Systematic evaluation and meta-analysis of the efficacy and safety of cinobufagin injection combined with western medicine in the treatment of liver cancer

Objective:To evaluate the clinical efficacy and safety of cinobufagin injection in the treatment of liver cancer.Methods:PubMed database,Embase database and Cochrane Library database,CNKI,Wanfang database,VIP database and Sinomed database were used to search for the randomized controlled trials of cinobufagin injection combined with Western medicine in the treatment of primary liver cancer.The retrieval time was from the establishment to December 15,2020.Two independent researchers conducted systematic screening,literature inclusion and quality assessment of the articles according to the inclusion criteria,respectively.Meta-analysis of the data was performed using RevMan 5.4 software.Results:A total of 30 studies with a total of 2355 patients were included.Compared with conventional western medicine treatment,the clinical effective rate of Hububutin injection combined with western medicine was significantly higher[RR=1.16,95%CI=(1.11,1.22),P<0.00001].It could effectively reduce the tumor size[RR=1.33,95%CI=(1.17,1.51),P<0.00001],prolong the survival time of patients[RR=1.41,95%CI=(1.31,1.52),P<0.00001],improve the quality of life[RR=1.37,95%CI=(1.19,1.57),P<0.00001],improve the liver function of patients[RR=-14.52,95%CI=(-16.15,-12.88),P<0.00001],and reduce the occurrence of adverse reactions[RR=0.94,95%CI=(0.85,1.42),P=0.25]such as bone marrow suppression[RR=0.44,95%CI=(0.31,0.62),P<0.00001].Conclusion:Cinobufagin injection combined with western medicine therapy can effectively improve the clinical symptoms of primary liver cancer,and the safety is good.However,the methodological quality of the included literature is low,which affects the objectivity of the outcome,and it still needs to be verified by multi-sample,multi-center,randomized double-blind controlled trial.

One new bufadienolide biotransformed from cinobufagin by Cunninghamella elegans

Cunninghamella elegans has been employed for the biotransformation of cinobufagin （1） to afford one metabolites. The structure of the transformation product has been characterized as 7β,12β-dihydroxylcinobufagin （2）. Product 2 is a new compound. In vitro cytotoxic activities of the biotransformation product and the substrate-cinobufagin have been assayed against HeLa; they all showed cytotoxic activities.

Intestinal Transport and Biotransformation of Resibufogenin and Cinobufagin in Chan Su via HPLC/APCI-MS^n

In vitro models of human colon carcinoma cell line(Caco-2 cell monolayer) and human intestinal bacteria were used to investigate the intestinal transport and biotransformation of resibufogenin and cinobufagin in Chan Su by HPLC/APCI-MSn. The experimental results of Caco-2 cell monolayer demonstrate that the apparent permeability coefficients(Papp) of resibufogenin and cinobufagin are higher than 10–6 cm/s, which indicates that both resibufogenin and cinobufagin have a good absorption in the small intestine. And the biotransformation result of human intestinal bacteria shows that resibufogenin has been transformed to 3-epiresibufogenin and cinobufagin has been transformed to 3-epicinobufagin, deacetylcinobufagin and 3-epideacetycinobufagin, respectively.

Cinobufagin, a bufadienolide from traditional Chinese medicine Bufo bufo gargarizans CANTOR, inhibits PC3 cell growth in vitro and in vivo

Objective:To explore the effects of cinobufagin (CBF),an active component of toad venom (Bufo bufo gargarizans CANTOR),on the proliferation and apoptosis of PC3 human prostate cancer cells in vitro and preliminarily investigate the mechanism of CBF in suppressing tumor cell growth in vivo.Methods:The effect of CBF on PC3 cells proliferation was detected using MTT assay.The morphological changes of PC3 cells were observed under an optical microscope.Colony formation assays were used to observe the CBF effect on colony formation by PC3 cells.PC3 cell apoptosis after treatment with CBF for 48 hours was monitored using flow cytometry.Furthermore,the effect of CBF on the expression of myeloid cell leukemia-1 (MCL-1) and related apoptotic proteins was examined using western blotting.A xenograft model was established in BALB/c nude mice to evaluate the effect of CBF on prostate cancer in vivo.Results:The MT-T assay results illustrated that PC3 cell proliferation was inhibited in vitro by CBF in a concentration-and time-dependent manner.Compared with the control group findings,CBF significantly inhibited the formation of PC3 cells (P =.005).Flow cytometry revealed that after treatment with 50 nM CBF for 48 hours,the apoptotic rate of PC3 cells was 41.97 (5.16)%,indicating that CBF could significantly induce its apoptosis (P =.003).In addition,optical and fluorescence microscopy uncovered remarkable inhibition of cell proliferation accompanied by morphologic changes.The western blotting result indicated that CBF obviously downregulated the expression of the anti-apoptotic protein MCL-1.Most importantly,ClBF reduced the carcinogenicity of PC3 xenografts in nude mice.Conclusion:CBF can inhibit the growth of PC3 cells both in vitro and in vivo and induce apoptosis of tumor cells.The corresponding mechanism may be correlated with the activation of caspase family proteins via MCL-1.

Crystal Structure and Anticancer Properties of Cinobufagin 3-Hemisuberate Methyl Ester

The title compound cinobufagin 3-hemisuberate methyl ester（1） was isolated from the venom of Bufo bufo gargarizans CANTOR. The crystal structure of 1, C35H48O9, was determined by single-crystal X-ray diffraction analysis. It belongs to orthorhombic, space group P212121 with a = 8.9338（3）, b = 16.2970（4）, c = 22.4019（6） , V = 3261.59（16） 3, Mr = 612.73, Z = 4, Dc = 1.248 g/cm3, μ = 0.725 mm-1, F（000） = 1320, S = 1.040, the final R = 0.0374 and wR = 0.0412 for 4458 unique reflections, of which 4088 were observed（I 〉 2σ（I））. In the solid state, short intermolecular C-H...O interactions involving a methine and the ester carbonyl group in cinobufagin moiety and a methyl in the suberate moiety linked adjacent molecules into a three-dimensional network. Detailed analysis of the 1H-NMR data showed that X-ray structure of 1 would be expected to closely resemble the solution conformation in chloroform. Compound 1 was inactive for the inhibition of PC3 and HepG2 cancer cells, but the parent compound cinobufagin showed potent inhibition with IC50 values of 0.145 and 5.48 μM, respectively, indicating that esterification at C（3） decreased the cytotoxic effect of 1.

Meta-analysis of Cinobufagin Capsules combined with chemotherapy in treating gastric cancer

Objective:To evaluate the efficacy and safety of Cinobufagin capsules combined with chemotherapy in the treatment of gastric cancer by Meta-analysis.Methods:Randomized controlled trials(RCTs)of Cinobufagin capsules combined with chemotherapy for gastric cancer included in Chinese database(VIP,CNKI,WanFang,CBM)and English database(PubMed,Cochrane Library,EMBase)were retrieved from the inception to June 2020.Data extraction and methodological quality evaluation were carried out for the included literature.The methodological quality assessment was based on Cochrane bias risk assessment tool.RevMan 5.3 and GRADEpro 3.6 were used for meta-analysis and evidence quality assessment,respectively.Results:12 trials with 830 patients were included in the review.Meta-analysis results showed that:the addition of Cinobufagin capsules for gastric cancer enhanced the objective remission rate(ORR)(RR=1.65,95%CI[1.41,1.93],P<0.00001);improved quality of Life Score(KPS score)(RR=1.77,95%CI[1.43,2.19],P<0.00001);reduced the incidence of nausea and vomiting(RR=0.46,95%CI[0.29,0.74],P=0.001),leukocyte toxicity(RR=0.40,95%CI[0.25,0.64],P<0.0001),and platelet toxicity(RR=0.45,95%CI[0.22,0.93],P=0.03);increased the level of CD4+(MD=6.99,95%CI[4.32,9.66],P<0.00001)was better than chemotherapy alone,with statistically significant differences.The quality of evidence is low by GRADE evaluation.Conclusion:In chemotherapy for gastric cancer,combined with Cinobufagin capsule has better efficacy and safety.

Clinical Study on TACE Combined with Elemene Injection and Cinobufagin Injection Respectively for Middle and Advanced Primary Hepatic Carcinoma

Objective: To observe and compare the differences in the clinical effect and the incidences of adverse reactions oftranscatheter arterial chemoembolization(TACE) combined with elemene injection and cinobufagin injection respectively for middleand advanced primary hepatic carcinoma. Methods: A total of 104 cases of patients with middle and advanced primary hepaticcarcinoma who were treated in the oncology department from August 2018 to January 2020 were included as the study objects, andwere randomly divided into two groups according to different treatment regimens, 52 cases in each group. Both groups were treatedwith TACE once;the cinobufagin injection group was given intravenous infusion with 500 mL of 5% glucose injection and 10 mLof cinobufagin injection once a day. The elemene injection group was given intravenous infusion with elemene injection of 0.4 geach time and once a day. Both groups were treated for two courses, 15 days of continuous treatment with a rest of 15 days beingone course. The clinical effect, the changes in the indexes of liver function including alanine amino transferase(ALT), aspartatetransaminase(AST) and total bilirubin(TBil), the scores of alpha-fetoprotein(AFP) and Karnofsky (KPS) and tumor volumes aswell as the difference in the incidences of adverse reactions between the two groups were observed and compared. Results: Thetotal clinical effective rate was 88.46% in the elemene injection group and 71.15% in the cinobufagin injection group, and thedifference was significant(P<0.05). After treatment, the levels of ALT, AST and TBil in serum in the two groups were significantlydecreased when compared with those before treatment, differences being significant(P<0.05). There was no significant differencebeing found in the comparison of the levels of ALT, AST and TBil in serum between the two groups (P>0.05). After treatment, thedecrease of AFP, tumor volume and the increase of KPS scores in the elemene injection group were significantly more than thosein the cinobufagin injection group, differences being significant (P<0.01). During treatment, there was no significant differencebeing found in the comparison of the total incidences of adverse reactions between the two groups(P>0.05). The adverse reactionsin the cinobufagin injection group were mainly nausea and vomiting, with higher incidence than that in the elemene injection group,the difference being significant (P<0.05). The adverse reactions in the elemene injection group were mainly pain at the injectionsite, with higher incidence than that in the cinobufagin injection group, the difference being significant (P<0.05). Conclusion: Thetherapy of elemene injection combined with TACE for middle and advanced primary hepatic carcinoma has better clinical effect thanthat of cinobufagin injection, but the occ.

A clinical study on the prevention of phlebitis caused by Cinobufagin using hand exercise combined with Jinhuang ointment

Objective:The aim of this study is to observe the clinical effect of hand exercise combined with jinhuang ointment on the prevention of phlebitis due to cinobufagin.Methods:A total of 90 cancer patients who have been receiving intravenous(Ⅳ)infusion of cinobufagin from May 2018 to June 2019 in the oncology department of our hospital were selected.They were then divided into three groups in a random manner,which include 30 cases in the control group,30 cases in the jinhuang ointment group,and 30 cases in the group of hand exercise combined with jinhuang ointment.The control group had a routine care before cinobufagin was infused from the first day of hospitalization.During the routine care mentioned previously,the jinhuang ointment group was given locally jinhuang ointment inunction,qd,and kept for 6 hours.Hand exercises were then done on the combined group excluding jinhuang ointment application,qd,at 10min.After treatment for 1 week,the phlebitis and pain incidences on the venipuncture site were then compared between the three groups.Results:After the 1-week treatment,the incidences of grade Ⅰ/Ⅱ phlebitis in the control group,the jinhuang ointment group,and the group of hand exercise combined with jinhuang ointment were 53.5%,23.3%,and 10%,respectively.The results have shown a significant decrease in the jinhuang ointment group and the combined group as in comparison with the control group(P=0.0169,P=0.0003).Even with the incidence of the combined group being lower than that of the jinhuang ointment group,no statistically significant difference(P=0.1659)was found.The incidences of grade Ⅲ/Ⅳ phlebitis in the control group,the jinhuang ointment group,and the group of hand exercise combined with jinhuang ointment were 23.3%,3.3%,and 3.3%,respectively.Both the jinhuang ointment group and the combined group had significantly lower results than that of the control group(P=0.0003,P=0.0227).There was no difference in the results of the combined group and the jinhuang ointment group.The incidences of pain in the control group,the jinhuang ointment group,and the group of hand exercise combined with jinhuang ointment were 56.7%,36.7%,and 20%,respectively.There was no significant difference in the results of the jinhuang ointment group and the control group(P=0.1205);the combined group has shown a significant decrease in the incidence compared with the control group(P=0.0035);the incidence of pain in the combined group was lower than that of the jinhuang ointment group,but without a statistical difference(P=0.1520).Conclusion:Hand exercise together with jinhuang ointment inunction can significantly reduce the incidence of phlebitis produced by cinobufagin.

非小细胞肺癌患者接受安罗替尼联合华蟾素治疗的疗效及安全性研究

目的分析安罗替尼联合华蟾素在非小细胞肺癌(Non-small Cell Lung Cancer,NSCLC)治疗中的效果。方法选取常熟市第二人民医院于2020年1月—2023年3月收治的109例NSCLC患者为研究对象,采用随机数表法分为对照组(n=54)、研究组(n=55)。对照组给予安罗替尼联合多西他赛注射液化疗,研究组给予安罗替尼联合华蟾素片化疗。比较两组症状缓解情况、肿瘤标志物水平、不良反应发生率。结果研究组症状缓解率高于对照组,肿瘤标志物水平低于对照组,差异有统计学意义(P均<0.05);研究组不良反应发生率(1.82%)低于对照组(11.11%),差异有统计学意义(χ^(2)=3.916,P<0.05)。结论安罗替尼和华蟾素联合治疗NSCLC效果显著,能够促进疾病缓解,降低肿瘤标志物水平,且不良反应发生风险较低,为NSCLC治疗提供了有效的治疗方案。

华蟾素对肝癌患者介入术后外周血Bcl-2和cyclinD1蛋白表达水平的影响

目的观察华蟾素对肝癌患者介入术后外周血Bcl-2和细胞周期蛋白(cyclinD1)表达水平的影响。方法本次研究对象在2020年5月—2021年5月于医院进行介入术治疗的肝癌患者中选择100例,随机分为两组,50例患者给予患者常规介入治疗(常规介入组),50例患者在常规化疗的基础上加用华蟾素治疗(华蟾素组)。检测患者的Bax、Bcl-2、cyclinD1水平及肝功能指标[甲胎蛋白(AFP)、谷丙转氨酶(ALT)、总胆红素(TBIL)、谷草转氨酶(AST)]、免疫功能指标[免疫球蛋白G(IgG)、自然杀伤(NK)细胞、T淋巴细胞亚群(CD_(4)^(+)、CD_(4)^(+)/CD_(8)^(+))],比较两组卡氏(KPS)评分、肝功能分级标准(Child-Pugh)评分,评价疗效,观察两组不良反应发生情况。结果华蟾素组患者治疗后的Bax水平高于常规介入组(P<0.05),Bcl-2、cyclinD1水平低于常规介入组(P<0.05);华蟾素组患者治疗后的AFP、TBIL、AST水平低于常规介入组(P<0.05),ALT水平高于常规介入组(P<0.05);华蟾素组患者治疗后的IgG、NK细胞、CD_(4)^(+)水平高于常规介入组(P<0.05),CD_(4)^(+)/CD_(8)^(+)水平低于常规介入组(P<0.05);华蟾素组患者治疗后的KPS评分高于常规介入组(P<0.05),Child-Pugh评分低于常规介入组(P<0.05);华蟾素组患者治疗有效率为92.00%,高于常规介入组的74.00%(P<0.05);华蟾素组患者的不良反应发生率为20.00%,低于常规介入组的44.00%(P<0.05)。结论华蟾素可以下调肝癌患者介入术后外周血Bcl-2和cyclinD1蛋白表达水平,上调Bax水平,促进肿瘤细胞凋亡;同时可以增强患者的肝功能和免疫功能,提高生活质量,改善预后;还能提高肝癌介入治疗疗效,降低发生不良反应的风险。

华蟾素调控PI3K/AKT通路逆转卵巢癌A2780/DDP细胞顺铂耐药的作用机制

目的研究华蟾素(CBG)对人卵巢癌细胞顺铂耐药的逆转作用和机制。方法A2780细胞株及其顺铂耐药细胞株A2780/DDP是临床常见的卵巢癌细胞,故选择这两种细胞株作为研究对象。通过CCK-8法检测细胞活力,平板克隆和5-乙炔基-2′脱氧尿嘧啶核苷(EdU)实验检测细胞的增殖能力,赫斯特染色(Hoechst)法观察细胞凋亡情况,细胞划痕实验和Transwell实验评估细胞的迁移和侵袭能力,Western blot和定量逆转录PCR(RT-qPCR)法检测磷脂酰肌醇3-激酶/蛋白激酶(PI3K/AKT)信号通路和上皮-间质转化(EMT)的相关蛋白和mRNA的表达差异。结果与A2780细胞相比,A2780/DDP细胞的耐药指数分别约为5.636、5.864、5.695,采用CBG(2、4、6 mg/ml)处理A2780/DDP耐药细胞后,逆转耐药指数分别为1.617、2.570、3.461。CBG呈浓度依赖性地上调细胞凋亡水平、抑制细胞的增殖、迁移和侵袭能力(P<0.05)。Western blot结果显示:与A2780细胞相比,对照组(A2780/DDP)细胞中P-PI3K/PI3K和P-AKT/AKT的蛋白水平相对比值以及N钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、蜗牛蛋白(Snail)的蛋白表达更高,E钙黏蛋白(E-cadherin)蛋白表达更低(t_(P-PI3K/PI3K)=8.115,t_(P-AKT/AKT)=17.62、t_(N-cadherin)=6.126、t_(Vimentin)=4.001、t_(Snail)=17.333、t_(E-cadherin)=4.620,P<0.01);随着CBG剂量升高,耐药细胞中的P-PI3K、P-AKT、N-cadherin、Vimentin、Snail的蛋白表达水平降低,而E-cadherin的蛋白表达量增加(F_(P-PI3K)=268.5、F P-AKT=190.5、F_(N-cadherin)=24.02、F_(Vimentin)=57.65、F_(Snail)=87.24、F_(E-cadherin)=135.8,P<0.05)。RT-qPCR结果显示:随着CBG浓度增加,PI3K、AKT、N-cadherin、Vimentin、Snail的mRNA表达水平随之降低,相反E-cadherin的mRNA表达水平逐渐升高(F PI3K=101.1、F_(AKT)=558.3、F_(N-cadherin)=86.97、F_(Vimentin)=105.9、F_(Snail)=85.71、F_(E-cadherin)=80.96,P<0.01)。结论CBG具有逆转卵巢癌A2780/DDP细胞株顺铂耐药的作用,其机制可能与CBG调控PI3K/AKT信号通路和抑制EMT发生有关。

人肝癌Huh7细胞上皮间充质转化与VEGFa/VEGFR2自分泌途径的相关性及华蟾素的调控作用

目的探究华蟾素(cinobufagin,CBG)注射液对人肝癌Huh7细胞血管内皮生长因子a(vascular endothelial growth factor a,VEGFa)/血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)自分泌信号通路及上皮间充质转化进程的影响。方法通过CCK-8实验、细胞划痕实验、Transwell侵袭实验和成球实验检测人肝癌Huh7细胞增殖、迁移、侵袭和成球能力;免疫荧光双标实验对人肝癌Huh7细胞VEGFa/VEGFR2进行共定位分析,ELISA法检测人肝癌Huh7细胞上清液中VEGFa水平,Western blot法检测人肝癌Huh7细胞中VEGFa/VEGFR2通路及上皮间充质转化相关蛋白表达水平。结果CBG注射液能显著抑制人肝癌Huh7细胞的增殖、迁移、侵袭和成球能力;人肝癌Huh7细胞可能存在VEGFa/VEGFR2自分泌途径;CBG注射液能抑制VEGFa的分泌,降低下游p-VEGFR2、p-PI3K、p-AKT及间充质标志物N-Cadherin、Vimentin、Snail蛋白的表达水平,升高上皮标志物E-Cadherin蛋白的表达水平。结论CBG注射液可能通过VEGFa/VEGFR2自分泌途径调控人肝癌Huh7细胞上皮间充质转化。

血小板增强肝癌血管生成及华蟾素的干预作用研究

目的:探讨血小板(PLT)对肝癌血管生成的影响以及华蟾素的干预作用。方法:先筛选PLT与肝癌细胞的适宜共孵育比例,制备条件培养基,确定华蟾素(CBG)的半数抑制浓度;然后,设对照组[人脐静脉内皮细胞(EC)+常规培养基]、串扰组[EC+CM_HP(HUH7与PLT串扰制备的条培)]、干预组(EC+CM_HP+CBG),采用划痕实验、小管形成实验、出芽实验和ELISA实验分别评估EC的迁移力、成管力、出芽能力以及共培养上清中血管内皮生长因子(VEGF)水平;运用Western blot检测VEGFR2、p-VEGFR2的表达,并采用抑制剂进行了反向验证。建立肝癌裸鼠皮下移植瘤模型,设Model组、Model+CBG组和Model+Apa组,通过Masson染色观察移植瘤的胶原表达情况,免疫荧光检测血管内皮标志物CD31和CD34的表达水平。结果:PLT∶HUH7=200时HUH7活力最强,HUH7与PLT串扰明显促进EC的增殖力(P<0.01)。与对照组相比,串扰组的迁移、成管、生芽能力增强,干预组的低于串扰组(P<0.01)。串扰组上清中VEGF的表达水平高于对照组,而干预组的低于串扰组(P<0.01)。串扰组p-VEGFR2蛋白的表达量较对照组显著增多,而干预组的表达量较串扰组少(P<0.01)。Model组可见大片胶原纤维沉积,CBG干预显著降低移植瘤组织胶原纤维沉积。Model组的肝癌移植瘤组织存在CD31和CD34表达,CBG干预显著降低肝癌移植瘤组织CD31和CD34的表达(P<0.01)。结论:PLT能增强肝癌的血管生成,CBG可能经由VEGF/VEGFR2通路抑制其成管能力。

华蟾素制剂在肿瘤患者的临床使用分析

目的分析住院肿瘤患者使用华蟾素制剂的应用特点,为华蟾素制剂的合理使用提供参考。方法采用PASSPharmAssist系统调取潍坊市人民医院2020年1月1日至2023年8月31日使用华蟾素制剂的住院患者数据,使用Excel 2016软件对人口学特征、诊断信息和具体使用情况进行统计分析。结果共纳入463例患者,使用华蟾素制剂629例次,调查发现肿瘤患者中男性[平均年龄(63.00±9.70)岁,57.02%]多于女性[平均年龄(59.11±10.83)岁,42.98%],平均年龄(61.31±10.37)岁,涉及前5位癌症种类为肺癌(44.45%)、结直肠癌(28.94%)、胃癌(9.51%)、食管癌(4.10%)、肝癌(3.02%),华蟾素制剂主要用药方案为联合化疗方案(36.57%),存在与免疫检查点抑制剂、靶向药物、镇痛药物以及放疗等方式联合应用230例,其中77例(33.48%)未出现ADR,119例(51.74%)为一般ADR,34例(14.78%)为严重ADR。临床实际应用中存在剂量过大的情况,用药疗程主要集中在10 d以内。结论华蟾素制剂主要用于肺癌以及消化道癌化疗前和化疗时用药,目前其临床疗效和作用机制需进一步验证和阐明,缺乏用药疗程的统一资料,应警惕剂量过大引发的心脏毒性。

华蟾素联合阿帕替尼治疗晚期胃癌二线化疗失败患者疗效观察及对免疫失衡、血清肿瘤标志物及相关血清学指标的影响

目的:观察华蟾素联合阿帕替尼治疗晚期胃癌二线化疗失败患者的近期疗效及对外周血辅助性T细胞(Th) 17/调节性T细胞(Treg)免疫失衡、血清肿瘤标志物及相关血清学指标的影响。方法:选择82例二线化疗失败的晚期胃癌患者作为研究对象,以随机数字表法分为观察组和对照组各41例。对照组予阿帕替尼治疗,观察组在对照组基础上联合华蟾素治疗,4周为1个疗程,连续治疗3个疗程。比较2组近期疗效及毒副反应情况,以及治疗前后血清肿瘤标志物[糖类抗原(CA) 72-4、CA19-9、CA242、癌胚抗原(CEA)]水平、相关血清学指标[血管内皮生长因子A (VEGF-A)、表皮生长因子受体(EGFR)、基质金属蛋白酶-9 (MMP-9)]及外周血Th17、Treg水平。结果:观察组客观缓解率为36.59%,对照组为24.39%,2组比较,差异无统计学意义(P>0.05);观察组疾病控制率为73.17%,对照组为51.22%,2组比较,差异有统计学意义(P<0.05)。治疗后,2组血清CA72-4、CA19-9、CA242、CEA水平均较治疗前降低(P<0.05),且观察组上述4项水平均低于对照组(P<0.05)。治疗后,2组外周血Th17、Treg表达水平和Th17/Treg比值均较治疗前降低(P<0.05),且观察组上述3项水平均低于对照组(P<0.05)。治疗后,2组血清VEGF-A、EGFR、MMP-9水平均较治疗前降低(P<0.05),且观察组上述3项水平均低于对照组(P<0.05)。观察组胃肠道反应、高血压、白细胞减少、血小板减少、蛋白尿、乏力、手足综合征的发生率均低于对照组,且2组白细胞减少、血小板减少、乏力发生率比较,差异有统计学意义(P<0.05)。结论:华蟾素联合阿帕替尼治疗二线化疗失败的晚期胃癌能有效下调患者血清肿瘤标志物和VEGF-A、EGFR、MMP-9水平,纠正外周血Th17/Treg免疫失衡,提高近期疗效,并能在一定程度上减少阿帕替尼引起的毒副反应。

Identification of Cinobufagin and Resibufogenin as Inhibitors of Enterovirus 71 Infection

In this paper, cinobufagin and resibufogenin were found to inhibit enterovirus 71（EV71） infection in vitro in cell viability and plaque reduction assays. The 50% inhibitory concentrations（lCs0） of einobufagin and resibufoge- nin were （10.94±2.36） and （218±31） nmol/L, respectively, the 50% cytotoxic concentrations（CCs0） of them were （1277±223） and （1385±254） nmol/L, respectively, and the anti-EV71 selectivity index（SI50） of cinobufagin was 116.7, which are promisingly developed into drug. Using a VP1 detection assay and a constructed reporter luciferase, we found that cinobufagin and resibufogenin disrupted the synthesis of EV71 protein. However, neither of them inhibited EV71 RNA replication. Our study suggests that cinobufagin and resibufogenin are the promising candidates that should he fllrther investigated for the treatment of EV71 caused disease.

Novel Cinobufagin Oxime Ether Derivatives as Potential Na＋/K＋-ATPase Inhibitors： Synthesis, Biological Screening and Molecular Docking

Some cinobufagin oxime ether derivatives as potential Na＋/K＋-ATPase inkibitors were synthesized by following the side chain of istaroxime. These compounds inhibit Na＋/K＋-ATPase in a dose-dependent manner. Compound 3c with an oxyethylamine side chain that is the same as that of istaroxime showed the most potent inhibi- tion, which was stronger than compound 3a with only hydroxyoxime moiety at C3 and compound 3b with a methy-lated hydroxyoxime moiety. Molecular docking was used to explore the binding modes of the target compounds with Na＋/K＋-ATPase, which suggested that the longer ethyl amine group at C3 oxime moiety of compound 3c could make stronger interaction with Na＋/K＋-ATPase via intermolecular charge-charge and H-bond interaction as compared with other derivatives.

华蟾素胶囊联合新辅助化疗方案治疗HER2阳性乳腺癌临床研究

目的:探讨华蟾素胶囊联合新辅助化疗方案治疗HER2阳性乳腺癌的临床效果。方法:纳入2022年2月—2023年2月于南昌市人民医院就诊的86例HER2阳性乳腺癌患者,使用随机数字表法将患者分为观察组和对照组,各43例,对照组予以曲妥珠单抗+帕妥珠单抗+紫杉类联合铂类新辅助化疗方案,观察组在对照组新辅助化疗方案基础上加用华蟾素胶囊治疗。比较两组临床疗效、肿瘤退缩分级、血清相关指标[血清癌胚抗原(CEA)、糖类抗原153(CA153)、组织多肽特异抗原(TPS)、免疫球蛋白(Ig)A、Ig M和T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+))]、生活质量[乳腺癌患者生活质量测定量表中文版(FACT-B)]及不良反应。结果:两组客观缓解率(51.16%vs 41.86%)、疾病控制率(81.40%vs74.42%)比较,差异均无统计学意义(P>0.05);两组肿瘤退缩分级比较,差异无统计学意义(P>0.05);治疗后,两组CEA、CA153、TPS水平均较治疗前降低,观察组均低于对照组(P<0.05);治疗后,观察组Ig A、Ig M、CD3^(+)、CD4^(+)水平均高于对照组(P<0.05);两组治疗前后CD8^(+)水平比较,差异无统计学意义(P>0.05);治疗后,观察组生理状况、情感状况、社会/家庭状况、附加关注评分均高于对照组,差异均有统计学意义(P<0.05);观察组头痛头晕、消化道不适、心脏毒性反应、肝肾损伤及骨髓抑制发生率均低于对照组(P<0.05)。结论:华蟾素胶囊联合新辅助化疗方案治疗HER2阳性乳腺癌,可在一定程度上抑制肿瘤进展,改善患者免疫失衡及生活质量水平,降低化疗毒副反应。

华蟾素通过铁死亡途径抑制上皮性卵巢癌细胞增殖、迁移与侵袭

目的:铁死亡在包含卵巢癌在内的多种癌症进展中发挥重要作用,是癌症干预的潜在策略。华蟾素(cinobufagin,CINO)已在多种癌症中发挥抗肿瘤作用,而其对上皮性卵巢癌中的作用及其对铁死亡的影响并不清楚。本研究旨在探究CINO对上皮性卵巢癌恶性生物学行为的影响,并初步探索其铁死亡的相关机制。方法:梯度浓度(0、0.1、1.0、10.0 mg/mL)CINO处理卵巢癌细胞株A2780和SKOV3,采用5-乙炔基-2’-脱氧尿苷(5-ethynyl-2’-deoxyuridine,EDU)染色法检测增殖;Transwell法检测细胞迁移与侵袭;FerroOrange染色评估细胞内Fe2+含量;酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测细胞内4-羟基壬烯醛(4-hydroxynonenal,4-HNE)水平;二氢乙锭(dihydroethidium,DHE)探针试剂盒法检测细胞内活性氧(reactive oxygen species,ROS)水平;蛋白质印迹法检测溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和转铁蛋白受体1(transferrin receptor 1,TFR1)表达。采用铁死亡抑制剂Ferrostatin-1(Fer-1)和铁离子螯合剂甲磺酸去铁胺(deferoxamine mesylate,DFO)预处理A2780和SKOV3细胞,再给予10.0 mg/mL CINO处理,分别检测细胞增殖、迁移与侵袭情况。结果:CINO能剂量依赖性抑制A2780细胞增殖、侵袭及迁移,增加细胞内Fe2+及4-HNE水平,降低SLC7A11表达,并增加TFR1表达(均P<0.05)。Fer-1和DFO可部分逆转CINO对卵巢癌细胞增殖、迁移与侵袭的抑制作用(均P<0.05)。结论:CINO抑制卵巢癌细胞的增殖、迁移与侵袭,其作用可能与其通过调节SLC7A11/TFR1信号通路增加卵巢癌细胞铁死亡有关。

华蟾素调控HIF-1α/VEGF通路逆转结肠癌HCT15/5-FU细胞耐药的体外研究

目的探讨华蟾素(CINO)联合5-氟尿嘧啶(5-FU)对人源结肠癌(CRC)耐药细胞株HCT15/5-FU的逆转作用,明确缺氧诱导因子(HIF-1α)/血管内皮生长因子(VEGF)通路在逆转结肠癌化疗耐药中的调控作用。方法MTT法检测细胞耐药性及耐药指数的变化,流式细胞术评价细胞凋亡的情况,划痕实验和Transwell法检测细胞迁移和侵袭能力的变化。Western blot检测上皮间充质转化(EMT)相关蛋白及HIF-1α/VEGF通路相关蛋白的表达。结果与HCT15细胞相比,HCT15/5-FU的耐药指数约为8.720。CINO联合5-FU作用后,能够明显提升HCT15/5-FU细胞的药物敏感性,降低耐药指数,剂量依赖性地上调细胞凋亡水平、抑制细胞迁移和侵袭能力;Western blot结果显示CINO联合5-FU作用后能够抑制EMT及HIF-1α/VEGF通路的活性。结论体外研究显示,华蟾素具有逆转结肠癌5-FU耐药的作用,其机制可能与调节HIF-1α/VEGF通路抑制EMT、血管生成有关。