Analysis of the tumor microenvironment and mutation burden identifies prognostic features in thymic epithelial tumors

Thymic epithelial tumors (TETs) are one of the rarest adult malignancies in the anterior mediastinum. Thymic carcinomas (TCs) are less prevalent among TETs, but they are more clinically aggressive. Immunotherapy has emerged as a promising therapeutic approach for refractory TETs, even though chemotherapy remains the conventional treatment for the advanced disease. However, limited attention has been paid to the features of the tumor microenvironment (TME) which might provide clinically relevant information and guide treatment regimen design. Especially, to date, there have been only a few studies focusing on the differences between the TME and genomic features preserved by TETs and TCs. We analyzed the TME and genomic characteristics of TETs using RNA sequencing and whole-exome sequencing, finding that distinct characteristics of TME in different pathogenic subtypes of TETs. According to those findings, we found that thymic carcinomas had significantly lower expression of HMGB1, a pro-inflammatory cytokine-related gene, than thymomas, and low HMGB1 expression was linked to a poor prognosis. Additionally, higher mutation burdens were significantly associated with the later stage and more advanced pathological types. Thymoma patients with lower mutation burdens tended to relapse within 3 years. In summary, different characteristics of TME and genomic features between thymoma and thymic carcinoma were associated with clinical outcomes of TETs and presented promisingly predictive value for efficacy and toxicity of immunotherapy.     

初中生心理问题早期干预的前瞻性研究

【目的】观察初中生心理卫生问题早期干预的效果。【方法】应用自编儿童心理卫生调查问卷、症状自评量表(Symptom Checklist 90,SCL-90)、父母养育方式评价量表(Egma Minnen av Bardndosnauppforstran,EMBU),以整群抽样的方法,抽取社区所属的中学1 585人为研究对象,随机选取其中一所中学841人为干预组,另一所中学744人为对照组,对干预组进行为期2年的综合干预,前瞻性观察心理卫生问题的干预效果。【结果】干预前干预组检出139人,阳性率16.53%;对照组117人,阳性率15.74%,两组比较差异无显著性(χ2=0.19,P>0.05)。干预前两组父母养育方式量表各因子得分比较,差异无显著性(P>0.05)。经过对干预组进行2年综合干预后,对两组再重新进行问卷调查,评估干预效果。干预组心理卫生问题检出阳性率明显下降至9.23%;对照组检出阳性率为16.50%,两组相比差异有非常显著性(χ2=18.41,P<0.01)。干预组在干预后父母养育方式量表各因子得分与对照组比较,差异有显著性(P<0.05)。【结论】对初中生的心理卫生问题进行早期干预,有助于减少心理问题的发生。     

铂类耐药性卵巢上皮癌患者卵巢上皮中VEGF-C表达的研究

目的:探讨铂类耐药性卵巢上皮癌患者卵巢上皮癌组织中VEGF-C蛋白的表达。方法:应用免疫组织化学染色法检测43例卵巢上皮癌组织中VEGF-C蛋白的表达,同时检测其在中-高分化的卵巢癌组织中的单层、复层及癌巢的表达情况。结果:VEGF-C蛋白在中-高分化的卵巢癌组织中的单层、复层及癌巢的表达无显著性差异(P>0.05)。在耐药复发组和为复发组的癌组织中VEGF-C蛋白表达有显著性差异(P<0.05)。耐药复发组中单层上皮、癌巢中存在VEGF-C过表达。结论:VEGF-C蛋白在卵巢癌组织中的表达与组织学分级不具有明显相关性。VEGF-C蛋白在耐药组卵巢组织存在过表达,VEGF-C蛋白的检测对提示可能出现铂类耐药有重要价值。     

中俄双边口岸地区鼠形动物监测名录

目的于2006至2009年间对中俄黑龙江、内蒙古双边11个口岸地区的鼠形动物进行本底调查。方法 5 m夹夜法、弓形夹法及电弧捕鼠器捕捉法。结果编著中俄双边11个口岸鼠形动物名录。结论该名录为中俄口岸公共卫生及鼠传疾病的防治提供本底资料。     

广义“水气病”范畴之我见

张仲景的《伤寒杂病论》中首次提出了"水气病"的病名,文章通从其中的水气病与其相关疾病入手,在检索古今文献的基础上,结合历代医家对水气病的认识,对"水气病"病名、源流、病因病机、相关疾病等几方面进行概括梳理,并探讨广义水气病之范畴,以期拓宽水气病临床治疗的思维空间,为水气病的理论与临床研究奠定基础或提供借鉴。     

肺总顺应性的容量反应性研究

目的 探讨肺总顺应性(C)的容量反应性以及高血压对其反应能力的影响。方法 选择广东省广州市红十字会医院行择期平卧位手术的患者65例为研究对象,记录患者机械通气后不同时点[5 min(t₁)、10 min(t₂)、15 min(t₃)、20 min(t₄)、25 min(t₅)、30 min(t₆)]的C、气道平台压(p_plat)、气道峰压(p_peak)、心率(HR)、平均动脉压(MAP)、心排血量(CO)、每搏量变异度(SVV)、每搏输出量(SV)。计算不同时间段[T₁(5～10 min)、T₂(>10～15 min)、T₃(>15～20 min)、T₄(>20～25 min)、T₅(>25～30 min)]患者C、SV、SVV的变化量(△C、△SV、△SVV)。采用日本科林动脉硬...     

反油酸对人脐静脉平滑肌细胞活力的影响

<正>平滑肌细胞是血管壁中膜的主要细胞成分,是动脉粥样硬化(atherosclerosis,As)和血管成形术后再狭窄(restenosis after angioplasty,RAA)等病理过程形成和发展的重要部位,对血管壁的完整性和紧张性的调节也起重要作用,其异常增殖及形态学转变将导致动脉粥样硬化过程     

SRS-FISH: A high-throughput platform linking microbiome metabolism to identity at the single-cell level

One of the biggest challenges in microbiome research in environmental and medical samples is to better understand functional properties of microbial community members at a single-cell level. Single-cell isotope probing has become a key tool for this purpose, but the current detection methods for determination of isotope incorporation into single cells do not allow high-throughput analyses. Here, we report on the development of an imaging-based approach termed stimulated Raman scattering–two-photon fluorescence in situ hybridization (SRS-FISH) for high-throughput metabolism and identity analyses of microbial communities with single-cell resolution. SRS-FISH offers an imaging speed of 10 to 100 ms per cell, which is two to three orders of magnitude faster than achievable by state-of-the-art methods. Using this technique, we delineated metabolic responses of 30,000 individual cells to various mucosal sugars in the human gut microbiome via incorporation of deuterium from heavy water as an activity marker. Application of SRS-FISH to investigate the utilization of host-derived nutrients by two major human gut microbiome taxa revealed that response to mucosal sugars tends to be dominated by Bacteroidales, with an unexpected finding that Clostridia can outperform Bacteroidales at foraging fucose. With high sensitivity and speed, SRS-FISH will enable researchers to probe the fine-scale temporal, spatial, and individual activity patterns of microbial cells in complex communities with unprecedented detail.

With the rapid advances in both genotyping and phenotyping of single cells, bridging genotype and phenotype at the single-cell level is becoming a new frontier of science (1). Methods have been developed to shed light on the genotype–metabolism relationship of individual cells in a complex environment (2, 3), which is especially relevant for an in-depth understanding of complex microbial communities in the environment and host-associated microbiomes. For functional analyses of microbial communities, single-cell isotope probing is often performed in combination with nanoscale secondary ion mass spectrometry (NanoSIMS) (4–7), microautoradiography (MAR) (8, 9), or spontaneous Raman microspectroscopy (10–12) to visualize and quantify the incorporation of isotopes from labeled substrates. These methods can be combined with fluorescence in situ hybridization (FISH) using ribosomal ribonucleic acid (rRNA)-targeted probes (13), enabling a direct link between metabolism and identity of the organisms. In addition, Raman-activated cell sorting has been recently developed using either optical tweezers or cell ejection for downstream sequencing of the sorted cells (14–16). While these approaches have expanded the possibilities for functional analyses of microbiome members (17), all of the aforementioned methods suffer from extremely limited throughput. Consequently, only relatively few samples and cells per sample are typically analyzed in single-cell stable isotope probing studies, hampering a comprehensive understanding of the function of microbes in their natural environment.To overcome the limited throughput of Raman spectroscopy, coherent Raman scattering microscopy based on coherent anti-Stokes Raman scattering (CARS) or stimulated Raman scattering (SRS) has been developed (18, 19). Compared with CARS, the SRS signal is free of the electronic resonance response (20) and is linear to molecular concentration, thus permitting quantitative mapping of biomolecules (21, 22). Both CARS and SRS microscopy have successfully been applied for studying single-cell metabolism in eukaryotes (23–26). In a label-free manner, SRS imaging has led to the discovery of an aberrant cholesteryl ester storage in aggressive cancers (27, 28), lipid-rich protrusions in cancer cells under starvation (29), and fatty acid unsaturation in ovarian cancer stem cells (30) and more recently, in melanoma (31, 32). CARS and SRS have also been harnessed to explore lipid metabolism in live Caenorhabditis elegans (33–36). Combined with stable isotope probing, SRS microscopy has allowed the tracing of glucose metabolism in eukaryotic cells (37, 38) and the visualization of metabolic dynamics in living animals (25). Recently, SRS was successfully applied to infer antibiotic resistance patterns of bacterial pure cultures and heavy water (D2O) metabolism (39). Yet, SRS microscopy has not been adapted for studying functional properties of members of microbiomes as SRS itself lacks the capability of identifying cells in a complex community.Here, we present an integrative platform that exploits the advantages of SRS for single-cell stable isotope probing together with two-photon FISH for the identification of cells in a high-throughput manner. To deal with the challenges in detecting low concentrations of metabolites inside small cells with diameters around 1 µm, we have developed a protocol that maximizes the isotope label content in cells and exploits the intense SRS signal from the Raman band used for isotope detection.Conventionally, FISH is performed separately by one-photon excited fluorescence microscopy (40). To enhance efficiency, we developed a system that implements highly sensitive SRS metabolic imaging with two-photon FISH using the same laser source. These efforts collectively led to a high-throughput platform that enables correlative imaging of cell identity and metabolism at a speed of 10 to 100 ms per cell. In comparison, it takes about 20 s to record a Raman spectrum from a single cell in a conventional spontaneous Raman FISH experiment (41, 42).Our technology enabled high-throughput analysis of single-cell metabolism in the human gut microbiome. In the human body, microbes have been shown to modulate the host’s health (43, 44). Analytical techniques looking into their activities and specific physiologies (i.e., phenotype) as a result of both genotype and the environment provide key information on how microbes function, interact with, and shape their host. As a proof of principle, we used stimulated Raman scattering–two-photon fluorescence in situ hybridization (SRS-FISH) to track the incorporation of deuterium (D) from D2O into a mixture of two distinct gut microbiota taxa. Incorporation of D from D2O into newly synthesized cellular components of active cells, such as lipids and proteins, occurs analogously to incorporation of hydrogen from water during the reductive steps of biosynthesis of various cellular molecules (10, 45, 46). Importantly, D incorporation from D2O has been shown to be reliable to track metabolic activity of individual cells within complex microbial communities in response to the addition of external substrates (10, 17, 47). When microbial communities are incubated in the presence of D2O under nutrient-limiting conditions, individual cells display only minimal activity and only minor D incorporation (11, 17, 47). In contrary, when cells are stimulated by the addition of an external nutrient, cells that can metabolize this compound become active and incorporate D into macromolecules, which lead to the presence of C-D bonds into the cell’s biomass. Consequently, D incorporation from D2O can be combined with techniques able to detect C-D signals, such as Raman-based approaches, and to track metabolic activity at the single-cell level in response to a variety of compounds. Here, we show that SRS-FISH enables fast and sensitive determination of the D content of individual cells while simultaneously unveiling their phylogenetic identity. We applied this technique to complex microbial communities by tracking in situ the metabolic responses of two major phylogenetic groups of microbes in the human gut (Bacteroidales and Clostridia spp.) and of a particular species within each group to supplemented host-derived nutrients. Our study revealed that 1) Clostridia spp. can actually outperform Bacteroidales spp. at foraging on the mucosal sugar fucose and shows 2) a significant interindividual variability of responses of these major microbiome taxa toward mucosal sugars. Together, our results demonstrate the capability of SRS-FISH to unveil the metabolism of particular microbes in complex communities at a throughput that is two to three orders of magnitude higher than other metabolism identity bridging tools, therefore providing a valuable multimodal platform to the field of single-cell analysis.     

Extremely high serum CA19-9 level along with elevated D-dimer in assisting detection of ruptured ovarian endometriosis

BackgroundTo clarify clinical importance of serum CA19-9, CA-125, and plasma D-dimer (D-D) levels in detecting spontaneously ruptured ovarian endometriosis (OE).Materials and MethodsWe retrospectively examined 173 patients with endometriosis out of 735 cases of OE between 2013 and 2019. Among these, 21 cases were diagnosed as “spontaneously ruptured” after surgery, while the remaining cases were unruptured. Venous blood was collected pre-operatively to detect CA19-9, CA-125, and D-D levels. A receiver operating characteristic curve analysis was applied to test clinical value of each marker.ResultsAmong the 21 patients with ruptured OE, 16 had a history of pelvic cysts, 19 claimed sudden onsets of lower abdominal pain, and fluid accumulation were detected in cul-de-sac in only six participants by ultrasound. For serological investigation, both CA19-9 and D-D were significantly elevated in the ruptured OE group (343.09 ± 367.67 U/ml vs. 36.84 ± 40.01 U/ml, 3.39 ± 4.90 mg/L vs. 0.43 ± 0.29 mg/L, both p < .0001). The area under curve (AUC) value for the combination of CA19-9 and D-D was 0.975 (95% CI, 0.939 − 0.993), with specificity of 96.69%, and sensitivity of 85.71%. The combination of CA-125, CA19-9 and D-D showed the highest AUC value that up to 0.976 (95% CI, 0.940–0.993), with sensitivity of 95.24%, and specificity of 87.50%.ConclusionThe combination of CA19-9 and D-D can be chosen as an effective and economical indicators to identify patients with spontaneously ruptured OE in pre-operation assessment. However, from the perspective of differential diagnosis, the combination of CA-125, CA19-9 and D-D is the best choice.

Key messages

• Taking into account the economic effect, the combination of CA19-9 and D-D can be chosen as an effective indicators to identify patients with spontaneously ruptured OE in pre-operation assessment.
• From the perspective of differential diagnosis, the combination of CA-125, CA19-9 and D-D is the best choice to identify patients with spontaneously ruptured OE.