Relationship between MLH1, PMS2, MSH2 and MSH6 gene-specific alterations and tumor mutational burden in 1057 microsatellite instability-high solid tumors

Microsatellite instability-high (MSI-H) and tumor mutational burden (TMB) are predictive biomarkers for immune-checkpoint inhibitors (ICIs). Still, the relationship between the underlying cause(s) of MSI and TMB in tumors remains poorly defined. We investigated associations of TMB to mismatch repair (MMR) protein expression patterns by immunohistochemistry (IHC) and MMR mutations in a diverse sample of tumors. Hypothesized differences were identified by the protein/gene affected/mutated and the tumor histology/primary site. Overall, 1057 MSI-H tumors were identified from the 32 932 tested. MSI was examined by NGS using 7000+ target microsatellite loci. TMB was calculated using only nonsynonymous missense mutations sequenced with a 592-gene panel; a subset of MSI-H tumors also had MMR IHC performed. Analyses examined TMB by MMR protein heterodimer impacted (loss of MLH1/PMS2 vs. MSH2/MSH6 expression) and gene-specific mutations. The sample was 54.6% female; mean age was 63.5 years. Among IHC tested tumors, loss of co-expression of MLH1/PMS2 was more common (n = 544/705, 77.2%) than loss of MSH2/MSH6 (n = 81/705, 11.5%; P < .0001), and was associated with lower mean TMB (MLH1/PMS2: 25.03 mut/Mb vs MSH2/MSH6 46.83 mut/Mb; P < .0001). TMB also varied by tumor histology: colorectal cancers demonstrating MLH1/PMS2 loss had higher TMBs (33.14 mut/Mb) than endometrial cancers (20.60 mut/Mb) and other tumors (25.59 mut/Mb; P < .0001). MMR gene mutations were detected in 42.0% of tumors; among these, MSH6 mutations were most common (25.7%). MSH6 mutation patterns showed variability by tumor histology and TMB. TMB varies by underlying cause(s) of MSI and tumor histology; this heterogeneity may contribute to differences in response to ICI.     

Use of biomimetic microtissue spheroids and specific growth factor supplementation to improve tenocyte differentiation and adaptation to a collagen-based scaffold in vitro

Tenocytes represent a valuable source of cells for the purposes of tendon tissue engineering and regenerative medicine and as such, should possess a high degree of tenogenic differentiation prior to their use in vivo in order to achieve maximal efficacy. In the current report, we identify an efficient means by which to maintain differentiated tenocytes in vitro by employing the hanging drop technique in combination with defined growth media supplements. Equine tenocytes retained a more differentiated state when cultured as scaffold-free microtissue spheroids in low serum-containing medium supplemented with l-ascorbic acid 2-phosphate, insulin and transforming growth factor (TGF)-β1. This was made evident by significant increases in the expression levels of pro-tenogenic markers collagen type I (COL1A2), collagen type III (COL3A1), scleraxis (SCX) and tenomodulin (TNMD), as well as by enhanced levels of collagen type I and tenomodulin protein. Furthermore, tenocytes cultured under these conditions demonstrated a typical spindle-like morphology and when embedded in collagen gels, became highly aligned with respect to the orientation of the collagen structure following their migration out from the microtissue spheroids. Our findings therefore provide evidence to support the use of a biomimetic microtissue approach to culturing tenocytes and that in combination with the defined growth media described, can improve their differentiation status and functional repopulation of collagen matrix.     

滋阴益肾法治疗特发性血小板减少性紫癜疗效及对患者血清IL-4、IL-6和免疫功能的影响

目的: 探讨滋阴益肾法治疗特发性血小板减少性紫癜(ITP)的疗效及对血清IL-4、IL-6、TNF-α及免疫功能的影响。方法:选取ITP患者84例,随机数字表法分为对照组和观察组,各42例。对照组采用口服泼尼松片治疗,观察组在对照组用药基础上给予滋阴益肾法治疗。比较两组临床疗效,治疗前后IL-4、IL-6、TNF-α、外周血T淋巴细胞亚群(CD₃⁺、CD₄⁺、CD₈⁺、CD₄⁺ /CD₈⁺)及免疫球蛋白水平变化情况。结果:两组治疗后中医症候积分明显降低,观察组优于对照组(P<0.05)。观察组总有效率95.24%高于对照组78.57%(P<0.05); 两组治疗后PLT计数均明显升高、PAIgG表达水平降低,观察组更明显(P<0.05); 两组治疗前IL-4、IL-6、TNF-α及外周血T淋巴细胞亚群水平比较差异无统计学意义(P>0.05); 治疗后CD₃⁺、CD₄⁺、CD₄⁺ /CD₈⁺均升高,IL-4、IL-6、TNF-α、CD₈⁺均降低(P<0.05); 观察组治疗后CD₃⁺、CD₄⁺、CD₄⁺ /CD₈⁺均高于对照组,IL-4、IL-6、TNF-α、CD₈⁺均低于对照组(P<0.05)。治疗后观察组的血清IgA和IgG水平明显高于对照组(P<0.05),而血清IgM水平差异无统计学意义(P>0.05)。两组不良反应情况比较无统计学差异(P>0.05)。结论: 滋阴益肾法可显著降低ITP患者血清IL-4、IL-6等因子水平,提高患者免疫能力。     

电子线照射对小鼠胰岛素生长因子相关蛋白表达的影响

目的：探讨电子线对胰岛素生长因子-1家族蛋白表达的影响，为研究辐射对肝脏的早期损伤提供依据。方法：采用电子线照射小鼠，照射剂量为1、2、4 Gy，用Western blot法检测照射后12、48和72 h小鼠肝脏中胰岛素生长因子1受体（IGF-1R）、胰岛素生长因子结合蛋白1（IGFBP1）、胰岛素生长因子结合蛋白4（IGFBP4）和胰岛素生长因子1（IGF-1）蛋白表达水平的改变。结果：照射剂量为1 Gy时，与对照组相比，IGF-1蛋白在照射后48 h时表达增加，IGFBP1、IGFBP4和IGF1R蛋白的表达在照射后12、48和72 h均表达减少。照射剂量为2 Gy时，IGF-1和IGFBP4蛋白在照射后48 h时表达增加，其余时间点表达减少。IGFBP1蛋白在照射后12 h时表达增加，48和72 h时表达减少。IGF1R蛋白在照射后3个时间点均呈现为表达减少。照射剂量4 Gy时，肝脏中IGF-1、IGFBP1和IGFBP4蛋白均表达减少，IGF1R蛋白在照射后12 h时表达增加，48、72 h时均表达减少（均为P < 0.05）。结论：电子线照射后小鼠肝脏中胰岛素生长因子-1家族蛋白多以下调表达为主，与前期基因表达的结果相一致，有可能作为反映放射性工作人员早期肝脏损伤的生物标志物之一。     

脑卒中后偏瘫肩痛患者生活质量的影响因素研究

目的 探讨脑卒中后偏瘫肩痛患者生活质量的直接和间接影响因素，以期为临床症状管理提供参考信息。 方法 采用一般资料调查表、脑卒中后偏瘫肩痛症状评估表、简易疲乏量表、医院焦虑抑郁量表、Fugl-Meyer上肢功能评分量表和脑卒中生活质量专用量表对205例脑卒中后偏瘫肩痛患者进行调查。结果 偏瘫肩痛患者的生活质量总分为（144.48±27.69），整体处于中等水平。生活质量的主要影响因素是肩痛困扰、肩痛频度、肩外旋、疲乏程度和抑郁，可以解释总变异的63.3%。路径分析结果显示，疲乏程度、肩痛频度和肩痛困扰可直接影响患者的生活质量（β_(疲乏程度-生活质量)=-0.252，P<0.001；β_(肩痛频度-生活质量)=-0.147，P<0.001；β_(肩痛困扰-生活质量)=-0.317，P<0.001）；疲乏程度和肩痛频度亦可通过肩痛困扰来间接影响患者的生活质量（ β_(疲乏程度-肩痛困扰-生活质量)=-0.066，P<0.001；β_(肩痛频度-肩痛困扰-生活质量)=-0.064，P<0.001）。结论 临床上应同时关注脑卒中患者疲乏和偏瘫肩痛症状，关注抑郁和疲乏对症状体验的协同作用，提高医护人员症状管理的效率，从而提高脑卒中偏瘫肩痛患者的生活质量。     

翁舌汤灌肠治疗大肠湿热型溃疡性结肠炎临床研究

目的:观察翁舌汤灌肠治疗大肠湿热型溃疡性结肠炎的疗效及其对患者血清炎症因子水平的影响。方法:将80例大肠湿热型溃疡性结肠炎患者按随机数字表法分为观察组和对照组各40例。观察组给予翁舌汤灌肠治疗,对照组给予美沙拉嗪栓肛门塞入治疗,两组均连续治疗8周。比较两组患者的临床疗效、肠黏膜征象积分和血清炎症因子水平。结果:治疗后,观察组和对照组总有效率分别为95.00%、70.00%,两组疗效比较,差异有统计学意义(P<0.05)。治疗后,观察组患者炎症增生、黏膜溃疡、黏膜糜烂、黏膜水肿、黏膜充血积分均下降,与治疗前比较,差异有统计学意义(P<0.05);且观察组各项积分均低于对照组,差异有统计学意义(P<0.05)。治疗后,两组血清白细胞介素-8(IL-8)、肿瘤坏死因子-β(TNF-β)水平均降低,血清白细胞介素-10(IL-10)水平均升高,与治疗前比较,差异均有统计学意义(P<0.05);且观察组血清IL-8、TNF-β水平明显低于对照组,血清IL-10水平明显高于对照组,差异有统计学意义(P<0.05)。结论:翁舌汤灌肠治疗大肠湿热型溃疡性结肠炎疗效较好,可改善患者肠黏膜征象积分,调节血清炎症因子水平。     

糖尿病肾病内热证与肾功能及炎症因子的相关性研究

目的:观察糖尿病肾病不同分期内热证与肾功能及炎症指标的相关性。方法:收集202例糖尿病肾病患者的临床资料,采用酶联免疫吸附法检测微炎症指标,包括超敏C反应蛋白(high-sensitivity C-reactive protein,HS-CRP)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),进行统计分析。结果:与非内热证组比较,中期、晚期内热证组24小时尿蛋白定量明显升高;晚期内热证组的血清肌酐明显升高,肾小球滤过率(glomerular filtration rate,GFR)明显降低;炎症因子CRP、IL-6、TNF-α随疾病进展均呈上升趋势;晚期内热证组与非内热证组比较,IL-6、TNF-α升高;中期HS-CRP与内热积分呈正相关,晚期TNF-α与内热积分呈正相关。结论:内热病机贯穿于糖尿病肾病的始终,与糖尿病肾病肾功能以及疾病进展密切相关;同时,内热积分与炎症因子HS-CRP、TNF-α的表达存在一定的相关性,这种相关性在疾病中期、晚期表现更加明显。