Pneumocystis jirovecii diagnosed by next-generation sequencing of bronchoscopic alveolar lavage fluid: A case report and review of literature

BACKGROUND The advent of molecular targeted agents and immune checkpoint inhibitors has greatly improved the treatment of advanced renal cell carcinoma(RCC), thus significantly improving patient survival. The incidence of rare drug-related adverse events has gained increased attention.CASE SUMMARY We report a patient with advanced RCC treated with multiple lines of molecular targeted agents and immune checkpoint inhibitors, who developed a pulmonary infection after treatment with everolimus in combination with lenvatinib. Determining the pathogenic organism was difficult, but it was eventually identified as Pneumocystis jirovecii by next-generation sequencing(NGS) of bronchoscopic alveolar lavage fluid(BALF) and successfully treated with trimethoprim-sulfamethoxazole.CONCLUSION Rare pulmonary infections caused by molecular targeted agents are not uncommon in clinical practice, but their diagnosis is difficult. Evaluating BALF with NGS is a good method for rapid diagnosis of such infections.

某高校新生焦虑现状及与人格特征的关系

目的了解某高校新生焦虑现状及影响因素,探讨焦虑与人格特征的关系。方法运用整群随机抽样的方法,选取某高校576例新生为研究对象,采用焦虑自评量表(SAS)和艾森克人格问卷(EPQ)进行心理健康状况调查。结果新生的焦虑检出率为10.24%,SAS的平均得分高于全国成年人常模(P<0.05)。焦虑组学生表现出敌对、内向、情绪不稳定的人格特征;相关分析显示学生的SAS标准分与EPQ-P、EPQ-N因素呈正相关,与EPQ-E、EPQ-L因素呈负相关(P<0.05);Logistic回归分析显示学生的专业、高考成绩、父亲的文化程度以及不稳定的情绪因素是焦虑的主要影响因素(P<0.05)。结论该高校新生焦虑水平较高,学生的专业、高考成绩、父亲文化程度及人格特征等因素影响焦虑的发生。

Insulin-like growth factor binding protein-7 induces activation and transdifferentiation of hepatic stellate cells in vitro

AIM:To investigate the role of insulin-like growth factor binding protein-7 (IGFBP-7) in the activation and transdifferentiation of hepatic stellate cells (HSC) in vitro.METHODS:Rat HSC-T6 cells were cultured in separate dishes and treated with various concentration of transforming growth factor (TGF)-β1,IGFBP-7 or antiIGFBP-7 antibody for 24 h.The supernatant or a cytoplasm suspension was obtained from cultured HSC,followed by transfer of cells to form cell-coated dishes.Immunocytochemistry and Western blotting were used to analyze the expression of IGFBP-7 induced by TGF-β1 and the level of fibronectin,collagen and α-smooth muscle actin (SMA).The pro-apoptotic effect of antiIGFBP-7 antibody was determined by flow cytometry.RESULTS:Immunocytochemistry and Western blotting revealed that the expression of IGFBP-7 in TGF-β1 treated HSC was significantly up-regulated compared to that in the control group.In addition,fibronectin,collagen and α-SMA also showed enhanced expression in accordance with the transdifferentiation process in a dose-dependent manner to some extent.Moreover,flow cytometry suggested that anti-IGFBP-7 antibody induced apoptosis of activated HSC,which is responsible for the development of liver fibrosis,and may represent a novel pathway and target for therapeutic intervention.CONCLUSION:IGFBP-7 showed increased expression in activated HSC and played an important role in the activation and transdifferentiation process of HSC.AntiIGFBP-7 antibody may ameliorate liver fibrogenesis.

Insulin-like growth factor binding protein related protein 1 knockdown attenuates hepatic ?brosis via the regulation of MMPs/TIMPs in mice

Background: Previous research suggested that insulin-like growth factor binding protein related protein 1(IGFBPrP1), as a novel mediator, contributes to hepatic fibrogenesis. Matrix metalloproteinases(MMP) and tissue inhibitors of metalloproteinases(TIMP) play an essential role in hepatic fibrogenesis by regulating homeostasis and remodeling of the extracellular matrix(ECM). However, the interaction between IGFBPrP1 and MMP/TIMP is not clear. The present study was to knockdown IGFBPrP1 to investigate the correlation between IGFBPrP1 and MMP/TIMP in hepatic fibrosis. Methods: Hepatic fibrosis was induced by thioacetamide(TAA) in mice. Knockdown of IGFBPrP1 expression by ultrasound-targeted microbubble destruction-mediated CMB-shRNA-IGFBPrP1 delivery, or inhibition of the Hedgehog(Hh) pathway by cyclopamine treatment, was performed in TAA-induced liver fibrosis mice. Hepatic fibrosis was determined by hematoxylin and eosin and Sirius red staining. Hepatic expression of IGFBPrP1, α-smooth muscle actin( α-SMA), transforming growth factor β 1(TGF β1), collagen I, MMPs/TIMPs, Sonic Hedgehog(Shh), and glioblastoma family transcription factors(Gli1) were investigated by immunohistochemical staining and Western blotting analysis. Results: We found that hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I were increased longitudinally in mice with TAA-induced hepatic fibrosis, concomitant with MMP2/TIMP2 and MMP9/TIMP1 imbalance and Hh pathway activation. Knockdown of IGFBPrP1 expression, or inhibition of the Hh pathway, reduced the hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I and re-established MMP2/TIMP2 and MMP9/TIMP1 balance. Conclusions: Our findings suggest that IGFBPrP1 knockdown attenuates liver fibrosis by re-establishing MMP2/TIMP2 and MMP9/TIMP1 balance, concomitant with the inhibition of hepatic stellate cell activation, down-regulation of TGF β1 expression, and degradation of the ECM. Furthermore, the Hh pathway mediates IGFBPrP1 knockdown-induced attenuation of hepatic fibrosis through the regulation of MMPs/TIMPs balance.

IGFBPrP1 induces liver fibrosis by inducing hepatic stellate cell activation and hepatocyte apoptosis via Smad2/3 signaling

AIM:To investigate the role and mechanism of insulinlike growth factor binding protein-related protein 1(IGFBPrP1)in the development of liver fibrosis.METHODS:An in vitro model using hepatic stellate cell(HSC)-T6 cells and an in vivo model of rat liver overexpressing IGFBPrP1 were established using an IGFBPrP1-expressing recombinant adenovirus.The expression of IGFBPrP1 was examined by immunofluorescence,and the expression of collagen?Ⅰ?and fibronectin was mea-sured by real-time reverse transcription-polymerase chain reaction and Western blot analysis.The expression of Smad2/3 and p-Smad2/3 was examined by Western blot and immunohistochemistry.A shSmad3-expressing recombinant adenovirus(AdshSmad3)was designed and used to knockdown the Smad3 gene in HSC-T6 cells and rat liver fibrosis transfected with IGFBPrP1.The expression of collagen?Ⅰ,fibronectin,andα-smooth muscle actin(α-SMA)was determined by Western blot analysis and immunohistochemistry.Hepatocyte apoptosis was assessed using TUNEL assay.RESULTS:IGFBPrP1 overexpression induced collagen deposition and up-regulated the expression ofα-SMA and p-Smad2/3,and AdshSmad3 inhibited IGFBPrP1-stimulated p-Smad2/3 activation and the expression ofα-SMA,collagen?Ⅰ?and fibronectin in HSC-T6 cells.Similarly,increased hepatocyte apoptosis(38.56%±3.42%vs 0.24%±0.03%,P<0.05),α-SMA positive stained cells(29.84%±1.36%vs 5.83%±1.47%,P<0.05),and increased numbers of Smad3(35.88%±2.15%vs10.24%±1.31%,P<0.05)and p-Smad2/3 positive cells(28.87%±2.73%vs 8.23%±0.98%,P<0.05)were detected in the livers of IGFBPrP1-overexpressing rats compared with the control group.Moreover,AdshSmad3 reduced IGFBPrP1-stimulated Smad3 expression and attenuatedα-SMA expression(29.84%±1.36%vs 8.23%±1.28%,P<0.05),hepatocyte apoptosis(38.56%±3.42%vs 6.75%±0.52%,P<0.05),and both collagen?Ⅰ?and fibronectin deposition in the livers of AdIGFBPrP1-treated rats.CONCLUSION:IGFBPrP1 induces liver fibrosis by mediating the activation of hepatic stellate cells and hepatocyte apoptosis in a Smad3-dependent mechanism.

Interaction between insulin-like growth factor binding protein-related protein 1 and transforming growth factor beta 1 in primary hepatic stellate cells

BACKGROUND:We previously showed that insulin-like growth factor binding protein-related protein 1(IGFBPrP1) is a novel mediator in liver fibrosis.Transforming growth factor beta 1(TGFβ1) is known as the strongest effector of liver fibrosis.Therefore,we aimed to investigate the detailed interaction between IGFBPrP1 and TGFβ1 in primary hepatic stellate cells(HSCs).METHODS:We overexpressed TGFβ1 or IGFBPrP1 and inhibited TGFβ1 expression in primary HSCs for 6,12,24,48,72,and 96 hours to investigate their interaction and observe the accompanying expressions of α-smooth muscle actin(α-SMA),collagen I,fibronectin,and phosphorylated-mothers against decapentaplegic homolog 2/3(p-Smad2/3).RESULTS:We found that the adenovirus vector encoding the TGFβ1 gene(Ad TGFβ1) induced IGFBPrP1 expression while that of α-SMA,collagen I,fibronectin,and TGFβ1 increased gradually.Concomitantly,Ad IGFBPrP1 upregulated TGFβ1,α-SMA,collagen I,fibronectin,and p-Smad2/3 in a time-dependent manner while IGFBPrP1 expression was decreased at 96 hours.Inhibition of TGFβ1 expression reduced the IGFBPrP1-stimulated expression of α-SMA,collagen I,fibronectin,and p-Smad2/3.CONCLUSIONS:These findings for the first time suggest the existence of a possible mutually regulation between IGFBPrP1 and TGFβ1,which likely accelerates liver fibrosis progression.Furthermore,IGFBPrP1 likely participates in liver fibrosis in a TGFβ1-depedent manner,and may act as an upstream regulatory factor of TGFβ1 in the Smad pathway.

MicroRNA标志物在早期诊断室间隔缺损中的价值

目的 评价血浆microRNA(miRNA)在室间隔缺损(VSD)早期诊断中的临床应用价值以及作为VSD分子诊断标志物的可行性,并建立临床早期诊断模型。方法 采用以济南市儿童医院、泰安市妇幼保健院为基础的病例-对照研究方法开展本次研究。研究分为VSD组85例和对照组80例,分3个阶段进行检测:首先选取VSD组及其匹配对照组各3例,应用miRNA全基因组表达谱芯片进行初筛;然后扩大样本量(20例VSD组vs 15例对照组),对选定的8个有差异的miRNA采用实时荧光定量聚合酶链反应(qRT-PCR)进行芯片结果的验证;最后进一步扩大样本(62例VSD组vs 62例对照组)进行qRT-PCR检测,筛选先天性心血管病早期诊断的生物标志物,建立多指标的Logistic回归模型,并应用MedCalc软件进行受试者工作特征(ROC)曲线分析评价诊断模型的价值。结果 芯片杂交初筛结果显示,与对照组比较,有36个表达有差异的miRNA(P<0.05),两阶段大样本qRT-PCR验证,检测到5种血浆miRNA(let-7e-5p、miR-155-5p、miR-222-3p、miR-433-3p和miR-487b-5p)在VSD中的表达差异有统计学意义(P<0.05),可以选择作为具有VSD早期诊断意义的生物标志物。5种血浆miRNA的ROC曲线下面积(AUC)分别在0.678~0.832,其中miR-222-3p的ROC曲线下面积最大为0.832。建立多指标Logistic回归分析模型,5种血浆miRNAs联合模型可提高VSD的诊断效率,ROC曲线下面积达到0.955,敏感性和特异性分别为83.87%和95.16%;3种血浆miRNA(let-7e-5p、miR-155-5p、miR-222-3p)联合诊断模型,ROC曲线下面积达到0.910,敏感性和特异性分别为82.30%和90.30%,接近5个miRNA的诊断效率。结论 5种血浆miRNA作为VSD早期诊断标志物,具有一定的可信性。将其整合后可建立起多指标的联合诊断模型。联合3种血浆miRNA构建诊断模型可提高VSD的诊断效率,临床应用价值更大。

TBX5基因靶序列单核苷酸多态性与先天性心脏病的关系

目的探讨TBX5基因3'非翻译区靶序列SNPs与先天性心脏病(CHD)遗传易感性的关系,为CHD的分子机制研究提供线索。方法利用HaploView软件筛选TBX5基因3'非翻译区功能性单核苷酸多态性(SNP),并应用高分辨率熔解曲线检测TBX5基因3'非翻译区SNP。采用病例对照研究分析SNP位点与CHD的关联。结果HaploView软件筛选出TBX5基因3'非翻译区有意义的标签SNP rs883079,高分辨率熔解曲线法成功检测该SNP位点(rs883079)的G/G、G/A及A/A 3种基因型。TBX5基因rs883079(G>A)位点等位基因或基因型分布频率比较,差异有统计学意义(P<0.05);与CHD易感性关联,A是CHD的危险等位基因,A/A为CHD的危险基因型(P<0.05)。结论TBX5基因rs883079位点SNP与CHD有关系,rs883079(G>A)携带突变纯合子基因型A/A个体患CHD的危险性升高。

Gastrointestinal stromal tumor with multisegmental spinal metastases as first presentation:A case report and review of the literature

BACKGROUND Gastrointestinal stromal tumor(GIST)usually originates in the stomach,followed by the small intestine,rectum,and other parts of the gastrointestinal tract.The most common sites of metastasis are the liver and peritoneum,whereas spinal metastases from GIST are extremely rare.CASE SUMMARY We found a case of GIST with the first presentation of multilevel spinal metastases involving the thoracic and lumbar vertebrae.A 61-year-old Chinese man presented to our clinic because of pain in his lower back and hip for 10 d without cause.Subsequently,computed tomography(CT)and magnetic resonance imaging(MRI)revealed abnormal signals in the vertebral appendages of T12 and L4 accompanied by spinal canal stenosis,which was considered as tumor metastasis.As there were no metastases to vital organs,posterior thoracic and lumbar spinal decompression+adnexal mass resection+pedicle internal fixation was adopted to achieve local cure and prevent nerve compression.The results of histopathological studies were consistent with the metastasis of GIST.No local recurrence or new metastases were found at the 6-mo follow-up at the surgical site.The patient has no neurological symptoms at present.It is worth mentioning that a rectal mass was found and surgically removed 1 mo after the patient was discharged from hospital,and the pathological diagnosis of the mass was GIST.CONCLUSION By reviewing 26 previously reported cases of spinal metastasis in GIST,it was found that spinal metastasis of GIST has become more common in recent years,so the possibility of early spinal metastasis should be recognized.CT and MRI are of great value in the diagnosis of spinal metastatic tumors,and pathological biopsy is the gold standard for the diagnosis of metastatic tumors.It is safe and feasible to treat isolated spinal metastasis in GIST by excising metastatic masses,decompressing the spinal canal,and stabilizing the spine.

Submonolayer biolasers for ultrasensitive biomarker detection

Biomarker detection is key to identifying health risks.However,designing sensitive and single-use biosensors for early diagnosis remains a major challenge.Here,we report submonolayer lasers on optical fibers as ultrasensitive and disposable biosensors.Telecom optical fibers serve as distributed optical microcavities with high Q-factor,great repeatability,and ultralow cost,which enables whispering-gallery laser emission to detect biomarkers.It is found that the sensing performance strongly depends on the number of gain molecules.The submonolayer lasers obtained a six-order-of-magnitude improvement in the lower limit of detection(LOD)when compared to saturated monolayer lasers.We further achieve an ultrasensitive immunoassay for a Parkinson's disease biomarker,alpha-synuclein(α-syn),with a lower LOD of 0.32 pM in serum,which is three orders of magnitude lower than theα-syn concentration in the serum of Parkinson's disease patients.Our demonstration of submonolayer biolaser offers great potentials in high-throughput clinical diagnosis with ultimate sensitivity.

Monosaccharide analysis and fingerprinting identification of polysaccharides from Poria cocos and Polyporus umbellatus by HPLC combined with chemometrics methods

Objective:Poria cocos and Polyporus umbellatus are similar medicinal fungi in traditional Chinese medicines.A method for fingerprint analysis of monosaccharide composition of polysaccharides by HPLC combined with chemometrics methods has been developed for characterization and discrimination of them in this research.Methods:The polysaccharides were extracted by decocting in water,and then completely hydrolyzed with hydrochloride.Monosaccharides in the hydrolyzates were derivatized with 1-phenyl-3-methyl-5-pyrazolone(PMP)for HPLC analysis.More than 20 batches of P.cocos and P.umbellatus from different regions were analyzed.Results:The fingerprints of P.cocos showed five common characteristic peaks,which were identified by comparing with the reference substances.The five peaks corresponded to the derivatives of mannose,ribose,glucose,galactose,and fucose.At the same time,the fingerprints of P.umbellatus showed eight common characteristic peaks,of which seven were identified as the derivatives of mannose,ribose,rhamnose,glucose,galactose,xylose,and fucose.Moreover,the similarity of their fingerprints was respectively calculated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM published by China Pharmacopoeia Committee(Version 2004 A).And the data were further processed by hierarchical cluster analysis(HCA)and principal component analysis(PCA).The similarity evaluation and HCA indicated that there were no significant difference in P.cocos or P.umbellatus samples from different geographical regions,but PCA was performed to characterize the difference in monosaccharide constituents between P.cocos and P.umbellatus,and linear discriminant analysis(LDA)showed the overall correct classification rate was 100%.Conclusion:The fingerprint analysis method of monosaccharide composition of water-soluble polysaccharides can distinguish P.cocos and P.umbellatus,and can be applied for the authentication or quality control for P.cocos and P.umbellatus.

Macrophage-NLRP3 Inflammasome Activation Exacerbates Cardiac Dysfunction after Ischemic Stroke in a Mouse Model of Diabetes

Ischemic stroke is one of the leading causes of death worldwide.In the post-stroke stage,cardiac dysfunction is common and is known as the brain-heart interaction.Diabetes mellitus worsens the post-stroke outcome.Stroke-induced systemic inflammation is the major causative factor for the sequential complications,but the mechanism underlying the brain-heart interaction in diabetes has not been clarified.The NLRP3(NLR pyrin domain-containing 3) inflammasome,an important component of the inflammation after stroke,is mainly activated in M1-polarized macrophages.In this study,we found that the cardiac dysfunction induced by ischemic stroke is more severe in a mouse model of type 2 diabetes.Meanwhile,M1-polarized macrophage infiltration and NLRP3 inflammasome activation increased in the cardiac ventricle after diabetic stroke.Importantly,the NLRP3 inflammasome inhibitor CY-09 restored cardiac function,indicating that the Ml-polarized macrophage-NLRP3 inflammasome activation is a pathway underlying the brain-heart interaction after diabetic stroke.

Parasitism of Pieris rapae （Lepidoptera： Pieridae） by the endoparasitic wasp Pteromalus puparum （Hymenoptera： Pteromalidae）： Effects of parasitism on differential hemocyte counts, micro- and ultra-structures of host hemocytes

由 endoparasitic 黄蜂 Pteromalus puparum 的寄生状态(Hymenoptera：Pteromalidae ) 由使用仅仅它的联系毒液，能压制 Pieris rapae 的 immunal 回答(鳞翅目：Pieridae ) 。直到现在，然而，机制的当前的知识被限制了。到寄生状态的主人血球的反应用光和传播电子显微镜学(TEM ) 的联合被调查。五种血球类型， prohemocytes (PR ) ， granulocytes (GR ) ， plasmatocytes (请) ， oenocytoids (OE ) 和 coagulocytes (公司) ，从 unparasitized 被观察并且描绘并且寄生于 Pieris rapae 蛹。轻显微镜学显示出那 GR 并且请周围成为了更多并且在寄生状态以后反常地传播了，而血球的另外的类型的形状仍然保持未受影响。另外，当 OE 变得更小时， PR 并且请的尺寸变得更大。PR 的比例显著地在寄生状态和的以后增加了请在 43.9% 减少了，但是没有 GR 和 OE 的重要增加。TEM 证明除了公司的血球的所有类型在寄生状态以后被损坏到各种各样的度，特别导致电子不透明的细胞质和原子核，不平的 endoplasmic 蜂窝胃的更少房间细胞器，线粒体和泡。我们的结果由 P 显示那个寄生状态。puparum 影响微分血球计数和主人血球的结构，特别地为 GR 并且请，它可以是压制主人的 parasitoid 的主要原因细胞的有免疫力的回答。

Dexmedetomidine as an adjunct to peripheral nerve blocks in pediatric patients

Background Combining local anesthetics with dexmedetomidine(DEX)for nerve blocks has been tested and confirmed as an alternative technique to strengthen analgesia in adults,but the evidence supported this proposal in pediatrics was not convincing enough.Thus,we intend to conduct a meta-analysis to make an appraisal of the effects of DEX as a local anesthetic adjuvant on peripheral nerve blocks in children.Methods The analysis has been registered on https://www.crd.york.ac.uk/prospero/with the registration number CRD42021254029 on June 10th,2021.PubMed,Ovid MEDLINE,Embase and Cochrane central register were searched for the randomized controlled trials(RCTs)that are relevant to child/pain/DEX or block up to April 30,2021.Results Eight RCTs were included for data analysis.The pooled results showed a significantly reduced number of patients who required postoperative analgesics within DEX groups[odds ratio=0.1,95%confidence interval(CI)0.03-0.32,P<0.001].And DEX significantly prolonged the duration of analgesia of local anesthetics in the meantime[standardized mean difference(SMD)=4.93,95%CI 3.02-6.83,P<0.001].Nevertheless,adding DEX as an adjunct to local anesthetics would significantly decrease heart rate(SMD=-0.95,95%CI-1.45 to-0.45,P<0.001)and increase sedation score(SMD=1.6,95%CI 0.53-2.66,P=0.003)simultaneously.Conclusions Adding DEX as an adjunct to peripheral nerve blocks can prolong analgesia until postoperative 8 hours in pediatric patients.However,the benefits should be weighed against the increasing risk of bradycardia and sedation in critical children.

ZnGeP2:A near-infrared-activated photocatalyst for hydrogen production

In this work,we prepared ZnGeP2(ZGP)photocatalyst using single flat temperature zone(SFT)method in a vacuum quartz ampoule.The XRD,SEM,EDS,DRS and XPS were used to characterize the crystal structure,morphology,elemental content,optical absorption and band gap structure of ZGP.The results of photocatalytic hydrogen evolution and apparent quantum efficiency show that ZGP is a promising photocatalyst for hydrogen production both under visible and near-infrared light irradiation.In addition,it is also found that adding the common stabilizer H3PO2 and ultrasonic treatment can efficiently improve the photocatalytic activity and stability of ZGP.