Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

Bone organoids can simulate the complex structure and function of the bone tissues, which makes them a frontier technology in organoid researches. Bone organoids show a tremendous potential of applications in bone disease modeling, bone injury repair, and medicine screening. Although advancements have been made so far in constructing bone organoids with functional structures like mineralization, bone marrow, trabecular bone, callus, woven bone, etc, the researches in this field are confronted with numerous challenges such as lack of standardized construction strategies and unified evaluation criteria, which limits their further promotion and application. To standardize researches in bone organoids, the Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, the Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, the Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and the Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine organized related experts to formulate Expert consensus on the construction, evaluation, and application of bone organoids ( version 2024) based on an evidence-based approach. A total of 17 recommendations were put forth, aiming to standardize researches and clinical applications of bone organoids and enhance their value in scientific research and clinical practice.

Objective:To investigate the incidence of various non-motor symptoms (NMS) in early stage of Parkinson′s disease (PD) patients and the differences between the body-first and brain-first subtypes.Methods:A total of 121 patients with PD (Hoehn-Yahr stage 1-2) were recruited from PD Clinic, Department of Neurology, Beijing Hospital from January 2012 to January 2015. The general information and clinical features of the patients were collected. The minimal diagnostic criteria of parasomnias described in the International Classification of Sleep Disorders-Revised were used to diagnose rapid eye movement sleep behavior disorder (RBD).According to the sequence of RBD and motor symptoms, the patients were divided into 2 groups: body-first subtype and brain-first subtype. NMS was evaluated by the Non-Motor Symptom Questionnaire (NMSQuest). The clinical features and the incidence of various NMS were compared between the 2 groups. The Unified Parkinson′s Disease Rating Scale (UPDRS) was used to evaluate the severity of the disease, and its third part (UPDRS-Ⅲ) was used to evaluate the motor function of the patients. Hamilton Rating Scale for Depression (HAMD) and Hamilton Rating Scale for Anxiety (HAMA) were used to evaluate the depression and anxiety status of the patients. The sleep status of patients was assessed by Parkinson′s Disease Sleep Scale (PDSS). The quality of life of the patients was assessed by 39-item Parkinson′s Disease Questionnaire (PDQ-39).Results:Of all the patients, 49.59% (60/121) had the body-first subtype and 50.41% (61/121) had the brain-first subtype of PD. There was no significant difference in UPDRS-Ⅲ score between the 2 groups. The average number of NMS in all PD patients was 10.97±4.88. Body-first subtype patients had higher NMS incidence than brain-first subtype in difficulty in swallowing [46.7% (28/60) vs 23.0% (14/61), χ 2=7.507, P=0.006], nausea and vomiting [16.7% (10/60) vs 3.3% (2/61), χ 2=6.069, P=0.014], constipation [85.0% (51/60) vs 55.7% (34/61), χ 2=12.393, P<0.001], fecal incontinence [8.3% (5/60) vs 0 (0/61), χ 2=5.302, P=0.021], difficulty in remembering recent events [58.3% (35/60) vs 32.8% (20/61), χ 2=7.962, P=0.005], loss of interest [43.3% (26/60) vs 24.6% (15/61), χ 2=4.743, P=0.029], inattention [45.0% (27/60) vs 19.7% (12/61), χ 2=8.884, P=0.003], depression [55.0% (33/60) vs 34.4% (21/61), χ 2=5.181, P=0.023], intense vivid dreams [73.3% (44/60) vs 39.3% (24/61), χ 2=14.196, P<0.001] and restless legs [53.3% (32/60) vs 27.9% (17/61), χ 2=8.140, P=0.004]. The differences were significant. Body-first subtype and NMSQuest ( r=-0.489, P<0.001), UPDRS ( r=-0.189, P=0.038), HAMD ( r=-0.231, P=0.011), HAMA ( r=-0.298, P=0.001) and PDQ-39 scores ( r=-0.276, P=0.002) were negatively correlated. Body-first subtype and PDSS score was positively correlated. NMSQuest (Δ R2=0.265, P<0.001) was the main determinant of PDQ-39 score. Conclusions:PD patients are accompanied by various NMS, which is a major factor affecting the quality of life. Compared with brain-first subtype, body-first subtype might have more NMS burden and higher incidence rate in most NMS in early PD patients.

Objective To investigate the biomarkers for Parkinson disease(PD) by analyzing differentially expressed proteins in the serum of patients with PD. Methods A total of 24 patients with PD who were diagnosed in Beijing Hospital were enrolled,among whom 12 patients only had a medical history of PD(group P1) and the other 12 had underlying diseases such as hypertension and diabetes(group P2). A total of 24 individuals,matched for age and sex,who underwent physical examination were enrolled as control groups C1 and C2.Serum samples were collected,and after high-abundant proteins were removed,the label-free quantitative proteomics technique was used to measure the expression of proteins in serum. The double comparison method was used for comparison between groups P1 and C1 and between groups P2 and C2 to screen for differentially expressed proteins,and the proteins with a consistent changing trend(upregulation or downregulation) were identified as the differentially expressed proteins for PD,which were analyzed and interpreted by bioinformatics methods. Results Comparison between groups identified four differentially expressed proteins,i.e. PRG4,CFHR-3,ACTG1,and HIST2H2BF,among which PRG4 and CFHR-3 showed upregulated expression and ACTG1 and HIST2H2BF showed downregulated expression in the serum of PD patients,and a certain degree of interaction was observed. Conclusion Label-free quantitative proteomics can be used to identify the differentially expressed proteins in PD,which may have a certain value in the diagnosis of PD.

Unlike healthy, non-transformed cells, the proteostasis network of cancer cells is taxed to produce proteins involved in tumor development. Cancer cells have a higher dependency on molecular chaperones to maintain proteostasis. The chaperonin T-complex protein ring complex (TRiC) contains eight paralogous subunits (CCT1-8), and assists the folding of as many as 10% of cytosolic proteome. TRiC is essential for the progression of some cancers, but the roles of TRiC subunits in osteosarcoma remain to be explored. Here, we show that CCT4/TRiC is significantly correlated in human osteosarcoma, and plays a critical role in osteosarcoma cell survival. We identify a compound anticarin-β that can specifically bind to and inhibit CCT4. Anticarin-β shows higher selectivity in cancer cells than in normal cells. Mechanistically, anticarin-β potently impedes CCT4-mediated STAT3 maturation. Anticarin-β displays remarkable antitumor efficacy in orthotopic and patient-derived xenograft models of osteosarcoma. Collectively, our data uncover a key role of CCT4 in osteosarcoma, and propose a promising treatment strategy for osteosarcoma by disrupting CCT4 and proteostasis.

Objective:To investigate the characteristics and clinical related factors of Parkinson′s disease (PD) patients with subjective cognitive decline (SCD).Methods:Ninety-nine PD patients with normal cognitive function enrolled in Beijing Hospital from January to December 2018 were collected for the study. Patients with PD were divided into groups with ( n=57) and without ( n=42) SCD using the first question in Part 1 of the Unified Parkinson′s Disease Rating Scale (UPDRS). All patients were assessed by Montreal Cognitive Assessment (MoCA), modified Hoehn-Yahr grading, UPDRS, Hamilton Rating Scale for Depression (HAMD), Hamilton Rating Scale for Anxiety (HAMA), Parkinson′s Disease Sleep Scale, Ability of Daily Living Scale and 39-item Parkinson′s Disease Questionnaire (PDQ-39). Levodopa equivalent dose conversion was performed for patients taking anti-PD drugs. Patients′ self-reported years of formal education were collected. Results:The proportion of PD with SCD in this group was 57.58% (57/99). There were statistically significant differences in MoCA [28.00 (27.00, 29.00) vs 28.00 (27.00, 29.00) ,Z=-2.28, P=0.023], HAMD [6.00 (5.00, 8.50) vs 5.00 (2.00, 8.00), Z=-2.23, P=0.026], HAMA [7.00 (6.00, 11.00) vs 6.00 (3.00, 8.25) , Z=-2.70, P=0.007], PDQ-39-emotional health [2.00 (0, 5.00) vs 1.00 (0, 3.00), Z=-2.03, P=0.042] and PDQ-39-cognitive scores [4.00 (2.00, 5.00) vs 2.00 (0, 4.00), Z=-3.42, P=0.001] between PD with and without SCD groups. SCD was correlated with MoCA ( r=-0.23, P=0.022), HAMD ( r=0.23, P=0.025) and HAMA ( r=0.27, P=0.006) scores to varying degrees. When controlling for HAMD and HAMA scores, the correlation between SCD and MoCA scores ( r′=-0.18, P=0.084) was no longer existed. Conclusions:SCD is common in PD patients with normal cognitive function and is associated with poorer cognitive performance and more severe symptoms of depression and anxiety. In this group of patients, the relationship between SCD and affective symptoms may be greater than that of objective overall cognitive function, which is worthy of further studies.

Objective:To explore the feasibility of predicting TP53 mutation risk by immunohistochemical staining (IHC) pattern of P53 in Chinese diffuse large B-cell lymphoma (DLBCL) and its correlation with a prognostic difference.Methods:Between January 2021 and December 2021, 51 DLBCL cases at Beijing Boren Hospital were gathered. These cases had both IHC and next-generation sequencing (NGS) results. IHC classified the P53 protein expression pattern into a loss (<1% ) , diffuse (>80% ) , and heterogeneous (1% -80% ) . The sensitivity and specificity of the predicting TP53 mutation by IHC were assessed by comparing the results of the NGS, and the TP53 high mutation risk group included both loss and diffuse expression of P53. From June 2016 to September 2019, Peking University Cancer Hospital collected 131 DLBCL cases with thorough clinicopathological and follow-up data. From their tumor blocks, tissue microarray blocks were made for IHC evaluation of P53 expression pattern, and prognosis effect of P53 studies.Results:Among 51 cases with both IHC and NGS results, 23 cases were classified as TP53 high mutation risk (7 cases loss and 16 cases diffuse) , 22/23 cases were proved with mutated TP53 by NGS. Only 1 of the 28 cases classified as TP53 low mutation risk was proved with mutated TP53 by NGS. IHC had a sensitivity and specificity of 95.7% and 96.4% for predicting TP53 mutation. NGS identified a total of 26 TP53 mutations with a mutation frequency of 61.57% (13.41% -86.25% ) . In the diffuse group, 16 missense mutations and 2 splice mutations were detected; 6 truncating mutations and 1 splice mutation were detected in the loss group; 1 truncating mutation was detected in the heterogeneous group. Multivariate analysis demonstrated that TP53 cases with high mutation risk have impartial adverse significance for the 131 patients included in survival analysis ( HR=2.612, 95% CI 1.145-5.956, P=0.022) . Conclusion:IHC of P53 exhibiting loss (<1% ) or diffuse (>80% ) pattern indicated TP53 high mutation risk, IHC can predict TP53 mutation with high specificity and sensitivity. TP53 high mutation risk is an independent predictor for adverse survival.

Objective:To investigate the clinical characteristics of elderly patients with coronavirus disease 2019(COVID-19), in order to provide scientific evidence for the diagnosis and treatment of COVID-19 in elderly patients.Methods:Clinical data of 102 patients with COVID-19 admitted to the B11 East Ward of the Zhongfaxincheng campus and the E1-3 ward of the Guanggu Campus of Tongji Hospital affiliated to Huazhong University of Science and Technology in Wuhan from 1 February 2020 to 28 February 2020 were retrospectively collected and analyzed.Patients were categorized into 2 groups: the elderly group(≥60 years old)and the young and middle-aged group(<60 years old). Differences in epidemiological features, demographics, clinical symptoms, laboratory results and imaging findings between the two groups were retrospectively analyzed.Results:Among 102 patients with COVID-19, 58 were in the elderly group(≥60 years old), with a median age of 67.0(63.8, 71.0)years old, and 44 in the young and middle-aged group(<60 years old), with a median age of 47.5(38.0, 51.8)years old.There was no significant difference in gender ratio between the two groups( χ2=0.033, P=0.855). Of 102 patients, 42.0%(21/50)had close contact with an infected person, 14.0%(7/50)were from infection clusters, and 18.0%(9/50)had suspected hospital-acquired infections.Fever and cough remained the most common symptoms, but gastrointestinal symptoms such as nausea, poor appetite, diarrhea and muscle cramps were also warning signs.Fatigue and cough were the most common presenting symptoms in elderly male patients.Bilateral patchy infiltrates(57.9%, 22/38)and ground-glass opacities(42.1%, 16/38)were the main imaging features and 42.1%(16/38)patients had multiple areas of the lungs involved.Over 50% patients had increased levels of blood glucose, D-dimer, fibrinogen, C-reactive protein, procalcitonin, multiple cytokines and neutrophil-to-lymphocyte ratio, as well as decreased levels of albumin, hemoglobin, hematocrit, lymphocytes and serum calcium.Compared with the young and middle-aged group, the elderly group had higher rates of abnormality in levels of D-dimer and serum calcium( χ2=7.067 and 4.166, P=0.008 and 0.041). Conclusions:Fever and cough are the most common symptoms in elderly patients with COVID-19.Elderly patients with COVID-19 have multiple abnormalities in clinical laboratory test results, which show a certain level of specificity compared with young and middle-aged patients.

To investigate the effects of Mycobacterium tuberculosis heat shock protein 65(HSP65)on Treg/Th17 immune balance in ApoE-knockout(ApoE-/-)mice, ApoE-/- mice with a high-cholesterol diet were immunized with M. tuberculosis HSP65. Sera were obtained for measurement of anti-HSP65 antibodies by ELISA; the effect of administration of different antigens was investigated, respectively, using flow cytometry analysis on the number of CD4+CD25+Foxp3+Tregs and CD4+IL-17+ Th17; the production of cytokines(IL-10, TGF-β1, IL-17 and IL-21)by these cells were determined by ELISA; total plasma cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were detected by biochemical autoanalyzer. Atherosclerotic lesions were measured by lipid deposition stained with oil red O. The results demonstrated that the levels of anti-HSP65 IgG antibodies were increased significantly in Mycobacterium tuberculosis HSP65-treated ApoE-/- mice, revealed obvious decrease in Treg number, Treg related cytokines(IL-10, TGF-β1)levels and significant increase in Th17 number, Th17 related cytokines(IL-17 and IL-21)levels, the levels of TC, TG, HDL-C and LDL-C did not change between groups, while the atherosclerotic lesions significantly increased. Results indicate that M. tuberculosis HSP65 could interrupt the Th17/Treg immune balance in ApoE-/- mice, suggesting a potential role in the formation and progression of atherosclerosis.

Objective To observe the effect of different doses of sufentanil on intubation guided by fiber bronchoscope in diffi‐cult airways .Methods One hundred and eighteen patients with difficult airways who underwent tracheal intubation were randomly divided into 3 groups [groupⅠ(n=40) ,groupⅡ (n=42) ,and groupⅢ (n=36)] .First all group received 1 μg/kg dexmedetomi‐dine ,then groupⅠ ,groupⅡ and group Ⅲ received sufentanil 0 .1μg/kg ,0 .2 μg/kg ,0 .3μg/kg infusion in bolus respectively .Heart rate(HR) ,mean arterial pressure(MAP)and saturation of pulse oximetry(SpO2 )were recorded at 5 min after patients arriving at operation room (T0 ) ,after drug infusion(T1 ) ,the time of intubation and seeing the epiglottis (T2 ) ,1 min(T3 )and 5 min(T4 )after intubation .side effect was recorded throughout the process .Results HR at T2 and T3 in groupⅡ and group Ⅲ were significantly lower than those in group Ⅰ(P<0 .05) .MAP at T2 and T3 in groupⅡ and group Ⅲ were significantly lower than those in groupⅠ(P<0 .05) .SpO2 at T2 in group Ⅲ was significantly lower than those of groupⅠ and groupⅡ(P<0 .05) .Cough reflex in groupⅠwas much frequent than others .Only group Ⅲ had respiratory inhibition .Conclusion Intravenous sufentanil (0 .2 μg/kg)could in‐hibit effectively stress reaction from endotracheal intubation ,in which less side effects such as haemodynamic changes and respirato‐ry depression occur .