Objective:ANXA2 plays a crucial role in cancer metastasis, but its mechanism is not yet fully understood. Therefore, it is necessary to establish an ANXA2 gene knockout mouse model to provide an effective tool for subsequent studies on ANXA2-related mechanisms.Methods:A gene knockout mouse model was constructed using CRISPR/Cas9 technology. The model was validated through tissue DNA extraction followed by polymerase chain reaction (PCR), sequencing, and western blot to confirm ANXA2 genotype and protein expression. The successfully constructed models were divided into a model group and a wild-type (WT) group for the creation of a mouse tail vein injection Lewis lung carcinoma (LLC) metastasis model. Metastatic foci formation was monitored using in vivo imaging technology, and the survival rates of the two groups were compared. Results:An sgRNA sequence targeting the first exon of ANXA2 was designed, and 16 founder mice were obtained through microinjection. Through consanguineous hybridization, 30 homozygous offspring were ultimately acquired. After establishing the strains of the mouse model, mice were divided into the ANXA2 knockout group and the WT group, with 8 mice in each group. An LLC lung metastasis model was established in both groups. Compared with the WT group, the number of metastatic foci was significantly increased in the ANXA2 knockout group (7 vs. 1), and the fluorescence intensity was stronger in the WT group than in the knockout group ( P=0.002). Using the GEPIA2 database to analyze ANXA2 gene expression in tumor tissues and normal tissues of lung cancer patients, it was found that ANXA2 expression levels were significantly higher in lung cancer tumor tissues compared to normal tissues ( P＜0.05). The database included data from 478 lung cancer patients, and patients were stratified into high-expression and low-expression groups based on ANXA2 levels. Compared to the low-expression group, patients in the high-expression group exhibited significantly shorter disease-free survival and overall survival ( P＜0.05, respectively). The survival time of mice in the ANXA2 knockout group (median survival time, 43 days) was significantly longer compared to the WT group (median survival time, 26 days; P=0.017). Additionally, ANXA2 expression is significantly associated with the prognosis of lung cancer patients ( P=6.4e-14). Conclusions:ANXA2 is closely associated with cancer metastasis and holds potential as a new target for metastasis treatment. Further in-depth research will greatly facilitate the transition of ANXA2 from basic research to clinical application.

Objective:ANXA2 plays a crucial role in cancer metastasis, but its mechanism is not yet fully understood. Therefore, it is necessary to establish an ANXA2 gene knockout mouse model to provide an effective tool for subsequent studies on ANXA2-related mechanisms.Methods:A gene knockout mouse model was constructed using CRISPR/Cas9 technology. The model was validated through tissue DNA extraction followed by polymerase chain reaction (PCR), sequencing, and western blot to confirm ANXA2 genotype and protein expression. The successfully constructed models were divided into a model group and a wild-type (WT) group for the creation of a mouse tail vein injection Lewis lung carcinoma (LLC) metastasis model. Metastatic foci formation was monitored using in vivo imaging technology, and the survival rates of the two groups were compared. Results:An sgRNA sequence targeting the first exon of ANXA2 was designed, and 16 founder mice were obtained through microinjection. Through consanguineous hybridization, 30 homozygous offspring were ultimately acquired. After establishing the strains of the mouse model, mice were divided into the ANXA2 knockout group and the WT group, with 8 mice in each group. An LLC lung metastasis model was established in both groups. Compared with the WT group, the number of metastatic foci was significantly increased in the ANXA2 knockout group (7 vs. 1), and the fluorescence intensity was stronger in the WT group than in the knockout group ( P=0.002). Using the GEPIA2 database to analyze ANXA2 gene expression in tumor tissues and normal tissues of lung cancer patients, it was found that ANXA2 expression levels were significantly higher in lung cancer tumor tissues compared to normal tissues ( P＜0.05). The database included data from 478 lung cancer patients, and patients were stratified into high-expression and low-expression groups based on ANXA2 levels. Compared to the low-expression group, patients in the high-expression group exhibited significantly shorter disease-free survival and overall survival ( P＜0.05, respectively). The survival time of mice in the ANXA2 knockout group (median survival time, 43 days) was significantly longer compared to the WT group (median survival time, 26 days; P=0.017). Additionally, ANXA2 expression is significantly associated with the prognosis of lung cancer patients ( P=6.4e-14). Conclusions:ANXA2 is closely associated with cancer metastasis and holds potential as a new target for metastasis treatment. Further in-depth research will greatly facilitate the transition of ANXA2 from basic research to clinical application.

Objective:To evaluate the cerebral infarct volume and the nerve fiber connectivity between cortical and neurogenesis-related regions in the mouse model of reperfusion after middle cerebral artery occlusion (MCAO) by 11.7 Tesla(11.7 T) magnetic resonance imaging (MRI).Methods:MCAO models were established in SPF grade adult male C57BL/6 mice using the suture-occluded method.MRI scans were performed at 3 days before and 1 day after modeling.Infarct volumes were calculated, and nerve fiber tracking was performed on specific brain regions to analyze the nerve fiber number and the parameters of fractional anisotropy(FA), mean diffusivity(MD), axial diffusivity (AD)and radial diffusivity(RD). SPSS 26.0 was used for statistical analysis, and paired t test was used to compare the data before and after modeling. Results:(1) After MCAO-induced ischemia, the infarct volume was up to (35.11±17.57)mm 3, and the FA value of the infarct area was significantly reduced compared with that of before modeling( t=4.73, P<0.01). (2) At the anterior-posterior(AP): + 1.2 mm section, the results of fiber tracking showed that compared with before modeling, the number of fiber bundles originating from the dorsal horn of the lateral sub-ventricle zone(SVZ)to the cortex reduced ((92 584.20±14 751.00) vs (59 815.60±6 752.46), t=4.87, P<0.01), and the number of fiber bundles projected to the infarcted area reduced ((107 671.40±10 497.57) vs (61 658.60±10 178.21), t=6.43, P<0.01). FA, AD, MD, and RD values were all decreased in different degrees( t=3.38-6.43, all P<0.05). (3) At the AP: -3.8 mm section, the number of fiber bundles originating from the dorsal horn of the SVZ to the cortex decreased (after modeling(96 944.00±18 331.09), before modeling(58 767.80±16 445.25), t=2.99, P<0.05), and the values of FA, AD, MD and RD decreased after ischemia ( t=7.30, 5.05, 6.74, 4.13, all P<0.05). Conclusion:The ultra-high field strength of 11.7 T MRI can accurately detect the following results that the number of nerve fiber bundles from the SVZ to the cortex or infarct area are both significantly reduced, and diffusion tensor parameters are consistently changed in mice after 1 day of ischemia-reperfusion.

Objective:To explore the quality of life (QOL) and the related influencing factors of patients with early esophageal cancer after endoscopic submucosal dissection (ESD).Methods:A questionnaire survey was conducted in 167 early esophageal cancer patients who underwent ESD in Shanxi Province Cancer Hospital from January 2022 to July 2022. European Organization for Research and Treatment of Cancer Quality of Life Assessment Core Scale (EORTC QLQ-C30) and the Esophageal Cancer Supplementary Scale (EORTC QLQ-OES18) were used to compare QOL of patients with different clinical characteristics before surgery, 1 month after surgery and 6 months after surgery, And multiple logistic regression analysis was used to analyze the influencing factors of patients' QOL at 6 months after surgery.Results:EORTC QLQ-C30 showed that the scores of the patients' physical function, role function, and social function at 1 month and 6 months after surgery were lower than those before surgery, and the differences were statistically significant (all P < 0.05). The scores of dyspnea, constipation, nausea and vomiting, fatigue, and economic status in the symptom area were higher than those before surgery, and the differences were statistically significant (all P < 0.05). According to EORTC QLQ-OES18, the scores of difficulty in swallowing oral fluid, obstruction, poor eating initiative, dry mouth, and cough at 1 month and 6 months after surgery were higher than those before surgery, and the differences were statistically significant (all P < 0.05). The score of dysphagia at 1 month after surgery was higher than that before surgery, while the score at 6 months after surgery was lower than that before surgery, and the differences were statistically significant (all P < 0.05). The score of dyspepsia at 1 month and 6 months after surgery was lower than that before surgery, and the difference was statistically significant (all P < 0.05). Multivariate analysis showed that the lesion perimeter >1/2 perimeter (lesion perimeter >1/2 perimeter vs. lesion perimeter ≤ 1/2 perimeter: OR = 2.072, 95% CI 1.536-2.796, P < 0.05) and postoperative esophageal stricture dilatation (undergoing esophageal stricture dilatation or not: OR = 2.193, 95% CI 1.429-2.789, P < 0.05) were independent risk factors affecting the QOL of patients at 6 months after surgery. Conclusions:The QOL of early esophageal cancer patients after ESD is decreased compared with that before surgery, and the main manifestations include physical function, role function, social function, and symptom. The area of lesion and undergoing esophageal stricture dilatation or not are factors affecting the QOL of patients after surgery.

Objective:To investigate the abnormal patterns of spontaneous neural activity of patients with type 2 diabetes comorbid depression (T2DD) by using resting-state functional magnetic resonance imaging (rs-fMRI) fractional amplitude of low-frequency (fALFF) analysis, and determine the neuroimaging features of brain damage in T2DD patients.Methods:A perspective study was performed. Fifty-nine type 2 diabetes mellitus (T2DM) patients and 52 T2DD patients, admitted to and accepted treatment in Department of Endocrinology of our hospital from November 2017 to November 2020, were chosen; another 57 healthy controls matched with gender, age and education level, admitted to our hospital at the same time period were enrolled. Their clinical data, neuropsychological test and rs-fMRI data were collected; whole brain fALFF values were calculated, and fALFF values of different brain regions were compared in subjects of the 3 groups. Pearson correlation analysis was used to verify the correlations of fALFF values with clinical variables and neuropsychological scale scores.Results:The fALFF values in bilateral precuneus showed significant difference among the three groups ( P<0.05). The fALFF values in bilateral precuneus of the T2DD and T2DM groups were significantly lower than those in heathy control group ( P<0.05), and those in the T2DD group were lower than those in the T2DM group without significant difference ( P>0.05). Pearson correlation analysis showed that there were no correlations of fALFF values with clinical data and psychometric scale scores in T2DD group and T2DM group ( P>0.05). Conclusion:The abnormal patterns of spontaneous brain activity in the bilateral precuneus may be the neuroimaging markers of brain damage in T2DD patients.

Objective:To identify the clinical characteristics and pathogenic gene of a Chinese Han family with achromatopsia (ACHM).Methods:The method of pedigree investigation was adopted.A Chinese Han ACHM family was recruited in Peking Union Medical College Hospital form July 2010 to July 2019, including 5 members of 2 generations.There were 2 patients and 3 phenotypically normal individuals.The medical history was collected and comprehensive ophthalmic examinations were performed, including visual acuity, colour vision, color fundus photography, fundus autofluorescence (FAF), optical coherence tomography (OCT), visual field and electroretinogram (ERG).Genomic DNA was extracted from peripheral blood sample from the patients and family members.Pathogenic variant was screened by whole exome sequencing (WES) and verified by Sanger sequencing and co-segregation analysis.The variant was annotated with the 1000 Genomes, Human Gene Mutation Database (HGMD), ExAC, ClinVar and OMIM databases to detect the single nucleotide polymorphism and whether it had been reported previously.The pathogenicity of the variant was evaluated according to the standards and guidelines of the American College of Medical Genetics and Genomics (ACMG).This study adhered to the Declaration of Helsinki.The study protocol was approved by the Institutional Review Board of Peking Union Medical College Hospital (No.JS-2059).Written informed consent was obtained from the guardians of juvenile patients.Results:There was consanguinity between the proband's parents and this family was consistent with autosomal recessive inheritance.Both male patients presented the reduction of visual acuity accompanied with photophobia and color blindness since childhood.Barely visible foveal light reflex in fundus images, hypofluorescence of foveal areas in FAF images, foveal defect with disruption of ellipsoid zone and interdigitation zone in OCT images were found in both patients.Central scotoma with or without peripheral visual field defects was detected.Generally normal scotopic 0.01, 3.0 and 10.0 responses, decreased oscillatory potentials amplitudes, no photopic 3.0 and 30 Hz flicker responses were observed.No sign of progression was found during the 9-year follow-up.It was confirmed that both patients carried a novel homozygous disease-causing variant c. 947insA (p.Asn316Lysfs*46) in ATF6 gene.Their mother had the heterozygous variant.The unaffected brother did not carry the variant.This family was consistent with co-segregation.This variant was labeled as pathogenic according to the ACMG standards and guidelines. Conclusions:A novel variant c.947insA (p.Asn316Lysfs*46) in ATF6 gene is the pathogenic variant of this achromatopsia family.This is the first time that this variant has been reported.

Objective:To evaluate the effects of different doses of ulinastatin on lung function in patients undergoing total aortic arch replacement.Methods:One hundred and thirty five patients with acute Stanford type A aortic dissection of both sexes, aged 20-70 yr, with body mass index of 16.2-33.3 kg/m 2, of American Society of Anesthesiologist physical status Ⅳ, were divided into 3 groups ( n=45 each) using a random number table method: high-dose ulinastatin group (group H with total dose of 30 000 U/kg), low-dose ulinastatin group (group L with total dose of 20 000 U/kg) and control group (group C). In group H and group L, half of the total dose of ulinastatin was given after induction of anesthesia, the rest of the total dose was primed after being added to cardiopulmonary bypass (CPB) circuit, while normal saline 100 ml was given at the same time point in group C. After induction of anesthesia (T 0), and at 3, 6, 12, 24 and 48 h after the beginning of CPB (T 1-5), blood samples from the central vein were collected for determination of plasma concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6). The oxygenation index (OI) and alveolar-arterial partial pressure of oxygen difference (P A-aO 2) at T 0 and T 2-T 5, intraoperative blood loss and blood transfusion, postoperative mechanical ventilation time, length of intensive care unit (ICU) stay and the incidence of complications within 30 days after surgery were collected. Results:Compared with group C, the plasma concentrations of TNF-α and IL-6 were significantly at T 1-T 5, OI was increased, and P A-aO 2 was decreased at T 2, 3 in H and L groups ( P<0.05). There was no significant difference in the mechanical ventilation time, length of ICU stay and incidence of complications within 30 days after surgery among the 3 groups ( P>0.05). Conclusion:Ulinastatin can inhibit inflammatory responses and improve lung function in patients undergoing total aortic arch replacement, but it has no value for clinical outcomes.

OBJECTIVES: Radiotherapy is one of the main therapies for colorectal cancer, but radioresistance often leads to radiotherapy failure. To improve the radioresistance, we explore the effect of oligomycin A, the H METHODS: The effects of different concentrations of oligomycin A on the survival rate and glycolysis of HT29 colorectal cancer cells at different time points were investigated via MTT and glycolysis assay. siRNA-PFK1 was synthesized in vitro and transfected into HT29 cells. The effects of oligomycin A on radiosensitivity of HT29 colorectal cancer cells were measured via MTT and colony formation assay. Western blotting was used to detect the effect of oligomycin A on the expression of glycolytic enzyme PFK1. We compared difference between the effects of siRNA-PFK1 group and oligomycin A combined with siRNA-PFK1 group on cell survival and glycolysis. After 4 Gy X-ray irradiation, the effects of cell survival and glycolysis between the siRNA-PFK1 group and the oligomycin A combined with siRNA-PFK1 group were compared. RESULTS: Compared with the 0 μmol/L oligomycin A group, the cell survival rate of HT29 cells treated with 4 μmol/L oligomycin A was significantly increased ( CONCLUSIONS Oligomycin A can promote the radioresistance of HT29 colorectal cancer cells, which may be related to up-regulation of the PFK1 expression and increase of cell glycolysis.
Cell Line, Tumor ; Colorectal Neoplasms/genetics* ; HT29 Cells ; Humans ; Oligomycins/pharmacology* ; Radiation Tolerance

Objective:To access the genetic defects and clinical characteristics of patients with KCNV2-associated cone dystrophy. Methods:Three pedigrees with KCNV2-associated cone dystrophy were recruited in Peking Union Medical College Hospital from August 2017 to December 2019.Peripheral blood from each patient and their parents was collected, and genomic DNA was extracted.Targeted exome capture plus next-generation sequencing (NGS) was used to detect the candidate variants.Suspected causative variants were validated by Sanger sequencing and segregation analysis.Comprehensive ocular examinations were performed, including vision acuity, colour vision, fundus photography, fundus autofluorescence (FAF), optical coherence tomography (OCT), visual field and electroretinogram (ERG). This study was approved by the Institutional Review Board of Peking Union Medical College Hospital and adhered to the tenets of the Declaration of Helsinki.Written informed consent was obtained from each patient prior to any medical examination. Results:Three probands from three unrelated Chinese families were confirmed carrying biallelic KCNV2 disease-causing variants.Two patients harbored compound heterozygous variants and one patient with history of consanguinity was identified carrying homozygous variant.Five novel variants in the KCNV2 gene were identified, including p. T121M, p.R244C, p.C199Y, p.M250R and p. L171Pfs*201.All patients enrolled in this study were male with age of 25, 16 and 2 years old, respectively.Three affected individuals complained of vision loss and photophobia and two patients demonstrated reduced color perception and nystagmus.Macular discoloration (bull's eye maculopathy or gold foil macular reflex) was observed in fundus photographs.Macular hypofluorescence was illustrated in FAF imaging, which accompanying a broad hyperfluorescent ring surrounding the central atrophy or not.Macular thinning with loss of the inner segment ellipsoid zone was noted in OCT images, and the disruption was more profound in older patients.Central scotoma with or without peripheral visual field defects was observed in perimetry.Severe cone function loss and variable scotopic rod impairment were demonstrated in ERG, whereas a broad a-wave trough response to scotopic bright flash stimulation was noted. Conclusions:Patients with KCNV2-associated cone dystrophy show a characteristic ERG manifestation.ERG results and KCNV2 variants in Chinese patients differ from those in foreigners.