Objective To construct high-capacity ribosome display single-chain Fv library for selection of high affinity ScFv antibody.Methods We isolate human lymphocyte from peripheralblood(2 normal,3 gastric cancer,3 colonic cancer,1 pancreatic cancer,each 5 mL and 2 newborn,each 2 mL)and extract RNA for cloning whole human heavy chain and light chain gene by RT-PCR.VH and VL were rearranged randomly by SOEing(splicing by overlap extension,SOEing).Finally,the elements for in vitro screening such as T7 promoter and ribosome binding site were introduced while the SOEing products were amplified.Moreover,ribosome display template were verified by blue/white screening and further sequencing.Results We successfully constructed ribosome display ScFv library with a volume of 1.1?1013.Conclusion The construction of high-capacity ScFv library shed light on multiple therapeutic ScFv screening.

Objective To investigate the causes and risk factors of postpartum hemorrhage (PPH) in hepatitis B virus (HBV) infected parturient.Methods Retrospective analysis was performed on the 1021 HBV infected parturient from Shanghai Public Health Clinical Center from July 2005 to June 2011.The comparisons were done by chi-square test.Results Among 1021 cases of HBV infected parturient,868 (85.01％) were asymptomatic and the PPH rate was 2.76％ (24/868) ;the remaining 153 cases (14.99％) were chronic active hepatitis B and the PPH rate was 16.99％(26/153).The difference between two groups was statistically significant (x2 =56.541,P＜0.01).The total incidence rate of PPH was 4.89％ (50/1021) and 17 cases (34.00％) were postpartum hemorrhage＞1000mL.The causes of PPH included uterine inertia (30/50,60.00％),abnormal placenta (11/50,22.00％),dysfunction of coagulation (5/50,10.00％) and lesion of birth canal (4/50,8.00％).The risk factors of PPH included delivery mode (x2 =6.528,P=0.038),abortion times (x2 =16.269,P=0.000),delivery times (x2 =6.990,P=0.008),ALT levels (x2=56.541,P=0.000) and HBV DNA (x2 =64.706,P=0.000).Conclusions The main causes of PPH in HBV infected parturient include uterine inertia,abnormal placenta,lesion of birth canal and dysfunction of blood coagulation.PPH is correlated with abortion times,delivery times,delivery mode,liver function and HBV DNA.The incidence of PPH in parturient with chronic active hepatitis B is higher than asymptomatic parturient.

Aim To investigate the effect of BMS- 345541 on the repair of DNA DSBs induced by VP-16 in AML cells and its possible mechanism. Methods The effects of BMS-345541 on the sensitivity of AML cells to VP-16 were determined by MTT. Flow cytome-try ( FCM) was applied to test the level of DNA dam-age, cell cycle progression and apoptosis in AML cells. High content analysis ( HCA) was used to verify the amount ofγ-H2AX,p-ATM,RAD51 in AML cells. Results BMS-345541 could significantly inhibit the proliferation of AML cells induced by VP-16 . BMS- ＜br> 345541 increased the amount of RAD51 foci and p-ATM foci in AML cells treated with VP-16 after 6 hours , which led to increased numbers of cells in the G2/M phases of the cell cycle,then induced apoptotic cell death. Conclusion BMS-345541 sensitizes AML cells to VP-16 via selective inhibition of homologous recombinational repair of DNA double-strand breaks.

Objective To describe the clinical features and imaging characteristics of nodular splenic sarcoidosis. Methods We describe a patient with splenic sarcoidosis and review the related medical literature, the etiology, symptomatology, pathology, diagnosis, differential diagnosis, management and prognosis of splenic sarcoidosis. Results The etiology of this rare disease remains unknown. Symptoms are scanty and usually mild; computed tomography usually reveals splenomegaly or the presence of multiple nodules, confusing with metastatic tumor in spleen. On histopathologic examination, sarcoid produces noncaseating granulomas. Sarcoid is typically treated only when symptomatic. Oral corticosteroids is the most important method of treatment in patients with progressive loss of organ functions. Prognosis has closed relationship with early clinical manifestation. Conclusion Splenic sarcoidosis is rare and often misdiagnosis as other diseases.

10.3969/j.issn.1000-3606.2013.06.019

Objective The researchers systematically retrieved,evaluated,and summarized the best evidence for the care of nebulized inhalation in adult patients on mechanical ventilation,to provide a basis for standardizing the care of nebulized inhalation in patients on mechanical ventilation.Methods We systematically searched the domestic and foreign databases to collect the evidence on the literature related to nebulization therapy for mechanical ventilation in adults.The time for the retrieval is from the inception of databases until February 2023.There were 3 researchers who evaluated the quality of the literature,and extracted and summarized the evidence based on this subject.Results A total of 19 articles were obtained in database.42 pieces of evidence were summarized,covering pre-assessment of nebulized inhalation,preparation before nebulized inhalation,medication management,selection and standardized use of nebulization devices,respiratory machine mode and parameter settings,equipment management during nebulized inhalation,evaluation of effect,management of adverse reactions,disposal of materials and environment after nebulized inhalation and management of nebulized inhalation for respiratory infectious diseases.Conclusion This study summarized the best evidence for nebulized inhalation nursing in adult patients with mechanical ventilation,so as to provide a reference of standardized nebulized inhalation therapy for such patients,which is conducive to ensuring the safety of patients.

Objective:To investigate the expression of heterogeneous nuclear ribonucleoprotein A2B1 (HNRNPA2B1) in human esophageal cancer tissues and its clinical significance.Methods:Single-cell data for esophageal cancer were downloaded from the Gene Expression Omnibus (GEO) database (GSE160269 dataset, last updated on November 29, 2020) to analyze the expression of HNRNPA2B1. Transcriptional sequencing data for esophageal cancer from The Cancer Genome Atlas (TCGA) database, including the fragments per kilobase of transcript per million mapped reads (FPKM) quantitative data (173 samples, consisting of 162 esophageal cancer tissues and 11 adjacent normal tissues), and survival data in the phenotype category were downloaded. Analysis of FPKM quantitative data from the TCGA database for esophageal cancer was performed. The top 250 genes most correlated with HNRNPA2B1 were selected and the R4.3.0 clusterProfiler package was used to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses on the selected gene set. FPKM quantitative data from the TCGA database for esophageal cancer were imported into the CIBERSORTx website to obtain immune cell abundance scores, and the correlation between HNRNPA2B1 and the degree of immune cell infiltration was analyzed. The clinicopathological data of patients from esophageal cancer tissue microarrays including 114 cases of esophageal squamous cell carcinoma tissues and 66 cases of adjacent normal tissues were collected. The patients underwent surgery from January 2006 to December 2008, and the follow-up period extended until July 2015. Cytokeratin (CK) and HNRNPA2B1 expression in esophageal cancer tissue microarrays were detected by using multi-color immunohistochemical (mIHC) staining, and multispectral tissue imaging was conducted. The R4.3.0 survival package and survminer package in TCGA database were used to calculate the optimal cut-off value of HNRNPA2B1 expression and the proportion of CK + HNRNPA2B1 + cells in tissue microarrays was used to calculate the cut-off value of HNRNPA2B1 expression based on which patients were categorized into high and low expression groups. The overall survival (OS) of both groups was compared and the factors influencing OS were analyzed by using the Cox proportional hazards model. Results:In the GSE160269 dataset of single-cell data for esophageal cancer, the expression level of HNRNPA2B1 in tumor epithelial cells was higher than that in normal epithelial cells, and HNRNPA2B1 was highly expressed in various immune cell subtypes. The high expression level of HNRNPA2B1 was positively correlated with regulatory T cells, naive B cells and memory CD4 + T cells. GO enrichment analysis revealed that HNRNPA2B1 was primarily involved in the biological process of nuclear division, cellular components were mainly enriched in chromosomal regions, and molecular functions were mainly enriched in ATP hydrolysis activity. KEGG enrichment analysis indicated that HNRNPA2B1 was primarily involved in biological processes such as the cell cycle, spliceosome, and DNA replication. Results from mIHC and multispectral tissue imaging demonstrated that CK was predominantly expressed in the cell membranes of tumor cells and normal esophageal epithelial cells, while HNRNPA2B1 was primarily expressed in the nuclei of tumor cells and normal esophageal cells. The expression level of HNRNPA2B1 in esophageal cancer tissues was higher than that in the normal paracancerous tissues ( U = 2 984.00, P < 0.05). Results of tissue microarrays and the survival analysis on the data in the TCGA database indicated that esophageal cancer patients with low HNRNPA2B1 expression had a better OS compared to those with high expression (both P < 0.05). Cox multivariate regression analysis revealed that age ( HR = 1.919, 95% CI: 1.158-3.182, P = 0.011), TNM stage ( HR = 2.404, 95% CI: 1.374-4.207, P = 0.002), T stage ( HR = 2.349, 95% CI: 1.150-4.789, P = 0.019), and the expression of HNRNPA2B1 in tumor epithelial cells ( HR = 2.160, 95% CI: 1.280-3.647, P = 0.004) were independent factors influencing OS in esophageal cancer patients. Conclusions:The high expression of HNRNPA2B1 protein in esophageal cancer tissues may play a role in the developement and progression of esophageal cancer, serving as a crucial biological indicator for prognostic assessment of esophageal cancer.

Objective To compare the positioning accuracy between personalized polyurethane foam with wing boards and negative pressure vacuum bag in radiotherapy for lung cancer using kilovoltage cone beam computed tomography(CBCT). Methods Thirty-nine patients with lung cancer who received chest radiotherapy in our hospital from 2015 to 2016 were enrolled as subjects. In those patients, 20 were immobilized by negative pressure vacuum bags(VB group)and the others by personalized polyurethane foam with wing boards(PF group).CBCT images were acquired weekly and registered with planning CT images. Setup errors in the left-right, superior-inferior, and anterior-posterior directions, three-dimensional(3D) displacement vector,and setup time were recorded. The margin of the planning target volume(PTV)was calculated using the Van Herk formula(2.5∑+0.7σ). Between-group comparison was made by paired t test. Results The PF group had a significant smaller setup error in the y-direction than the VB group (2.54±1.79 vs.3.19±2.14 mm,P=0.03),while there were no significant differences in setup errors in the x-or z-direction between the two groups(1.80± 1.48 vs. 1.90± 1.41 mm, P=0.46;2.14± 1.75 vs. 2.25± 1.75 mm,P=0.35). There were no significant differences in rotational setup errors in the Rx-,Ry-,or Rz-direction between the two groups(1.15°±0.76°vs. 1.15°±0.85°, P=0.50;0.71°±0.60°vs. 0.72°±0.43°, P=0.45;0.62°±0.54° vs. 0.46°±0.30°,P=0.06). The PTV margins in the x?,y?,and z?directions were expanded by 5.56, 8.57, and 7.02 mm, respectively, in the PF group, and by 5.62, 9.27, and 7.23 mm,respectively,in the VB group. The proportion of patients with 3D displacement vectors larger than 5 mm was 40% in the PF group and 45% in the VB group.Conclusions For patients undergoing radiotherapy for lung cancer,personalized polyurethane foam with wing boards can,to a certain extent,reduce the setup error in the superior-inferior direction and PTV margin expansion.[Key words] Lung neoplasms/radiotherapy; Polyurethane foam; Vacuum bag; Setup errors;Margin

Background: and Purpose Previous studies suggested increased visit-to-visit variability of total cholesterol (TC) is associated with stroke. This study aimed to investigate the associations of various lipids measurements variability and the risk of stroke and stroke type (ischemic and hemorrhagic stroke). Methods: Fifty-one thousand six hundred twenty participants in the Kailuan Study without history of myocardial infarction, stroke, and cancer who underwent three health examinations during 2006 to 2010 were followed for incident stroke. Variability in TC, triglycerides, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) measurements were measured using the coefficient of variation (CV), standard deviation (SD), variability independent of the mean (VIM), and average real variability (ARV). Results: During a median of 6.04 years of follow-up, 1,189 incident stroke (1,036 ischemic and 160 hemorrhagic stroke) occurred. In the multivariable-adjusted model, the hazard ratio (HR) comparing participants in the highest versus lowest quartile of CV of HDL-C were 1.21 (95% confidence interval [CI], 1.02 to 1.45; P for trend=0.013) for ischemic stroke. The highest quartile of CV of LDL-C was associated with 2.17-fold risk of hemorrhagic stroke (HR, 2.17; 95% CI, 1.25 to 3.75; P for trend=0.002) compared with the lowest quartile. We did not observe any significant association between TC and triglycerides variability with any of stroke. Consistent results were obtained when calculating variability index using SD, VIM, or ARV. Conclusions These findings suggest the high visit-to-visit HDL-C and LDL-C variability were associated with an increased incidence of ischemic and hemorrhagic stroke, respectively.

OBJECTIVE: To explore the genetic basis for a child featuring global developmental disorder with epilepsy. METHODS: A child who had presented at Guangzhou Women and Children's Medical Center in July 2022 was selected as the study subject. Clinical data was collected. Potential variant was detected by whole exome sequencing (WES). Candidate variant was validated by Sanger sequencing and bioinformatic analysis. RESULTS: The child, a three-year-old ethnic Zhuang Chinese girl, had presented with global developmental disorder and epilepsy, for which rehabilitation therapy was ineffective. Genetic testing revealed that she has harbored a homozygous c.821T>C (p.Leu274Pro) missense variant of the PIGW gene, for which both of her parents and sister were heterozygous carriers. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as variant of uncertain significance. CONCLUSION The homozygous c.821T>C (p.Leu274Pro) variant of the PIGW gene probably underlay the onset of disease in this child. Above finding has enriched the mutational spectrum of the PIGW gene.
Child, Preschool ; Female ; Humans ; Computational Biology ; Developmental Disabilities ; Epilepsy/genetics* ; Genetic Testing ; Homozygote