Objective:To evaluate the role of ubiquitin-specific peptidase 22 (USP22) in myocardial ischemia-reperfusion (I/R) injury in diabetic mice.Methods:Seventy-eight SPF male C57BL/6 mice, aged 6-8 weeks, were divided into 6 groups using a random number table method: sham operation group (Sham group, n=12), type 1 diabetes mellitus + sham operation group (T1D+ Sham group, n=12), myocardial I/R injury group (I/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury group (DI/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury + empty vector group (DI/R+ V group, n=15), and type 1 diabetes mellitus + myocardial I/R injury + USP22 overexpression group (DI/R+ U group, n=15). Type 1 diabetes mellitus was induced by intraperitoneal injection of streptozotocin-citrate buffer. Myocardial I/R was induced by ligation of the left coronary artery. At 1 day before developing the myocardial I/R injury model, DI/R+ U group and DI/R+ V group received an intramyocardial injection of USP22 overexpression plasmid or empty vector plasmid, respectively. At 24 h of reperfusion, cardiac function was assessed using the echocardiography to measure the left ventricular ejection fraction and left ventricular fractional shortening. The mice were then sacrificed, and their hearts were harvested for measurement of the myocardial infarct size, for microscopic examination of pathological changes (using HE staining) and for determination of the apoptosis rate (TUNEL staining), reactive oxygen species(ROS) activity (DHE staining), and USP22 expression (by Western blot, immunofluorescence, and immunohistochemistry). Proteomic analysis was performed to identify downstream proteins regulated by USP22, and protein-protein interactions were investigated using co-immunoprecipitation. Results:Compared with Sham group, the cardiac function indices were significantly decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was down-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of myocardial infarct size was significantly increased, the cardiac function indices were decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was aggravated in DI/R group. Compared with DI/R+ V group, the percentage of myocardial infarct size was significantly decreased, the cardiac function indices were increased, the apoptosis rate of myocardial cells and ROS activity were decreased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was alleviated in DI/R+ U group. The results of proteomics combined with co-immunoprecipitation experiments showed an interaction between calponin 1 and USP22. Conclusions:During myocardial I/R injury in diabetic mice, USP22 may act as an endogenous protective mechanism, and calponin 1 might be a downstream mechanism through which USP22 exerts its protective effects.

Objective:To evaluate the clinical value of the frozen section assessment of surgical margin in preserving urinary continence and erectile function during robotic radical prostatectomy (RARP).Methods:From Feb. 2022 to Jul 2023, 11 patients underwent RARP using intrafascial dissection technique and frozen section assessment in Beijing United Family Hospital. The median age was 64(61, 69)years, the median PSA was 8.0(6.0, 11.2)ng/ml, and the median IIEF-5 was 17.5(10.3, 22.0). All the patients were diagnosed with prostate cancer and PSMA-PET/CT showed negative for metastasis. The da Vinci Si platform, 4-arm, transabdominal anterior approach was applied in all cases. The prostate specimen was removed from the umbilical incision and submitted to the pathology department for frozen section assessment. If the frozen section assessment showed cancer in the surgical margin, a secondary resection of the ipsilateral periprostatic fascia and neurovascular bundle (NVB) was performed. Totally, 3 cases underwent secondary resection, who underwent unilateral NVB-sparing, and the other 8 cases underwent bilateral NVB-sparing surgery. In follow-up, the PSA, urinary continence and erectile function was assessed and recorded.Results:All the surgeries of 11 cases were successful, without convertion to open surgery. The median operation time was 260(225, 285)minutes, the median blood loss was 200(100, 300)ml, the median hospital stay was 4(3, 5)days. There was no over Clavien Dindo Ⅱ perioperative complications.The median catheter indwelling time was 12(10, 14) days, with 10 cases achieving full continence upon catheter removal. The median follow-up time was 10 months. The median PSA level at 3 months, 6 months and 9 months postoperatively was 0.017(0.006, 0.170)ng/ml, 0.015(0.006, 0.390)ng/ml, and 0.007(0.006, 0.650)ng/ml respectively. The median IIEF-5 at 3 months, 6 months and 9 months postoperatively was 0.017(0.006, 0.170)ng/ml, 0.015(0.006, 0.390)ng/ml, 0.007(0.006, 0.650)ng/ml respectively.The median IIEF-5 at 3 months, 6 months and 9 months post-operation was 5.0(4.5, 11.0), 7.5(4.3, 17.5), and 9.5(6.0, 17.5)respectively.Conclusions:The RARP combined with frozen section assessment of surgical margin has excellent safety profile and can achieve favorable clinical value in tumor control, NVB preserving, urinary continence and erectile function.

Herein, we define the role of ferroptosis in the pathogenesis of diabetic cardiomyopathy (DCM) by examining the expression of key regulators of ferroptosis in mice with DCM and a new ex vivo DCM model. Advanced glycation end-products (AGEs), an important pathogenic factor of DCM, were found to induce ferroptosis in engineered cardiac tissues (ECTs), as reflected through increased levels of Ptgs2 and lipid peroxides and decreased ferritin and SLC7A11 levels. Typical morphological changes of ferroptosis in cardiomyocytes were observed using transmission electron microscopy. Inhibition of ferroptosis with ferrostatin-1 and deferoxamine prevented AGE-induced ECT remodeling and dysfunction. Ferroptosis was also evidenced in the heart of type 2 diabetic mice with DCM. Inhibition of ferroptosis by liproxstatin-1 prevented the development of diastolic dysfunction at 3 months after the onset of diabetes. Nuclear factor erythroid 2-related factor 2 (NRF2) activated by sulforaphane inhibited cardiac cell ferroptosis in both AGE-treated ECTs and hearts of DCM mice by upregulating ferritin and SLC7A11 levels. The protective effect of sulforaphane on ferroptosis was AMP-activated protein kinase (AMPK)-dependent. These findings suggest that ferroptosis plays an essential role in the pathogenesis of DCM; sulforaphane prevents ferroptosis and associated pathogenesis via AMPK-mediated NRF2 activation. This suggests a feasible therapeutic approach with sulforaphane to clinically prevent ferroptosis and DCM.

Objective:To compare biomechanical stabilities between screw-plate fixation and non-cannulated screw fixation for Lisfranc ligament injury by a 3-D finite element analysis.Methods:A 3-D model of a healthy foot was developed from computed tomography images. The 1st and the 2nd dorsal tarsometatarsal ligaments and Lisfranc ligament were cut in the 3-D model of a healthy foot to establish a Lisfranc ligament injury model, in which screw-plate fixation (with 2 locking plates and 8 standard screws and one non-cannulated screw) and non-cannulated screw fixation (with 3 non-cannulated screws) for Lisfranc ligament injury were simulated respectively. Finite element analyses were carried out by Abaqus 6.14 software after loads were added in the 3-D models of screw-plate fixation and non-cannulated screw fixation for Lisfranc ligament injury. The overall stress-strain nephogram, the stress distribution and displacement of the foot bone, and the stress distribution on the internal fixation system were compared between the 2 kinds of models.Results:Under the same load, the stress of the whole screw-plate fixation was concentrated on the fixators, and the stress of the non-cannulated screws was also greater than that of the bones. In both models, the strain of the whole foot led to arch collapse, especially in the medial column. The maximum stress on the screw holes in the medial and middle columns in the screw-plate fixation model was 39.91 MPa, smaller than that in the non-cannulated screw fixation model (53.13 MPa). The relative displacement of the first metatarsal joint in the screw-plate fixation model was 8.515 × 10 -1 mm, much greater than that in the non-cannulated screw fixation model (3.893 × 10 -1 mm). Stress concentration was observed in both models. The stress of the screw-plate system was concentrated on the fibular side of the middle section of the plate used to fix the first tarsometatarsal joint, decreasing towards both ends. The maximum stress of the non-cannulated screws was located in the middle of the medial column screw for fixation of the first tarsometatarsal joint, significantly greater than those of the both ends. The maximum stress of the screw-plate system was 239.5 MPa, smaller than that of the non-cannulated screws (256.8 MPa). Conclusions:Non-cannulated screw fixation demonstrates a greater biomechanical stability for Lisfranc ligament injury than screw-plate fixation. However, the former may have a higher risk of screw breakage because it bears a greater stress.

Objective:To observe any regulatory effect of a pulsed electromagnetic field (PEMF) on A2A adenosine receptors (A2ARs) in the nucleus pulposus of rats with intervertebral disc degeneration (IDD), and to explore any combination with the A2ARs′ agonist-mediating ROS/PI3K/Akt signaling pathway.Methods:Fifty Sprague-Dawley rats were randomly divided into a control group, an intervertebral disc degeneration group (the model group), an A2AR agonist CGS-21680 treatment group (the agonist group), a PEMF group and a PEMF combined with CGS-21680 treatment group (the observation group). IDD was modeled in all except the rats in the control group. 100μL of CGS-21680 (100μg/kg) was injected into the L 5-6 intervertebral discs of the agonist group, while the PEMF group was given 30 minutes of PEMF intervention daily for 14 days at 1.5mT and 75Hz with a pulse width of 150μs. The observation group was injected with CGS-21680 and then given the same PEMF intervention. Primary nucleus pulposus cells from each group (50ng/mL) were cultured and the expressions of 8-OHDG, SOD, MDA and ROS were detected by immunohistochemistry, immunofluorescence or with an ELISA kit. The A2AR, PI3K, AKT and p-AKT protein levels were detected using western blotting. Results:The nucleus pulposus cells and the annulus fibrosus were obviously wrinkled, necrotic and broken in the model group but the annulus fibrosus was intact and the nucleus pulposus was almost normal in the observation group. Compared with the model group, the levels of SOD and A2AR, PI3K, p-AKT and AKT protein were higher in the agonist, PEMF and observation groups, while the expressions of MDA, ROS and 8-OHDG were weaker. The ROS level in the observation group was significantly lower than that in the agonist and PEMF groups, and the phosphorylation level of p-AKT in the observation group was significantly higher than in the agonist and PEMF groups. The average levels of SOD, A2AR, PI3K, p-AKT and AKT protein in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly higher than the IL-1β group′s average, while the average levels of MDA, ROS and 8-OHDG were significantly lower. The ROS levels in the observation group were significantly lower than in the agonist and PEMF groups, while the A2AR protein content and p-AKT phosphorylation in the observation group were significantly greater. The average Bax levels in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly lower than that in the IL-1β group, and the expression of Bcl-2 was significantly increased. There was significantly less apoptosis observed in the observation group than in the agonist and PEMF groups, while the expression of Bcl-2 was significantly higher.Conclusions:PEMF plays an anti-oxidative stress role by up-regulating A2AR activity and reducing ROS generation. Treatment with PEMF and A2AR agonist could further activate the phosphorylation of PI3K/Akt, down-regulate Bax and up-regulate Bcl-2, thus inhibiting the apoptosis of nucleus pulposus cells and alleviating the malignant progression of IDD.

Objective:To explore a medical big data algorithm to screen the core indicators in clinical database that can be used to evaluate the prognosis of elderly patients with pneumonia.Methods:Based on the clinical database of a Beijing Chaoyang Hospital Consortium Chaoyang Emergency Ward in Beijing Chaoyang Hospital, Capital Medical University, patients with pulmonary infection were selected through the big data retrieval technology. According to the prognosis at the time of discharge, they were divided into death group and survival group. The general data of patients were collected, including gender, age, blood gas and laboratory indices. A computer language called Python was used to make batch calculations of key indicators that affect mortality in elderly patients with pneumonia. Logistic regression analysis was used to analyze the relationship between laboratory indicators and patients' prognosis. Receiver operating characteristic curve (ROC curve) was drawn to analyze the predictive value of screening method for patients' prognosis.Results:A total of 265 patients were included in the study, 64 died and 201 survived. The data of the first detection indexes of each patient after admission were collected, and 23 key indicators with significant differences were selected from 472 indicators: blood routine indicators ( n = 7), blood gas indicators ( n = 3), tumor markers indicators ( n = 3),coagulation related indicators ( n = 4), and nutrition and organ function indicators ( n = 6). ① The key indicators of blood gas in patients died of pneumonia: Cl - was 97-111 mmol/L in 51.6% (33 cases) of patients, lactic acid (Lac) was 0.5-2.5 mmol/L in 81.2% (52 cases) of patients, and H + was 0-46 mmol/L in 87.5% (56 cases) of patients. ② The key indicators of blood routine of patients died of pneumonia: hemoglobin count (Hb) of 46.9% (30 cases) patients was 80-109 g/L, the eosinophils proportions (EOS%) in 67.2% (43 cases) patients was 0.000-0.009, the lymphocytes proportions (LYM%) in 51.6% (33 cases) patients was 0.00-0.09, the red blood cell count (RBC) in 50.0% (32 cases) patients was (3.0-3.9)×10 12/L, the white blood cell count (WBC) in 54.7% (35 cases) patients was (0.0-9.9)×10 9/L, and the red blood cell volume distribution width coefficientof variability (RDW-CV) in 48.4% (31 cases) patients was 10.0%-14.9%, serum C-reactive protein (CRP) was 0.0-49.9 mg/L in 48.4% (31 cases) patients. ③ The key indicators of tumor markers in patients died of pneumonia: 76.6% (49 cases) of patients had negative free prostate specific antigen/total prostate specific antigen (FPSA/TPSA, the ratio was 0), 92.2% (59 cases) had cytokeratin 19 fragment (CYFRA21-1) between 0.0-11.0 μg/L, and 75.0% (48 cases) had carbohydrate antigen 125 (CA125) between 0-104 kU/L.④ The key coagulation indexes of patients died of pneumonia: 68.8% (44 cases) of patients had activated partial thromboplastin time (APTT) of 57-96 s, 73.4% (47 cases) of patients had D-dimer of 0-6 mg/L, 93.8% (60 cases) of patients had thrombin time (TT) of 14-22 s, and 89.1% (57 cases) of patients had adenosine diphosphate (ADP) inhibition rate of 0%-53%. ⑤ Nutrition and organ function key indicatorsin patients died of pneumonia: 92.2% (59 cases) of brain natriuretic peptide (BNP) in patients with 0, 46.9% (30 cases) of patients had prealbumin (PA) of 71-140 mg/L, 90.6% (58 cases) of the patients with uric acid (UA) for 21-41 μmol/L, 75.0% (48 cases) of the patients with albumin (Alb) to 10-20 g/L, 93.5% (60 cases) of patients had albumin/globulin ratio (A/G ratio) of 0-0.9, 84.4% (54 cases) of the patients with lactate dehydrogenase (LDH) from 0-6.68 μmol/L·s -1·L -1. ⑥ Logistic regression analysis and ROC curve analysis: Logistic regression analysis showed that PA and Lac were the prognostic factors. PA could reduce the risk of death by 0.9%, Lac could increase the risk of death by 69.4%; the area under ROC curve (AUC) between laboratory indicators and the prediction effect of death prediction model for patients' prognosis was 0.80, which showed that the classification effect was better, and this study model could better predict the prognosis of elderly patients with pneumonia. Conclusion:By using big data technology, 23 core indicators for evaluating the prognosis of elderly patients with pneumonia can be screened from the clinical database of emergency ward, which provides a new perspective and method for clinical evaluation of the prognosis of elderly patients with pneumonia.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is the fourth-leading cause of cancer-related deaths worldwide. HCC is refractory to many standard cancer treatments and the prognosis is often poor, highlighting a pressing need to identify biomarkers of aggressiveness and potential targets for future treatments. Kinesin family member 2C (KIF2C) is reported to be highly expressed in several human tumors. Nevertheless, the molecular mechanisms underlying the role of KIF2C in tumor development and progression have not been investigated. In this study, we found that KIF2C expression was significantly upregulated in HCC, and that KIF2C up-regulation was associated with a poor prognosis. Utilizing both gain and loss of function assays, we showed that KIF2C promoted HCC cell proliferation, migration, invasion, and metastasis both in vitro and in vivo. Mechanistically, we identified TBC1D7 as a binding partner of KIF2C, and this interaction disrupts the formation of the TSC complex, resulting in the enhancement of mammalian target of rapamycin complex1 (mTORC1) signal transduction. Additionally, we found that KIF2C is a direct target of the Wnt/β-catenin pathway, and acts as a key factor in mediating the crosstalk between Wnt/β-catenin and mTORC1 signaling. Thus, the results of our study establish a link between Wnt/β-catenin and mTORC1 signaling, which highlights the potential of KIF2C as a therapeutic target for the treatment of HCC.
Adult ; Aged ; Animals ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition/genetics* ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins/metabolism* ; Kinesins/metabolism* ; Liver Neoplasms/pathology* ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplasm Staging ; Prognosis ; Protein Binding ; RNA, Small Interfering/metabolism* ; Survival Analysis ; Tumor Burden ; Wnt Signaling Pathway ; Xenograft Model Antitumor Assays ; beta Catenin/metabolism*

Objective:To explore the effect of combining pulsed electromagnetic field (PEMF) stimulation with A 2A adenosine receptor agonist CGS-21680 on apoptosis and inflammation of nucleus pulposus cells in cases of intervertebral disc degeneration. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a sham operation group (Sham group), an intervertebral disc degenerative disease group (Model group), an A 2A adenosine receptor agonist CGS-21680 group (Agonist group), and a group in which PEMF was combined with CGS-21680 (Observation group). The rat model of intervertebral disc degeneration (IDD) was established in all other groups than the sham operation group. The rats in the Agonist group were injected with 100μL of CGS-21680 (100μg/kg) at the L 5-6 intervertebral disc. The Observation group was injected with CGS-21680 similarly but then received 14 conse-cutive days of PEMF stimulation (30min/time). The Sham and Model groups were injected with the same amount of normal saline solution. Eight weeks later, HE staining was used to evaluate the pathological changes in the intervertebral disc tissues. The expression of type II collagen was determined by immunohistochemistry. The content and mRNA expression of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined using enzyme-linked immunosorbent assays and reverse transcription-polymerase chain reactions (RT-PCRs). The protein and mRNA levels of A 2A, NLRP3 and caspase-3 were determined by western blotting and RT-PCR. Results:The degeneration in the nucleus pulposus and annulus fibrosus in the Model group was significant, while significantly less shrinkage, necrosis and fibrous annulus rupture was observed in the Observation group. Compared with Model and Agonist groups, the positive expression of Col Ⅱ in the nucleus pulposus, A 2AR protein levels and relative expression of its mRNA had all increased significantly in the Observation group, while the average levels and mRNA expression of pro-inflammatory cytokines IL-6 and TNF-α had decreased significantly. The average protein level and mRNA expression of NLRP3 and caspase-3 in the intervertebral disc tissues of the Observation group were significantly lower than in the Model and Agonist groups. Conclusions:Combining pulsed electromagnetic field stimulation with A 2A adenosine receptor agonist CGS-21680 can inhibit the apoptosis of nucleus pulposus cells, alleviate disease response and delay IDD by up-regulating the activity of A 2A receptors and down-regulating the expressions of pro-inflammatory factors IL-6, TNF-α, NLRP3 and caspase-3 in nucleus pulposus cells.

OBJECTIVE： To provide reference for the establishment of quality standard of Tussilago farfara formula granules. METHODS： TLC method was used for qualitative identification of tussilagone in T. farfara formula granules. The content of tussilagone in T. farfara formula granule was determined by HPLC. The determination was performed on a Thermo ODS Hypersil C18 column with the mobile phase consisted of methanol-water （85 ∶ 15， V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 220 nm， the column temperature was 25 ℃. Sample size was 20 μL.  RESULTS： TLC spots of tussilagone were clear and well-separated， without interference from negative control. The linear range of tussilagone was 1.39-27.75 μg/mL （r＝0.999 9）. The limits of quantification and detection were 0.153 87 and 0.051 42 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were lower than 2％. The recoveries were 97.12％-103.96％ （RSD＝2.60％， n＝6）. CONCLUSIONS： The method is simple， accurate and reproducible， and suitable for quality control of T. farfara formula granules.

Objective To investigate the efficacy of radiotherapy,as well as the prognostic factors of survival in patients with extracranial oligometastases. Methods A total of 164 patients who underwent intensity-modulated radiotherapy ( IMRT ) of the extracranial oligometastases, from January 2013 to December 2016, were enrolled in the study. The short-term efficacy, local control rate, overall survival, progression free survival and adverse effects of treatment were observed. Results Short-term efficacy was assessed within the first 1-3 months after the end of radiotherapy. The objective response rate (CR+PR) was 78. 7% and the short-term efficacy is mainly related to the T stage of primary tumor ( P＝0. 004).Until the last follow-up,all patients with 1-,2-and 3-year LC were 89. 8%,82. 5% and 74. 9% respectively.Univariate analysis showed that the influencing factors of LC include tumor size and gross tumor volume dose ( all P＜0. 05),multivariate analysis found no significant influence factors. The 1-,2- and 3-year OS were 83. 4%, 69. 6% and 54. 6% respectively. Univariate and multivariate analysis showed that the primary tumor sources, metastasis organs,whether synchronous or adjuvant chemotherapy and short-term efficacy were independent prognostic factors in patients of OS ( P＜0. 05 ).Main toxicity-associated events were grade 1-2 acute reactions,with only 6 patients experiencing grade 3 toxicity;no grade≥4 toxic reactions or treatment-related deaths occurred. Conclusions Radiation therapy for the treatment of extracranial oligometastases can achieve good curative effect,is well-tolerated and has low toxicity.