Objective:To study the antitumor activity of zeylenone in colorectal cancer and its related regulatory mechanism.Methods:Human colorectal cancer cells(DLD-1 and HCT116)and control cell(HcoEpic)were treated with different concentrations of zeylenone(0,2.5,5,10 and 20 μmol/L)for 48 h.Cell viability of DLD-1,HCT116 and HcoEpic were determined by CCK-8 kit.TUNEL staining was used to evaluate cell apoptosis.Caspase-3 activity and LDH level were measured by ELISA.Relative levels of M1 and M2 polarization markers,p-PI3K/PI3K and p-AKT/AKT were detected by Western blot and qRT-PCR.DLD-1 cells were subcuta-neously injected into the armpit of nude mice to establish a mouse xenograft tumor model.Intraperitoneal dose of zeylenone given to nude mice was 30 mg/kg,administered once every two days.After 2 weeks,the effect of zeylenone on growth of colorectal cancer tumors was detected.Results:Zeylenone inhibited cell activity and promoted cell apoptosis in colorectal cancer cells.Additionally,zeylenone stimulated M1-polarization and inhibited M2-polarization of tumor-associated macrophages(P<0.05).PI3K/AKT signaling pathway was inhibited by zeylenone in colorectal cancer cells(P<0.05).PI3K/AKT signaling pathway activator(740 Y-P)attenuated the effect of zeylenone on colorectal cancer cells.In mouse xenograft model,zeylenone inhibited the growth of colorectal cancer tumors(P<0.05).Conclusion:Zeylenone can inhibit colorectal cancer cell activity,promote colorectal cancer cell apoptosis,and promote M1 po-larization of tumor-associated macrophages by regulating PI3K/AKT signaling pathway,ultimately inhibiting the progression of colorec-tal cancer.

Objective:To investigate differences in bacterial and fungal microbiome between infected nails and healthy nails among patients with onychomycosis.Methods:Nail scraping samples were collected from infected nails and healthy nails of 31 patients with onychomycosis, who visited Dalian Dermatosis Hospital from August 2020 to July 2021. The total DNA of nail microbiota was extracted, and the V3-V4 regions of the bacterial 16S rDNA gene and the fungal internal transcribed spacer (ITS) region were amplified and sequenced using Illumina technology. The USEARCH and mothur softwares were used for data cluster analysis to obtain the operational taxonomic units (OTUs) , Wilcoxon rank sum test was used to analyze α diversity, analysis of similarities (ANOSIM) was performed to analyze β diversity, linear discriminant analysis of effect size (LEfSe) was performed to evaluate the species difference.Results:Among the 31 patients with onychomycosis, 16 were males and 15 were females. According to the age, they were divided into young group (18 - 35 years old, 10 cases) , middle-aged group (36 - 60 years old, 11 cases) , and elderly group (over 60 years old, 10 cases) . As the α-diversity analysis revealed, the infected nail group showed significantly decreased Shannon index ( W = 290, P = 0.007) , but significantly increased Simpson index ( W = 663, P = 0.010) compared with the healthy nail group, suggesting that the diversity and evenness of bacterial communities were lower in the infected nail group than in the healthy nail group; however, there was no significant difference in the diversity of fungal communities between the infected nail group and healthy nail group. The β-diversity analysis based on the unweighted-UniFrac distance matrix showed no significant difference in the fungal or bacterial community composition between the infected nail group and healthy nail group (bacterial communities: R = 0.0052, P = 0.331; fungal communities: R = 0.0036, P = 0.337) ; the β-diversity analysis based on the weighted-UniFrac distance matrix showed significant differences in the abundance of bacterial and fungal communities between the two groups (both P = 0.001) . In terms of the species composition, the bacterial flora with significantly decreased abundance in the infected nail group compared with the healthy nail group included Bacteroidetes, Proteobacteria, Betaproteobacteria, Burkholderiales, Ralstonia, Sphingomonas and Streptococcus, while the abundance of Bacilli, Bacillales and Staphylococcus was significantly higher in the infected nail group than in the healthy nail group. Compared with the healthy nail group, the fungal flora with significantly increased abundance in the infected nail group included Eurotiomycetes, Onygenales, Leotiomycetes-ord-incertae-sedis, Arthrodermataceae, Periconia, Erysiphe, Tilletiopsis, Trichophyton, Erysiphe cruciferarum, Trichophyton rubrum, Malassezia sympodialis, while the abundance of Saccharomycetes, Saccharomycetales, Saccharomycetaceae, Dothioraceae, Candida and Alternaria was significantly lower in the infected nail group than in the healthy nail group. Conclusion:The diversity and abundance of bacterial communities significantly differed between infected nails and healthy nails among patients with onychomycosis, while only the abundance of fungal communities differed between the two groups, and perhaps there was correlations between some bacterial and fungal communities.

Objective:To evaluate the safety and efficacy of endoscopic resection in the treatment of patients with primary non-ampullary duodenal early cancer.Methods:A total of 20 cases with primary non-ampullary duodenal early cancer receiving endoscopic resection were collected from Jan 2015 to Dec 2019 at the Department of Endoscopy, the First Affiliated Hospital of Shandong First Medical University.Results:The size of lesions ranged from 0.3-2.5 cm (0.9±0.5)cm.The size of removed membrane samples ranged from 1.5-3.5 cm (2.5±0.7)cm. The edges were all negatiue pathologically. Duodenal perforation occurred in 2 cases, and all were successfully clipped by endoscopy. The follow-up time was from 4-42 months (20.4±11.4)m and no recurrence was found.Conclusion:Endoscopic resection is a safe and effective method for primary non-ampullary duodenal early cancer.

BACKGROUND: Targeted therapy for patients with driver genes positive and immunotherapy for patients with driver gene-negative but high programmed death ligand 1 (PD-L1) expression are the standards of first-line treatment for patients with advanced non-small cell lung cancer (NSCLC). The treatment options for patients with driver gene positive and high PD-L1 expression are still worth exploring. METHODS: The characteristics of 315 patients with NSCLC were identified to analyze the clinicopathological characteristics of patients with driver gene positive and high PD-L1 expression, and the efficacy of targeted therapy. RESULTS: Among the 315 patients, the total positive rate of driver genes was 62.2%, and the high PD-L1 expression rate (≥50.0%) was 11.2%. The proportion of patients with driver gene positive and high PD-L1 expression was 10.7%. PD-L1 was highly expressed in patients with epidermal growth factor receptor (EGFR) mutation, KRAS mutation, ALK fusion, BRAF mutation, and MET 14 exon skip mutation, the proportions were 7.8% (11/141), 18.2% (4/22), and 23.1%, (3/13), 50.0% (2/4) and 100.0% (1/1) respectively. EGFR mutation positive with PD-L1 high expression was mainly in patients with stage IV lung adenocarcinoma. KRAS mutation positive with PD-L1 high expression was mainly in patients with a history of smoking. Among them, two patients were followed in detail for targeted therapy, who with ALK fusion-positive and PD-L1 high expression (90.0%), EGFR L858R mutation and PD-L1 high expression (70.0%) respectively. The total OS of the patients was 5 months, 2 months. CONCLUSIONS The high PD-L1 expression rate in NSCLC patients with different driver gene mutations was variable, which maybe correlated with distinct clinicopathological characteristics. Patients with sensitive mutations and high PD-L1 expression may be less benefit from targeted therapy and have poor prognosis.

BACKGROUND: Immunotherapy represented by immune checkpoint inhibitors (ICIs) has been widely used in the treatment of lung cancer. There are controversies in clinical practice for patients with advanced non-small cell lung cancer (NSCLC) and high programmed cell death-ligand 1 (PD-L1) expression receiving ICIs monotherapy or combination chemotherapy. METHODS: This study retrospectively analyzed the clinical data of 49 patients with advanced NSCLC and high PD-L1 expression. Immunohistochemistry was performed with 22C3 antibody, and the expression level of PD-L1 was evaluated according to tumor proportion score (TPS). Objective response rate (ORR) and progression free survival (PFS) were compared by groups of different clinical characteristics. RESULTS: ORR of monotherapy and combination therapy group was 47.1% (8/17) and 43.8% (14/32), respectively, without statistical difference (P=0.825). The median PFS of monotherapy and combination therapy group was 8.0 months and 6.8 months, respectively, without statistical difference (P=0.502). Statistical analysis of predictors of immunotherapy for the patients showed first-line immunotherapy had better ORR than subsequent immunotherapy (12/19, 63.2% vs 10/30, 33.3%, P=0.041), however no difference in PFS. And there were no differences in ORR or PFS among groups of age, gender, smoking status, performance status (PS), pathological type, tumor size and tumor-node-metastasis (TNM) stage. CONCLUSIONS The therapeutic effect is similar between ICIs monotherapy and combination chemotherapy for patients with advanced NSCLC and high PD-L1 expression. ORR of first-line immunotherapy was better in patients with advanced NSCLC and high PD-L1 expression. The optimal treatment for this population remains further prospective clinical studies.

BACKGROUND: Immune checkpoint inhibitor monotherapy is reported to have little effect in advanced non-small cell lung cancer (NSCLC) patients with driver oncogenes. However, recent studies have shown that some patients with driver genes are still benefit from combination immunotherapy after tyrosine kinase inhibitors (TKIs) drug resistance. The purpose of this study was to analyze the efficacy of posterior line immunotherapy in NSCLC patients with epidermal growth factor (EGFR) sensitive mutation, and to evaluate the value of immunotherapy in posterior line therapy in patients with advanced EGFR mutation. METHODS: A total of 27 patients with EGFR mutation diagnosed in Beijing Chest Hospital, Capital Medical University from June 2018 to November 2020 were collected. After the progress of targeted therapy, they had received programmed cell death protein 1 (PD-1) checkpoint inhibitor combined with chemotherapy and anti-angiogenic drug therapy. RESULTS: Of the 27 advanced NSCLC patients, 19 cases (70.4%) did not have T790M mutation. There were 8 cases (29.6%) with T790M point mutation. The total objective response rate (ORR) was 40.7%. Kaplan-Meier survival analysis showed that there was no statistically significant difference among different EGFR mutations (χ²=4.15, P=0.230). But progression-free survival (PFS) was significantly longer in patients without T790M mutation than in patients with T790M mutation (9.2 mon vs 3.3 mon, χ²=2.808, P=0.041), and the same trend was observed in patients with overall survival treated with the PD-1 inhibitor (12.2 mon vs 7.3 mon, χ²=3.22, P=0.062). ORR of patients without T790M was significantly better than that with T790M (52.63% vs 12.5%, P=0.045). CONCLUSIONS Patients with EGFR mutation can benefit from later-line combined immunotherapy. The patients with T790M mutation in the population of EGFR mutation had the worst effect of immunotherapy in the later line. Therefore, the follow-up treatment and whole-course management of these patients need to explore better treatment strategies to improve the benefit.

Objective:To evaluate the sternocleidomastoid muscle hardness in patients with tension-type headache (TTH) by acoustic radiation force impulse (ARFI), and to explore its clinical value in evaluating TTH.Methods:Thirty-five patients with TTH admitted to our hospital from March 2019 to March 2020 and 35 subjects accepted physical examination at the same time period were enrolled in this study. The thicknesses of sternocleidomastoid muscles were measured by two-dimensional ultrasound. On this basis, shear wave velocity (SWV) of sternocleidomastoid muscles was quantitatively evaluated by ARFI. The correlation between thicknesses of sternocleidomastoid muscles and headache was further analyzed.Results:The thickness of sternocleidomastoid muscles, SWV in transverse and longitudinal sections, visual analogue scale (VAS) scores in patients with TTH during headache and after massage were significantly higher than those in the heathy subjects ( P<0.05). Meanwhile, SWV in transverse and longitudinal sections, VAS scores decreased significantly in TTH after massage, which showed significant difference as compared with those during headache ( P<0.05). During headache, the thickness of sternocleidomastoid muscles was negatively correlated with VAS scores ( r=-0.397, P=0.002), SWV in longitudinal section of sternocleidomastoid muscles was positively correlated with VAS scores ( r=0.643, P=0.000). Conclusions:The thickness and hardness of sternocleidomastoid muscle are closely related with TTH. ARFI can quantitatively evaluate the thickness and hardness of sternocleidomastoid muscle.

Objective: To explore the molecular mechanism of miR-195-5p targeting FGF2 to inhibit the proliferation, apoptosis, invasion and migration of endometrial cancer HEC-1B cells. Methods: After culture and transfection, HEC-1B cells were divided into 4 groups: HEC-1B group, miR-195-5p mimic group, pLV-FGF2 group and miR-195-5p+FGF2 group. The expressions of miR-195-5p and mRNA levels of FGF2 were detected by qRT-PCR. The targeted relationship of miR-195-5p and FGF2 was verified by luciferase assay. The protein expression of FGF2 was examined by Western blotting; Proliferation of HEC-1B cells was measured by CCK-8; Apoptosis was tested by flow cytometry; HEC-1B cell invasion was detected by transwell, and migration was measured by scratch assay. Results: Compared with HEC-1B group, the expression of miR-195-5p in miR-195-5p mimic group was elevated while FGF2 mRNA level was declined (all P<0.01). Luciferase assay indicated that FGF2 was a target of miR-195-5p. Compared with HEC-1B group, the protein level of FGF2 in miR-195-5p mimic group was decreased, and the protein levels of FGF2 in pLV-FGF2 group were enhanced (P<0.01). The protein levels of FGF2 in miR-195-5p+FGF2 group were lower than that in pLV-FGF2 group (all P<0.01). The proliferation in miR-195-5p mimic group was lower than HEC-1B group (P<0.01), while the proliferation in pLV-FGF2 group was higher than that in HEC-1B group (all P<0.01). Compared with HEC-1B group, apoptosis in miR-195-5p mimic group was increased, and apoptosis in pLV-FGF2 group was decreased (P<0.01); moreover, apoptosis in miR-195-5p+FGF2 group was higher than that in pLV-FGF2 group (P<0.01). Compared with HEC-1B group, the number of invasive cells per field and the rate of wound healing in miR195-5p mimic group were decreased, while those in pLV-FGF2 group was enhanced (P<0.01); moreover, the number of invasive cells per field and the rate of wound healing in miR-195-5p+FGF2 group was lower than those in pLV-FGF2 group (all P<0.01). Conclusion: miR-195-5p inhibits proliferation, invasion and migration and promotes apoptosis of endometrial cancer HEC-1B cells by targeting FGF2, and could be used as a treatment target of endometrial cancer.

Objective Exploring theoretical teaching model of chronic diseases management of preventive medicine undergraduates,to provide feasible suggestions to improve the theoretical teaching of the chronic disease management ability of preventive medicine specialty in China so that graduates can better adapt to chronic diseases management work.Methods On the basis of reading a large number of relevant literature both at home and abroad,the research team designed questionnaires,and conducted a questionnaire survey on 190 respondents who engaged in chronic diseases management or teaching in central China.The content includes the understanding of the importance of training chronic disease management ability in undergraduate education of preventive medicine and the constitution and training mode of undergraduates' chronic disease management ability.EpiData 3.1 software was used to input survey data,SPSS 23.0 software was used for statistical description analysis,and the usage ratio and component ratio were used for statistical description analysis.Results The survey found that more than 50％ of the respondents believed that training students with chronic disease management should focus on prevention,intervention services and health promotion ability,and chronic disease modules need to be added to undergraduate courses in preventive medicine.Conclusions preventive medicine undergraduates need to be improved,and medical colleges should change teaching model to increase undergraduates' ability of chronic diseases management.

Background and objective Non-small cell lung cancer (NSCLC) has been transformed from the treatment according to histological type to genotype treatment model. The epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) genes are the most important drivers in lung cancer. The aim of this study is to explore the clinical characteristics and prognostic factors of patients with advanced NSCLC with different genotypes. Methods We retrospectively reviewed the clinical data of 553 advanced NSCLC patients with EGFR mutations and ALK positive who were hospitalized in the Beijing Chest Hospital from July 2004 to December 2015, and the independent prognostic factors of pa-tients were analyzed by Cox proportional hazards regression model. Results The clinical data of 553 patients (227 with EGFR mutations, 58 with ALK positive, 2 with EGFR and ALK co-mutation and 266 with wild-type) with advanced NSCLC were enrolled in this study. The median survival time of 227 patients with EGFR mutations was 28.7 mo (95%CI: 22.160-35.240), and the performance status (PS) score (0-1) (HR=4.451; 95%CI: 2.112-9.382; P<0.001) and EGFR-tyrosine kinase inhibitors(TKIs) targeted therapy (HR=2.785; 95%CI: 1.871-4.145; P<0.001) were the independent prognostic factors for the survival of patients harboring EGFR mutations. The median survival time of 58 patients with ALK positive was 15.5 mo (95%CI:10.991-20.009), and treatment with crizotinib (P=0.022) was the independent influence factor for the survival of ALK positive patients. The median survival time of 266 patients with wild-type was 12.1 mo (95%CI: 10.660-13.540), and the PS score (0-1) (HR=2.313; 95%CI: 1.380-3.877; P=0.001) and treatment with chemotherapy (HR=1.911; 95%CI: 1.396-2.616; P<0.001) were the independent prognostic factors for the survival of wild-type patients. Conclusion The prognosis of patients with advanced NSCLC is associated with genetic mutation, and targeted therapy has a improvement on survival for patients with EGFR mutations or ALK rearrangement.