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.

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 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.

Purpose To discuss the clinical pathological features, diagnosis and differential diagnosis, treatment and prognosis of pri-mary breast diffuse large B-cell lymphoma ( DLBCL) . Methods 7 cases of primary breast DLBCL were collected, their clinical path-ological characteristics and immunophenotypes were also observed, and the treatment and prognostic factors were discussed. Results All of the 7 patients were women, aged 28~75 years, with the median 51 years. 5 cases involved left breast, 2 cases were located in the right breast. Painless mammary mass and ipsilateral axillary lymphadenopathy were the commonest clinical manifestations. On mi-croscopic observation, tumor cells were large to medium-sized which characterized as diffuse infiltration between the lobules of mamma-ry gland, around the duct, interstitial and fat tissue, some were single file cord pattern. 1 case was ALK-1 positive DLBCL, 6 cases were DLBCL of the non special type. The immunophenotype showed 5 cases were of non-GCB, 2 cases were of GCB type. Ki-67 index were from 60% to 95%. According to Ann Arbor staging, 2 cases were stage I EA, 5 cases were stage II EA. IPI score:4 cases with 0 score, 3 cases with 1 score. Of 7 patients, one case was modified radical mastectomy, 4 cases were lumpectomy, 2 cases were diag-nosed by core needle biopsy. 2 patients died without chemotherapy, 5 patients were chemotherapy or radiotherapy for 4 to 6 cycles ( CHOP/R-CHOP) , 4 patients received complete response, one died. Patients were followed up from 1 to 114 months, the median fol-low-up time was 18 months. The 1 year OS rate was 57. 1%, 5 years OS rate was 14. 3%. Conclusion The diagnosis of primary breast DLBCL is confirmed by pathological biopsy and immunohistochemical markers. The immunophenotype was mainly non-GCB type. Comprehensive treatment including chemotherapy and radiotherapy is appropriate. The prognosis should be comprehensively eval-uated by multiple factors.

Objective To identify the specific genotype and analyze clinical features of spinocerebellar ataxias (SCAs) pedigree in the region of Yunnan Province.Methods Fourteen SCAs pedigrees and 183 blood samples of the family members were collected between January 2011 and July 2014 from Department of Neurology,First Affiliated Hospital of Kunming Medical College.Polymerase chain reaction (PCR) amplification,agarose gel electrophoresis and DNA sequencing technologies were utilized to identify the specific genotype of SCAs pedigree.Presymptomatic tests were carried out and the clinical features and genetic test results of patients were carefully analyzed.Results SCA3 was the most common subtype of SCAs in the Han nationality of Yunnan region.Nine of the 14 families were SCA3,only one family was SCA2.Additionally,there were four SCAs families that remained indeterminate.The patients with di-allele mutations (46/77) of SCA3 gene had early onset,rapid progression and serious clinical symptoms.Hereditary SCA3 and autonomic dominant polycystic kidney disease can happen simultaneously in a family.The proband SCA3 gene' s CAG repeat number is 28/76,and repetitions of the mutation allele are in all range.The PKD1 gene exon 23 is found to be in abnormal sequence.Conclusions SCA3 is the most common subtype of SCAs in the Han population of Yunnan region.There are 15/46 incomplete penetrance nutation and 46/77 di-allele mutations.It is possible that di-allele mutations make the disease worse and accelerate clinical course progression.SCA3 and polycystic kidney disease can uncommonly happen simultaneously in a family,which perhaps suggests there are interactions between the two disease-virulence genes.