Rationale: Cytokine storm is now considered to be a systemic inflammatory response, but local cytokine storm may exist in systemic diseases of the blood system. Monitoring of regional cytokine storm is an important clue for the diagnosis of systemic diseases. Patient concerns: A 72-years-old male presented to our hospital with multiple serosal effusion without solid mass or enlarged lymph nodes. We found that the level of cytokines in ascites was tens to hundreds of times higher than that in plasma, mainly IL-6 and IL-8. Diagnoses: The patient was diagnosed with multiple serous effusion, hemophagocytic syndrome, B-cell lymphoma, Epstein–Barr virus infection, and hypoproteinemia. Interventions: During hospitalization, the patient was treated with 5 courses of R-CVEP therapy and supportive treatment. Outcomes: After the first R-CVEP regimen, the patient’s condition was evaluated as follows: hemophagocytic syndrome improved: no fever; Serum triglyceride 2.36 mmol/L; Ferritin 70.70 ng/L; no hemophagocyte was found in the bone marrow; the lymphoma was relieved, ascites disappeared, and bone marrow cytology showed: the bone marrow hyperplasia was reduced, and small platelet clusters were easily seen. Bone marrow flow cytometry showed that lymphocytes accounted for 13.7%, T cells increased for 85.7%, CD4/CD8 = 0.63, B cells decreased significantly for 0.27%, and NK cells accounted for 10.2%. Blood routine returned to normal: WBC 5.27 × 109/L, HB 128 g/L, PLT 129 × 109/L; Epstein–Barr virus DNA < 5.2E + 02 copies/mL; correction of hypoproteinemia: albumin 39.7 g/L. Lessons: Cytokines in ascites are significantly higher than those in plasma by tens to hundreds of times, suggesting that “regional cytokine storms” may cause serosal effusion.

The purpose of the present study was to evaluate cimigenol (Cim) treatment effects to cell proliferation by breaking bone marrow stromal cells (BMSCs) through C-X-C chemokine receptor type 4 (CXCR4)/stromal cell-derived factor-1α (SDF-1α) pathway. MV-4-11 and U937 cell lines were used. The present study was divided into two parts. First, the cell lines were divided into normal control (NC), BMSC (cells co-cultured with BMSCs), BMSC + DMSO, BMSC + Low (treated with 5 mg/ml Cim), BMSC + Middle (treated with 10 mg/ml Cim), BMSC + High (treated with 20 mg/ml Cim). In the second step, the cell lines were divided into NC, BMSC, BMSC + BL8040 (treated with BL8040 which inhibits CXCR4), BMSC + Cim and BMSC + Cim + BL8040. EdU positive cell numbers were measured by EdU assay and apoptosis rate by flow cytometry and TUNEL assay. Relative gene and protein expression was measured by reverse transcription-quantitative PCR and western blotting assay. BMSCs were able to protect proliferation of cancer cells and decreased cell apoptosis compared with the NC group (P<0.001, respectively). With Cim supplement, the cell proliferation was decreased with cell apoptosis increasing compared with NC group (P<0.001 respectively). However, the anti-tumor effects of Cim were not significantly different from the BL8040 treated groups (P<0.001, respectively). In conclusion Cim decreased acute myeloid leukemia cells protected by BMSCs through the CXCR4/SDF-1α pathway.

PURPOSE:Cimicifuga dahurica (C. dahurica), which has been used in traditional oriental medicine for a long period, was reported to exert extensive antitumor activity, but the effect and molecular biological mechanism of C. dahurica on multiple myeloma (MM) has not been elaborated. Tumor-associated macrophages (TAMs) exhibit a sustained polarization between tumor killing M1 subtype and tumor supporting M2 subtype. And a lower ratio of M1/M2 is associated with tumor angiogenesis, proliferation and invasion. We explored the inhibitory effect of the aqueous extract of the root of C. dahurica (CRAE) on tumor growth by reprogramming macrophage polarization in the tumor microenvironment.METHODS:Mice bearing SP2/0 multiple myeloma were treated with CRAE. Western blotting (WB), immunohistochemistry (IHC) and immunofluorescence staining were utilized to assess tumor growth and TAM populations. Macrophages were depleted by injection of clodronate liposomes to determine and measure the role of CRAE as an anti-tumor agent by targeting macrophages. To simulate tumor microenvironment, MM cells H929 and TAMs were co-cultured using the transwell co-culture system. By using CRAE as an immunoregulator in M2-like macrophages, we analyzed CRAE-treated macrophage-associated surface markers and cytokines by flow cytometry and WB.RESULTS:The results indicated that CRAE treatment could reduce tumor burden of MM mice and a high degree of M1-like macrophages infiltration was detected in tumor tissues. In vitro co-culture system, CRAE significantly promoted the polarization of M2 to M1 phenotype, which led to the increase in apoptosis of myeloma cells. It was found that the M1 polarization induced by CRAE depended on the TLR4-MyD88-TAK1-NF-κB signal transduction.CONCLUSION:This study elucidated the anticancer mechanism of the aqueous extract of C. dahurica (CRAE) through reprogramming macrophage polarization and highlighted that CRAE could act as a potential novel option for cancer immunotherapy.

多发性骨髓瘤(multiple myeloma,MM)是一种以骨髓中单克隆浆细胞恶性增殖为特征的浆细胞疾病,是目前全球第二高发的血液系统肿瘤.骨痛是MM常见的临床症状之一,大约2/3的MM患者因骨痛就诊,80％的MM患者确诊时即出现骨质疏松及病理性骨折等骨骼病变,即多发性骨髓瘤骨病(multiple myeloma bone disease,MBD)[1].

Solasonine, the main active ingredient of Solanum nigrum L ., has been reported to exert extensive antitumor activity. However, the antitumor effects in acute monocytic leukemia and the exact mechanisms involved are unknown. In this study, we investigated the role of solasonine on inhibiting the progression of acute monocytic leukemia. Our findings showed that solasonine inhibited the proliferation of acute monocytic leukemic cell lines (THP-1 and MV4-11) in vitro . Solasonine promoted apoptosis and induced cell cycle arrest in the G2/M phase. Analysis of RNA-seq data suggested that solasonine correlated with increased expression of genes in the AMPK/FOXO3A pathway. Inhibition of AMPK with compound C followed by treatment with solasonine showed that solasonine reduced apoptosis, caused less cell cycle arrest, and inactivated the AMPK/FOXO3A axis in THP-1 and MV4-11 cells. Solasonine also inhibited tumor growth by the activation of the AMPK/FOXO3A axis. In conclusion, solasonine inhibited the progress of acute monocytic leukemia in vitro and in vivo and triggered the apoptosis and cell cycle arrest in the G2/M phase by upregulating the AMPK/FOXO3A pathway.

Shengma Biejia decoction (SMBJD), a traditional Chinese formula recorded in the Golden Chamber , has been widely used for the treatment of malignant tumors. However, its underlying molecular targets and mechanisms are still unclear. This study showed that SMBJD inhibited tumor growth and stimulated hemogram recovery significantly in a multiple myeloma xenograft model. Western blot and immunohistochemistry assays of tumor tissues showed that SMBJD reduced the ratio of autophagy-related proteins LC3-II/LC3-I, while P62 and apoptosis-related proteins cleaved caspase-3/caspase-3 and Bax/Bcl-2 were upregulated. In vitro experiments demonstrated the time-dependent and dose-dependent cytotoxicity of SMBJD on multiple myeloma cell lines H929 and U266 through MTT assays. The LC3-II/LC3-I ratio and number of GFP-LC3 puncta showed that SMBJD inhibited the autophagy process of H929 and U266 cells. Moreover, both SMBJD and 3-methyladenine (3-MA) caused a decrease in LC3-II/LC3-I, and SMBJD could not reverse the upregulation of LC3-II/LC3-I caused by bafilomycin A1 (Baf-A1). Furthermore, the results of annexin V-FITC and propidium iodide double staining demonstrated that SMBJD treatment induced the apoptosis of H929 and U266 cells. These data prove that SMBJD inhibits autophagy and promotes apoptosis in H929 and U266 cells. The results also show that rapamycin could reduce the rate of SMBJD-induced apoptosis in H929 and U266 cells, at a concentration which had no effect on apoptosis but activated autophagy. In addition, analysis of the mechanism indicated that levels of phosphorylated ERK and phosphorylated mTOR were increased by treatment with SMBJD in vivo and in vitro . These results indicate that SMBJD, an old and effective herbal compound, could inhibit the viability of H929 and U266 cells and induce autophagy-mediated apoptosis through the ERK/mTOR pathway. Thus, it represents a potential therapy strategy for multiple myeloma.

Background Tumor associated macrophages (TAMs), a kind of inflammatory cells in the tumor microenvironment, are crucial for the occurrence and development of various tumors which increased the expression of CD163. Nevertheless, not much has been established regarding soluble CD163 and its connection to tumor diagnosis. In this case, a meta-analysis was conducted to determine the tumor diagnostic importance of serum sCD163. Methods In order to assess the correlation between sCD163 and the overall survival (OS) or progression-free survival (PFS) among tumor patients, a systematic perusal of literature published until June 2020 was conducted. Relevant data were primarily obtained from papers that have the following qualifications: 1) a confidence interval (CI) of 95%; 2) a report of the hazard ratios; and, 3) pooled by means of the Mantel-Haenszel random-effect representation. Results For the final meta-analysis, eight papers comprised of 1,236 cases were involved. Through pooled investigation, it was determined that a correlation exists between elevated serum sCD163 and worse OS (HR = 2.24, 95% CI: 1.50-3.35, P < 0.001) and PFS (HR = 3.90, 95% CI: 2.33-6.52, P < 0.001) among tumor cases. Subgroup analysis stratified by medium age at diagnosis demonstrated that patients over 60 years old with high sCD163 had worse OS (HR 2.28, 95% CI: 1.58-3.29, P < 0.001) than under 60 (HR 1.43, 95% CI: 1.15-1.77, P = 0.001). Subgroup analysis revealed that analysis method and medium age at diagnosis were the potential source of heterogeneity. Conclusions Overall, diagnosis of tumor cases can be adversely determined through substantial sCD163 levels. Consequently, it is encouraged that extensive researches regarding the rates of cancer survival be accomplished.

老年急性髓系白血病患者群体具有较为显著的特殊性,多数患者合并各种基础疾病,发病常伴各种染色体不良预后核型以及多种耐药基因表达,造成患者的治疗效果及耐受性低下.我们首次提出节拍式化疗结合升麻鳖甲汤加减的中西医结合治疗方案,并根据子午流注疗法,强调患者状态调整、适时服药,同时把中医体质思想渗透到辨证论治体系中,从而指导合理科学用药.在多年临床实践中,该治疗方案使广大患者获益,延缓疾病进展,延长生存周期.

目的 研究升麻鳖甲汤加减方在体外对人急性髓系白血病(acute myeloid leukemia,AML)细胞株HL-60以及人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖的影响;随后以人急性髓系白血病细胞HL-60的培养上清对人脐静脉内皮细胞HUVEC进行诱导培养,并加入不同浓度的升麻鳖甲汤加减方进行干预,检测人脐静脉内皮细胞HUVEC迁移、趋化、管腔形成能力的变化,从而揭示升麻鳖甲汤加减方对人急性髓系白血病细胞诱导的脐静脉内皮细胞迁移、趋化、管腔形成的影响.方法 体外培养HL-60以及HUVEC细胞,待细胞进入对数生长期后进行相应实验;首先将不同浓度升麻鳖甲汤加减方在体外分别用于干预人急性髓系白血病细胞HL-60和人脐静脉内皮细胞HUVEC,在干预时间达48 h后,采用MTT法分别检测不同浓度升麻鳖甲汤加减方对HL-60以及HUVEC细胞增殖能力的影响;根据MTT实验的数据提示,选择适宜终浓度的升麻鳖甲汤加减方与HL-60细胞培养上清进行混合制备含药条件培养基用于干预HUVEC细胞,并分别运用划痕迁移、Transwell小室趋化以及Matrigel胶小管形成等实验方法观察HUVEC细胞经干预后的迁移、趋化效应以及成管能力的变化.结果 不同浓度的升麻鳖甲汤加减方干预48 h后能够分别显著抑制HL-60以及HUVEC细胞的增殖(P＜0.01),并且这种增殖抑制效应呈现出明显的时间-浓度依赖性,并且在HL-60和HUVEC两种细胞之间存在差异性细胞毒作用,即对HUVEC细胞的增殖抑制显著高于HL-60细胞(P＜0.01);不同终浓度的升麻鳖甲汤加减方对由HL-60细胞上清诱导培养的HUVEC细胞的迁移运动能力均有不同程度的抑制作用(P＜0.05),并且细胞迁移率随着药物浓度的增加而下降;细胞Transwell小室趋化实验结果显示,不同终浓度的升麻鳖甲汤加减方可显著降低HL-60细胞上清诱导培养的HUVEC细胞的趋化性(P＜0.05);HUVEC细胞小管形成连接点、分支数量等均明显低于阳性对照组(P＜0.01),小管形成抑制率随着药物浓度的增加而上升,且HUVEC细胞生成小管的连接点及分支数量与药物浓度呈负相关(P＜0.01).结论 升麻鳖甲汤加减方可抑制HL-60及HUVEC细胞的增殖,并存在差异细胞毒作用,提示该复方可能具有抑制急性髓系白血病肿瘤血管形成的效应,而这种效应的产生可能与其抑制肿瘤血管内皮细胞的迁移、趋化及成管能力等相关.

Context ShengMaBieJia decoction (SMBJD) is used to treat solid and hematological tumours; however, its anti-angiogenesis activity remains unclear. Objective This study verified the anti-angiogenic effects of SMBJD in vitro and in tumour-bearing acute myeloid leukaemia (AML) mouse models. Materials and methods In vivo, the chicken chorioallantoic membrane (CAM) and BALB/c null mouse xenograft models were treated with SMBJD (0, 2, 4, and 8 mg/mL) for 48 h and for 2 weeks, respectively. Anti-angiogenic activity was assessed according to microvessel density (MVD) and immunohistochemistry (IHC) targeting CD31 and VEGFR2. In vitro, proliferation viability, migratory activity and tube formation were measured. Western blots and polymerase chain reaction (PCR) assays were used to examine the levels of PI3K, Akt, and VEGF. Results HPLC analyses revealed the active constituents of SMBJD such as liquiritin, cimifugin, ferulic, isoferulic, and glycyrrhizic acids. In vitro, SMBJD treatment decreased cellular migration, chemotaxis, and tube formation at non-cytotoxic concentrations (2, 4, and 8 mg/mL) in a time- and dose-dependent manner. The dosage of less than IC20 is considered safe. In vivo, CAM models exhibited a decrease in MVD, and the tissues of xenografted mice possessed reduced CD31 and VEGFR2 expression. Conditioned media (CM) from AML cells (HL60 and NB4 cells) treated with non-cytotoxic doses of SMBJD inhibited chemotactic migration and tube formation in vitro. Both CM (HL60) and CM (NB4) exhibited downregulated expression of PI3K, Akt, and VEGF. Discussion and conclusions SMBJD inhibited angiogenesis in AML through the PI3K/AKT pathway, which might be combined with targeted therapy to provide more effective treatment.

目的 探讨升麻鳖甲汤加减方治疗急性髓系白血病的可能机制.方法 采用MTT法观察0、2、4、6、8、10 mg/ml浓度的升麻鳖甲汤加减方干预HL-60细胞24h、48h、72h时细胞增殖抑制率,筛选最佳实验时间和药物浓度.取对数生长期HL-60细胞5×105个/ml,按每孔2ml体积种入6孔培养板中.分别加入筛选浓度的升麻鳖甲汤加减方设为升麻鳖甲汤加减方低、中、高剂量组,另设对照组加入等体积完全培养基,继续培养筛选得出的最佳实验时间后收集各组细胞.采用Heochest 33258染色、流式细胞术观察各组细胞凋亡率,检测细胞周期分布、线粒体膜电位及细胞凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3(caspase-3)、半胱氨酸天冬氨酸蛋白酶8 (caspase-8)、半胱氨酸天冬氨酸蛋白酶9(caspase-9)、多聚二磷酸腺苷核糖聚合酶(PARP)表达,MAPK信号通路相关蛋白p38丝裂原活化蛋白激酶(p38 MAPK)、细胞外信号调节激酶(ERK)、c-Jun氨基末端激酶(JNK)蛋白表达.结果 筛选得出24h为最佳作用时间,升麻鳖甲汤加减方高、中、低剂量组干预浓度分别为4、6、8 mg/ml.升麻鳖甲汤加减方各剂量组均不同程度抑制人急性髓系白血病细胞HL-60增殖(P＜0.05);Heochest 33258染色和Annexin V/PI检测结果均显示,升麻鳖甲汤加减方各剂量组可呈浓度依赖性的增加细胞凋亡率,同时升高线粒体膜电位(P＜0.05);升麻鳖甲汤加减方各剂量组可呈浓度依赖性阻滞细胞周期升高凋亡相关蛋白表达以及MAPK信号通路相关蛋白水平(P＜0.05).结论 升麻鳖甲汤加减方可通过阻滞HL-60细胞细胞周期、诱导其线粒体膜电位的丢失引发细胞凋亡,从而抑制其增殖,可能与调控MAPK信号通路有关.

The objective was to investigate the effects of tenacigenin B on apoptosis of human acute myeloid leukemia cell lines HL-60, NB-4, and K562 and the underlying mechanisms. The inhibitory effect of tenacigenin B on the proliferation of human acute myeloid leukemia cell lines HL-60, NB-4 and K562 was observed by MTT assay; the effect of tenacigenin B on the morphology of apoptotic nuclei in these cell lines was detected using Hoechst 33258 staining; the effect of tenacigenin B on the apoptosis rate and mitochondrial membrane potential of these cell lines was determined by flow cytometry; expression of apoptosis-related proteins, including caspase-3, caspase-8, caspase-9, and PARP, and p38, p-p38, ERK1/2, pERK1/2, JNK1/2, and p-JNK1/2 in HL-60 cells was determined by Western blot after treated with enacigenin B for 24 h; and a nude mouse xenograft model was constructed by subcutaneous injection of HL-60 cells to evaluate the antitumor effect of tenacigenin B in vivo. Tenacigenin B inhibited the proliferation of human acute myeloid leukemia cell lines and induced cell apoptosis in a concentration- and time-dependent manner, and caspase-3, caspase-8, and caspase-9 were activated with cleaved PARP. In addition, the phosphorylation of p38, JNK1/2 and ERK1/2 proteins was also up-regulated to different degrees. Further inhibition assay confirmed that the activation of caspase induced by tenacigenin B was reversed by inhibiting the MAPK pathway. In vivo assay showed that tenacigenin B effectively inhibited the growth of transplanted tumors. In conclusion, tenacigenin B induces apoptosis of acute myeloid leukemia cells via caspase-dependent pathways, and upstream MAPK signaling pathway is involved in the regulation of this biological process.

Background: The aim of this study was to examine the effects and mechanisms of tenacigenin B in lymphoma treatment by in vitro and in vivo experiment. Material/Methods: Raji cells were treated by difference methods. Measuring the cell proliferation of difference groups was done by MTT assay; cell apoptosis and cell cycle of difference groups were evaluated by flow cytometer; relative mRNA expression was evaluated by real-time polymerase chain reaction (RT-PCR), and relative protein expressions were measured by western blot assay in an in vitro study. In an in vivo study, we used a nude mice model to explore the anti-tumor effects and mechanism of tenacigenin B. Cell apoptosis was measured by TUNEL assay; relative protein expressions were evaluated by immunohistochemistry assay, and relative mRNA expression was evaluated by RT-PCR. In addition, the blood components of difference groups were measured. Results: Compared with the Normal control group, the cell proliferation rate was significantly downregulated, with cell apoptosis significantly increasing with G1 phase in the Drug group and the si-Aurora-A group (P<0.05, respectively). The PTEN, PI3K, AKT, P53, and P21 mRNA and protein expressions of the Drug group, the si-Aurora-A group, and the si-Aurora-A+Drug group were significantly different (P<0.01, respectively), The tumor volume and weight of the Drug group, the si-Aurora-A group, and the si-Aurora-A+Drug group were significantly suppressed compared with the Normal group (P<0.01, respectively). The positive apoptosis cell number in the Drug group, the si-Aurora-A group, and si-Aurora-A+Drug group were increased compared with that of Normal group (P<0.01, respectively). Conclusions: Tenacigenin B had anti-tumor effects on lymphoma via regulation of Aurora-A in vitro and in vivo.

目的:系统评价联合复方黄黛片的方案用于治疗急性早幼粒细胞白血病的疗效及安全性.方法:计算机检索Cochrane图书馆、EMBase、Medline、PubMed、中国生物医学文献数据库、维普、万方、中国知网数据库,查找与复方黄黛片治疗急性早幼粒细胞白血病相关的随机和半随机对照研究,检索时限为建库至2017年12月.根据纳入及排除标准对文献进行筛选,采用Cochrane Reviewer's Handbook 5.0评价标准和工具对纳入的研究进行方法学质量评价,对纳入的文献进行数据提取,采用RevMan 5.2软件进行统计分析.共计纳入10个随机对照研究,共498例患者.结果:复方黄黛片治疗急性早幼粒细胞白血病与目前常用治疗方案比较,在完全缓解、达到完全缓解的时间、早期病死率等方面比较,差异无统计学意义(P＞0.05),各指标的RR及其95％ CI分别1.64(0.53,5.05)、2.91(-4.55,10.38)、0.98(0.39,2.44);同时也对药物安全性进行了对比,在肝功能异常、胃肠反应、皮疹方面比较,两组之间差异亦无统计学意义(P＞0.05),各评价指标的RR及其95％ CI分别为0.98(0.39,2.44)、1.08(0.74,1.57)、0.75(0.38,1.48);在凝血功能异常方面,其RR及95％ CI分别为0.12(0.02,0.56),差异具有统计学意义(P＜0.05).结论:复方黄黛片治疗急性早幼粒细胞白血病并未提高治疗有效性或减少治疗相关肝功能异常、胃肠反应、皮疹的作用;但可降低患者治疗相关凝血功能异常发生率.

By expounding the etiological factors and pathogenesis of non-infectious diarrhea from modem medicine and traditional medicine,the author discussed the therapies and suggested that the treatments of modem and traditional medicine had their own features and we should take both their advantages so as to treat non-infectious diarrhea and carry out more feature therapy.