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.

血液高RDW和高N-MID OC/β-CTX比值在老年女性椎体骨折中的诊断价值

目的 探讨血液红细胞分布宽度(RDW)及骨钙素N段中分子片段(N-MID OC)/Ⅰ型胶原羧基端肽β特殊序列(β-CTX)比值联合检测对老年女性骨质疏松椎体骨折(OPVF)的诊断价值。方法 纳入2021年1月—2021年12月我院≥60岁女性骨质疏松住院患者786例,分为无骨折组(A组,378例)和OPVF组(B组,388例)。N-MID OC、β-CTX、NMID-OC/β-CTX比值、红细胞计数(RBC)、红细胞压积(HCT)、血红蛋白(Hb)、红细胞平均体积(MCV)、平均血红蛋白浓度(MCHC)、平均血红蛋白量(MCH)、红细胞分布宽度(RDW-CV)、钙(Ca)、磷(P)及镁(Mg)。结果 与A组相比,B组Ca、P、Hb、HCT、MCV、MCH、MCHC含量降低,RDW-CV、N-MID OC/β-CTX含量增加,差异有统计学意义(均P<0.05)。OPVF与β-CTX、Ca、P呈负相关,与RDW-CV、N-MID OC/β-CTX呈正相关(均P<0.05)。骨折患者RDW与N-MID OC、β-CTX、Ca、P、Mg呈负相关(均P<0.05)。RDW、N-MID-OC/β-CTX为OPVF的风险因素(均P<0.05),OR分别为1.38、1.005。二者含量越高,椎体骨折风险越高,最高层是最低层的约3倍和2倍,OR为2.818,1.734。二者联合检测可提高对OPVF的诊断性能,其AUC敏感性、特异性、阳性预示值、阴性预示值分别为0.64、42.78%、82.01%、70.9%、58.3%。结论 血液高RDW和高N-MID OC/β-CTX比值与老年女性骨质疏松性椎体骨折明显相关,是其危险因素,可用于辅助诊断及预测其发生。

β-CTX、t-P1NP及N-MID-OT在绝经后女性骨质疏松性骨折风险评估中的应用

目的探讨β胶原特殊序列(β-crosslaps,β-CTX)、总1型胶原氨基端延长肽(t-P1NP)及N端中段骨钙素(N-MID-OT)在绝经后女性骨质疏松性骨折风险评估中的应用价值。方法回顾性分析2020年2月至2022年12月解放军总医院第六医学中心收治的102例绝经后骨质疏松(postmenopausal osteoporosis,PMOP)患者的临床资料,将其作为研究组,并根据是否发生骨折将其分为PMOP骨折组(39例)与PMOP未骨折组(63例),另选100名健康体检者作为对照组。记录所有研究对象的β-CTX、t-P1NP及N-MID-OT水平并进行比较,用ROC曲线评价β-CTX、t-P1NP及N-MID-OT在绝经后女性骨质疏松性骨折风险评估中的应用价值。结果研究组的平均体重、体质量指数、股骨颈骨密度(bone mineral density,BMD)、左髋总和BMD及L 1~4总和BMD均明显低于对照组(P<0.05);而两组研究对象的平均年龄、平均身高及绝经年龄等比较差异均无统计学意义(P>0.05)。研究组的β-CTX、t-P1NP及N-MID-OT均明显高于对照组(t值依次为12.688、37.430、26.599,P<0.05)。β-CTX+t-P1NP+N-MID-OT联合检测用于预测PMOP的AUC值为0.978,敏感度为98.04%,特异度为97.00%,表明β-CTX+t-P1NP+N-MID-OT三指标联合检测用于预测PMOP的效能更高。PMOP骨折组的β-CTX、t-P1NP及N-MID-OT均明显高于PMOP未骨折组(t值依次为6.078、16.363、12.227,P<0.05)。β-CTX+t-P1NP+N-MID-OT联合检测用于评估PMOP骨折风险的AUC值为0.939,敏感度为94.87%,特异度为95.24%,表明β-CTX+t-P1NP+N-MID-OT联合检测用于评估PMOP骨折风险的效能更高。结论PMOP患者β-CTX、t-P1NP及N-MID-OT水平均明显升高,而PMOP骨折患者β-CTX、t-P1NP及N-MID-OT水平升高更为明显,且β-CTX+t-P1NP+N-MID-OT联合检测可显著提高对PMOP预测及骨折风险评估效能,值得借鉴。

UPLC-Q-TOF/MS-based metabolomics reveals modulatory effects of Mesona chinensis Benth polysaccharide in liver injury mice induced by cyclophosphamide

The main purpose of this study was to investigate the improvement effect of Mesona chinensis Benth polysaccharide(MP)on cyclophosphamide(CTX)induced liver injury in mice.To explore metabolic profile of liver tissue and feces among normal group,CTX-induced group and MP management group based on metabolomics method by using UPLC-Q-TOF/MS.The results showed that MP could alleviate liver injury and promote the production of short chain fatty acids(SCFAs),with the best dose of 200 mg/kg·body weight(bw).The principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLSDA)scores plots of the liver and feces samples showed a clear separation among normal,model and highdose of MP(MPH).There were 18 endogenous metabolites in liver and 29 endogenous metabolites in feces,which were mainly involved in 8 metabolic pathways:taurine and hypotaurine metabolism,phenylalanine metabolism,α-linolenic acid metabolism,tricarboxylic acid(TCA)cycle,phenylalanine,tyrosine and tryptophan biosynthesis,arachidonic acid metabolism,sphingolipid metabolism as well as tryptophan metabolism.Moreover,a common metabolite arachidonic acid was observed in liver and feces samples.These endogenous metabolites may be considered to be MP’s response to liver protection.It will help to further understand the mechanism of MP and provide a basis for further research.

基于药效团模型筛选CTX-M-14型超广谱β-内酰胺酶抑制剂

【目的】开发针对头孢噻肟-水解酶-14(CTX-M-14)型超广谱β-内酰胺酶(extended-spectrumβ-lactamase,ESBLs)的抑制剂,用以缓解细菌耐药带来的严重危害。【方法】使用DiscoveryStudio Visualizer(DS Visualizer)构建基于受体结构的药效团模型(structure-based pharmacophore,SBP)和基于配体共同特征的定性药效团模型(common feature pharmacophore generation,HIPHOP),并将验证后的模型作为查询条件对ZINC数据库进行以CTX-M-14蛋白为靶标的虚拟筛选,得到拟合分数良好的中药单体成分甘草酸(glycyrrhizic acid,GL),对其进行相关作用力分析、分子动力学模拟和结合自由能计算,分析甘草酸与CTX-M-14蛋白的结合模式、结合能力及稳定性;最后通过联合抑菌试验及酶动力学试验考察甘草酸的抗菌增敏活性、抑酶作用及抑酶方式。【结果】中药单体成分甘草酸主要与CTX-M-14蛋白活性中心多个氨基酸残基形成氢键和范德华作用力,两者的对接分数与结合自由能分别为-10 kcal/mol及-22.06 kcal/mol;甘草酸与头孢噻肟钠联用呈协同作用(FICI≤0.5);甘草酸可竞争性抑制β-内酰胺酶对底物抗生素的水解作用,对头孢噻肟钠的抑酶保护率可达58.53%,与克拉维酸接近(60.98%)。【结论】中药单体甘草酸可与CTX-M-14蛋白稳定结合,通过竞争性抑制β-内酰胺酶对底物抗生素的水解作用,提高多重耐药大肠杆菌E320和重组蛋白阳性菌BL-21对头孢噻肟钠的敏感性,实现抗生素的减量增效。

Treatment outcome analysis of bevacizumab combined with cyclophosphamide and oxaliplatin in advanced pseudomyxoma peritonei

BACKGROUND Pseudomyxoma peritonei(PMP)is a rare peritoneal malignant tumor syndrome.Cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy is its standard treatment.However,there are few studies and insufficient evidence regarding systemic chemotherapy of advanced PMP.Regimens for colorectal cancer are often used clinically,but there is no uniform standard for late-stage treatment.AIM To determine if bevacizumab combined with cyclophosphamide and oxaliplatin(Bev+CTX+OXA)is effective for treatment of advanced PMP.The primary study endpoint was progression-free survival(PFS).METHODS Retrospective analysis was conducted on the clinical data of patients with advanced PMP who received Bev+CTX+OXA regimen(bevacizumab 7.5 mg/kg ivgtt d1,oxaliplatin 130 mg/m2 ivgtt d1 and cyclophosphamide 500 mg/m2 ivgtt d1,q3w)in our center from December 2015 to December 2020.Objective response rate(ORR),disease control rate(DCR)and incidence of adverse events were evaluated.PFS was followed up.Kaplan-Meier method was used to draw survival curve,and log-rank test was used for comparison between groups.Multivariate Cox proportional hazards regression model was used to analyze the independent influencing factors of PFS.RESULTS A total of 32 patients were enrolled.After 2 cycles,the ORR and DCR were 3.1%and 93.7%,respectively.The median follow-up time was 7.5 mo.During the follow-up period,14 patients(43.8%)had disease progression,and the median PFS was 8.9 mo.Stratified analysis showed that the PFS of patients with a preoperative increase in CA125(8.9 vs 2.1,P=0.022)and a completeness of cytoreduction score of 2-3(8.9 vs 5.0,P=0.043)was significantly longer than that of the control group.Multivariate analysis showed that a preoperative increase in CA125 was an independent prognostic factor for PFS(HR=0.245,95%CI:0.066-0.904,P=0.035).CONCLUSION Our retrospective assessment confirmed that the Bev+CTX+OXA regimen is effective in second-or posterior-line treatment of advanced PMP and that adverse reactions can be tolerated.A preoperative increase in CA125 is an independent prognostic factor of PFS.

广东省腹泻仔猪大肠杆菌blaCTX-M-65与mcr-1基因共传播特征

【目的】调查分析广东省某生猪养殖场腹泻仔猪的大肠杆菌耐药情况以及同时携带blaCTX-M-65与mcr-1两种耐药基因的阳性大肠杆菌的分子特征与其共同传播特征,为耐药性监测及风险防控提供科学依据。【方法】从广东省肇庆市某生猪养殖场采集腹泻仔猪直肠棉拭子样品90份,并进行大肠杆菌分离鉴定;采用PCR方法检测产超广谱β-内酰胺酶(ESBLs)与mcr-1基因,通过测序确定CTX-M亚型;采用琼脂二倍稀释法和微量肉汤稀释法测定分离菌对12种抗菌药物的最小抑菌浓度(MIC);使用脉冲场凝胶电泳(PFGE)及多位点序列分型(MLST)分析菌株间的遗传背景;通过接合转移方法确定质粒水平转移的能力;使用复制子分型、S1-PFGE、Southern blotting方法检测质粒类型、耐药基因的定位及定位质粒的大小;通过三代测序及序列分析确定质粒的重组过程。【结果】共分离鉴定86株大肠杆菌,检测到44株携带CTX-M型ESBLs基因,其中blaCTX-M-55基因最流行(n=16,36.4%),其次是blaCTX-M-65(n=10,22.7%)、blaCTX-M-27(n=8,18.2%)、blaCTX-M-15(n=5,11.4%),还检测出blaCTX-M-79(n=2,4.5%)、blaCTX-M-14(n=2,4.5%)和blaCTX-M-24(n=1,2.3%)。10株blaCTX-M-65基因阳性菌有8株同时携带mcr-1基因。这8株大肠杆菌均表现为多药耐药,包括氨苄西林、阿莫西林、头孢噻呋、头孢喹肟、黏菌素等多种抗生素。8株菌共分为6种ST型、7个PFGE谱型。接合转移试验发现,有6株菌的blaCTX-M-65和mcr-1基因发生了共转移,且blaCTX-M-65与mcr-1基因均位于IncHI2型(253 kb)质粒上。其中1株菌在接合转移的过程中,其所携带的一个253 kb IncHI2质粒与一个69 kb IncFⅡ质粒发生融合,形成一个大小323 kb的融合质粒。对融合质粒进行三代测序解析,结果表明,在接合转移的过程中IS26介导了两个不同大小质粒的重组。【结论】IncHI2质粒是介导blaCTX-M-65和mcr-1基因共同传播的主要媒介,IS26通过与靶位点GTTTCACT的整合导致IncHI2质粒与IncFⅡ质粒融合。质粒重组扩大了大肠杆菌的耐药谱,加速了耐药基因的传播,促使多药耐药现象更严重,对公共卫生安全构成威胁,本研究为阐明多重耐药大肠杆菌的传播机制提供了参考依据。

新疆维吾尔自治区塔额垦区牛源大肠杆菌耐药性、CTX-M基因携带情况与毒力基因检测

[目的]了解新疆维吾尔自治区塔额垦区牛源大肠杆菌耐药性及CTX-M基因和毒力基因携带情况。[方法]从塔额垦区某牛场采集16份牛腹泻粪样,通过选择性培养、革兰染色镜检、16S rDNA PCR扩增及测序进行大肠杆菌分离鉴定。利用PCR法对牛源大肠杆菌分离株进行分子分型(系统发育群和脂多糖核心型)以及耐药基因、CTX-M基因亚型、毒力基因检测;分别使用K-B纸片法、CTX与TCL双纸片法进行药物敏感性试验和产ESBLs大肠杆菌鉴定;采用结晶紫染色法半定量检测菌株的生物被膜形成能力。[结果]根据菌落生长特征、革兰染色特性及16S rDNA测序结果,从16份牛腹泻粪样中分离鉴定出16株大肠杆菌,分离率为100%;系统发育群主要为B1群(14/16,87.50%),脂多糖核心型多为R1型(15/16,93.75%)。分离菌株对青霉素、利福平、复方新诺明表现出较强耐药性,耐药率分别为100%、68.75%、68.75%;对多黏菌素、替加环素、美罗培南敏感,耐药率均为0;有12株(75.00%)具有多重耐药表型;有11株(68.75%)产ESBLs;有15株(93.75%)可形成生物被膜。16株大肠杆菌中共检出17种耐药基因,CTX-M、ant(6′)、sul1等8种耐药基因的检出率为100%;有14株(87.50%)为CTX-M-1G基因亚型;共检出12种毒力基因,iroN、ompA、yijP等5种毒力基因的检出率为100%。[结论]新疆维吾尔自治区塔额垦区牛源大肠杆菌CTX-M基因检出率较高,携带多种耐药基因和毒力基因,耐药性较强,对该地区牛健康养殖存在潜在威胁,应加强该地区CTX-M型大肠杆菌的流行情况监控。

基于血清PINP和β-CTX水平分析唑来膦酸联合阿法骨化醇在绝经后骨质疏松症治疗中的价值

目的基于血清Ⅰ型前胶原氨基端前肽(PINP)和β-Ⅰ型胶原交联羧基末端肽(β-CTX)水平分析唑来膦酸联合阿法骨化醇在绝经后骨质疏松症(PMOP)治疗中的价值。方法选取2023年7月~12月收治的500例PMOP患者作为研究对象,根据随机数字表法分为单一治疗组(使用唑来膦酸治疗)和联合治疗组(唑来膦酸联合阿法骨化醇治疗),每组250例。比较两组治疗效果,腰椎L_(1~4)、股骨颈及股骨转子骨密度(BMD)以及骨代谢指标[PINP、β-CTX、骨钙素(BGP)、碱性磷酸酶(ALP)]。结果联合治疗组治疗总有效率高于单一治疗组(98.00%vs.92.40%,P<0.05);治疗后,联合治疗组腰椎L_(1~4)、股骨颈及股骨粗转子BMD均较单一治疗组高,差异有统计学意义(P<0.05);治疗后,联合治疗组血清PINP、β-CTX、BGP、ALP水平均低于单一治疗组,差异有统计学意义(P<0.05);血清PINP、β-CTX、BGP水平与不同部位BMD均呈负相关性(P<0.05)。结论唑来膦酸联合阿法骨化醇治疗PMOP疗效确切,且血清PINP、β-CTX水平与BMD具有显著相关性,有望用于临床评估PMOP治疗效果。

Moxibustion at ST36 activates peritoneal macrophages in CTX-induced immunosuppression in mouse model via IFN-γ

Background:To explored whether moxa cone moxibustion can reduce peritoneal inflammation by increasing the content of peritoneal macrophages and B cells via interferon-gamma.Methods:The mice were randomly divided into three groups with six mice in each group:the control group,model group,and moxibustion group,and the model was established in mice using cyclophosphamide.In the moxibustion group,the mice received moxa cone moxibustion at Zusanli(ST36)for 7 days.Analysis of Peritoneal cell were detected by flow cytometry and immunofluorescence,the protein expression level in the peritoneal fluid were measured with mouse cytokine antibody arrays and verified by enzyme linked immuno sorbent assay test,and RNA-Sequencing was used for peritoneal cell RNA analysis.Results:Our results showed that moxa cone moxibustion could reduce the loss of large peritoneal macrophages and B1 cells(P<0.05).With the cytokine array analysis and enzyme linked immuno sorbent assay test of peritoneal fluid,we found that IFN‐γwas up-regulated in moxibustion group(P<0.05).There were 169 genes were down-regulated in the model group and up-regulated in the moxibustion group while 19 genes that were up-regulated in the model group and down-regulated in the moxibustion group via RNA-sequencing.Kyoto Encyclopedia of Genes and Genomes pathway analysis of 188 intersect differentially expressed genes were found that the top 3 pathways with the highest enrichment of up-regulated genes included Hematopoietic cell lineage,Inflammatory bowel disease and Malaria.The differentially expressed genes visualization protein-protein interaction network shows the top 10 genes including Ifng,Grb2,CCR7,CTLA4,CXCR5,Foxp3,kit,PRF1,CD5 and klrg1.Conclusion:These findings showed that moxa cone moxibustion can alleviate chemotherapy-induced diarrhea by reducing the loss of large peritoneal macrophages and B1 cells in the peritoneal cavity,possibly through up-regulating inflammatory bowel disease signaling pathway via interferon-gamma to regulate the survival and function of large peritoneal macrophages and B1 cells.

健脾益气活血汤治疗糖尿病并骨质疏松的疗效及对25-(OH)D、β-CTX、Pref-1的影响

目的:探讨健脾益气活血汤治疗糖尿病(diabetes mellitus,DM)并骨质疏松(osteporosis,OP)的疗效及对25羟维生素D[25⁃(OH)D]、β-骨胶原交联(β-CTX)、前脂肪因子-1(Pref-1)的影响。方法:选择2021年6月至2022年6月在河北省沧州中西医结合医院治疗的DM合并OP患者206例,应用随机数表法将其分为对照组(常规治疗)及观察组(联合健脾益气活血汤)各103例,对比2组疗效、治疗前后中医证候积分、骨密度(BMD)、血清25-(OH)D、β-CTX、Pref-1水平及不良反应。结果:观察组总有效率高于对照组(P<0.05)。治疗后,2组腰背疼痛、无力/肌肉关节刺痛、口干舌燥积分、β-CTX、Pref-1水平降低(P<0.05),且观察组低于对照组(P<0.05)。2组血清25-(OH)D水平,L 2~4、股骨颈、股骨大转子BMD均升高(P<0.05),且观察组高于对照组(P<0.05)。2组不良反应发生率差异无统计学意义(P>0.05)。结论:健脾益气活血汤治疗DM并OP可有效改善患者症状和血清25-(OH)D、β-CTX、Pref-1水平,调节骨代谢,增加BMD,提高疗效。

Effect of Rituximab Versus Mycophenolate Mofetil or Cyclophosphamide as Control in Lupus Nephritis:A Meta-Analysis

Objective:To evaluate the effects of rituximab versus mycophenolate mofetil or cyclophosphamide as control in lupus nephritis by meta-analysis.Methods:A systematic search was carried out up to January 2022,obtaining 7 studies involving 645 participants with lupus nephritis at the commencement of the investigation;198 of them were treated with rituximab,while 447 were treated with mycophenolate mofetil or cyclophosphamide.We determined the odds ratio(OR)and mean difference(MD)with 95%confidence index(CI)to compare rituximab’s efficacy to that of mycophenolate mofetil or cyclophosphamide as control in lupus nephritis using random-or fixed-effects model by dichotomous or continuous techniques.Results:The rituximab group showed significantly higher complete renal remission rate(OR=2.52;95%CI 1.30-4.91,P=0.006)and total renal remission rates(OR=2.22;95%CI 1.36-3.63,P=0.001)than the control group.However,there was no significant difference in terms of end Systemic Lupus Erythematosus Disease Activity Index(SLEDAI)score(MD-1.16;95%CI-2.88-0.57,P=0.19),proteinuria(MD-0.31;95%CI-0.70-0.09,P=0.013),and serum creatinine(MD 0.01;95%CI-0.04-0.07,P=0.64)between the rituximab group and the control.Conclusion:Rituximab exhibited significantly greater complete renal remission rate and total renal remission rates,with no significant difference in terms of shorter-end SLEDAI,proteinuria,and serum creatinine,compared with the control in individuals with lupus nephritis.

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.

Effects of low dose pre-irradiation on the toxicity of cyclophosphamide

Objective:The aim of the study was to investigate the effects as well as the possible mechanisms of low dose γ-ray pre-irradiation on hepatic damage,DNA damage of peripheral lymphocytes and genetic material damage caused by high dosage of cyclophosphamide(CTX).Methods:Kunming strain male mice were randomly divided into five groups:control group,sham-irradiated group,low dose irradiation group(LDR group),cyclophosphamide chemotherapy group(CTX group) and low dose irradiation combined with chemotherapy group(LDR + CTX group).Having being raised for one week,all the mice were implanted subcutaneously with S180 cells in the left inguen(control group excluded).On days 8 and 11,mice of LDR and LDR + CTX groups were given 75 mGy whole-body γ-irradiation,30 h later mice of CTX and LDR + CTX groups were injected i.p.3.0 mg cyclophosphamide.All the mice were sacrificed on day 13.DNA damage of the peripheral lymphocytes was analyzed using single cell gel electrophoresis(SCGE);ALT activity,total protein(TP) and albumin(ALB) of the plasma were analyzed using automatic biochemistry analyzer;MDA content,SOD and GSH-PX activity of the hepatic homogenate were analyzed using chromometry;genetic material damage was analyzed using micronucleus frequency(MNF) of polychromatoerythrocytes(PCE) in bone marrow.Results:1.Differences of MDA contents,SOD and GSH-PX activity of hepatic homogenate between 5 groups had notable statistical significance(P < 0.01);in control group MDA content was the lowest,SOD and GSH-PX activity were the highest,while in CTX group MDA content was the highest,SOD and GSH-PX activity were the lowest;compared with CTX group MDA content decreased significantly(P < 0.01) and SOD and GSH-PX activity increased significantly(P < 0.05) in LDR + CTX group.2.Differences of ALT activity of plasma between 5 groups had no statistical significance(F = 1.262,P > 0.05).Differences of TP and ALB of plasma between 5 groups had statistical significance(F = 12.879 and 6.336 respectively,P < 0.01);TP and ALB in control group were higher than those of other groups and compared with sham-irradiated group,TP and ALB in LDR group elevated significantly(P < 0.05).3.Differences of DNA damage of peripheral lymphocytes had notable statistical significance(F = 6.383,P < 0.01);DNA damage in control group was the lightest,while DNA damage in CTX group was the severest;compared with CTX group,DNA damage in LDR + CTX group was much lighter(P < 0.05).4.MNF of PCE between 5 groups had remarkable significance(F = 179.652,P < 0.01);compared with control group and sham-irradiated group,MNF in CTX group increased significantly(P < 0.01);compared with CTX group,MNF in LDR + CTX group had a tendency of decline,which had no statistical significance(P > 0.05).Conclusion:1.CTX can damage the hepatic tissue through oxidative stress;75 mGy γ-irradiation before CTX chemotherapy can induce activities of anti-oxidative enzymes,promote elimination of free radicals,so as to alleviate the damaging effects of oxidative stress to hepatic tissue caused by high-dose chemotherapy.2.A 75 mGy γ-irradiation before CTX chemotherapy has no obvious effect on ALT activity of plasma,but may have protective effect on the protein synthesis function of liver.3.High-dose CTX chemotherapy can cause DNA damage of peripheral lymphocytes;75 mGy γ-irradiation before chemotherapy may have certain protective effect on DNA damage.4.CTX has potent mutagenic effect,can cause significant increase of MNF of PCE;75 mGy γ-ray pre-irradiation did not show obvious protection against genetic toxicity of high-dose CTX chemotherapy.

Protective Effect of phytolacca Acinosa Polysaccharides Ⅰ(PAP-Ⅰ)on Hematopoiesis in Cyclophosphamide-treated and 60￣Co-irradiated Mice

The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.

经皮椎体后凸成形术治疗椎体压缩性骨折瘦素、脂联素及CTX-Ⅰ水平分析

目的 探讨经皮椎体后凸成形术(PKP)治疗椎体压缩性骨折瘦素、脂联素及Ⅰ型胶原蛋白C末端异物肽(CTX-Ⅰ)水平。方法 选取2019年6月至2022年5月南京中医药大学附属南京市中西医结合医院收治的92例椎体压缩性骨折患者为研究对象,根据治疗方式的不同分为对照组45例[予经皮椎弓根螺钉内固定术(PPSF)]、观察组47例(予PKP治疗);比较两组瘦素、脂联素和CTX-Ⅰ水平、分析观察组不同临床特征与病理特征的瘦素、脂联素和CTX-Ⅰ水平、采用多元Logistic回归分析影响观察组瘦素、脂联素和CTX-Ⅰ水平的危险因素,比较两组临床疗效及并发症情况。结果 观察组血清瘦素水平高于对照组,脂联素和CTX-Ⅰ水平均低于对照组,差异具有统计学意义(P<0.05);观察组不同年龄段、有无骨折病史、术前骨折椎体数和术中骨水泥量之间瘦素水平比较差异有统计学意义(P<0.05);不同性别、不同年龄段、有无骨折病史和术中骨水泥量之间脂联素水平比较差异有统计学意义(P<0.05);不同性别、不同年龄段、有无骨折病史和术中骨水泥量之间CTX-Ⅰ水平比较差异有统计学意义(P<0.05);经多元Logistic回归分析可知,女性、年龄>62岁、有骨折病史、术前骨折椎体数≥2个及术中骨水泥量使用1~5 mL为影响瘦素、脂联素和CTX-Ⅰ水平的危险因素(P<0.05);观察组临床总有效率(95.74%)高于对照组(82.22%),差异具有统计学意义(P<0.05);观察组并发症总发生率(2.13%)比对照组(13.33%)低,差异具有统计学意义(P<0.05)。结论 经PKP治疗能有效提高椎体压缩性骨折患者的临床疗效,有效改善患者血清瘦素、脂联素及CTX-Ⅰ水平,通过检测上述三指标水平可对PKP术后治疗提供参考依据。

Protective effect of ascorbic acid on cyclophosphamide induced testicular gametogenic and androgenic disorders in male rats

Aim:To study the detrimental effects of cyclophosphamide on the testicular androgenic and gametogenic activities through endocrine inhibition and/or induction of oxidative stress in male albino rats and to evaluate the protective effect of ascorbic acid.Methods:The testicular△^(5),3β-hydroxysteroid dehydrogenase(HSD),17β-HSD,peroxidase and catalase activities along with the levels of malondialdehyde(MDA)and conjugated dienes in testicular tissue were measured for the evaluation of testicular oxidative stress.The plasma testosterone(T)level was measured by immunoassay.Various germ cells at stageⅦof spermatogenic cycle were quantified from testicular stained sections.Results:Cyclophosphamide treatment results in a significant inhibition in the testicular△^(5),3β-HSD and 17β-HSD activities,a decrease in plasma T level and a diminution in the counts of various germ cells.Moreover,this treatment was also associated with a significant inhibition of the peroxidase and catalase activities along with high levels of MDA and conjugated dienes in the testis.All these changes were reversed by ascorbic acid co-administration.Conclusion:Cyclophosphamide treatment at the dosage used caused testicular gametogenic and androgenic disorders as well as induced testicular oxidative stress that can be reversed by ascorbic acid co-administration.