Assessment of oral ciprofloxacin impaired gut barrier integrity on gut bacteria in mice

Gut bacteria and gut barrier plays important roles in body homeostasis. Ciprofloxacin (CPFX) is widely used to treat bacterial infections. However, whether high dosage of CPFX has side effects on gut barrier integrity is still unclear. Our results indicated that the High CPFX treatment (1 mg/ml) caused weight loss, nervousness, anorexia, and increased apoptosis cells in gut, but less influence was observed in the Low CPFX group (0.2 mg/ml). Meanwhile, the High CPFX treatment impaired tight junction molecules Ocln/ZO-1 level and down-regulated antibacterial genes expression (reg3γ, pla2g2α and defb1). Further, the High CPFX treatment increased pro-inflammatory cytokine IL-1β in intestinal tract, decreased IL-17A of duodenum but increased IL-17A of colon at day 37. In addition, the gut bacterial diversity and richness behaved significantly loss regarding CPFX treatment, especially in the High CPFX group during the experiment. Indole exhibited sharply decline in both Low and High CPFX groups at day 7, and the High CPFX mice needed longer time on restoring indole level. Meanwhile, CPFX treatment strongly decreased the concentrations of butyric acid and valeric acid at day 1. Correlation analysis indicated that the linked patterns between the key bacteria (families Bacteroidales_S247, Ruminococcaceae and Desulfovibrionaceae) and metabolites (indole and butyric acid) were disturbed via the CPFX treatment. In conclusion, the High CPFX treatment impaired the gut barrier with the evidence of reduced expression of tight junction proteins, increased apoptosis cells and inflammatory cells, decreased the bacterial diversity and composition, which suggesting a proper antibiotic-dosage use should be carefully considered in disease treatment.     

干扰素在恶性淋巴瘤自体移植后早期干预的应用

目的:观察非霍奇金淋巴瘤（non-Hodgkin's lymphoma，NHL）患者自体造血干细胞移植（autologous hematopoietic stem cell transplantation，ASCT）术后应用重组人α-2b干扰素（α-2b IFN）进行早期干预治疗的临床疗效。方法:选取18例行ASCT的NHL患者为研究对象，移植前疾病评估均未达到完全缓解（complete remission，CR），试验组血象恢复后给予IFN 3 000 000 U次/隔日干预治疗，3个月后停用；对照组未行干扰素干预治疗，分析总体疗效及两组对比的生存情况。结果:随访中位时间为34（10~50）个月，患者中位生存时间为37（31~45）个月，3年总体无进展生存（progressive free survival，PFS）、总生存（overall survivial，OS）分别为54.7%、66.8%。ASCT后试验组1年内无疾病复发，2年内复发率为12.5%；对照组1年内复发率为20%，2年内复发率为30%。结论:NHL患者在ASCT后给予重组人α-2b IFN早期干预治疗，患者耐受性好，可能降低移植后早期复发率。     

从干细胞方向研究针灸作用机制的研究进展

针灸是我国传统医学的重要组成部分，其疗效得到世界范围广泛认可，对其作用机制研究不断扩展、深入。干细胞自上世纪60年代被发现以来，其基础研究以及临床应用迅速发展，对组织的修复、再生或替代治疗产生了积极的影响。近年来，国内外也有研究报道试图从针灸对干细胞的影响方面揭示针灸的治疗机制，就此内容检索相关文献，从脑病、脊髓损伤、关节炎等不同疾病中针灸对干细胞的影响，以及不同针灸方法对干细胞的影响等方面进行综述，以期为针灸的临床应用和基础研究提供相关思路。     

GPER-induced signaling is essential for the survival of breast cancer stem cells

G protein-coupled estrogen receptor-1 (GPER), a member of the G protein-coupled receptor (GPCR) superfamily, mediates estrogen-induced proliferation of normal and malignant breast epithelial cells. However, its role in breast cancer stem cells (BCSCs) remains unclear. Here we showed greater expression of GPER in BCSCs than non-BCSCs of three patient-derived xenografts of ER⁻/PR⁺ breast cancers. GPER silencing reduced stemness features of BCSCs as reflected by reduced mammosphere forming capacity in vitro, and tumor growth in vivo with decreased BCSC populations. Comparative phosphoproteomics revealed greater GPER-mediated PKA/BAD signaling in BCSCs. Activation of GPER by its ligands, including tamoxifen (TMX), induced phosphorylation of PKA and BAD-Ser118 to sustain BCSC characteristics. Transfection with a dominant-negative mutant BAD (Ser118Ala) led to reduced cell survival. Taken together, GPER and its downstream signaling play a key role in maintaining the stemness of BCSCs, suggesting that GPER is a potential therapeutic target for eradicating BCSCs.     

Efficacy and Safety of the Modified EPOCH Regimen (Etoposide,Vincristine, Doxorubicin,Carboplatin, and Prednisolone) for Adult T-cell Leukemia/Lymphoma: A Multicenter Retrospective Study

BackgroundWe retrospectively analyzed patients with untreated aggressive adult T-cell leukemia/lymphoma who received the modified EPOCH (mEPOCH) regimen.Patients and MethodsPatients received up to 6 mEPOCH cycles. Etoposide (50 mg/m²/day), doxorubicin (10 mg/m²/day), and vincristine (0.4 mg/m²/day) were each given as a continuous 96-hour infusion on days 1 to 4. Prednisolone (40 mg/m²/day) was given intravenously or orally on days 1 to 4 and then tapered and stopped on day 7, and carboplatin (dose calculated for each patient individually using Calvert’s formula according to a target under the curve of 3 mg/mL/min) was given as a 2-hour intravenous infusion on day 6.ResultsIn 103 patients, overall response rate and complete response rate were 58% and 25%, respectively. With a median follow-up of 8.9 months, the median survival time was 9.8 months (95% confidence interval, 7.2-13.9 months). The median progression-free survival (PFS) was 4.2 months (95% confidence interval, 3.4-5.7 months). Patients who completed ≥ 4 cycles experienced significantly better overall survival and PFS compared with those who completed < 4 cycles. Twenty-eight patients underwent allogeneic hematopoietic stem cell transplantation after mEPOCH and demonstrated significantly prolonged overall survival and PFS compared with those who did not undergo transplantation.ConclusionThe mEPOCH regimen is effective with tolerable adverse effects and may be an alternative treatment option for adult T-cell leukemia/lymphoma.     

High infiltration of mast cells is associated with improved response to adjuvant chemotherapy in gallbladder cancer

Recent studies have reported that tumor‐infiltrating mast cells (TIM) play an important role in tumor regression, but the effect of TIM in gallbladder cancer (GBC) remains unclear. The present study aims to investigate the prognostic value of TIM in GBC patients and its responsiveness to gemcitabine‐based adjuvant chemotherapy (ACT). A total of 298 GBC patients from Zhongshan Hospital were recruited for this study. TIM infiltration was measured by immunohistochemical staining. Accumulation of TIM is significantly associated with prolonged overall survival in GBC patients. The benefit from gemcitabine‐based ACT was superior among patients with high infiltration of TIM with GBC. Multivariate analysis identified TIM infiltration as an independent prognostic factor for overall survival. A heatmap showed that TIM‐activated gene signatures were positively correlated with CD8+ T cells' gene signatures. Gene set enrichment analysis (GSEA) suggested that TIM was related to multiple T cell‐related processes and signaling pathways, including the interferon gamma signaling pathway and the leukocyte migration signaling pathway. It was confirmed that CD8+ T cell infiltration was positively correlated with high TIM infiltration in tissue microarray (TMA), suggesting that TIM infiltration was linked to the immune surveillance in GBC. TIM can be used as an independent prognostic factor and a predictor of therapeutic response of gemcitabine‐based ACT in GBC patients, which may mediate immune surveillance by recruiting and activating CD8+ T cells in GBC.     

A prognostic signature based on immune-related genes for cervical squamous cell carcinoma and endocervical adenocarcinoma

Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) is the fourth commonest female malignancy worldwide. CESC progresses in immune-microenvironment mainly composed of infiltrating immune and stromal cells. Here, we performed an integrated analysis incorporating the expression profiles from the Cancer Genome Atlas (TCGA) database and scores of immune and stromal cells calculated by Estimation of Stromal and Immune cells in Malignant Tumours using Expression data (ESTIMATE) algorithm. A two-gene signature (CD1C and CD6 genes) was established to predict the prognosis of CESC. Based on this signature, patients were divided into the high- and low-risk groups, and this signature showed good prognostic performance according to the results of Kaplan-Meier analysis and receiver operating characteristic (ROC) analysis in train set and two validation sets. A nomogram was built for evaluating the clinical applicability of this signature. In addition, based on Tumor Immune Estimation Resource (TIMER) database, 2 hub genes showed negative correlations with tumor purity and positive correlations with infiltrating levels of immune filtrating cells. What’s more, we propose new treatment strategies for the two prognostic subtypes. Low- risk patients were found presenting with a higher level of immune checkpoint molecules and showing higher immunogenicity in immunophenoscore (IPS) analysis, which indicated a better response for immunotherapy. Meanwhile, estimated by Genomics of Drug Sensitivity in Cancer (GDSC) database, the high-risk patients showed sensitive responses to five chemotherapy drugs. Finally, 10 candidate small-molecule drugs for CESC were defined. In summary, the CD1C-CD6 signature can accurately predict the prognosis of CESC.     

营养及化疗对肿瘤患者血清微量元素和免疫功能的影响

目的 探讨肿瘤患者化疗后血清微量元素及免疫功能的变化，以及肠内营养支持对上述测定指标的影响。方法 　 选择 2018 年 3 月至 5 月进入武汉市中心医院化疗的肿瘤患者 19 例，年龄为 18~70 岁；KPS 评分≥ 60。随机分为两组： 9 例患者化疗期间行肠内营养干预，10 例患者未进行营养干预。在化疗前后分别测定血清微量元素和外周血淋巴细胞亚 群，并进行统计学分析。结果 未经营养干预的患者化疗后血清锰含量比化疗前显著减低（P=0.004），化疗后血清铅呈 现临界水平降低（P=0.057）；行营养干预的患者血清锌含量显著降低（P=0.013），外周血中NK 细胞比例在化疗后呈 临界水平降低（P=0.059），而 CD4+/CD8+ 之比在化疗后呈临界水平升高（P=0.057）。结论 化疗及营养干预对血清微 量元素浓度有一定的作用；营养干预对外周血T 细胞的分布有一定的影响，表现为NK 细胞比例在化疗后有降低趋势， CD4+/CD8+ 比值有升高趋势。由于病例数较少以及营养干预时间较短，因此有必要增加病例数以及延长营养干预时间并 做进一步深入研究。     

基于灰色关联度分析和聚类分析的3种滇产重楼抗肝癌作用谱效关系探索

目的:研究长柱重楼、滇重楼和南重楼的HPLC指纹图谱与其抗肝癌作用的谱效关系,为明确重楼抗肝癌作用的物质基础提供实验依据。方法:采用HPLC建立3种重楼提取物的指纹图谱,流动相乙腈(A)-水(B)梯度洗脱(0～10 min,20%A; 10～20 min,20%～25%A; 20～30 min,25%～30%A; 30～40 min,30%～35%A; 40～50 min,35%～40%A; 50～60 min,40%A; 60～75 min,40%～45%A; 75～80 min,45%～60%A),流速0. 9 m L·min~(-1),检测波长203 nm;利用噻唑蓝(MTT)比色法测定3种重楼提取物对肝癌HepG2细胞的增殖抑制作用,计算半数抑制浓度(IC_(50));运用聚类分析(HCA)和灰色关联度分析(GRA)研究3种重楼指纹图谱和抗肝癌作用的关系,找出对抗肝癌作用贡献较大的成分。结果:在3种重楼的HPLC指纹图谱中,确定其中11个色谱峰为共有峰。作用时间72 h时长柱重楼、滇重楼、南重楼的IC_(50)分别为148. 33,178. 87,208. 09 mg·L~(-1),其中长柱重楼的抗肝癌活性最强。灰色关联度结果显示,滇重楼共有峰中关联度较高的为1～10号峰,长柱重楼共有峰关联度较高的为1～7号峰,南重楼共有峰中关联度较高的为1～4,6～10,N1号峰,与IC_(50)关联度均0. 7。各重楼变量的聚类分析结果显示,可与IC_(50)聚为一类的色谱峰的关联度均 0. 7。结论:建立了3种重楼的HPLC指纹图谱,重复性良好。3种重楼中的1～4,6和7号色谱峰对抗肝癌药效贡献最大。     

盐霉素对鼻咽癌CNE-2细胞放射敏感性的影响

目的:体外实验研究盐霉素对鼻咽癌CNE-2细胞放射敏感性的影响。方法:通过CCK-8法检测盐霉素作用后鼻咽癌CNE-2细胞的增殖情况。利用克隆集落形成实验观察鼻咽癌细胞成活情况，单击多靶模型拟合剂量存活曲线，计算放射增敏指数（SER）；Chou-Talalay数学模型绘制联合指数（CI）曲线，判断盐霉素和放射的作用关系。利用γ-H2AX焦点形成检测DNA分子损伤情况。结果:盐霉素能够抑制鼻咽癌细胞的增殖，并且具有明显的时间和剂量依赖性；盐霉素作用鼻咽癌细胞的IC50值在24 h时为17.81 μmol/L，48 h时为3.98 μmol/L。根据单击多靶模型拟合细胞的存活曲线，可知在鼻咽癌细胞CNE-2中盐霉素浓度为0.1 μmol/L和0.5 μmol/L时SER分别为1.19和1.21，均大于1.0，表明盐霉素对鼻咽癌细胞具有明显的放射增敏作用。制作联合指数（CI）曲线，可知在盐霉素浓度为0.1 μmol/L时，放射剂量为2 Gy时，CI值≈1，盐霉素和放射对CNE-2的作用接近相加作用，放射剂量为4、6、8 Gy时，CI值均＜1，二者为协同作用；在盐霉素浓度为0.5 μmol/L时，放射剂量为2、4、6、8 Gy时，CI值均＜1，二者均为协同作用。相对于照射组，盐霉素+照射组能增加DNA损伤数目。结论:盐霉素能提高鼻咽癌细胞的放射敏感性，为一种潜在的放射增敏药物。