Serum Exosomal microRNA-27-3p Aggravates Cerebral Injury and Inflammation in Patients with Acute Cerebral Infarction by Targeting PPARγ

Inflammation - Acute cerebral infarction (ACI) possesses high mortality. Exosomes present in serum have potential application value in ACI diagnosis. This study investigated the mechanism of serum...     

Rho相关激酶抑制剂Y27632对小鼠腔前卵泡体外发育的作用

目的 探讨Rho相关激酶（ROCK）抑制剂Y27632在小鼠腔前卵泡体外发育中的作用。 方法 取出生12.5 d的小鼠卵巢,机械法收集单枚腔前卵泡,采用超低吸附96孔板模拟3D环境对其进行培养。设置对照组和含有Y27632的实验组,通过形态学观察、卵泡直径的变化、成熟卵泡的数量、成熟卵母细胞纺锤体的变化等来评估卵泡的整体发育情况。采用Real-time PCR技术检测颗粒细胞中卵泡刺激素受体（FSHR）、卵母细胞中骨形态发生蛋白15（BMP15）、生长分化因子9（GDF9）等以及凋亡相关基因（Bad、Bax和Caspase-3）的表达情况,同时,用细胞存活染料检测卵泡发育过程中颗粒细胞的凋亡情况。 结果 ROCK抑制剂Y27632对卵泡直径增长和基本发育指标（存活数、成腔率和成熟率）无明显影响（P>0.05）,但对照组卵母细胞成熟后纺锤体组装出现异常。培养8 d后,与对照组相比,实验组颗粒细胞特异性基因FSHR上调;卵母细胞特异性基因BMP15和GDF9上调,而叉头框O3（FoxO3)下调;凋亡基因Bax、Bad和Caspase3表达下调,实验组卵泡内的颗粒细胞凋亡明显少于对照组。 结论 小鼠卵泡体外发育过程中,ROCK抑制剂Y27632能够抑制颗粒细胞的凋亡,防止卵母细胞纺锤体组装异常,进而提高卵泡体外发育质量。     

Highly sensitive and specific detection of hepatitis B virus DNA and drug resistance mutations utilizing the PCR-based CRISPR-Cas13a system

ObjectivesUndetectable or low-level hepatitis B virus (HBV) DNA and drug resistance mutations in patients may increase the risk of HBV transmission or cause active viral replication and other clinical problems. Here, we established a highly sensitive and practical method for HBV and drug resistance detection using a polymerase chain reaction (PCR) -based CRISPR-Cas13a detection system (referred to as PCR-CRISPR) and evaluated its detection capability using clinical samples.MethodsSpecific CRISPR RNAs (crRNAs) are designed for HBV DNA detection and YMDD (tyrosine-methionine-aspartate-aspartate) variant identification. The HBV DNA was detected in 312 serum samples for HBV diagnosis using quantification PCR (qPCR) and PCR-CRISPR. Additionally, 424 serum samples for YMDD testing were detected by qPCR, direct sequencing, and our assay.ResultsUsing PCR-CRISPR, one copy per test of HBV DNA was detected with HBV-1 crRNA in 15 min after PCR amplification. Consistent results with qPCR were observed for 302 samples, while the remaining 10 samples with low-level HBV DNA were detectable by PCR-CRISPR and droplet digital PCR but not by qPCR. PCR-CRISPR diagnosed all 412 drug-resistant samples detected by the YMDD detection qPCR kit and direct sequencing, as well as the other 12 drug-resistant samples with low-level HBV DNA undetectable by qPCR and direct sequencing.ConclusionsWe developed a novel PCR-CRISPR method for highly sensitive and specific detection of HBV DNA and drug resistance mutations. One copy per test for HBV DNA and YMDD drug resistance mutations could be detected. This method has wide application prospects for the early detection of HBV infection, drug resistance monitoring and treatment guidance.     

CD8+ T cell survival in lethal fungal sepsis was ameliorated by T-cell-specific mTOR deletion

Lethal fungal sepsis causes high morbidity and mortality in intensive care patients. Fungal infections have an immunological basis, and it has been shown in recent studies that decreased CD8⁺ T-cell count in fungal infections is related to prognosis, while the underlying mechanism is still unclear. Here, a lethal fungal sepsis model induced by candidemia was created and we found a decreased CD8⁺ T-cell count and exaggerated apoptosis. Simultaneously, expression of light chain (LC)3B in CD8⁺ T cells increased, along with increased autophagosomes and accumulation of p62 in infected mice. We regulated the activity of the mammalian target of rapamycin (mTOR) pathway using T-cell-specific mTOR/ TSC1 deletion mice. We observed increased number of autophagosomes and expression of LC3B in CD8⁺T cells after T-cell-specific mTOR knockout, while accumulation of p62 was not ameliorated, and there was no increase in the number of autolysosomes. Apoptosis rate and expression of BIM, a pro-apoptotic gene, decreased in CD8⁺ T cells in mTOR-deletion mice but increased in TSC1-deletion mice. Our results showed increased CD8⁺ T-cell death in spleen of lethal fungal sepsis mice, and decreased expression of mTOR ameliorated CD8⁺ T-cell survival. mTOR may be a possible target to reverse CD8⁺ T-cell immune dysfunction in lethal fungal sepsis.     

膝内翻患者全膝关节置换术后保留轻度内翻对临床效果的影响

目的 评价膝内翻患者全膝关节置换术后保留轻度内翻对临床效果的影响。方法 对2016年3月至2019年11月于武汉大学人民医院因膝内翻膝关节骨性关节炎行TKA治疗的93例（93膝）患者的临床资料进行回顾性分析。其中,男17例,女76例;年龄71 ～ 87岁,平均77岁。按患者术后下肢力线角度将患者分为中立组、轻度内翻组、严重内翻组。用ROM、HSS、AKS评分量表评价患者膝关节功能。结果 患者随访时间15 ～ 47个月,平均（24.4±8.9）个月,ROM术前平均（49.9±6.7）°,术后末次随访中立组、轻度内翻组、严重内翻组分别为平均（111.4±5.3）°、（112.2±5.0）°、（103.1±2.7）°,HSS评分术前平均（52.4±3.0）分,术后末次随访中立组、轻度内翻组、严重内翻组分别为平均（84.2±3.0）分、（87.1±2.6）分、（81.6±1.9）分,AKS功能评分术前平均（49.8±2.8）分,术后末次随访中立组、轻度内翻组、严重内翻组分别为平均（73.2±5.3）分、（80.1±3.0）分、（72.6±1.5）分,AKS活动评分术前平均（60.5±3.7）分,术后末次随访中立组、轻度内翻组、严重内翻组分别为平均（86.0±2.6）分、（89.9±2.0）分、（80.6±2.9）分,三组术后HSS、AKS评分均较术前有所提高,差异有统计学意义。结论 从短期随访来看,膝内翻患者全膝关节置换术后保持3° ～ 6°的轻度内翻会带来更好的临床疗效,但目前还不能判断内翻对齐和翻修率之间是否存在关联。     

Allogeneic Vγ9Vδ2 T-cell immunotherapy exhibits promising clinical safety and prolongs the survival of patients with late-stage lung or liver cancer

Vγ9Vδ2 T cells are promising candidates for cellular tumor immunotherapy. Due to their HLA-independent mode of action, allogeneic Vγ9Vδ2 T cells can be considered for clinical application. To apply allogeneic Vγ9Vδ2 T cells in adoptive immunotherapy, the methodology used to obtain adequate cell numbers with optimal effector function in vitro needs to be optimized, and clinical safety and efficacy also need to be proven. Therefore, we developed a novel formula to improve the expansion of peripheral γδ T cells from healthy donors. Then, we used a humanized mouse model to validate the therapeutic efficacy of expanded γδ T cells in vivo; furthermore, the expanded γδ T cells were adoptively transferred into late-stage liver and lung cancer patients. We found that the expanded cells possessed significantly improved immune effector functions, including proliferation, differentiation, and cancer cell killing, both in vitro and in the humanized mouse model. Furthermore, a phase I clinical trial in 132 late-stage cancer patients with a total of 414 cell infusions unequivocally validated the clinical safety of allogeneic Vγ9Vδ2 T cells. Among these 132 patients, 8 liver cancer patients and 10 lung cancer patients who received ≥5 cell infusions showed greatly prolonged survival, which preliminarily verified the efficacy of allogeneic Vγ9Vδ2 T-cell therapy. Our clinical studies underscore the safety and efficacy of allogeneic Vγ9Vδ2 T-cell immunotherapy, which will inspire further clinical investigations and eventually benefit cancer patients.     

Tree-based exploration of the optimization objectives for automatic cervical cancer IMRT treatment planning

Objective:To develop and evaluate a practical automatic treatment planning method for intensity-modulated radiation therapy (IMRT) in cervical cancer cases.Methods:A novel algorithm named as Optimization Objectives Tree Search Algorithm (OOTSA) was proposed to emulate the planning optimization process and achieve a progressively improving IMRT plan, based on the Eclipse Scripting Application Programming Interface (ESAPI). 30 previously treated cervical cancer cases were selected from the clinical database and comparison was made between the OOTSA-generated plans and clinical treated plans and RapidPlan-based (RP) plans.Results:In clinical evaluation, compared with plan scores of the clinical plans and the RP plans, 22 and 26 of the OOTSA plans were considered as clinically improved in terms of plan quality, respectively. The average conformity index (CI) for the PTV in the OOTSA plans was 0.86 ± 0.01 (mean ± 1 standard deviation), better than those in the RP plans (0.83 ± 0.02) and the clinical plans (0.71 ± 0.11). Compared with the clinical plans, the mean doses of femoral head, rectum, spinal cord and right kidney in the OOTSA plans were reduced by 2.34 ± 2.87 Gy, 1.67 ± 2.10 Gy, 4.12 ± 6.44 Gy and 1.15 ± 2.67 Gy. Compared with the RP plans, the mean doses of femoral head, spinal cord, right kidney and small intestine in the OOTSA plans were reduced by 3.31 ± 1.55 Gy, 4.25 ± 3.69 Gy, 1.54 ± 2.23 Gy and 3.33 ± 1.91 Gy, respectively. In the OOTSA plans, the mean dose of bladder was slightly increased, with 2.33 ± 2.55 Gy (versus clinical plans) and 1.37 ± 1.74 Gy (vs RP plans). The average elapsed time of OOTSA and clinical planning were 59.2 ± 3.47 min and 76.53 ± 5.19 min.Conclusion:The plans created by OOTSA have been shown marginally better than the manual plans, especially in preserving OARs. In addition, the time of automatic treatment planning has shown a reduction compared to a manual planning process, and the variation of plan quality was greatly reduced. Although improvement on the algorithm is warranted, this proof-of-concept study has demonstrated that the proposed approach can be a practical solution for automatic planning.Advances in knowledge:The proposed method is novel in the emulation strategy of the physicists’ iterative operation during the planning process. Based on the existing optimizers, this method can be a simple yet effective solution for automated IMRT treatment planning.     

DIP-microhaplotypes: new markers for detection of unbalanced DNA mixtures

International Journal of Legal Medicine - The identification of a suspect in a degraded and unbalanced DNA mixture has been a challenge for the standard short tandem repeat polymorphisms (STR)...     

Deletions and duplications of 42 Y chromosomal short tandem repeats in Chinese Han population

International Journal of Legal Medicine - Genotypes of 42 Y chromosome STR (Y-STR) loci were analyzed for a sample of 1420 unrelated males and 1160 father-son pairs from a Chinese Han population....     

不同性别老年脑出血患者血脂水平与预后的关系研究

目的 探讨血脂水平与不同性别老年脑出血患者90 d临床预后的关系。 方法 本研究数据来源于多中心、前瞻性、观察性队列登记研究——北京地区以病因为基础的 脑出血医疗质量评价与微创手术治疗技术研究（登记号：2011-2004-03）,从数据库中筛选2014年12 月-2016年9月连续纳入的经头颅CT确诊为急性期脑出血患者的临床资料进行回顾性分析。收集患者 的性别、年龄、NIHSS评分、GCS评分、血肿体积、出血部位等临床资料,以及白细胞和血小板计数、血 脂水平等实验室检查资料。以90 d mRS>2分定义为预后不良,分别比较男性和女性不同预后患者血 脂水平的差异,并应用logistic回归分析血脂水平对不同性别患者发病90 d预后的影响。 结果 本研究最终纳入212例脑出血患者,平均年龄73.4±6.5岁,男性126例（59.4%）,女性86 例（40.6%）。单因素分析结果显示,在男性患者中,预后不良组年龄（P =0.038）、入院时NIHSS评 分（P <0.001）、空腹血糖（P =0.014）、HDL-C水平（P =0.010）、血肿体积（P =0.003）及出血破入脑 室患者比例（P =0.015）高于预后良好组,入院时GCS评分（P <0.001）低于预后良好组;在女性患者 中,预后不良组年龄（P =0.031）、入院时NIHSS评分（P <0.001）及血肿体积（P =0.023）高于预后良好 组,入院时GCS评分（P <0.001）、TG（P =0.016）及非高密度脂蛋白胆固醇（non-high density lipoprotein cholesterol,non-HDL-C）水平（P =0.020）低于预后良好组。logistic回归分析结果显示,对于男性患者, 高龄（OR 1.119,95%CI 1.027～1.219,P =0.010）、入院时高NIHSS评分（OR 1.373,95%CI 1.188～1.586, P <0.001）和出血破入脑室（OR 3.471,95%CI 1.112～10.832,P =0.032）是90 d预后的独立危险因素; 对于女性患者,入院时高NIHSS评分（OR 1.254,95%CI 1.078～1.459,P =0.003）是90 d预后的独立危 险因素,高水平的non-HDL-C是90 d预后的保护性因素（OR 0.978,95%CI 0.961～0.996,P =0.014）。 结论 血脂水平对老年脑出血患者的临床预后预测价值存在性别差异。高水平的non-HDL-C是老年 女性脑出血患者90 d预后的保护性因素。