目的：筛选果蝇Zeste基因增强子同源物2（EZH2）基因上游miRNA及lncRNA，分析其在胃癌细胞中的表达并验证其间的靶向关系，探讨它们对胃癌细胞增殖、迁移和凋亡的影响。方法：通过ENCORI、miRDB和Target Scan数据库查询并分析、筛选EZH2上游miRNA（has-miR-450b-5p），ENCORI数据库和DAINA数据库筛选has-miR-450b-5p上游lncRNA（lncRNA NEAT1），预测hsa-miR-450b-5p、lncRNA NEAT1与EZH2之间的结合位点，双荧光素酶报告基因实验验证hsa-miR-450b-5p与lncRNA NEAT1的结合关系。采用qPCR和WB法检测lncRNA NEAT1和EZH2在正常胃黏膜细胞（GES-1）与胃癌细胞（MGC-803、SGC-7901和MKN-28）中的表达量。按转染物的不同将MGC-803和SGC-7901细胞分为hsa-miR-450b-5p-mimic组、mimic-NC组、si-NEAT1组和si-NC组，转染36~48 h后qPCR法验证过表达及敲减效果；通过qPCR、WB法检测观察过表达hsa-miR-450b-5p对细胞中lncRNA NEAT1和EZH2 mRNA、蛋白表达的影响，以及敲减lncRNA NEAT1对hsa-miR-450b-5p和EZH2 mRNA表达的影响；CCK-8法、划痕愈合实验和流式细胞术分别检测敲减EZH2或敲减lncRNA NEAT1对细胞增殖、迁移和凋亡能力的影响。结果：生物信息学分析筛选获得EZH2上游miRNA和lncRNA为has-miR-450b-5p和lncRNA NEAT1，双荧光素酶报告基因实验验证了两者间存在靶向关系。lncRNA NEAT1和EZH2 mRNA、蛋白在胃癌细胞中均呈高表达（均P<0.05）。与mimic-NC组相比，hsa-miR-450b-5p-mimic组MGC-803、SGC-7901细胞中miR-450b-5p水平均显著升高，而EZH2 mRNA、蛋白和lncRNA NEAT1的表达量均显著降低（P<0.05或P<0.01）；与si-NC组相比，si-NEAT1组MGC-803、SGC-7901细胞中lncRNA NEAT1和EZH2 mRNA的表达量均显著降低（均P<0.01），SGC-7901细胞中hsa-miR-450b-5p表达量显著升高（P<0.05）。敲减EZH2或敲减lncRNA NEAT1后，MGC-803、SGC-7901细胞的增殖、迁移能力均显著降低（均P<0.01）。结论：lncRNA NEAT1 和EZH2在胃癌细胞中均呈高表达，lncRNA NEAT1可通过hsa-miR-450b-5p促进EZH2的表达并提高胃癌MGC-803和SGC-7901细胞的增殖和迁移能力。

Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.

Tumor-associated macrophages (TAMs) are the predominant immune cells in the tumor microenvironment (TME). They have been shown to play an important immunosuppressive role in the development of TME and promote tumor immune escape, growth and metastasis. It is a current research hotspot to regulate the functional polarization of TAMs through trained immunity (metabolic reprogramming, epigenetic remodeling) to affect the occurrence and development of tumors. Therefore, in-depth research in this field not only presents a more comprehensive perspective on the pathogenesis of immune-mediated diseases, but also can provide new strategies for clinical anti-tumor immunotherapy. This paper outlines the origin of TAMs and the phenotypes and mechanisms of TAMs polarization, discusses the mechanisms by which metabolic reprogramming and epigenetic remodeling regulate TAMs, summarizes the regulation of TAMs activation and polarization by them, and provides an overview of the progress in TAMs at the current stage of clinical practice, hoping to provide reference for the development of new immunoprevention and treatment strategies.

OBJECTIVE To design and synthesize novel α-naphthylthiol amino acid ester lysine specific demethylase 1(LSD1) inhibitors, evaluate their inhibitory activity with selectivity against LSD1, and explore their binding mechanism through molecular docking and dynamics simulation. METHODS Based on the binding mode of hit compound 3a with LSD1, the α- naphthyl mercapto amino acid ethyl ester small molecule compound were designed by fixing the planar hydrophobic naphthyl ring in the structure, while introducing hydrophilic amino fragment, and they were prepared through a multi-component one-pot cascade reaction. All the compounds were evaluated for their inhibitory activity against LSD1 at concentrations of 5.0 and 1.0 μmol·L–1 using the LSD1 screening platform of research group. The most potent compound was tested for its IC50 value and enzyme selectivity over MAO-A and MAO-B, and its binding mode was investigated through molecular docking and dynamics simulation. RESULTS A total of 13 compounds were obtained, all of which exhibited significant inhibitory effects on LSD1. Among them, nine compounds showed an inhibitory rate of over 50.0% against LSD1 at a concentration of 1.0 μmol·L–1, while compound 3l displaying the best activity with an IC50 value of 0.17 μmol·L–1, 174 times higher than the positive control. It also showed excellent selectivity towards MAO-A and MAO-B. Molecular docking and dynamics simulations indicated that compound 3l inhibited the activity of LSD1 through multiple interactions. CONCLUSION The structures of α-naphthylthiol amino acid ester can serve as lead compounds or active fragments, laying a solid foundation for the subsequent design of LSD1 inhibitors based on structure-oriented drug design.

Retinopathy of prematurity(ROP)is a retinal vascular proliferative disease that occurs in preterm low-birth-weight infants and is the leading cause of blindness in children worldwide.Currently,with the intensive study of ROP,it has been found that retinal neovascularization can spontaneously regress at any time in the early stages of the disease.Therefore,early screening,diagnosis and intervention are particularly important in children with ROP.However,the exist-ing screening methods have problems such as long screening times and poor cooperation of infants and children,which may lead to missed diagnoses,thus more reliable prediction methods are needed.This article briefly describes the screening and prediction methods for ROP at home and abroad in recent years,and systematically analyzes the predictive value of different types of biomarkers for ROP,with a view to providing a basis for establishing a reasonable prediction model for ROP.

The symptoms of pulmonary nodules are insidious,with inflammatory nodules,inflammatory granuloma,early invasive cancer and lung cancer,and the clinical differential diagnosis is still difficult.Regular CT follow-up observation of most pulmonary nodules provides a"window period"for TCM Intervention in pulmonary nodules.From the aspects of external cold attacking the lung,dense cold and humid geographical environment,cold diet,summer air conditioning,etc.,this paper considers that the soaking of cold pathogenic factors is the basic cause of the formation of pulmonary nodules,and cold phlegm are the basic pathogenesis of pulmonary nodules.The clinical manifestations of cold phlegm in pulmonary nodules are summarized from the two actual situations that can be distinguished from clinical symptoms and no symptoms.It is proposed that Mahuang Fuzi Xixin Decoction and Sanzi Yangqin decoction are the basic formulas,Discussion on the treatment of pulmonary nodules by warming yang and dispelling cold to cure the root cause,eliminating phlegm and softening hard mass to treat the symptoms;Improve the ability of TCM diagnosis and treatment of pulmonary nodules.

Objective : To investigate the effect of lncRNA SNHG16 (SNHG16) on the proliferation and migration of gastric cancer cells and its molecular regulatory mechanism. Methods : The expression of SNHG16 in gastric cancer tissue was retrieved based on online database and the downstream target gene of SNHG16 was screened. The interaction between SNHG16 and miR⁃195 ⁃5p was verified by bioinformatics analysis and double luciferase reporter gene experiment. The expression of SNHG16 , miR⁃195 ⁃5p and MYB in gastric cancer cells (HGC⁃27 , MKN⁃28) was detected by real⁃time fluorescent quantitative PCR (qRT⁃PCR) ; SNHG16 was knocked down to detect the expression of miR⁃195 ⁃5p or MYB ; The expression of MYB was detected by overexpression of miR⁃195 ⁃5p;Western blot analysis was used to detect the protein expression level of MYB in each group. Knock down SNHG16 or overexpress miR⁃195 ⁃5p , and the proliferation and migration ability of gastric cancer cells (HGC⁃27 , MKN⁃28) were detected through cell proliferation and scratch test. Results : The expression of LncRNA SNHG16 increased in gastricancer tissues and gastric cancer cells (HGC⁃27 , MKN⁃28 , MKN⁃45 , NCI⁃N87) . The results of double luciferase reporter gene experiment showed that psiCHECK2⁃SNHG16⁃WT and miR⁃195⁃5p mimic were transfected into HGC⁃27 at the same time , significantly inhibiting the luciferase activity of HGC⁃27 cells. The results of qRT PCR and WB experiments showed that knocking down SNHG16 in HGC⁃27 and MKN⁃28 cells upregulated miR⁃195⁃5p and inhibited the expression of MYB at the transcription and translation levels ; Overexpression of miR⁃195 ⁃5p in HGC⁃27 and MKN⁃28 cells inhibited MYB expression. The results of CCK8 proliferation test and cell scratch test showed that knocking down SNHG16 or overexpressing miR⁃195 ⁃5p could inhibit the proliferation and migration of HGC⁃27 and MKN⁃28 cells. Conclusion LncRNA SNHG16 can regulate the expression of MYB in gastric cancer cells through miR⁃195 ⁃5p , and the high expression of SNHG16 can promote the proliferation and migration of gastric cancer cells.

Diabetes mellitus （DM） is a metabolic disease mainly characterized by chronic hyperglycemia and has multiple etiologies. The complications of DM， such as coronary atherosclerosis， nephropathy， foot disease and cardiac dysfunction， have high morbidity， disability rate and mortality. DM and its complications have a long course of disease and are easy to relapse， which are difficult to be cured， seriously affecting people's life and health. NOD-like receptor pyrin domain-containing 3 （NLRP3） inflammasome is an important component of inflammatory response and innate immune system. The inflammatory cascade induced by NLRP3 activation is involved in the occurrence and development of DM as well as its complications by releasing inflammatory factors， damaging endothelial cells and affecting metabolic stress. Therefore， as the core of the inflammatory response， NLRP3 may provide a new target for the treatment of DM and its complications. Traditional Chinese medicine plays a key role in the treatment of DM and its complications， and has a regulatory effect on NLRP3. Thus it has become a novel research strategy to prevent and treat DM and its complications via modulating NLRP3. However， at present， there are relatively scattered reports and a lack of systematic review on the role of traditional Chinese medicine in the treatment of DM and its complications from the perspective of NLRP3. As a result， this paper reviewed domestic and foreign literature in recent years and conducted the discussion from two aspects： the influence of NLRP3 on the occurrence and development of DM and its complications， and the progress of traditional Chinese medicine in intervening in DM and its complications through NLRP3. This paper provided reference for the research on the regulation of NLRP3 and a new direction for the treatment of DM and its complications.

Objective:To investigate the molecular mechanism of VRC01 resistance in HIV-1 subtype B′ strains isolated from a patient (DRVI01) with broadly neutralizing antibody (bNAb).Methods:Sequences of the HIV-1 subtype B′ strains isolated from patient DRVI01 were compared with those of HIV-1 subtype B′ strains that were isolated at the same time but sensitive to VRC01 antibody. Key amino acids that might affect the neutralization of VRC01 were selected according to literature reports. Effects of the selected amino acids on VRC01 neutralization were verified by site-directed mutation and sequence exchange of membrane proteins from different patients.Results:Single-point mutations of E279D and R282K in LoopD region and N460A and N463Q in V5 region reversed the viral sensitivity to VRC01 neutralization. Combined mutations in two or three above-mentioned sites significantly increased the viral sensitivity to VRC01 antibody compared with single-point mutations. Contrary to literature reports, the glycosylation site mutation of N276 had no influence on the viral sensitivity to VRC01.Conclusions:HIV-1 subtype B′ strains isolated from patient DRV01 with bNAb carried the mutations of D279 and K282 in LoopD region and N460 and N463 in V5 region, resulting in resistance to VRC01 antibody.

Objective:To explore the application effect of the caregiver self-management support project in main caregivers of adolescents with epilepsy.Methods:Using the convenient sampling method, the main caregivers of adolescents with epilepsy who were enrolled before implementation of the caregiver self-management support project of the First Affiliated Hospital of Zhengzhou University from January to August 2019 were included in the control group ( n=101) , and they were given routine follow-up. The main caregivers of adolescents with epilepsy who were enrolled after the implementation of the project from September 2019 to June 2020 were included in the study group ( n=101) . Before and after the intervention, the Connor-Davidson Resilience Scale (CD-RISC) , Simplified Coping Style Questionnaire (SCSQ) , Zarit Burden Interview (ZBI) , Life Satisfaction Index A (LSIA) and Medical Outcome Study Social Support Survey (MOS-SSS) were used to compare the psychological resilience, coping style, quality of life and social support of caregivers between the two groups. Finally, a total of 98 caregivers in the study group and 97 caregivers in the control group completed the study. Results:After the intervention, the scores of dimensions of CD-RISC, positive coping score of SCSQ, LSIA score and dimensions scores of MOS-SSS of main caregivers in the study group were higher than those in the control group, while the scores of negative coping of SCSQ and ZBI were lower than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The caregiver self-management support project helps to strengthen the psychological resilience of the main caregivers of adolescents with epilepsy, enhance their active coping style and social support and improve their quality of life.