AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.

Objective To investigate the expression of the ubiquitination enzyme UBE2S in different cell types in hepatocellular carcinoma(HCC)microenvironment and its impact on proliferation and stemness of HCC cells.Methods TCGA and CPTAC database were used to analyze the transcriptional and promoter methylation levels and protein expressions of UBE2S in HCC.Specific expression patterns of UBE2S,intercellular communication and key transcription factors in different cell types were analyzed based on single-cell sequencing data from TISCH website.We further examined UBE2S expressions in clinical samples of HCC tissues,HCC cells and T cells using immunohistochemistry and immunofluorescence staining.We also tested the effects of UBE2S knockdown on stemness of HCC-LM3 and HepG2 cells using clone formation experiments and sphere formation assay.Results Analysis based on TCGA database suggested significant overexpression of UBE2S in both paired and non-paired tumor tissues(P<0.001),and its transcriptional level increased with tumor grades.The methylation level of UBE2S promoter was significantly decreased in HCC(P<0.001),and its transcription level increased obviously in HCC with TP53 mutation(P<0.001).Analysis of CPTAC database also demonstrated overexpression of UBE2S protein in HCC tissues(P<0.001).Three prognostic models suggested that HCC patients with high UBE2S expression had poorer prognosis(P<0.001).Single-cell sequencing data analysis revealed high expressions of UBE2S in T cells and high intensities of interaction between endothelial cells,epithelial cells and fibroblasts in HCC microenvironment.Immunohistochemistry and immunofluorescence staining demonstrated high UBE2S expressions in clinical samples of HCC tissues,HCC cells and T cells.In HCC-LM3 and HepG2 cells,UBE2S knockdown significantly inhibited cell clone formation and tumor sphere formation(P<0.05).Conclusion UBE2S is highly expressed in T cells in HCC microenvironment in close correlation with a poor prognosis.High UBE2S expression promotes the stemness of HCC cells.

Objective:To investigate the effect of decarbromodiphenyl ether(BDE-209)exposure on the migration ability of triple-negative breast cancer(TNBC)cells and to explore the underlying mechanism.Methods:Human TNBC MDA-MB-231 cells were divided into blank control group and BDE-209 exposure groups(treated with 0.02,0.20,2.00,20.00 and 200.00 ng/mL BDE-209 in high glucose DMEM).Extracellular vehicles(EVs)secreted by MDA-MB-231 cells were isolated by differential ultracentrifugation.Transmission electron microscopy(SEM),nanoparticle tracking analysis(NTA)and Western blotting were performed to characterize the EVs.The effect of the EVs induced by BDE-209 exposure(EVs-BDE-209)on the migration and invasion of MDA-MB-231 cells was detected by wound-healing assay and Transwell test.qRT-PCR was used to measure the miR-221 level in EVs-BDE-209.The expression of MMP9 in MDA-MB-231 cells was determined by Western blotting.Results:Compared with the blank control,BDE-209 exposure increased the tumor cell-derived EVs in dose-dependent manner.The MDA-MB-231 cells co-cultured with EVs released by 200.00 ng/mL BDE-209 exposure showed an 86%increase in cell migration rate,a 1.32-fold higher number of membrane-penetrating cells,a 2.71-fold higher expression level of miR-221,and a 1.62-fold higher expression level of MMP9 compared with the blank control group(all P<0.05).While transfection with anti-miR-221 antibody to decrease miR-221 level in EVs significantly reversed the increased invasion ability of the MDA-MB-231 cells treated with EVs-BDE-209.Conclusion:BDE-209 exposure may promote metastasis potential of MDA-MB-231 cells via EVs-BDE-209 transmitted miR-221.

Objective To investigate the expression of the ubiquitination enzyme UBE2S in different cell types in hepatocellular carcinoma(HCC)microenvironment and its impact on proliferation and stemness of HCC cells.Methods TCGA and CPTAC database were used to analyze the transcriptional and promoter methylation levels and protein expressions of UBE2S in HCC.Specific expression patterns of UBE2S,intercellular communication and key transcription factors in different cell types were analyzed based on single-cell sequencing data from TISCH website.We further examined UBE2S expressions in clinical samples of HCC tissues,HCC cells and T cells using immunohistochemistry and immunofluorescence staining.We also tested the effects of UBE2S knockdown on stemness of HCC-LM3 and HepG2 cells using clone formation experiments and sphere formation assay.Results Analysis based on TCGA database suggested significant overexpression of UBE2S in both paired and non-paired tumor tissues(P<0.001),and its transcriptional level increased with tumor grades.The methylation level of UBE2S promoter was significantly decreased in HCC(P<0.001),and its transcription level increased obviously in HCC with TP53 mutation(P<0.001).Analysis of CPTAC database also demonstrated overexpression of UBE2S protein in HCC tissues(P<0.001).Three prognostic models suggested that HCC patients with high UBE2S expression had poorer prognosis(P<0.001).Single-cell sequencing data analysis revealed high expressions of UBE2S in T cells and high intensities of interaction between endothelial cells,epithelial cells and fibroblasts in HCC microenvironment.Immunohistochemistry and immunofluorescence staining demonstrated high UBE2S expressions in clinical samples of HCC tissues,HCC cells and T cells.In HCC-LM3 and HepG2 cells,UBE2S knockdown significantly inhibited cell clone formation and tumor sphere formation(P<0.05).Conclusion UBE2S is highly expressed in T cells in HCC microenvironment in close correlation with a poor prognosis.High UBE2S expression promotes the stemness of HCC cells.

Objective:To evaluate the clinical effect of microbotox injection of botulinum toxin type A in improving the fine lines of lower eyelid skin and achieving the rejuvenation of the eye area.Methods:The clinical data of patients with mildly sagging lower eyelid skin and fine wrinkles admitted to Facial and Neck Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College; Deparment of Plastic and Cosmetic Surgery, Guangyuan Central Hospital; Department of Dermatology, People’s Hospital of Qingshen County; Department of Plastic and Cosmetic Surgery, Dujiangyan People’s Hospital; Devision of Women’s Healthcare, Panzhihua Maternal and Child Hospital; Chengdu Badachu Cosmetic Hospital from August 2022 to January 2023 were analyzed retrospectively. Using the microbotox injection method, the botulinum toxin type A with a diluted concentration of 8 U/ml was micro-injected into the dermis layer or between the dermis and the superficial muscle layer at an interval of 0.6 cm, and a single point injection of 0.2 U. The injection range was the anatomical distribution range of the orbicularis oculi muscle. After treatment, patients were followed up to investigate patient satisfaction, which was divided into very satisfied, satisfied, dissatisfied and very dissatisfied, and to calculate subjective satisfaction (percentage of the number of very satisfied and satisfied cases in the total number of cases). At the same time, a independent physician used the global aesthetic improvement scale (GAIS) to evaluate the effect based on the patient’s photos before treatment and during follow-up. 5 indicates as worse than before, 4 as no improvement, 3 as were slight improvement, 2 as moderate improvement, and 1 as significant improvement. SPSS 24.0 software was used for descriptive analysis of the data.Results:A total of 23 patients were enrolled, including 1 male and 22 females, aged (36.4±2.5) years old. The follow-up was 1-6 months, with an average of 1.4 months, and the subjective satisfaction of patients was 100% (23/23). The result of the independent physician evaluation were as follows: the overall cosmetic improvement score of the observers was 1.26±0.54, 100% (23/23) of the patients reached the standard of obvious or moderate improvement. In terms of complications, 2 patients presented local epidermis bruises after injection, who recovered within 1 week; 1 patient complained of pain after injection, which was significantly relieved 12 hours later. No ectropion, symblepharon, facial palsy, dry eyes, or other complications were observed.Conclusion:Application of botulinum toxin type A microbotox injection can improve periorbital wrinkles and improve skin texture at the same time, and the therapeutic effect is definite.

Objective:To evaluate the clinical effect of microbotox injection of botulinum toxin type A in improving the fine lines of lower eyelid skin and achieving the rejuvenation of the eye area.Methods:The clinical data of patients with mildly sagging lower eyelid skin and fine wrinkles admitted to Facial and Neck Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College; Deparment of Plastic and Cosmetic Surgery, Guangyuan Central Hospital; Department of Dermatology, People’s Hospital of Qingshen County; Department of Plastic and Cosmetic Surgery, Dujiangyan People’s Hospital; Devision of Women’s Healthcare, Panzhihua Maternal and Child Hospital; Chengdu Badachu Cosmetic Hospital from August 2022 to January 2023 were analyzed retrospectively. Using the microbotox injection method, the botulinum toxin type A with a diluted concentration of 8 U/ml was micro-injected into the dermis layer or between the dermis and the superficial muscle layer at an interval of 0.6 cm, and a single point injection of 0.2 U. The injection range was the anatomical distribution range of the orbicularis oculi muscle. After treatment, patients were followed up to investigate patient satisfaction, which was divided into very satisfied, satisfied, dissatisfied and very dissatisfied, and to calculate subjective satisfaction (percentage of the number of very satisfied and satisfied cases in the total number of cases). At the same time, a independent physician used the global aesthetic improvement scale (GAIS) to evaluate the effect based on the patient’s photos before treatment and during follow-up. 5 indicates as worse than before, 4 as no improvement, 3 as were slight improvement, 2 as moderate improvement, and 1 as significant improvement. SPSS 24.0 software was used for descriptive analysis of the data.Results:A total of 23 patients were enrolled, including 1 male and 22 females, aged (36.4±2.5) years old. The follow-up was 1-6 months, with an average of 1.4 months, and the subjective satisfaction of patients was 100% (23/23). The result of the independent physician evaluation were as follows: the overall cosmetic improvement score of the observers was 1.26±0.54, 100% (23/23) of the patients reached the standard of obvious or moderate improvement. In terms of complications, 2 patients presented local epidermis bruises after injection, who recovered within 1 week; 1 patient complained of pain after injection, which was significantly relieved 12 hours later. No ectropion, symblepharon, facial palsy, dry eyes, or other complications were observed.Conclusion:Application of botulinum toxin type A microbotox injection can improve periorbital wrinkles and improve skin texture at the same time, and the therapeutic effect is definite.

AIM: To explore whether Agkistrodon Halys venom antitumor component-I (AHVAC-I) affects the migration of gastric cancer cells by human primary gastric cancer-associated fibroblast (GCAFs). METHODS: Tissue block culture and trypsin digestion were used to separate and culture human primary gastric cancer-associated fibroblasts (GCAFs); the GCAFs-CM

Objective To investigate the effect of different treatment regimens guided by magnetic resonance angiography (MRA) and diffusion weighted imaging (DWI) mismatch on the outcomes of patients with mild ischemic stroke caused by acute middle cerebral artery (MCA) M1 segment occlusion. Methods From January 2013 to February 2018, the clinical data of patients with mild ischemic stroke caused by acute MCA M1 segment occlusion and admitted to the Department of Neurology, the Affiliated Hospital of Yangzhou University were analyzed retrospectively. Mild stroke was defined as the National Institutes of Health Stroke Scale (NIHSS) score ≤5, and the MRA-DWI mismatch was defined as MCA M1 segment occlusion confirmed by MRA and the DWI-Alberta Stroke Program Early Computed Tomography Score ≥6. According to the clinical decision, they were divided into endovascular treatment group and intravenous thrombolytic therapy group. The primary outcome measure was the modified Rankin Scale score at 90 days after onset, ≤2 was defined as good outcome. The secondary outcome measure was the incidence of symptomatic intracranial hemorrhage (sICH) within 7 days after treatment and the mortality rate at 90 d. Multivariate logistic regression analysis was used to determine the independent effects of different treatment regimens on outcomes. Results A total of 38 patients were enrolled, 19 (50. 00％) in the intravenous thrombolytic therapy group, and 19 in the endovascular treatment group (50. 00％, including 5 patients with intratracheal thrombectomy after intravenous thrombolysis); 27 patients had good outcomes (71. 05％) and 11 had poor outcomes (28. 95％). Except for total cholesterol level, there were no significant differences in demography, vascular risk factors, and all baseline clinical data between the endovascular treatment group and the intravenous thrombolytic therapy group. The rate of good outcome in the endovascular treatment group was significantly higher than that in the intravenous thrombolytic therapy group (89. 47％ vs. 2. 63％; P = 0. 029), and there was no significant difference between the incidence of sICH within 7 days (15. 79％ vs. 5. 26％; P = 0. 604) and 90-day mortality (0％ vs. 10. 53％; P = 0. 486). The proportion of patients who underwent endovascular treatment in the good outcome group was significantly higher than that in the poor outcome group (62. 96％ vs. 18. 18％; P = 0. 029). Multivariate logistic regression analysis showed that endovascular treatment was an independent predictor of good outcome (odds ratio 0. 103, 95％ confidence interval 0. 015-0. 714; P = 0. 021). Conclusion Endovascular treatment is an independent predictor of good outcome in patients with mild ischemic stroke caused by acute MCA M1 segment occlusion.

Objective To explore the set-up errors of position fixation with simple perforated foam pad in colorectal cancer radiotherapy and their causes as well as the improvement measures.Methods Sixty cases of color-ectal cancer patients undergoing radiotherapy were collected.Prone position was adopted with the position fixed by perforated foam pad and membrane.CBCT imaging was taken weekly before treatment,setup errors were corrected if necessary.Errors throughout the course of treatment for each patient were recorded and compared between patients, followed by analyzing the reasons of errors.Results Before calibration,the maximum errors on the direction of left and right (X-axis),front and back (Z),upside and downside (Y)in these 60 patients were 0.5cm,0.9cm and 0.7cm respectively with the average errors of (0.22 ±0.03)cm,(0.38 ±0.03)cm and (0.27 ±0.04)cm respec-tively.Conclusion The method of applying perforated foam pad and thermoplastic mask in colorectal cancer radio-therapy is currently the most commonly used in the clinical position fixation techniques,by which the small intestine, bladder and other pelvic tissues can be well protected.Due to poor comfort prone position,it is necessary to further improve perforated foam pad through improving the quality of the foam pad,thus improving patients comfort and posi-tioning repeatability and reducing position fixation errors.

Purpose To investigate the clinicopathlogic characteristic, diagnosis and histogenesis of pancreatic solid-pseudopapillary tumors ( SPPT) . Methods Combined with relevant literature, the clinical history, histopathological features and immunohistochemi-cal characteristics were analyzed in 11 cases of SPPT. Results There were 10 female patients and only 1 male in total 11 cases, aged from1 7 to 60 years (mean 33). The sizes of tumors were from 3. 2 to 10. 0 cm. Histologically, they were composed of papillary and microcysticsolid structures. Pseudopapillary with a fibrovascular core was remarkable. Immunohistochemically, the tumors expressed EMA (1/11), vimentin (10/11), NSE (11/11), Syn (7/11), CgA (1/11), CD56 (11/11), CD10 (11/11), PR (9/11), CD99 (9/11),α-AT (11/11),β-catenin (11/11), E-cadherin (11/11), Cyclin D1 (11/11), c-Myc (11/11). 6 patients were followed up for a period of 20 to 112 months, and they were all alive and had no recurrence and metastasis. Conclusions SPPT is a tumor with low malignancy of the pancreas that most frequently affect young females. SPPT may be derived from multipotent stem cells and closely related withβ-catenin signaling pathway. Pathological morphology and immunohistochemistry are very important to the diag-nosis and differential diagnosis of SPPT.