Objective To explore the mechanism of ubiquitin specific peptidase 21 (USP21) increasing the stability of forkhead box protein M1 (FOXM1) and promoting M2 polarization of macrophages in endometriosis (EM). Methods Eutopic endometrial stromal cells (EESC) collected from patients and normal endometrial stromal cells (NESC) from routine health examiners were cultured in vitro, and the expression levels of USP21 and FOXM1 were detected using RT-qPCR and Western blot. EESCs were co-cultured with macrophages. M1 polarization markers of interleukin 6 (IL-6) and CXC chemokine ligand 10 (CXCL10) and M2 polarization markers of CD206 and fibronectin 1 (FN1) were tested using RT-qPCR. M2 marker CD206 was further detected by flow cytometry. IL-6, tumor necrosis factor-alpha (TNF-α), IL-10, and transforming growth factor-beta (TGF-β) levels in cell supernatant were detected by ELISA. Co-immunoprecipitation was used to assess the interaction between USP21 and FOXM1, and the ubiquitination level of FOXM1. FOXM1 protein stability was detected through cycloheximide (CHX) assay. Results USP21 and FOXM1 expression levels in the EESC group were significantly increased compared with those in the NESC group; compared with the NESC + M0 group, the EESC + M0 group showed no significant difference in the expression of M1 polarization markers (IL-6 and CXCL10), but increased expression of M2 polarization markers (CD206 and FN1), along with notably increased number of M2 macrophages; there was no significant difference in IL-6 and TNF-α levels, but increased levels of IL-10 and TGF-β in the cell supernatant. The above findings indicated that the deubiquitinase USP21 was highly expressed in EM, promoting M2 polarization of macrophages. Knocking down USP21 or FOXM1 can inhibit M2 polarization of EM macrophages. USP21 interacted with FOXM1 in EESC, leading to a decrease in FOXM1 ubiquitination level and an increase in FOXM1 protein stability. Overexpression of FOXM1 reversed the inhibitory effect of knocking down USP21 on M2 polarization of EM macrophages. Conclusion The deubiquitinase USP21 interacts with FOXM1 to increase the stability of FOXM1 and promote M2 polarization of EM macrophages.
Humans ; Forkhead Box Protein M1/genetics* ; Female ; Macrophages/cytology* ; Endometriosis/genetics* ; Ubiquitin Thiolesterase/genetics* ; Cells, Cultured ; Endometrium/metabolism* ; Ubiquitination ; Adult ; Interleukin-10/metabolism* ; Interleukin-6/metabolism* ; Protein Stability ; Stromal Cells/metabolism*

Objective·To explore the role of advanced platelet-rich fibrin(A-PRF)in osteochondral regeneration.Methods·Bone-marrow mesenchymal stem cells(BMSCs)and knee joint chondrocytes were obtained from New Zealand rabbits.A-PRF was obtained by low-speed centrifugation of the heart blood of rabbits.The histological structure of A-PRF was observed by an optical microscope.The release of growth factors in A-PRF was detected by ELISA,including platelet-derived growth factor,transforming growth factor-β,insulin-like growth factor,vascular endothelial growth factor,epidermal growth factor and fibroblast growth factor.A-PRF's cytotoxicity and capability for promoting the proliferation of rabbit BMSCs were detected by live/dead double staining and MTT methods.The effect of A-PRF on the gene expression of type Ⅱ collagen,aggrecan,alkaline phosphatase(ALP)and osteocalcin(OCN)in rabbit BMSCs was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Transwell chambers were used to determine the effect of A-PRF on the migration ability of rabbit BMSCs and the chondrocytes.Rabbit knee osteochondral defect models were established,and 18 rabbits were randomly divided into 3 groups.The A-PRF group(n=6)was implanted with A-PRF in the defect,the A-PRF+BMSCs group(n=6)was implanted with rabbit BMSCs on A-PRF,and the control group(n=6)did not undergo implantation.The rabbits were sacrificed 12 weeks after surgery and the knee joint specimens were stained with hematoxylin-eosin(H-E),toluidine blue and safranin O/fast green.Based on the surface morphology and histology of the knee joints,the International Cartilage Repair Society(ICRS)scoring system was used for macroscopic and histological scoring.Results·A-PRF had a loose network structure and can slowly release growth factors.No cytotoxicity to rabbit BMSCs was observed after adding A-PRF,and the the capability for promoting the proliferation of rabbit BMSCs was significantly increased at 24,48 and 72 h after adding A-PRF(all P<0.05).Chondrogenesis-related gene Ⅱ collagen and aggrecan,as well as osteogenesis-related genes ALP and OCN were significantly up-regulated(all P<0.05).After adding A-PRF,the migration abilities of rabbit BMSCs and chondrocytes were significantly enhanced(both P<0.05),and the migration ability of rabbit BMSCs was significantly higher than that of chondrocytes(P=0.025).The joint surface morphology in the rabbit knee joint defect models was observed.It can be seen that the defects in the A-PRF group and the A-PRF+BMSCs group were basically restored,while the the defects in the control group were only covered by soft tissue.In the ICRS macroscopic score,there was no statistical difference between the A-PRF group and the A-PRF+BMSCs group,but the scores of the two groups were all significantly higher than those of the control group(all P<0.05).According to the histological results,both the A-PRF group and the A-PRF+BMSCs group formed osteochondral repair,but the cartilage in the A-PRF group was more mature,while the control group formed fibrous repair.In the ICRS histological score,there was no statistical difference between the A-PRF group and the A-PRF+BMSCs group,but the scores of both the groups were significantly higher than those of the control group(both P<0.05).Conclusion·Autologous A-PRF has good biocompatibility and the capability for promoting the proliferation of BMSCs.It can promote the repair of cartilage and subchondral bone both in vitro and in vivo.

Objective:To develop a risk prediction lineogram of neooperative atrial fibrillation in patients with esophageal cancer.Methods:The clinical data of 1 509 patients undergoing esophageal cancer surgery admitted to the department of esophageal surgery of our hospital from December 2019 to April 2022 were gathered, and they were divided into two layers according to whether they had new atrial fibrillation after surgery. In each layer, they were randomly divided into training set and test set in a ratio of 7∶3. In the training population, the multi-factor logistic regression method was used to establish the prediction model, and the line graph of the prediction model was drawn. The ROC curve and calibration curve were drawn to assess the differentiation ability and calibration ability of the prediction model. The test set population is used to validate the prediction model. Results:A total of 1 509 patients with esophageal cancer were included in the study, and the incidence of new atrial fibrillation after surgery was 247 patients(16.4%). A total of 1 039 patients(68.9%) were enrolled in the training set. The multivariate logistic regression model indicated that age, gender, BMI, pulmonary infection, the use of invasive ventilator, and the need for additional drainage of fluid accumulation were the influencing factors for new postoperative atrial fibrillation. The AUC of the training set prediction model under ROC curve was 0.775(95% CI: 0.737-0.812, P<0.001), indicating that the model has high predictive discrimination ability. Calibration curve and Hosmer- Lemeshow test results P=0.796, indicating that the model has good consistency of prediction ability. There were 470 subjects(31.1%) in the test set. The results showed that the AUC of the prediction model under the ROC curve was 0.773(95% CI: 0.719-0.826, P<0.001), indicating that the prediction model still has a high discriminative ability in the test set population. Conclusion:Patients with age, gender, BMI, pulmonary infection, the use of invasive ventilator, and the need for additional drainage of effusion are at higher risk of new atrial fibrillation after surgery. The timely prediction, prevention and management of POAF are crucial to improve the prognostic quality of postoperative patients with esophageal cancer by constructing clinical prediction models.

Objective     To search for the key microRNAs (miRNAs) involved in myocardial fibrosis in hypertrophic cardiomyopathy, and to further explore the mechanisms involved in the regulation of myocardial fibrosis. Methods    Forty-two patients with hypertrophic cardiomyopathy diagnosed and treated surgically in West China Hospital of Sichuan University from January 2014 to June 2018 were selected, including 29 males and 13 females, with a median age of 46 (15-69) years. In the myocardial tissue of patients with hypertrophic cardiomyopathy with different degrees of fibrosis, miRNAs with significantly different expression were screened and further verified at the cellular level. By regulating the expression of the target miRNAs, the expressions of fibrosis-related proteins and target genes were detected respectively. Finally, the target-binding relationship was verified by dual-luciferase reporter gene detection. Results    miR-484 was up-regulated in severely fibrotic myocardial tissue and activated cardiac fibroblasts. After cardiac fibroblasts were activated by TGF-β1, the expression of miR-484 was significantly up-regulated, the expression of fibrosis-related proteins (CollagenⅠ, α-SMA) increased, and the expression of the target gene HIPK1 decreased (P<0.05). After inhibiting the expression of miR-484 by transfection of miR-484 antagomir, the expression of fibrosis-related proteins decreased, while expression of HIPK1 was up-regulated (P<0.05). The detection of dual luciferase reporter gene showed that the luciferase activity of the transfected WT-miRNA-484 mimics group was lower than that of the control group (P<0.05). Conclusion    miR-484 is a pro-fibrotic miRNA that participates in the process of myocardial fibrosis by negatively regulating the expression of HIPK1. It can be used as a regulatory target to provide a therapeutic strategy for myocardial fibrosis.

OBJECTIVE: To investigate the effect of body mass index (BMI) on the short-term effectiveness of high tibial osteotomy (HTO) in the treatment of varus knee arthritis. METHODS: The clinical data of 84 patients (84 knees) with varus knee arthritis treated with HTO between May 2016 and August 2020 were retrospectively analyzed. According to BMI, the patients were divided into normal group (32 patients in group A, BMI<25 kg/m ²), overweight group (27 patients in group B, BMI>30 kg/m ²), and obese group (25 patients in group C, BMI>30 kg/m ²). The BMI of groups A, B, and C were (23.35±0.89), (26.65±1.03), and (32.05±1.47) kg/m ², respectively. There was no significant difference ( P>0.05) in gender, age, surgical side, disease duration, and preoperative Hospital for Special Surgery (HSS) score, visual analogue scale (VAS) score, knee range of motion, and hip-knee-ankle angle (HKA) between groups. The operation time, intraoperative dominant blood loss, and the decrease of hemoglobin on the 3rd day after operation were recorded and compared between groups. The improvement of knee joint function and pain status were evaluated by knee joint HSS score, knee range of motion, and VAS score before and after operation, and measuring the HKA of patients on X-ray film. During the follow-up, the X-ray films of the knee joint were reexamined to observe the position of the internal fixator and the healing of osteotomy. RESULTS: All patients completed the operation successfully and were followed up 8-40 months (mean, 19.3 months). There was no significant difference in follow-up time, operation time, intraoperative dominant blood loss, and the decrease of hemoglobin on the 3rd day after operation between groups ( P>0.05). No operative complications such as severe vascular or nerve injury occurred. After operation, deep venous thrombosis of lower extremities occurred in 1 case in groups A and B respectively, and fat liquefaction of surgical incision occurred in 2 cases in group C. There was no significant difference in the incidence of perioperative complications between groups (3.1% vs. 3.7% vs. 8.0%) ( P=0.689). During the follow-up, there was no bone nonunion, plate fracture or loosening. At last follow-up, HSS score, VAS score, knee range of motion, and HKA significantly improved in the 3 groups when compared with those before operation ( P<0.05), but there was no significant difference in the differences of the above indexes between groups before and after operation ( P>0.05). CONCLUSION BMI does not affect the short-term effectiveness of HTO in the treatment of varus knee arthritis. HTO can be selected for overweight and obese patients after standard medical treatment is ineffective.
Humans ; Osteoarthritis, Knee/surgery* ; Body Mass Index ; Overweight ; Retrospective Studies ; Treatment Outcome ; Knee Joint/surgery* ; Obesity/complications* ; Osteotomy ; Blood Loss, Surgical

Background: Domestic yaks are the most important livestock species on the Qinghai-Tibetan Plateau. Adult female yaks normally breed in the warm season (July to September) and enter anestrous in the cold season (November to April). Nevertheless, it is unclear how ovarian activity is regulated at the molecular level. Objectives: The peculiarities of yak reproduction were assessed to explore the molecular mechanism of postpartum anestrus ovaries in yaks after pregnancy and parturition. Methods: Sixty female yaks with calves were observed under natural grazing in Haiyan County, Qinghai Province. Three yak ovaries in pregnancy and postpartum anestrus were collected. RNA sequencing and quantitative proteomics were employed to analyze the pregnancy and postpartum ovaries after hypothermia to identify the genes and proteins related to the postpartum ovarian cycle. Results: The results revealed 841 differentially expressed genes during the postpartum hypoestrus cycle; 347 were up-regulated and 494 genes were down-regulated. Fifty-seven differential proteins were screened: 38 were up-regulated and 19 were down-regulated. The differential genes and proteins were related to the yak reproduction process, rhythm process, progesterone-mediated oocyte maturation, PI3K/AKT signaling pathway, and MAPK signaling pathway categories. Conclusions Transcriptome and proteomic sequencing approaches were used to investigate postpartum anestrus and pregnancy ovaries in yaks. The results confirmed that BHLHE40, SF1IX1, FBPX1, HSPCA, LHCGR, BMP15, and ET-1R could affect postpartum hypoestrus and control the state of estrus.

Objective:To investigate the psychological stress and related factors of community medical staff in Shanghai during infectious public health emergencies.Methods:Random cluster sampling, questionnaires combining general demographic information, self-rating anxiety scale(SAS), self-rating depression scale(SDS)and yale-brown obsessive compulsive scale(YB)were administrated to investigate the psychological stress of medical staff from 20 community healthcare centers in 10 Shanghai districts.Results:A total of 696 valid questionnaires were collected. 17.36% of the staff had SAS scores exceeding the boundary value. Nurses, staff with per capita monthly family income<5 000 yuan, or with below-average family relations presented higher SAS scores. 28.19% of the staff had SDS scores exceeding the boundary value. Staff with 5-10 years seniority, nurses, with per capita monthly family income<5 000 yuan, and with poor family relations presented higher SDS scores. 6.68% of the staff had YB score≥16. Nurses presented a higher YB score.Conclusions:Some community healthcare center staff experienced psychological stress during public health emergencies.Factors associated with psychological stress, such as 5-10 years′ seniority, nurses, per capita monthly family income<5 000 yuan, below-average or poor family relations, deserve more attention in order to prevent from or alleviate harmful psychological stress.

Objective: The aim of this study was to explore the correlation between the clinicopathological characteristics of colorectal adenocarcinoma and the growth pattern, tumor budding, and CD8+T lymphocyte infiltration in anterior invasive margins, and to assess their value as prognostic indicators. Methods: Paraffin embedded samples were collected from 126 patients with primary colorectal adenocarcinoma who underwent surgical resection in Zhangye People's Hospital Affiliated to Hexi College from January 2008 to December 2019. A total of 126 pathological sections were stained by immunohistochemistry. Anti-cytokeratin antibodies were used to mark tumor cell budding and anti-CD8 antibody markers T lymphocytes were evaluated. Results: In colorectal adenocarcinoma, infiltrative growth patterns and high-grade tumor budding in invasive margins were significantly associated with pathological stage of tumor size (pT) (P=0.029 and P<0.001, respectively), pathological stage of lymph node metastasis (pN) (P<0.001 and P=0.023, respectively) and vessel infiltration (P<0.001 and P<0.001, respectively). Furthermore, high-grade CD8+T lymphocyte infiltration was associated with the absence of lymph node metastases (P=0.050). Conclusions: Infiltrative growth patterns and high-grade tumor budding in colorectal adenocarcinoma invasive margins were correlated with patient prognosis. Importantly, these two features are easily detectable (with the help of pan-cytokeratin immunohistochemistry staining), in a reproducible manner. Therefore, we propose that they could be used as prognostic indicators in colorectal adenocarcinoma patients.

Objective:To investigate the relationship between self-management ability and self-efficacy in coronary heart disease (CHD) patients, and the influential factors.Methods:From April to June 2016, a questionnaire survey was conducted among 236 patients with coronary heart disease selected by simple random sampling method in the outpatient clinic of Pengpu Community Health Service Center; and the Coronary Artery Disease Self-management Scale and self-efficacy questionnaire were used in the survey. The correlation between scores of the two scales was analyzed; the scores were compared between patients with percutaneous coronary intervention (PCI) and those without PCI, between patients with myocardial infarction (MI) and those without MI. Multiple linear regression was performed to elucidate the influencing factors of self-management ability and self-efficacy of CHD patients.Results:The mean scores of self-management ability and self-efficacy were (75.5±15.5) and (24.8±10.8), respectively. The self-management ability was significantly correlated with self-efficacy in CHD patients(r=0.398-0.683, P<0.05). The self-management score of patients with PCI was higher than those without PCI[ (80.4±14.9) vs.(72.5±15.1), t=3.38, P<0.01]. The scores of self-management[(85.0±16.4) vs.(72.9±14.2), t=5.14, P<0.01]and self-efficacy[(29.3±12.0) vs. (23.6±10.2), t=3.39, P<0.01] of patients with MI were higher than those without MI. Multiple linear stepwise regression analysis indicated that age over 65, history of MI, history of PCI, history of smoking and self-efficacy score were all factors influencing self-management behavior in patients with CHD ( R=0.735; R2=0.540; F=26.451, P<0.01). Smoking history and self-management score were the influencing factors of self-efficacy in patients with CHD ( R=0.690; R2=0.476; F=20.425, P<0.01). Conclusion:The self-efficacy of patients with CHD is closely related to their self-management behavior, so we should strengthen their self-efficacy and improve their self-management level by comprehensive measures, particularly for patients without PCI.

Objective: To investigate the subjective well-being of gynecologic tumor patients and analyze the influencing factors. Methods: Investigated a total of 411 gynecologic tumor patients by using the a general information questionnaire, Memorial University of Newfoundland Scale of Happiness (MUNSH), Social Support Rating Scale(SSRS) and Medical Coping Modes Questionnaire(MCMQ). Results: The total scores of SWB was (32.20±9.54) points, at a moderate level. The influencing factors of subjective well-being of gynecologic tumor patients included: type of tumor, complication, education, the way of payment for medical treatment, average income per person in family, social support and medical coping modes. Furtherly, average income per person in family, social support and the resigntion dimension of medical coping modes were the main factors. Conclusions The SWB of gynecologic tumor patients need to be improved. To increase the SWB of the patients, the family, society and medical staff should give them more emotional, informational and other kinds of support, and try to minimize negative coping modes such as resigntion.