[摘 要] 目的：基于噬菌体展示靶向锚定蛋白文库，以人白细胞抗原G（HLA-G）为靶点筛选锚定序列作为HLA-G结合蛋白（HGBP）并评价其功能。方法：利用TCGA、GTEx等数据库分析肿瘤组织中HLA-G表达与临床预后和免疫浸润的相关性。利用噬菌体展示靶向锚定蛋白文库对HLA-G胞外片段进行生物淘选并随机挑取单克隆进行测序。通过ELISA、免疫荧光染色鉴定优势噬菌体克隆的功能。利用原核体系生产纯化HGBP，通过ELISA、表面等离子共振（SPR）、免疫荧光染色法评价其亲和力及肿瘤特异性结合能力。结果：生物信息学分析发现，HLA-G在肿瘤组织中普遍呈高表达，其与临床总生存期、免疫细胞浸润水平具有相关性（P < 0.05）。5轮噬菌体文库筛选后获得优势克隆，ELISA及免疫荧光染色结果均显示，优势噬菌体对HLA-G阳性细胞结合显著强于阴性细胞（P < 0.05, P < 0.001）。经纯化生产的HGBP与HLA-G之间的亲和力可达17 nmol/L，ELISA结果显示，HGBP与HLA-G分子的结合显著（P < 0.05）；免疫荧光染色结果表明，HGBP能与HLA-G阳性细胞特异性结合（P < 0.01）。结论：经噬菌体展示文库筛选出的HGBP对HLA-G具有高亲和力，可特异性结合肿瘤细胞表达的HLA-G。

Objective To investigate the effect of 5-Aza-CdR on Notch1 pathway and neural regeneration and to explore the effects of 5-Aza-CdR on learning memory ability in mice by exploring active avoidance behavior.Methods Sixty 6～8-week-old SPF-grade ICR male mice were divided into two groups.5-Aza-CdR was administered to one group of mice via lateral ventricular injection,while the control group was injected with bovine serum albumin.Notch1 and HES1 mRNA and protein expression levels were detected by Real-time PCR and Western blot 24 hours after injection;5-bromo-2'-deoxyuridine-positive cells were observed by laser confocal microscopy,and Notch1 expression in hippocampal dentate gyrus was viewed with laser confocal microscopy.Notch1 methylation changes were detected by ethylation-specific PCR,and learning and memory behaviors of mice were assessed by passive avoidance tests and shuttle avoidance assays.Results Injection of 5-Aza-CdR increased hippocampal Notch1 pathway activity,promoted neuronal regeneration in the DG region,decreased methylation levels in the Notch1 promoter region,and enhanced the ability of mice to perform active avoidance behavior.Conclusions The effect of 5-Aza-CdR on active avoidance behavior may be related to the influence of hippocampal neural regeneration through the Notch 1 pathway.

Objective To evaluate the scientificity,transparency and applicability of Chinese guidelines and expert consensuses on nursing published in 2022,in order to improve the quality of guidelines and consensuses.Methods Databases including Medline,Embase,Web of Science,CBM,CNKI,WanFang database,Chinese Medical Journal,and related websites were electronically searched,as well as China Hong Kong,Macao and Taiwan medical journals,to collect Chinese guidelines and expert consensuses on nursing from January to December 2022.STAR tool was used to evaluate the quality of each guidelines and consensuses by 3 assessors independently.Total score,scoring rate of each domain and item were adopted to analyze the outcomes.Results A total of 3 guidelines and 33 expert consensuses were included.The total guidelines and expert consensuses STAR score(33.5±14.3).The quality of guidelines and consensuses was low.The quality of guidelines was moderate with average score of 55.1,and the quality of consensuses was low with average score of 31.5.The included guidelines and consensuses had a highest score rate(52.4％)in the domain of recommendation.Among 39 items of STAR tool,the top 4 items including listing participants and institutions,explaining additional instructions for implementation,describing consensus method,and listing references for recommendations had a high score rate of 100％,83.3％,77.8％,75.0％respectively.However,the items of registration,providing registration information,protocols being searched on public platforms and explaining the role of funding had a low score rate,urgent need for attention and upgrading.Conclusion The overall quality of the Chinese guidelines and expert consensuses on nursing published in 2022 was low.As a medical and nursing practice guidance document,the quality of guidelines and expert consensuses should be improved by encouraging registration,strengthening management of interest conflict,enhancing the rigor of guideline developing process,and expanding the dissemination.

Background and purpose:Rectal cancer occurs on the inner wall of the rectum,and metastasis-associated in colon cancer 1(MACC1)can promote drug resistance in colon cancer cells.This study aimed to investigate the effect of MACC1 on oxaliplatin resistance in rectal cancer and its mechanism.Methods:Biosignal analysis of MACC1 and TCF7 expressions in rectal cancer tissues and signaling pathways enriched for MACC1 were carried out.The expressions of MACC1 and TCF7 in rectal cancer cells and rectal cancer oxaliplatin-resistant cells were detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),cell viability was detected by cell counting kit-8(CCK-8),and cell proliferation was detected by cell colony formation assay.ECAR values,lactate production and glucose consumption were detected by Seahorse Biosciences XF96.Western blot was used to detect the expressions of glycolysis-related proteins.Dual luciferase reporter gene and ChIP were used to validate the regulatory relationship between MACC1 and TCF7.Normality test and homogeneity of variance analysis was performed on the data of each group by GraphPad Prism 8.0.If it conformed to normal distribution,one-way ANOVA or t-test would be used for inter group comparison,otherwise the comparison between groups would be conducted using Wilcoxon rank sum test.Using a two-sided test,P＜0.05 was considered statistically significant.Results:MACC1 and TCF7 expressions were upregulated in rectal cancer,and knockdown of MACC1 significantly inhibited the viability of rectal cancer drug-resistant cells and the IC50 values of different concentrations of oxaliplatin treatments,as well as reduced the proliferation ability of rectal cancer oxaliplatin-resistant cells.Overexpression of MACC1 was able to promote the proliferation and glycolytic capacity of rectal cancer cells.Further studies revealed that the transcription factor TCF7 existed in the upper reaches of MACC1.Besides,this study analyzed the data from Cancer Genome Atlas Program(TCGA),and the result showed that knockdown of TCF7 was able to attenuate the promotional effect of overexpression of MACC1 on the glycolytic capacity and oxaliplatin resistance of rectal cancer cells.Conclusion:This study demonstrated that the TCF7/MACC1 axis could promote aerobic glycolysis and thus oxaliplatin resistance in rectal cancer cells.The findings suggest that targeting the TCF7/MACC1 axis or inhibiting the aerobic glycolysis pathway may be a novel therapeutic approach to inhibit oxaliplatin resistance in rectal cancer.

Objective:To explore the effect of fully automatic image segmentation of adenoid and nasopharyngeal airway by deep learning model based on U-Net network. Methods:From March 2021 to March 2022, 240 children underwent cone beam computed tomography（CBCT） in the Department of Otolaryngology, Head and Neck Surgery, General Hospital of Shenzhen University. 52 of them were selected for manual labeling of nasopharynx airway and adenoid, and then were trained and verified by the deep learning model. After applying the model to the remaining data, compare the differences between conventional two-dimensional indicators and deep learning three-dimensional indicators in 240 datasets. Results:For the 52 cases of modeling and training data sets, there was no significant difference between the prediction results of deep learning and the manual labeling results of doctors（P>0.05）. The model evaluation index of nasopharyngeal airway volume: Mean Intersection over Union（MIOU） s （86.32±0.54）%; Dice Similarity Coefficient（DSC）: （92.91±0.23）%; Accuracy: （95.92±0.25）%; Precision: （91.93±0.14）%; and the model evaluation index of Adenoid volume: MIOU: （86.28±0.61）%; DSC: （92.88±0.17）%; Accuracy: （95.90±0.29）%; Precision: （92.30±0.23）%. There was a positive correlation between the two-dimensional index A/N and the three-dimensional index AV/（AV+NAV） in 240 children of different age groups（P<0.05）, and the correlation coefficient of 9-13 years old was 0.74. Conclusion:The deep learning model based on U-Net network has a good effect on the automatic image segmentation of adenoid and nasopharynx airway, and has high application value. The model has a certain generalization ability.
Child ; Humans ; Adolescent ; Adenoids/diagnostic imaging* ; Image Processing, Computer-Assisted/methods* ; Pharynx ; Cone-Beam Computed Tomography ; Nose

Objective:To discuss the application of virtual endoscopy in the diagnosis of adenoid hypertrophy and the morphologic classification of adenoid. Methods:The clinical data of 97 children with adenoid hypertrophy admitted to Department of Otorhinolaryngology Head and Neck Surgery, Shenzhen University General Hospital from July 2022 to December 2022 were collected. The virtual endoscopic reconstruction of the nasopharynx was performed by cone beam computed tomography. The results of virtual endoscopic adenoid size measurement were compared with the results of nasopharyngeal CT median sagittal position and nasopharyngeal endoscopy. Virtual endoscopic classification of adenoid based on the size of the adenoids and their relationship with the torus tubarius. Results:The t-test results of the size of adenoids measured by virtual endoscopy and nasopharyngeal CT were t=1.699 and P=0.093, and the results of intra-group correlation coefficient（ICC） analysis were ICC=0.921 and P<0.01. The proportion of adenoids measured by virtual endoscopy and nasopharyngeal CT was highly consistent. The t-test results of the size of adenoids measured virtual endoscopy and nasopharyngeal endoscopy were t=1.543 and P=0.15, and the results of intra-group correlation coefficient（ICC） analysis were ICC=0.900 and P<0.01. The proportion of adenoids measured by virtual endoscopy and nasopharyngeal endoscopy was highly consistent. Among the 97 children, the morphological classification results of adenoids were 48 cases of overall hypertrophy type, 47 cases of central bulge type, and 2 cases of flat thickening type. Conclusion:The diagnosis of adenoid hypertrophy by virtual endoscopy has high accuracy, which not only avoids the invasive operation of traditional nasopharyngeal endoscopy, but also can observe the adenoid condition and its relationship with the torus tubarius from multiple angles. And, the morphological classification of adenoids using virtual endoscopy has guiding significance for perioperative preparation.
Child ; Humans ; Adenoids/surgery* ; Nasopharynx/diagnostic imaging* ; Adenoidectomy ; Endoscopy/methods* ; Hypertrophy/surgery*

The maxillofacial region has multiple functions such as breathing, language, and facial expressions. Children's maxillofacial development is a complex and long process, which is affected by many factors such as genetics, diseases, bad habits and trauma. Early detection, early diagnosis, and early treatment are important concepts in children's maxillofacial management. Digital technology medicine is an emerging technology based on medical imaging and anatomy that has emerged in recent years. The application of this technology in the field of clinical medicine will undoubtedly bring great benefits to children's maxillofacial management. This article summarizes the research on digital technology in children's maxillofacial management, and focuses on the research on children's malocclusion, children's OSA, cleft lip and palate and other related diseases.
Humans ; Child ; Cleft Lip/surgery* ; Cleft Palate/surgery* ; Digital Technology ; Respiration

The extremely low bioavailability of oral paclitaxel (PTX) mainly due to the complicated gastrointestinal environment, the obstruction of intestinal mucus layer and epithelium barrier. Thus, it is of great significance to construct a coordinative delivery system which can overcome multiple intestinal physicochemical obstacles simultaneously. In this work, a high-density PEGylation-based glycocholic acid-decorated micelles (PTX@GNPs) was constructed by a novel polymer, 9-Fluorenylmethoxycarbonyl-polyethylene glycocholic acid (Fmoc-PEG-GCA). The Fmoc motif in this polymer could encapsulate PTX via π‒π stacking to form the core of micelles, and the low molecular weight and non-long hydrophobic chain of Fmoc ensures the high-density of PEG. Based on this versatile and flexible carriers, PTX@GNPs possess mucus trapping escape ability due to the flexible PEG, and excellent intestine epithelium targeting attributed to the high affinity of GCA with apical sodium-dependent bile acid transporter. The in vitro and in vivo results showed that this oral micelle could enhance oral bioavailability of PTX, and exhibited similar antitumor efficacy to Taxol injection via intravenous route. In addition, oral PTX@GNPs administered with lower dosage within shorter interval could increase in vivo retention time of PTX, which supposed to remodel immune microenvironment and enhance oral chemotherapy efficacy by synergistic effect.

Objective To investigate the effects of microRNA(miR)-432-5p on the proliferation,metasta-sis and radiosensitivity of gastric cancer cells and the targeted regulatory relationship with DEAD-box helicase 41(DDX41)in this process.Methods The expressions of miR-432-5p and DDX41 in different gastric cancer cells were detected by quantitative real-time PCR(qRT-PCR).HGC-27 cells were transfected and divided into NC group(not transfected with plasmid),miR-NC group(transfected with miR-432-5p mimics negative con-trol),miR-432-5p group(transfected with miR-432-5p mimics),miR-432-5p+pcDNA-NC group(transfection miR-432-5p and pcDNA-DDX41 negative control)and miR-432-5p+pcDNA-DDX41 group(transfection miR-432-5p mimics and pcDNA-DDX41).After 48 h,qRT-PCR was used to detect the expression of miR-432-5p and DDX41 in each group of cells,MTT assay was used to detect the proliferation activity of each group of cells,Transwell assay was used to detect the migration ability of each group of cells,colony formation assay and flow cytometry were used to detect the radiosensitivity of each group of cells.The dual luciferase reporter gene assay was used to verify the targeting relationship between miR-432-5p and DDX41.The tumor growth of nude mice was detected,and the expression of DDX41 protein in tumor tissues was detected by Western blot.Results Compared with normal cells,cancer of the stomach cells miR-432-5p levels were decreased(P<0.05),DDX41 mRNA levels were increased(P<0.05).Compared with NC group and miR-NC group,the cell survival rate and DDX41 mRNA level in miR-432-5p group were decreased,the number of migrating cells was decreased(P<0.05),the miR-432-5p level was increased,and the radiosensitivity was enhanced(P<0.05).Overexpression of DDX41 reversed the effects of miR-432-5p on proliferation,migration and radiosensitivity of gastric cancer cells(P<0.05).Compared with NC group and miR-NC group,the volume and weight of tumor tissue and the expression of DDX41 protein in miR-432-5p group were significantly decreased(P<0.05).Con-clusion miR-432-5p through targeted cut DDX41 to inhibit the proliferation and metastasis of gastric cancer cells,enhance cell radiation sensitivity.

Objective To explore the clinical features and genetic characteristics of epilepsy and neurodevelopmental disorders caused by SLC6A1 gene mutation. Methods The clinical data of a patient with SLC6A1 gene mutation from Tongji Hospital,HuaZhong University of Science and Technology was collected. The related literatures were reviewed to summarize the characteristics of SLC6A1 gene mutation and the clinical phenotype. Results A 2 years and 6 months old girl was enrolled in the study.Her first attack happened at the age of 20 months and leg shaking,standing instability and wrestling accompanied by frequent blinking were observed. Cognitive development was impaired,especially the language. Electroencephalography (EEG) was abnormal with slow background rhythm and extensive high amplitude slow wave spike slow wave during the sleep and awake period. Brain MRI scan showed bilateral frontal lobe speckled lesions with high intensity on T2 Flair sequence. Gesell assessment scale showed development quotient 64,and development age 21 months. Children autism assessment scale suggested that there was no obvious autism performance. Whole exome-sequencing study identified a heterozygous variant of c. 889G>A (p. Gly297Arg) in SLC6A1 gene. Both her mother and grandmother of the child carried this mutation. Summarizing the previous literatures,the main clinical characteristics related to SLC6A1 gene mutation were epilepsy (absence,atonic,myoclonus and myoclonus-atonic),developmental retardation,cognitive impairment and autism or autism-like manifestations. The variants of SLC6A1 gene included missense mutation,nonsense mutation,frameshift mutation,splicing mutation,and chromosome microdeletion. Conclusion The main SLC6A1 variant was missense variant. The main clinical features of patients with SLC6A1 gene mutations were epilepsy and cognition impairment. The relationship between genotype and phenotype needs further study. Valproic acid is the first-line drug and PBA is the potential effective drug.