Establishment of a Mouse Thrombocytopenia Model Induced by Cyclophosphamide

An experiment was conducted to compare the effects of two mouse thrombocytopenia models induced by cyclophosphamide at two different administration routes to determine a proper cyclophosphamide administration route that could cause stable thrombocytopenia. A suitable drug dosage that could induce thrombocytopenia in mouse efficiently with the definite administration route was then investigated. BALB/c mice were randomly divided into Normal, Model A and Model B groups. To Model A, 200 mg/kg of cyclophosphamide was given by vena caudalis injection as first dose and 30 mg/kg as maintenance dose by intraperitoneal injection at the following 6 days. To Model B, 150 mg/kg of cyclophosphamide was given by subcutaneous injection once a day for consecutive 3 days. All groups were under investigation for 15 days. The result suggested that a decrease in the number of blood platelets of Model B at the 7th day were significantly than that of Normal. Other platelet related indices like platelet distribution width, mean platelet volume and platelet-large cell ratio of Model B increased significantly in comparison with those of Normal group. The platelets count was reduced but fluctuated greatly, and more than half of the mice died in Model A. Therefore, subcutaneous injection of cyclophosphamide for 3 days was used for the cyclophosphamide dosage test. BALB/c mice were randomly divided into Normal, cyclophosphamide low dose （100 mg/kg）, medium dose （120 mg/kg） and high dose （140 mg/kg） groups. All groups were under investigation for 11 days. Though all 3 dosages successfully initiated thrombocytopenia as the platelets number dropped at the 7th day, the low dose was considered to be a suitable one that was of high efficacy and low toxicity. Thus, BALB/c mice challenged by subcutaneous injection of cyclophosphamide 100 mg/kg per day for 3 consecutive day is one simple, feasible and stable mouse thrombocytopenia model that could be used for pharmacodynamic test of the drugs which are supposed to have platelets increasing effect.

基于线粒体Cyt b基因序列的4个黄尾鲴养殖群体遗传多样性分析

黄尾鲴(Xenocypris davidi)是浙江省自然水域增殖放流的主要鱼类,为了解人工繁育对黄尾鲴遗传多样性的影响,利用线粒体DNA细胞色素b基因(Cyt b)对4个养殖群体的遗传多样性进行研究,旨在为黄尾鲴增殖放流策略制定和实施提供基础数据。结果显示,在编码Cyt b的1 140 bp序列中,检测到157个变异位点,界定了21种单倍型,其中长兴、双浦、八里店和醴陵群体的单倍型数目分别为10个、11个、7个和2个,单倍型多样性介于0.226~0.794,核苷酸多样性介于0.006 14~0.023 86。除醴陵群体遗传多样性较低外,其余3个养殖群体的遗传多样性具有高单倍型数和高核苷酸多样性的特点。4个黄尾鲴养殖群体间的遗传距离为0.018 74~0.092 74,遗传分化指数为0.808 63(P<0.01),其中长兴和八里店群体的分化程度较低,双浦和醴陵群体的分化程度较高,且遗传变异主要发生在群体间。本研究结果可从分子水平为黄尾鲴的资源保护和人工增殖放流提供参考依据。

基于Cyt b基因的江苏省湖泊鳙群体遗传多样性和遗传结构研究

为了解江苏省湖泊鳙群体遗传资源现状,科学地开展鳙增殖放流,采用线粒体Cyt b基因,分析了6个湖泊群体、223个个体的遗传多样性和遗传结构。结果表明,223条Cyt b序列共有10个变异位点,定义9个单倍型,单倍型多样性(Hd)和核苷酸多样性(Pi)分别为(0.725±0.017)和(0.00191±0.00080),整个鳙群体呈现出高Hd和低Pi的遗传多样性模式。6个群体的单倍型多样性为(0.625±0.077)~(0.771±0.032),核苷酸多样性为(0.00159±0.00027)~(0.00221±0.00011)。分子方差分析结果显示,遗传变异主要来源于群体内部(97.17%),总遗传分化系数(Fst)为0.0283。群体间Fst为-0.0258~0.0907,遗传距离为0.0016~0.0022,表明6个群体间遗传分化水平较低。中性检验结果和错配分布分析表明,鳙群体保持稳定,未经历群体扩张。指出,鳙群体遗传多样性较低,遗传结构趋于同质化,需要采取措施,提高湖泊鳙群体遗传多样性。

Immune enhancement effect of an herb complex extract through the activation of natural killer cells and the regulation of cytokine levels in a cyclophosphamide-induced immunosuppression rat model

Objective: To investigate the effects of a herb complex extract(HCE) prepared from Cornus officinalis Sieb. Et Zucc., Eriobotrya japonica Lindley, and olive leaves on immune response of mouse spleen NK cells in vitro and in vivo analysis. Methods: The activity of natural killer(NK) cells was measured in splenocytes and YAC-1 cells. Mice were immunosuppressed using cyclophosphamide(5 mg/kg body weight). Three different doses of HCE(200, 400, and 800 mg/kg body weight) and red ginseng extract(800 mg/kg body weight) which was used as standard immunomodulatory herb were administered orally for 4 weeks. The body weight, dietary, water intake, organs(liver, thymus, and spleen) weight, completed blood count, and cytokines(tumor necrosis factor alpha, interferon gamma, and interleukin-2) production was measured. Results: At the maximum concentration of HCE, the activity of NK cells was increased by 48.5%. HCE increased liver, spleen, and thymus weights without altering numbers of white blood cells, lymphocytes, and neutrophils in a cyclophosphamide-induced immunosuppression rat model. However, HCE recovered the inhibited cytokine expression; HCE(800 mg/kg) increased cytokines levels. The results indicate the immune enhancement potential of this HCE. Conclusion: The HCE enhances immunity by increasing NK cell activity, regulating cytokine levels, and maintaining spleen weight. Therefore, it may be used as a potential immunity enhancer.

Sesamol Alleviates the Cytotoxic Effect of Cyclophosphamide on Normal Human Lung WI-38 Cells via Suppressing RAGE/NF-κB/Autophagy Signaling

Cyclophosphamide(CYL)is a chemotherapeutic medication commonly used in managing various malignancies like breast cancer or leukemia.Though,CYL has been documented to induce lung toxicity.Mechanism of CYL toxicity is through oxidative stress and the release of damage-associated molecular patterns(DAMPs).Sesamol(SES)is a natural antioxidant isolated from Sesamum indicum and its effect against CYL-induced lung toxicity is not studied yet.This study aims to inves-tigate whether SES could prevent any deleterious effects induced by CYL on lung using normal human lung cells,WI-38 cell line,without suppressing its efficacy.Cells were pretreated with SES and/or CYL for 24 h,then cell viability was estimated by MTS and trypan blue assays.The mode of cell death was determined by AO/EB staining.Additionally,caspase-3 level,oxidative stress,and inflammatory markers were evaluated by colorimetric and ELISA techniques.qRT-PCR was performed to evaluate RAGE,NF-κB,and Beclin-1 mRNA-expression.CYL-treated WI-38 cells developed a significantly increased cell death with enhanced oxidative and RAGE/NF-κb/Autophagy signaling,which were all attenuated after pretreatment with SES.Thus,we concluded that SES offered a protective role against CYL-induced lung injury via suppressing oxidative stress and RAGE/NF-κB/Autophagy signaling,which is a natural safe therapeutic option against CYL toxicities.

基于cyt b基因序列的花斑副沙鳅种群遗传多样性比较研究

为了解花斑副沙鳅Parabotia fasciata的种质资源现状,基于cyt b基因序列比较分析了长江水系4个野生群体和5个养殖群体的遗传多样性水平。结果显示,208个样本共检测到29个单倍型,38个多态位点。群体总体单倍型多态性较高(Hd=0.711),核苷酸多态性较低(Pi=0.00235),但野生群体(Hd=0.602,Pi=0.00374)的遗传多样性显著大于养殖群体(Hd=0.381,Pi=0.00091),说明养殖群体需要增加亲本资源。中性检验、错配分布和贝叶斯天际图结果表明,野生群体在历史上经历了种群扩张。赤水群体的单倍型最多,遗传多样性最高(Hd=0.836,Pi=0.00899),与其他群体遗传距离大,说明赤水河的花斑副沙鳅资源较丰富,且已形成相对独立、稳定的地理种群,种质优良,适合用作人工繁殖和新品系创制的种源。

基于cyt b基因序列的光唇鱼群体遗传多样性和遗传结构分析

为了解光唇鱼Acrossocheilus fasciatus自然分布的不同地理群体遗传多样性和遗传结构,基于cyt b基因序列,对浙江省的苕溪、金华江、曹娥江、椒江和安徽省的新安江、青弋江共6个水系的116尾光唇鱼进行分析。结果显示,116条cyt b基因序列定义了12个单倍型,光唇鱼6个群体的单倍型多样性为0~0.648,核苷酸多样性为0~0.004 35,除曹娥江群体呈现高单倍型多样性和低核苷酸多样性的特点外,其他群体均呈现出低单倍型多样性和低核苷酸多样性的特点。遗传分化指数分析显示,除苕溪与椒江群体间属中度遗传分化外,其余各群体间均属于极大的遗传分化(P<0.01)。基于单倍型序列构建的系统发育树显示,新安江、金华江、曹娥江、苕溪和椒江5个群体的单倍型混杂在一起聚为一支,青弋江群体不同单倍型聚为一支。遗传距离分析结果显示,青弋江群体与其他群体间的遗传距离均大于0.03。综合遗传距离、遗传分化指数和单倍型系统发育树结果,青弋江群体与其他群体间产生了明显的遗传分化。核苷酸错配分布和中性检验结果显示,光唇鱼近期并未经历种群扩张事件。本研究结果为光唇鱼种质资源保护与利用、增殖放流和良种选育等相关工作提供了理论依据。

基于线粒体Cyt b基因和D-loop区分析元江鲤和杞麓鲤群体遗传结构

本文利用线粒体Cyt b基因和D-loop区部分序列,分析元江鲤(Cyprinus carpio yuankiang)和杞麓鲤(Cyprinus carpio chilia)群体的遗传结构。结果表明,在两个鲤群体中均获得929 bp的Cyt b基因序列和650 bp的D-Loop区序列。两个群体间平均遗传距离为0.00595(Cyt b)、0.00852(D-loop)。Cyt b基因共检测出元江鲤3种、杞麓鲤15种单倍型,两个群体共享两个单倍型(C-Hap1和C-Hap3);D-loop区序列共检测出元江鲤3种、杞麓鲤6种单倍型,两个群体共享两个单倍型(D-Hap1和D-Hap2)。元江鲤和杞麓鲤群体单倍型多样性(Hd)分别为0.30±0.10(Cyt b)、0.25±0.10(D-loop)和0.89±0.04(Cyt b)、0.73±0.05(D-loop)。元江鲤和杞麓鲤群体核苷酸多样性(π)分别为0.12±0.04(Cyt b)、0.12±0.09(D-loop)和0.77±0.09(Cyt b)、0.87±0.07(D-loop),杞麓鲤遗传多样性水平明显高于元江鲤群体。分子方差分析(AMOVA)显示,两个群体间有明显的遗传分化,且遗传变异主要来源于群体间。系统发育分析显示,元江鲤群体单倍型较少,类型较单一,与杞麓鲤群体差异大,多数元江鲤样本单倍型属于华南鲤(Cyprinus carpio rubrofuscus)类型;多数杞麓鲤样本具特有单倍型,部分属于华南鲤、远东鲤(Cyprinus carpio haematopterus)类型。结果表明,杞麓鲤与元江鲤群体遗传分化明显,元江鲤线粒体遗传多样性较低。

小飞鼠Cyt b基因编码蛋白质生物信息学分析

对小飞鼠Cyt b基因编码的蛋白质进行生物信息学分析,预测小飞鼠Cyt b基因的功能和特性.研究结果表明:小飞鼠Cyt b编码的蛋白质为亲水蛋白质,由379个氨基酸组成,其中亮氨酸(Leu)所占比例最高,为15.8%;二级结构为无规则卷曲和α-螺旋;有2个N-糖基化位点,9个明显的跨膜螺旋结构域;小飞鼠与花鼠(Tamias sibiricus)遗传距离比较近.

Genotoxic and Cytotoxic Damage by Cyclophosphamide and Adriamycin as a Response to Treatment in Breast Cancer Patients: Pilot Study

The aim of this study was to evaluate the genotoxicity induced by cyclophosphamide-adriamycin treatment in breast cancer patients through the frequencies of Sister Chromatid Exchange (SCE), Replication Index (RI), Mitotic Index (MI) and Cell Proliferation Index (CPI) and to study the possible association between biomarkers of genotoxicity and the early response to treatment. The frequencies were obtained before and immediately after therapy from 17 patients with breast cancer (p < 0.001). Response to treatment was assessed after two years resulting in 12 patients in a state of remission. MI and CPI had high values after treatment in women with active cancers compared to those in a state of remission, however there were not significant differences. Conclusions: It is possible that MI y CPI biomarkers can serve as indicators for early assessment of treatment with cyclophosphamide-adriamycin. It should be noted that these are preliminary results and further study is necessary.

电针对慢性疼痛抑郁共病大鼠海马组织Cyt-C、Caspase-9表达的影响

目的 观察电针“合谷”“太冲”对慢性疼痛抑郁共病(CPDC)大鼠海马组织细胞色素C(Cyt-C)、胱天蛋白酶-9(Caspase-9)表达的影响,探讨电针治疗CPDC的可能机制。方法 60只SD大鼠随机分为假手术组、模型组、药物组和电针组,每组15只。采用左后足足底2次注射完全弗氏佐剂复制CPDC模型。电针组于双侧“合谷”“太冲”行电针干预,20 min/次,药物组予10 mg/kg度洛西汀混悬液灌胃,连续14 d。分别于首次注射后7、14、21、28 d检测机械缩足阈值及热缩足潜伏期,14、28 d检测悬尾不动时间和糖水偏好率,尼氏染色观察海马神经元形态及尼氏体数量,Western blot、免疫组化检测海马组织Cyt-C、Caspase-9蛋白表达。结果 与假手术组比较,模型组大鼠机械缩足阈值、热缩足潜伏期减少(P<0.01),悬尾不动时间增加(P<0.01),糖水偏好率下降(P<0.01),海马神经元排列稀松,神经元受损、丢失,尼氏体数量减少(P<0.01),海马神经元细胞凋亡率升高(P<0.01),海马组织Cyt-C、Caspase-9表达及cleaved-Caspase-9/Caspase-9比值升高(P<0.01);与模型组比较,药物组和电针组大鼠机械缩足阈值、热缩足潜伏期增加(P<0.01),悬尾不动时间减少(P<0.01),糖水偏好率升高(P<0.01),海马神经元丢失减少,细胞排列较整齐,核仁清晰,尼氏体数量增加(P<0.01),海马神经元细胞凋亡率降低(P<0.01),海马组织Cyt-C、Caspase-9表达及cleaved-Caspase-9/Caspase-9比值降低(P<0.01)。结论 电针“合谷”“太冲”可减轻CPDC模型大鼠疼痛及抑郁行为,发挥神经保护作用,其机制可能与抑制海马组织Cyt-C、Caspase-9蛋白表达,抑制海马神经元凋亡有关。

基于cyt b基因分析河南省4大水系红鳍原鲌种群的遗传多样性

选用长江、黄河、淮河、海河4大水系的红鳍原鲌Cultrichthys erythropterus样本共100尾,基于cyt b基因分析其种群的遗传多样性、遗传结构及种群历史动态等。结果显示:河南省4大水系红鳍原鲌种群整体单倍型多态性为0.909,整体核苷酸多样性为0.004 88,具有较高的遗传多样性;4个水系均有特有单倍型,这表明红鳍原鲌种群的遗传分化数值较大,遗传分化程度较高;通过构建和分析系统发育树、贝叶斯系统发生树和单倍型网络图,发现河南省4大水系红鳍原鲌种群间谱系分化不明显,且无明显的地理结构;AMOVA分析显示,种群内的变异占72.11%,组间的变异占4.63%,表明变异主要发生在种群内;Tajima’s D和Fu’s F_S分析结果表明,河南省红鳍原鲌种群近期未发生过种群扩张事件。本文研究和分析了河南省境内红鳍原鲌种群遗传多样性和遗传结构,为河南省红鳍原鲌种质资源的保护和合理开发利用提供了参考。

Damage effect and mechanisms of cyclophosphamide to human neuroblastoma SH-SY5Y cells

OBJECTIVE To investigate the damage effect and mechanisms of cyclophosphamide(CTX)and its active metabolite derivative 4-hydroperoxycyclophosphamide(4-HC)to human neuroblas⁃toma SH-SY5Y cells.METHODS SH-SY5Y cells were treated with CTX[0(cell control),0.01,0.1,1,5,10,20,40 and 80 mmol·L^(-1)]and 4-HC[0(cell control),0.01,0.1,1,5,10,20,40 and 80μmol·L^(-1)]for 48 h.Cell confluence and morphology were observed by the IncuCyte ZOOM system.Cell viability was assessed by CCK-8 assay.Lactate dehydrogenase(LDH)release was measured by LDH assay kit.SH-SY5Y cells were treated with CTX(0,1,5,10 and 20 mmol·L^(-1))and 4-HC(0,1,5,10 and 20μmol·L^(-1))for 48 h before cell proliferation was analyzed by 5-ethynyl-2′-deoxyuridine(EdU)staining assay.Immunofluorescence was employed to assess the levels of the DNA double-strand break markerγ-H2AX and to evaluate changes in mitochondrial membrane potential.SH-SY5Y cells were treated with CTX(0,1,5 and 10 mmol·L^(-1))and 4-HC(0,1,5 and 10μmol·L^(-1))for 48 h,and the alterations in glycolysis and oxidative phosphorylation levels were analyzed using the Seahorse XFe96 Analyzer.RESULTS Compared with the cell control group,cell confluence and cell viability were significantly reduced in the CTX and 4-HC groups(P<0.01),and the half-maximal inhibitory concentrations(IC50)for CTX and 4-HC were 4.44 mmol·L^(-1) and 4.78μmol·L^(-1),respectively.The release rate of LDH was signif⁃icantly increased while the percentage of EdU+cells was significantly reduced in the CTX and 4-HC groups(P<0.01).The percentage ofγ-H2AX+cells was significantly increased and mitochondrial membrane potential significantly decreased in the CTX and 4-HC group(P<0.05).Treatment with CTX and 4-HC resulted in reduced levels of maximum glycolytic capacity,glycolytic reserve,maximal respi⁃ration,and ATP production(P<0.05).CONCLUSION CTX and 4-HC exert significant cytotoxic effects on SH-SY5Y cells by disrupting cell membrane structure,impeding cell proliferation,and reducing cell viability.The mechanisms underlying these effects may involve intracellular DNA damage,disturbance of energy metabolism and mitochondrial dysfunction.

Geraniol alleviates cyclophosphamide-induced cardiotoxicity in mice

Objective:To explore the effect of geraniol on cyclophosphamide-induced cardiotoxicity.Methods:Mice were divided into five groups:the control group,the cyclophosphamide group(200 mg/kg cyclophosphamide,i.p.on day 7),the group treated with geraniol 100 and 200 mg/kg from day 1 to day 14,along with a single dose of cyclophosphamide on day 7,and the geraniol alone group(200 mg/kg geraniol from day 1 to day 14).At the end of the study,animals were sacrificed,and blood and heart were collected and analyzed for biochemical,histopathological,and immunohistochemical changes.Results:Treatment with 200 mg/kg geraniol significantly reduced the levels of cardiac injury markers,malondialdehyde,and inflammatory and apoptotic markers,while increasing antioxidant activities in mice with cyclophosphamide-induced cardiotoxicity.Moreover,it remarkably alleviated histopathological aberrations in cardiac tissue.Conclusions:Geraniol attenuates cyclophosphamide-induced cardiotoxicity via antioxidant,anti-inflammatory,and antiapoptotic effects.

Efficacy and safety of low-dose cyclophosphamide combined with lenvatinib, pembrolizumab and TACE for unresectable hepatocellular carcinoma:A single-center, prospective,single-arm clinical trial

Objective: Unresectable hepatocellular carcinoma(uHCC) continues to pose effective treatment options. The objective of this study was to assess the efficacy and safety of combining low-dose cyclophosphamide with lenvatinib, pembrolizumab and transarterial chemoembolization(TACE) for the treatment of uHCC.Methods: From February 2022 to November 2023, a total of 40 patients diagnosed with uHCC were enrolled in this small-dose, single-center, single-arm, prospective study. They received a combined treatment of low-dose cyclophosphamide with lenvatinib, pembrolizumab, and TACE. Study endpoints included progression-free survival(PFS), objective response rate(ORR), and safety assessment. Tumor response was assessed using the modified Response Evaluation Criteria in Solid Tumors(mRECIST), while survival analysis was conducted through KaplanMeier curve analysis for overall survival(OS) and PFS. Adverse events(AEs) were evaluated according to the National Cancer Institute Common Terminology Criteria for Adverse Events(version 5.0).Results: A total of 34 patients were included in the study. The median follow-up duration was 11.2 [95% confidence interval(95% CI), 5.3-14.6] months, and the median PFS(mPFS) was 15.5(95% CI, 5.4-NA) months.Median OS(mOS) was not attained during the study period. The ORR was 55.9%, and the disease control rate(DCR) was 70.6%. AEs were reported in 27(79.4%) patients. The most frequently reported AEs(with an incidence rate >10%) included abnormal liver function(52.9%), abdominal pain(44.1%), abdominal distension and constipation(29.4%), hypertension(20.6%), leukopenia(17.6%), constipation(17.6%), ascites(14.7%), and insomnia(14.7%). Abnormal liver function(14.7%) had the most common grade 3 or higher AEs.Conclusions: A combination of low-dose cyclophosphamide with lenvatinib, pembrolizumab, and TACE is safe and effective for u HCC, showcasing a promising therapeutic strategy for managing uHCC.

Parthenolide enhances the metronomic chemotherapy effect of cyclophosphamide in lung cancer by inhibiting the NF-kB signaling pathway

BACKGROUND Parthenolide(PTL),a sesquiterpene lactone derived from the medicinal herb Chrysanthemum parthenium,exhibits various biological effects by targeting NF-kB,STAT3,and other pathways.It has emerged as a promising adjunct therapy for multiple malignancies.AIM To evaluate the in vitro and in vivo effect of PTL on cyclophosphamide(CTX)metronomic chemotherapy.METHODS The cytotoxicity of PTL and CTX on Lewis lung cancer cells(LLC cells)was assessed by measuring cell activity and apoptosis.The anti-tumor efficiency was evaluated using a tumor xenograft mice model,and the survival of mice and tumor volume were monitored.Additionally,the collected tumor tissues were analyzed for tumor microenvironment indicators and inflammatory factors.RESULTS In vitro,PTL demonstrated a synergistic effect with CTX in inhibiting the growth of LLC cells and promoting apoptosis.In vivo,metronomic chemotherapy com-bined with PTL and CTX improved the survival rate of tumor-bearing mice and reduced tumor growth rate.Furthermore,metronomic chemotherapy combined with PTL and CTX reduced NF-κB activation and improved the tumor immune microenvironment by decreasing tumor angiogenesis,reducing Transforming growth factorβ,andα-SMA positive cells.CONCLUSION PTL is an efficient compound that enhances the metronomic chemotherapy effects of CTX both in vitro and in vivo,suggesting its potential as a supplementary therapeutic strategy in metronomic chemotherapy to improve the chemotherapy effects.

补骨脂素对环磷酰胺抑制小鼠骨髓间充质干细胞成骨分化的恢复作用

背景:补骨脂素具有很强的抗骨质疏松活性,可能对化疗导致的骨质疏松具有恢复作用。目的:探讨补骨脂素对环磷酰胺抑制小鼠骨髓间充质干细胞成骨分化的恢复作用及机制。方法:分离培养C57BL/6小鼠骨髓间充质干细胞,以MTT法检测补骨脂素对骨髓间充质干细胞活力的影响,以成骨诱导结合碱性磷酸酶染色确定补骨脂素对环磷酰胺抑制骨髓间充质干细胞成骨分化恢复作用的最佳剂量。采用RT-qPCR法检测补骨脂素干预不同时间点成骨分化标志基因Runx2、ALP、Osteocalcin、骨保护素及Wnt/β-catenin信号通路相关基因Wnt1、Wnt4、Wnt10b、β-catenin、c-MYC的mRNA表达;采用Western blot法检测补骨脂素干预不同时间点成骨特异性转录因子Runx2及Wnt/β-catenin信号通路相关基因Activeβ-catenin、DKK1、c-MYC、Cyclin D1的蛋白表达。结果与结论:(1)不同浓度补骨脂素对骨髓间充质干细胞活力没有显著影响;200μmol/L补骨脂素干预后对环磷酰胺诱导骨髓间充质干细胞成骨分化抑制的恢复作用最佳;(2)补骨脂素可以逆转环磷酰胺条件培养基导致的骨髓间充质干细胞成骨标志基因Runx2、ALP、Osteocalcin、骨保护素mRNA表达和Runx2蛋白表达的降低;(3)补骨脂素可以逆转环磷酰胺条件培养基导致的骨髓间充质干细胞Wnt/β-catenin通路相关基因Wnt4、β-catenin、c-MYC mRNA表达和Activeβ-catenin、c-MYC、Cyclin D1蛋白表达的降低以及DKK1蛋白表达的升高;(4)结果表明,环磷酰胺能抑制小鼠骨髓间充质干细胞成骨分化,补骨脂素对其具有恢复作用,且200μmol/L补骨脂素干预效果最佳,而这种保护作用可能与补骨脂素激活Wnt4/β-catenin信号通路有关。

赤眼鳟线粒体D-loop和Cyt b基因序列的对比分析

研究了长江和珠江野生赤眼鳟Squaliobarbus curriculus共4个群体24个个体的线粒体控制区(D-loop)和细胞色素b(Cyt b)基因序列,通过PCR扩增与测序,分别获得了长度为598 bp和752 bp的D-loop和Cyt b基因片段。分析表明,D-loop序列共定义了18个单倍型,存在34个简约信息位点,67个多态性位点,发生转换40次,颠换15次,插入/缺失14次。A、T、C、G平均含量分别为33.7%、35.1%、17.5%、13.7%。Cyt b序列共定义了18个单倍型,存在68个简约信息位点,72个多态性位点,发生转换60次,颠换15次,无插入/缺失位点。A、T、C、G平均含量分别为29.3%、26.4%、29.8%、14.5%。Cyt b除简约信息位点百分比大于D-loop外,多态位点百分比和单突变位点百分比均小于D-loop,说明D-loop具有较快的进化速率。D-loop序列的单倍型多样性(H)、平均核苷酸差异数(K)、核苷酸多样性(π)分别0.963 8、16.543 5和0.028 4,Cyt b的H、K和π分别为0.971 0、31.855 1和0.042 6,显示赤眼鳟具有较高的遗传多样性水平,且遗传变异主要存在于群体间。分析群体间的遗传距离和FST值,D-loop和Cyt b都一致表明:长江和珠江水系间存在明显的遗传分化,系统树和分子变异等级分析(AMOVA)也支持这一观点。同一水系群体间的遗传差异较小,Cyt b则更小;不同水系群体间的遗传差异较大,Cyt b则更大。故,Cyt b呈现出"小则越小,大则越大"的态势。分析其原因,可能与D-loop和Cyt b进化速率密切相关。

从Cyt b基因序列探讨鹿亚科动物的系统发生关系

鹿亚科共有 4个属 ,各属间和属内特别是鹿属内的系统进化关系存在疑义。根据Cytb基因序列分析 ,探讨了鹿亚科及中国鹿属、马鹿亚种的进化关系。结果分析表明 :现行分类系统中 ,斑鹿属可能并非单系发生 ,暗示应将豚鹿并入鹿属 ;麋鹿属与鹿属有较近的进化关系 ,也应并入鹿属 ;鹿属的进化地位有待进一步研究 ;归并后的鹿属为单系发生。中国马鹿各亚种在系统发生上是一单系群 ,其中马鹿天山亚种和阿尔泰亚种聚为最原始的一支。

雀形目15种鸟类CoⅠ与Cyt b基因序列的比较

对雀形目Passeriformes6科15种鸟类线粒体DNA的Cyt b基因全序列(1143bp)和CoⅠ基因部分序列(1176bp)进行了比较,结果显示Cyt b和CoⅠ基因序列的变异位点分别为454个和366个,简约信息位点为337个和303个,而且CoⅠ基因比Cytb基因略微保守,进化速率也较低。采用邻接法、最大简约法、最大似然法和贝叶斯法分别构建了CoⅠ和Cyt b基因两组数据集的分子系统发生树及其合一树,并对建树结果进行了比较分析。基于以上两点,本文认为CoⅠ基因比Cyt b基因更适合于确定雀形目科级阶元之间的系统发生关系,而且它也能够作为雀形目物种鉴定的分子标记,但在物种鉴定方面不如Cyt b基因稳定和准确。