHAQ-PBCA-NSin liver and heterotopic liver tumor was found to be 71.31±10. 49% of total amount of drug in animal body. It was also found that the content of ￣(3)H-DHAQ-PBCA-NS in liver was higher than that in liver tissue, and the content of ￣(3)H-DHAQ-PBCA-NS in annpit tumor was higher than that in armpit muscle tissue,but had no significant difference;It provides an ideal preparation for the DHAQ admini-stration.

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.

Study of Candida Albicans Vaginitis Model in Kunming Mice

The model of vaginal candidiasis in Kunming mice was constructed in order to search for the optima construction conditions and provide an economic animal model of Candida albicans (C. albicans) vaginitis. Estrogen benzoate (E2) was given to mice at different concentrations ranging from 0.0 to 0.05 mg/mouse (4 levels) beginning 72 h prior to vaginal inoculation, then mice were in- oculated intravaginally with various concentrations of stationary-phase C. albicans blastoconidia (ATCC90028) (5 levels) in 20 μL of phosphate-buffered saline (PBS) in each E2 level. General state, scores of genital pathology, the hyphae and vaginal fungal burden (CFU) in vaginal lavage fluid, the hydrops rate of uterus and vaginal tissues for pathological section in mice were observed and ob- tained at day 2, 4, 7, 14 and 21 after inoculation. The results showed the infection rate in mice was related to the dosage of E2 and concentration of C. albicans blastoconidia. Additionally there was better cross-effect between the two treated factors. The infection rate was about 80% on the day 4, and could reach 100% on the day 7 until the end of experiment after inoculated intravaginally in groups of E2I3, E2 0.025 mg/mouse injected hypodermically and inoculated intravaginally with 5×104 C. albicans blastoconidia, and large amount of hyphae and blastoconidia could be observe in superfi- cial layer tissue and canal of vaginal by PAS. From the results in our experiment it was concluded that E2I3 was the optima construction condition in kunming mice.

Prevention and Treatment of Chinese Herbal Preparation Yinqiaotiangan against Pathologic Model of Streptococcus suis Serotype II in Kunming Mice

[ Objectlve] This study aimed to investigate the antimicrobial effect of Chinese herbal preparation Yinqiaotiangan against Streptococcus suis serotype II in vivo. [ Method ] The prevention and treatment tests were conducted with Kunming mice weighing about 18 -22 g. In the prevention test, Kunming mice were inocu- lated with Streptococcus suis serotype II and simultaneously taken orally 0.5, 1.0 and 2.0 g/ml Chinese herbal preparation Yinqiaotiangan respectively for continu- ous 3 d, once a day; the incidence rate, mortality rate and protective rate were detected after 7 d. In the treatment test, Kunming mice were inoculated with Strepto- coccus suis serotype II to establish the Streptococcus suis serotype II pathogenic model, and then taken orally 0.5, 1.0 and 2.0 g/ml Chinese herbal preparation Yin- qiaotiangan respectively for continuous 3 d, twice a day; the mortality rate, cure rate and effective rate were detected after 7 d. [ Result ] Results of the prevention test showed that the protective rate in experimental groups was extremely significantly higher than that in control group （P 〈0.01 ）, while the incidence rate and mortality rate were extremely significantly lower than that in control group （P 〈0.01 ）. Results of the treatment test showed that the incidence rate in experimental groups was extremely significantly lower than that in control group （P 〈0.01 ）, the cure rate in 0.5 g/ml group was extremely significantly higher than that in 1.0 g/ml group and 2.0 g/ml group （P 〈 0.01 ）, the effective rate in 0.5 g/ml group was significantly higher than that in 1.0 g/ml group and 2.0 g/ml group （ P 〈 0.05 ）, with no significant difference from the positive group （P 〉 0.05 ）. [ Conclusion ] The pathologic model of Streptococcus su/s serotype II could be effec- tively prevented and treated by oral intake of low dose of Chinese herbal preparation Yinqiaotiangan in Kunming mice.

Establishment of Xenotransplantation Model of Human CN-AML with FLT3-ITD^(mut)/NPM1 in NOD/SCID Mice

Summary： Patients with FLT3-ITD^mmutt/NPM1- cytogenetically normal acute myeloid leukemia （CN-AML）, as high-risk molecular group in CN-AML, are associated with a worse prognosis than other CN-AML patients. It is beneficial to generate xenotransplantation model of FLT3-ITD^mut/NPM1- CN-AML to better understand the pathogenesis and therapeutic strategies of such AML subtype. The purpose of present study was to establish the xenotransplantation model in NOD/SCID mice with FLT3-ITD^mut/NPM1- CN-AML primary cells. The FLT3-ITD^mut/NPM1- CN-AML primary cells from 3 of 7 cases were successfully transplanted into NOD/SCID mice, and human CD45 positive cells were detected in the peripheral blood, spleen and bone marrow of mice by using flow cytometry. Infiltration of human leukemia cells in various organs of mice was observed by using immunohistochemistry. Gene analysis confirmed sustained FLT3/ITD mutation without NPM1 mutation in mice. By performing serial transplantation, it was found that characteristics of the leukemia cells in secondary and tertiary genera- tion models remained unchanged. Moreover, in vivo cytarabine administration could extend survival of NOD/SCID mice, which was consistent with clinical observation. In conclusion, we successfully estab- lished xenotransplantation model of human FLT3-ITD^mut/NPM1- CN-AML in NOD/SCID mice. The model was able to present primary disease and suitable to evaluate the curative effects of new drugs or therapy strategies.

Can outbred mice be used as a mouse model of mild cognitive impairment?

Deficits in spatial learning and memory are some of the earliest symptoms in mild cognitive impairment （MCI）. However, there are few valid MCI animal models available to evaluate putative therapeutic strategies. The aim of this study was to obtain a natural animal model of MCI. Outbred Kunming （aged 5 and 12.5 months） and ICR （7 and 12 months） mice were utilized in the present study. Morris water maze and radial six-arm water maze （RAWM） were simultaneously used to evaluate impaired spatial learning and memory in middle-aged mice （approximately 12 months of age）. Compared with younger mice in the respective groups, the middle-aged mice suffered visible impairment of spatial memory in the Morris water maze and RAWM, and mild spatial learning deficiency occurred in the RAWM study alone. Thus outbred Kunming and ICR mice could be utilized as a natural animal model for MCI, in particular for memory impairment studies.

BALB/c mice model of cytomegalovirus-induced myocarditis

Objective: A BALB/c mice model of cytomegalovirus-induced myocarditis was established. Methods: Twenty-five inbred female BALB/c mice free of murine cytomegalovirus(MCMV) infection (5 weeks old, 16-18 g),were infected with 1×10~4 PFU MCMV by the intraperitoneal (i.p.) administration. All experimental mice were sacrificed on day 3, 5, 7, 10,and 14 after i. p. administration. The hearts were removed under aseptic conditions, and were transected along the midline. Aliquots of hearts were handled with Bouin's fixative for histological examination. Residual hearts were immediately frozen in liquid nitrogen and stored at -80℃ until MCMV titre was determined by a plaque assay. Seurm cTnI level was assayed by ELISA. Results: MCMV in the heart was at extremely low level on day 3 after i. p. administration, reached to the peak on day 7-10, and then ran down. A mixed cellular infiltrate composed of polymorphonuclear neutrophils and mononuclear lymphocytes was observed on day 3, reaching to the peak on day 7-10 after MCMV infection, and was maintained for at least 3-4 months later. Seurm cTnI levels were elevated on day 3 after i.p. administration, reaching to the peak it day 7-10. Conclusion: The BALB/c mice model for cytomegalovirus-induced myocarditis was successfully established, that might make it possible to screen antiviral drugs for treating viral myocarditis and to investigate and evaluate the pathogenesis and prognosis of this disease.

Establishment of Experimental Model of Hepatic Schistosoma Japonicum Egg Granulomas in Mice

An experimental model of hepatic Schistosoma japonicum egg granuloma was established in C57BL/6 mice sensitized with soluble egg antigen(SEA) by direct injection of vital egg suspension into the spleen. The mice infected with cercariae of Schistosoma japonicum in abdominal skin were used for comparative studies. The results showed that morbidity of hepatic Schistosoma japonicum egg granulomas in the group sensitized with SEA was 100% and that the morphology, cellular constituents, developing process and the diameter and size of the egg granulomas in the group sensitized with SEA were similar to those of the group infected with cercariae. The authors suggest that this experimental model is a useful and appropriate tool for the study on egg granulomas of Schistosoma japonicum.

Type Ⅰ interferon receptor knockout mice as models for infection of highly pathogenic viruses with outbreak potential

Due to their inability to generate a complete immune response, mice knockout for type I interferon （IFN） receptors （Ifnar-/-） are more susceptible to viral infections, and are thus commonly used for pathogenesis studies. This mouse model has been used to study many diseases caused by highly pathogenic viruses from many families, including the Flaviviridae, Filoviridae, Arenaviridae, Bunyaviridae, Henipaviridae, and Togaviridae. In this review, we summarize the findings from these animal studies, and discuss the pros and cons of using this model versus other known methods for studying pathogenesis in animals.

Establishment of Retinoblastoma Model in NOD-SCID Mice and Study of Metastasis

Purpose: To establish model of retinoblastoma subcutaneously in NOD-SCID mice and study rules of formation and distribution of retinoblastoma metastasis.Methods: Retinoblastoma cells SO-RB50 were inoculated subcutaneously in NOD-SCID mice. Animal acts and tumor formation, growth and metastasis in NOD-SCID mice were observed. Primary and metastatic tumors were studied pathohistologically by HE and immunohistochemical staining.Results: The latent periods of tumor growth were 12～19 days and the taken rate of tumor was 100%. 32 days later, 5 NOD-SCID mice were found with tumors that had metastasized to areas mainly located in the abdominal cavity and the side of the kidney; the metastatic time of tumors in the mice also differed. The tumor cells of the primary nodules and the metastasis were similar with human retinoblastoma cells and positive in immunohistochemical staining of NSE.Conclusion: The subcutaneous model of retinoblastoma in NOD-SCID mice showed a high taken rate and a short latent period of tumor, which had a high metastatic rate and was the best model in research of behaviors of retinoblastoma at present.

绵羊肺炎支原体小鼠感染模型的建立

旨在探索小鼠作为绵羊肺炎支原体(Mycoplasma ovipneumoniae,Mo)实验室感染模型的可行性,筛选建立Mo感染小鼠模型的最佳方法。将60只BALB/c小鼠随机分为对照组、Mo感染组1、感染组2、感染组3和感染组4(n=12)。Mo感染组1小鼠分别滴鼻30μL和腹腔注射25μL 10^(8 )CCU·mL^(-1) Mo NJ01株菌液各1次,Mo感染组2和3小鼠分别滴鼻2和3次30μL 10^(8 )CCU·mL^(-1) Mo NJ01株菌液,Mo感染组4小鼠喉头喷雾50μL 10^(8 )CCU·mL^(-1) Mo NJ01株菌液2次,对照组用正常培养基处理。分别于感染后第0、7和14天称量小鼠体重。感染后第14天处死所有小鼠,采集血液和肺组织。通过HE染色法进行肺组织病理学检查、剖检进行肺组织病理评分、qPCR方法检测肺组织中Mo的载荷量和ELISA检测血清Mo IgG水平,确定小鼠感染模型是否建立成功及最佳建立方法。Mo感染组2和组4中小鼠体重第14天时分别比对照组显著降低17.2%(P<0.05)和21.6%(P<0.05);感染第13天,感染组2和组4小鼠出现死亡;感染后第14天剖检,Mo感染组2和组4小鼠肺部有炎症,肺病变平均评分分别为3.5和3.3,均显著高于Mo感染组1(P<0.05)和组3(P<0.05),而对照组小鼠无病变;组织病理学检查发现,Mo感染组小鼠肺可见不同程度的间质性肺炎,肺泡腔内有炎细胞浸润;对照组小鼠肺组织结构正常、完整,肺泡内未见明显细胞浸润。小鼠肺组织中Mo DNA拷贝数在Mo感染组1和组3中分别为10^(2.56)和10^(3.21)拷贝·g^(-1);感染组2和组4分别为10^(3.84)和10^(3.77)拷贝·g^(-1),且显著高于Mo感染组1(P<0.05)和组3(P<0.05),对照组为阴性。小鼠血清Mo抗体OD_(450 nm)值在Mo感染组1、组2、组3和组4分别为0.63、1.05、0.81和0.99,均显著高于对照组(P<0.05),且组2和组4均显著高于1组(P<0.05)。本研究通过1×10^(8 )CCU·mL^(-1)的Mo NJ01株菌液滴鼻2次或喉头喷雾2次均可成功建立Mo感染小鼠模型。为绵羊肺炎支原体的致病机制及防治策略研究奠定基础。

NADPH氧化酶4在1型糖尿病模型小鼠角膜病变中的致病作用及其机制

目的探讨还原型烟酰胺腺嘌呤二核苷酸氧化酶4(Nox4)在1型DM模型小鼠角膜病变中的致病作用及其可能机制。方法选择Nox4基因敲除(Nox4^(-/-))纯合子雄性小鼠40只,以鼠龄、性别匹配的野生型C57BL/6(Nox4^(+/+))小鼠120只作为对照。采用随机数字表法分别将2种小鼠随机分为DM组和非DM组,DM组小鼠采用链脲佐菌素腹腔内注射法构建1型DM模型。采用随机数字表法分别将Nox4^(+/+)小鼠DM组和非DM组分为普通饲料喂养小鼠和添加Nox4抑制剂GKT137831(GKT)饲料喂养小鼠。于DM造模后第16周采用酚红棉线法检测各组小鼠泪液分泌量;采用荧光素钠染色评分法评估角膜上皮完整性;采用激光扫描共聚焦显微镜观察角膜基质层神经纤维密度变化;采用CellROX荧光探针检测角膜上皮中活性氧簇(ROS)含量;采用免疫荧光染色法检测小鼠角膜上皮中E-Cadherin蛋白和核因子-κB(NF-κB)蛋白表达变化;采用角膜铺片TUBB3染色法检测角膜中央区神经纤维密度。结果Nox4^(+/+)小鼠DM组和非DM组泪液分泌量分别为(2.40±1.18)和(5.30±1.02)mm/min,差异有统计学意义(P<0.01);Nox4^(-/-)小鼠DM组泪液分泌量为(4.19±0.63)mm/min,明显多于Nox4^(+/+)小鼠DM组,差异有统计学意义(P<0.05);普通饲料喂养小鼠与GKT添加饲料喂养小鼠DM组泪液分泌量分别为(2.23±0.83)和(4.02±0.71)mm/min,差异有统计学意义(P<0.01)。与Nox4^(+/+)小鼠非DM组比较,Nox4^(+/+)小鼠DM组角膜荧光素染色评分显著升高,角膜神经纤维密度显著降低,角膜上皮中ROS荧光强度明显增强,E-Cadherin蛋白表达荧光强度减弱,NF-κB蛋白表达荧光强度增强。Nox4^(-/-)或GKT添加饲料喂养小鼠DM组与非DM组比较角膜上皮中ROS荧光增强,E-Cadherin蛋白表达荧光减弱。Nox4^(-/-)和GKT添加饲料喂养小鼠DM组角膜上皮细胞中NF-κB蛋白荧光强度均较弱,与非DM组强度一致。角膜铺片免疫荧光染色显示,Nox4^(+/+)小鼠DM组中TUBB3染色的神经纤维密度明显低于非DM组,Nox4^(-/-)或GKT添加饲料喂养小鼠DM组角膜基质层神经纤维与非DM组比较无明显减少。结论Nox4参与了糖尿病角膜病变的致病过程,其机制可能与氧化应激诱导ROS产物聚集,激活NF-κB介导的炎症反应有关。

肝郁气滞、肝胆湿热型胆囊胆固醇沉着症动物模型建立与评价

目的:初步建立并评价肝郁气滞、肝胆湿热证型的胆囊胆固醇沉着症小鼠模型,为深入研究该病的发病机制、病理变化,探索临床治疗方案提供模型支持。方法:48只C57BL/6J雄性小鼠,随机分为空白对照组(WT)、造模3周组(LD-3)、造模6周组(LD-6)、造模9周组(LD-9),每组12只。WT组给予普通饲料+正常饮用水喂养,造模组分别给予3、6、9周高脂高胆固醇饲料+正常饮用水喂养。在造模期间,每日观察小鼠一般情况,分别在3、6、9周麻醉后处死动物,眼眶取血,检测血清中总胆固醇(TC)、甘油三酯(TG)含量,取组织标本,采用尼罗红染色观察胆囊脂质沉积情况,红外光谱分析胆汁成分;胆固醇代谢指标:免疫组化法检测肝脏三磷酸腺苷结合盒转运体A组1(ABCA1)蛋白表达,进行模型评价。结果:与WT组相比,各组造模小鼠体重显著增加(P<0.05);血清TC水平显著升高(P<0.05),血清TG水平明显降低(P<0.05);胆囊脂质沉积显著,胆汁混浊度增加;肝脏ABCA1蛋白表达明显增加。结论:本研究成功建立小鼠胆囊胆固醇沉着症模型构建方法,符合临床特征。并成功建立肝郁气滞、肝胆湿热证型的疾病证候类型,可为后续针对胆囊胆固醇沉着症发病机制及治疗方法研究提供稳定的动物模型。

脾虚湿阻型银屑病病证结合小鼠模型的系统评价研究

目的构建脾虚湿阻型银屑病小鼠模型,并从多维度、多方向评价该模型,以期为中医药治疗脾虚湿阻型银屑提供研究支持。方法采用高脂饮食喂养建立脾虚湿阻证小鼠模型,外涂咪喹莫特乳膏建立银屑病小鼠模型,并二者结合构建病证结合小鼠模型。比较体质量、进食量和饮水量,评价脾虚湿阻证候指征。比较皮损面积、PASI评分、经皮水分丢失值、HE染色下皮肤病理学改变评价银屑病严重程度。流式细胞术检测各类细胞表达情况,评价炎症程度。观察脂肪指数、肝脏HE染色下肝脏病理学变化、RT-q PCR法检测肝脏和附睾脂肪的相关因子m RNA表达水平、Western Blot法检测皮肤ABCA1蛋白表达水平,评价脂代谢紊乱情况。结果与寻常型银屑病组小鼠比较,脾虚湿阻型银屑病组小鼠体质量升高(P<0.001),进食量下降(P<0.005),出现毛发油腻,精神萎靡等脾虚湿阻证候指征;经皮水分丢失值升高(P<0.001),PASI评分增加(P<0.001),皮肤HE染色病理结果提示表皮棘层肥厚、杵状增生等银屑病样表现;CD11^(bhigh)Ly6G+中性粒细胞、CD11b^(in)Ly6C^(high)单核细胞、CD11binCD11chigh经典树突细胞、F4/80-CD11c+树突状细胞表达上升(P<0.001);肝脏HE染色病理结果提示细胞空泡样变性,且皮下白色脂肪指数和附睾脂肪指数上升(P<0.005);肝脏FABP4、CD36 m RNA水平升高(P<0.005,P<0.001);附睾脂肪ABCA1和PPARγm RNA水平下降(P<0.05,P<0.01),皮肤ABCA1蛋白水平升高(P>0.05)。结论脾虚湿阻型银屑病小鼠模型可作为可靠的病证结合动物模型,为进一步探讨中医药治疗脾虚湿阻型银屑病提供参考。

石莼酶解提取物延缓皮肤衰老作用

目的:研究石莼酶解提取物对D-半乳糖致衰老模型小鼠的抗衰老作用。方法:按800 mg/kg·d进行颈背部皮下注射D-半乳糖6周,建立小鼠亚急性衰老模型;提取物和V_(C)给药28 d后,表征小鼠皮肤老化及氧化应激水平。结果:与水浸提物相比,石莼酶解提取物中可溶性总糖、蛋白质、还原糖、多酚固形物含量显著提高(P<0.05),且对羟自由基和DPPH自由基清除率及SOD活力提高5.72、7.95、13.26倍。动物实验显示,石莼酶解提取物显著改善衰老性皮肤GSH-Px、T-SOD、CAT、T-AOC活力及MDA、蛋白质羰基水平,缓解小鼠脏器萎缩;皮肤中胶原蛋白含量提高71.63%,Ⅰ/Ⅲ型胶原比例提高55.40%;显著提高机体抗氧化酶GSH-Px、T-SOD、CAT活力和T-AOC水平;抑制机体生物大分子的氧化降解,如降低蛋白质羰基和MDA水平(P<0.05)。结论:石莼酶解提取物可降低小鼠氧化应激水平,延缓小鼠皮肤衰老。

凋亡小分子探针在急性大脑中动脉栓塞和再通模型中的应用

目的探讨凋亡小分子探针CYS-F在急性大脑中动脉栓塞和再通模型中活体分子成像的可行性,分析其反映病变程度的能力。材料与方法阿霉素诱导Hela细胞凋亡,体外验证探针靶向能力;模拟临床,构建小鼠急性大脑中动脉栓塞模型(n=15)和再通模型(n=15),行磁共振血管成像评估靶血管情况;经尾静脉注射CYS-F探究探针分布情况。建模24 h后,T2WI评估病灶体积,近红外成像活体评估细胞凋亡情况。采用尼氏染色和c-fos染色比较两组的差异。结果CYS-F有较好的体外细胞凋亡靶向能力。建模后多普勒血流仪和磁共振血管成像显示,栓塞组成功栓塞大脑中动脉,再通组大脑中动脉复通;近红外成像显示栓塞组大脑中动脉区域荧光信号缺失。建模24 h后,T2WI显示再通组梗死灶体积率明显小于栓塞组(0.055±0.015比0.512±0.220;t=19.761,P<0.001)。栓塞组荧光强度较再通组强,靶背景比分别为1.215±0.162、0.731±0.085(t=10.252,P<0.001)。栓塞组可见大量激活的神经元细胞表达c-fos蛋白,尼氏染色可见大量细胞发生核固缩和裂解。结论小鼠急性大脑中动脉栓塞模型和再通模型贴近临床,应用CYS-F小分子探针可在体对细胞凋亡成像,反映梗死程度,且能在注射初期反映局部组织血流灌注情况。

Ⅱ型糖尿病模型小鼠粪便微生物多样性分析

【目的】揭示粪便微生物对Ⅱ型糖尿病的作用以及影响。【方法】以SPF级BKS-DB雄性小鼠为T2DM模型,通过测定对照组(CN组)和模型组(T2DM组)粪便微生物16SrRNA基因分析其群落结构与组成。【结果】门水平上,发现T2DM模型组小鼠粪便中蝴蝶菌门、弯曲杆菌门菌群丰度显著高于CN组。属水平中,T2DM组发现16个特有菌属,其中丰度占比最大的是库特氏菌属。在差异显著的前10个菌属中,9个菌属T2DM组菌群丰度显著高于CN组(P<0.05),仅有1个菌群丰度显著低于CN组。通过LEfSe分析,发现厌氧菌属、棒状杆菌属在T2DM模型组出现富集,而这两种菌属都存在于免疫力低下的人群中,与糖尿病呈正相关存在。【结论】在T2DM模型小鼠的粪便中,蝴蝶菌门、拟杆菌属等菌群出现高表达现象,同时,厌氧菌属、棒状杆菌属也出现了失衡,这可能导致了机体免疫力降低,正向影响糖尿病的发生。