Comparison of immune responses and intestinal flora in epicutaneously sensitized BALB/c or C57BL/6 mouse models of food allergy

Cutaneous exposure to food allergens through a disrupted skin barrier is recognized as an important cause of food allergy,and the cutaneous sensitized mouse model has been established to investigate relevant allergic disorders.However,the role of different genetic backgrounds of mice on immune responses to food allergens upon epicutaneous sensitization is largely unknown.In this study,two strains of mice,i.e.,the BALB/c and C57BL/6 mice,were epicutaneously sensitized with ovalbumin on atopic dermatitis(AD)-like skin lesions,followed by intragastric challenge to induce IgE-mediated food allergy.Allergic outcomes were measured as clinical signs,specific antibodies and cytokines,and immune cell subpopulations,as well as changes in intestinal barrier function and gut microbiota.Results showed that both strains of mice exhibited typical food-allergic symptoms with a Th2-skewed response.The C57BL/6 mice,rather than the BALB/c mice,were fitter for establishing an epicutaneously sensitized model of food allergy since a stronger Th2-biased response and severer disruptions in the intestinal barrier and gut homeostasis were observed.This study provides knowledge for selecting an appropriate mouse model to study food-allergic responses associated with AD-like skin lesions and highlights the role of genetic variations in the immune mechanism underlying pathogenesis of food allergy.

靶向高通量测序在非小细胞肺癌中的临床应用

目的:对8种与非小细胞肺癌(non-small cell lung cancer,NSCLC)个性化治疗高度相关的驱动基因进行检测,分析基因变异与临床病理特征的关系。方法:收集天津医科大学肿瘤医院2016年6月至2017年8月212例NSCLC患者样本,对EGFR、KRAS、BRAF、ALK、MET、ERBB2、ROS1、RET 8种基因进行高通量测序。结果:8种基因中EGFR基因变异率高达52.8%,其次为KRAS(8.5%)、ALK(8.0%)、ERBB2(6.1%)、MET(3.8%)、BRAF(1.4%)、RET(0.9%)、ROS1(0.9%),75%样本检出至少1个驱动基因变异,驱动基因变异间呈现强烈互斥。最常见的EGFR突变为19外显子缺失和L858R突变,EGFR T790M突变与前面两个突变位点伴随出现。19外显子缺失患者携带非EGFR T790M突变比例低于L858R突变患者携带非EGFR T790M突变比例(P=0.04)。15.2%EGFR突变伴EGFR扩增,携带EGFR扩增且EGFR突变率>40%患者比例高于无EGFR扩增且EGFR突变率>40%患者(P<0.01)。女性、不吸烟、腺癌患者易发生EGFR特别是EGFR敏感突变(P<0.01)。肺腺癌(P=0.013)、临床分期晚(P=0.048)、淋巴结转移(P=0.027)患者携带EGFR扩增比例高。男性(P=0.009)、左侧肺癌(P=0.048),吸烟患者(P=0.037)KRAS突变发生率较高。携带非KRAS突变、ALK融合的患者更年轻(P=0.005,P=0.031),而携带KRAS突变患者年龄较高(P=0.055)。结论:高通量测序可同时高效检测NSCLC患者中8种与靶向治疗相关驱动基因的变异谱,为临床医生的个体化诊疗提供参考,以多基因为基础的高通量测序为NSCLC诊疗提供更多的可能性。

Benefit of everolimus as a monotherapy for a refractory breast cancer patient bearing multiple genetic mutations in the PI3K/AKT/mTOR signaling pathway

A postmenopausal patient with a diagnosis of estrogen receptor(ER)(+), progesterone receptor(PR)(+), and human epidermal growth factor receptor-2(HER2)(-) breast cancer was reported. The patient refused surgery and was resistant to conventional chemotherapy regimens. Computed tomography and the circulating tumor cell test indicated that the patient's tumor burden increased rapidly even after several chemotherapy sessions. Multiple genetic aberrances in the phosphatidylinositol3-kinases(PI3 K) signaling pathway were detected via next-generation sequencing(NGS)-based liquid biopsy, including a p. G1007 R missense mutation in exon 21 of PIK3 CA(33.61%), a p.L70 fs frameshift mutation in exon 3 of phosphatase and tension homolog deleted on chromosome ten(PTEN)(49.14%), and a p. D1542 Y missense mutation in exon 32 of mammalian target of rapamycin(m TOR)(1.66%). Therefore, only the m TOR inhibitor everolimus was administered to the patient. Partial remission(PR) was observed after 2 months, and sustained stable disease(SD) was observed after a year and a half. Subsequent sequencing showed that the mutation ratio of PIK3 CA decreased to 4.17%, and that the PTEN and m TOR mutations disappeared, which revealed the significant curative effect of everolimus. We report the first case of successful monotherapy treatment using everolimus in a patient with advanced breast cancer bearing mutations in genes involved in the PI3 K/ARK/m TOR signaling pathway. The success of this case highlights the invaluable clinical contribution of NGS-based liquid biopsy, as it successfully provided an optimal therapeutic target for the patient with advanced breast cancer.

Characterization of integrons and novel cassette arrays in bacteria from clinical isloates in China,2000-2014

Rapid dissemination of antibiotic resistance genes among bacterial isolates is an increasing problem in China. Integron, a conserved DNA sequence, which is carried on episomal genetic structures, plays a very important role in development of antibiotic resistance. This systematic analysis was based on MEDLINE and EMBASE databases. We summarized the distribution and proportion of different types of gene cassette arrays ofintegrons （including class 1, 2, 3 and atypical class 1 integron） from clinical bacteria isolates in China. Fifty-six literatures were included in this study. Most of the strains were Gram-negative bacteria （94.1%, 7,364/7,822） while only 5.9% strains were Gram- positive bacteria. Class 1 integrons were detected in 54.2% （3956/7295） Gram-negative strains, aadA2 was the most popular gene cassette array detected from 60 Gram-positive bacteria while dfrA 17-aadA5 were detected in 426 Gram- negative bacteria. This study identified 12 novel gene cassette arrays which have not been previously found in any species. All the novel gene cassette arrays were detected from Gram-negative bacteria. A regional characteristic of distribution of integrons was presented in this study. The results highlight a need for continuous surveillance of integrons and provide a guide for future research on integron-mediated bacteria resistance.

甲状腺良恶性结节的基因变异差异比较

目的:应用二代测序和生物信息学方法探究甲状腺癌与良性结节的基因变异差异,在分子层面对甲状腺良恶性结节进行鉴别。方法:收集2014年12月至2018年5月天津医科大学肿瘤医院187例甲状腺癌患者的甲状腺结节以及81例癌旁正常甲状腺样本的DNA和RNA,根据B超和细针穿刺活检/病理学结果将样本分为癌组织和可疑良性结节。通过二代测序技术进行基因变异检测。利用生物信息学方法,对检测结果进行分析。结果:共检出突变位点476个,累计突变基因40个,主要包括驱动基因、DNA损伤修复基因、免疫相关基因、染色体重塑相关基因和激素相关基因等。突变类型包括错义突变、框移突变、插入/缺失突变和无义突变。甲状腺癌的基因变异特征与可疑良性结节存在显著性差异(均P<0.05),其中BRAF、ZNF717、TERT、GGT1、CHEK2、OTUD4、RET蛋白激酶域的突变占比和基因融合的发生频率均显著高于可疑良性结节,而且RET蛋白激酶域突变和GGT1突变均与甲状腺癌淋巴结转移呈正相关(均P<0.05)。检出融合类型14例,包括CCDC6-RET、NCOA4-RET、ETV6-NTRK3和TPM3-NTRK1共4种融合类型。结论:甲状腺癌的基因变异特征与可疑良性结节存在显著性差异,虽然癌组织和可疑良性结节在部分基因的突变占比上无显著性差异,但是在癌组织中BRAF、TERT、ZNF717、GGT1、CHEK2、OTUD4、RET、PI3K/Akt通路、AMPK通路相关基因以及融合基因的突变占比均显著高于可疑良性结节,上述变异基因可以作为潜在的生物标志物,在分子水平方面有效的区分恶性甲状腺结节和良性甲状腺结节。

Rapid response of brain metastasis to crizotinib in a patient with KLC1-ALK fusion and MET gene amplification positive non-small cell lung cancer:a case report

Non-small cell lung cancer(NSCLC) ranks as the leading cause of cancer-related death in the world. Brain metastasis(BM) is a common complication of NSCLC, with 25%–40% of patients developing BM during the course of the disease. A significant strategy of local disease control in the central nervous system is radiation therapy. With the development of precision medicine,the concept of treating lung cancer BM has gradually changed. In this case, we performed a surgical procedure to obtain enough tumor tissue for the detection of the target gene and other related experiments after the patient was informed. Finally, we found that the patient had both hepatocyte growth factor receptor(MET) gene amplification and kinesin light chain 1-anaplastic lymphoma kinase fusion(KLC1-ALK) through next-generation sequencing and showed sensitivity to the targeted therapy of crizotinib. The patient exhibited good response. Our case was successful and underwent targeted therapy with the guidance of precise diagnosis.

Skeleton-aware implicit function for single-view human reconstruction

The aim is to reconstruct a complete and detailed clothed human from a single-view input.Implicit function is suitable for this task because it represents fine shape details and varied topology.Current methods,however,often suffer from artefacts such as broken or disembodied body parts,missing details,or depth ambiguity due to the ambiguity and complexity of human articulation.The main issue observed by the authors is structureagnostic.To address these problems,the authors fully utilise the skinned multi-person linear(SMPL)model and propose a method using the Skeleton-aware Implicit Function(SIF).To alleviate the broken or disembodied body parts,the proposed skeleton-aware structure prior makes the skeleton awareness into an implicit function,which consists of a bone-guided sampling strategy and a skeleton-relative encoding strategy.To deal with the missing details and depth ambiguity problems,the authors’body-guided pixel-aligned feature exploits the SMPL to enhance 2D normal and depth semantic features,and the proposed feature aggregation uses the extra geometry-aware prior to enabling a more plausible merging with less noisy geometry.Additionally,SIF is also adapted to the RGB-D input,and experimental results show that SIF outperforms the state-of-the-arts methods on challenging datasets from Twindom and Thuman3.0.

Correlation between serum fructosamine and hyperglycemia in patients with acute cerebrovascular disease

BACKGROUND:Diabetes mellitus is one of the risk factors in patients with acute cerebral disease,and always leads to stroke or get it worse.There is often a high level of blood glucose in those patients with diabetes mellitus and cerebral disease,but it is hard to distinguish from both kinds of hyperglycemia.Serum fructosamine is said to be correlated with blood glucose.OBJECTIVE:To explore the relationship between serum fructosamine and blood glucose in patients with acute cerebrovascular disease.DESIGN:A case-controlled study.SETTINGS:Department of Clinical Laboratory,Health Department for Cadres and Department of Neurology of Affiliated Hospital,Qingdao University Medical College.PARTICIPANTS:Forty-eight inpatients and outpatients with cerebrovascular diseases were selected from the Department of Neurology,Affiliated Hospital of Qingdao University Medical College from December 2004 to April 2005.All the patients were confirmed with CT and MRI.There were 25 patients with diabetes mellitus secondary cerebrovascular diseases,who met the diagnostic standards of diabetes mellitus set by WHO,including 12 males and 13 females with an average of(60±8)years old,the course of diabetes mellitus ranged from 1 to 21 years..The other 23 patients had no diabetes mellitus(without diabetes mellitus group),including 14 males and 9 females with an average of(62±6)years old.Meanwhile,another 50 healthy physical examinees in the hospital were selected as control group,including 26 males and 24 females with the average age of(62±5)years old.Informed content was obtained from all the participants.METHODS:Venous blood was drawn from all the participants,and content of blood glucose was assayed by means of glucose oxidase,and the concentration of serum fructosamine was determined by nitroblue tetrazolium colorimetric method.Comparison between groups was performed by the analysis of variance and q test,and the correlation was tested by linear regression analysis.MAIN OUTCOME MEASURES:①Comparison of blood glucose and serum fructosamine among the groups;②Correlation between serum fructosamine and blood glucose in patients with diabetes mellitus secondary cerebrovascular diseases and those without diabetes mellitus.RESULTS:All the 48 patients with cerebrovascular disease and 50 healthy subjects were involved in the analysis of results.①Contents of blood glucose and serum fructosamine:There were obvious differences in the contents of blood glucose and serum fructosamine among the diabetes mellitus group,without diabetes mellitus group and control group(F=577.7,115.1,P<0.01).The content of serum fructosamine in the diabetes mellitus group[(4.25±1.35)mmol/L]was obviously higher than those in the control group and without diabetes mellitus group[(1.65±0.27),(1.96±0.25)mmol/L,q=1.47,1.30,P<0.01],whereas there was no significant difference between the without diabetes mellitus group and control group(P>0.05).The content of blood glucose was obviously higher in the patients with and without diabetes mellitus groups[(15.80±2.13),(9.50±1.78)mmol/L]than in the control group[(4.56±0.77)mmol/L,q=1.86,2.46,P<0.01],also markedly higher in the with diabetes mellitus group than in the without diabetes mellitus group(q=1.42,P<0.01).②Results of correlation analysis:The content of serum fructosamine was positively correlated with the level of fasting blood glucose in the patients with diabetes mellitus secondary cerebrovascular diseases(r=0.603,P<0.01).But there was no relationship between serum fructosamine and fasting blood glucose in the patients without diabetes mellitus(r=0.357,P>0.05).CONCLUSION:The contents of blood glucose and serum fructosamine were obviously different among the diabetes mellitus group,without diabetes mellitus group and control group.There are closer relations between serum fructosamine and blood glucose in patients with diabetes mellitus secondary cerebral disorders,which are not observed in the patients without diabetes mellitus.Fructosamine is significant in differentiating the reasons for the increased blood glucose in patients with acute cerebrovascular disease.

Identification of Resistance of Powdery Mildew and Stripe Rust of Winter Wheat Strains in Xinjiang

Wheat powdery mildew and stripe rust are the major diseases in wheat producing area in Xinjiang.To obtain wheat germplasm resources and varieties resistant to powdery mildew and rust,36 high-generation stable strains of Xinjiang winter wheat were evaluated using the method of natural inducement from 2018 to 2020.A total of 5 strains with high resistance to powdery mildew,4 strains with slow stripe rust and 1 strain with resistance to powdery mildew and adult plant slow stripe rust were obtained.And the parental combination of disease-resistant varieties was analyzed.These studies will provide theoretical basis for the breeding of resistant wheat varieties in Xinjiang.

Enhancing prime editing efficiency and flexibility with tethered and split pegRNAs

Dear Editor,Most human genetic diseases arise from mutations such as insertion,deletion,or point mutations(Landrum et al.,2016).CRISPRCas system has been repurposed to correct pathogenic mutations in a variety of genetic diseases(Choi et al.,2022).There are many concerns about using CRISPR-mediated double-stranded DNA breaks(DSBs)for therapeutic purposes,primarily due to off-targeted mutations(Kosicki et al.,2018).Nevertheless,base editing cannot correct deletions,insertions,or some point mutations such as transversion mutations.Prime editing has its advantages of precisely correct point mutations,small insertions,or deletions in animal cells(Anzalone et al.,2019)and plants(Lin et al.,2020).However,prime editing efficiency varies among genomic sites or cell types(Chen et al.,2021;Nelson et al.,2022).

A novel microRNA miR-MTCO3P38 inhibits malignant progression via STAT3/PTTG1/MYC in hepatocellular carcinoma

To the Editor,Hepatocellular carcinoma(HCC)is a malignant tumor with the fourth highest incidence and the third highest mortality among all tumor sites in China.1 Finding new diagnostic markers and therapeutic molecules may be of great value in the treatment of HCC and the improvement of prognosis.

High expression of EPO gene using un-dhfr negative cell

Erythropoietin （EPO） genomic gene was cloned and its expression vector pOP13/EPO was constructed. CHOK12 cell was transfected by this vector using lipofectin method. A stable expression cell strain C10 cell with the EPO production at 160IU/d in 10\+6 cells were obtained at 400 μg/mL G418. Based on the C10 cell, another vector pHY/dhfr （dihydrofolate reductase） that carries a dhfr gene and a selecting marker of hygromycin B resistant gene was transferred to this cell. Several cell clones were obtained at 200 μg/mL hygromycin B. These cell clones that can express both EPO gene and exogenous dhfr gene were selected under the progressively increased concentration to 1 μmol methotrexate（MTX）. Some high EPO expression cell clones were obtained, the highest expression was 2 400 IU/d in 10\+6 cells, 15 times higher than that without MTX pressure. Then, a method of EPO high expression by using undhfr negative cell was primarily established. EPO bioactivity was found by using TF1 cell.