High-Speed Centrifugation Efficiently Removes Immunogenic Elements in Osteochondral Allografts.

OBJECTIVES: The current clinical pulse lavage technique for flushing fresh osteochondral allografts (OCAs) to remove immunogenic elements from the subchondral bone is ineffective. This study aimed to identify the optimal method for removing immunogenic elements from OCAs. METHODS: We examined five methods for the physical removal of immunogenic elements from OCAs from the femoral condyle of porcine knees. We distributed the OCAs randomly into the following seven groups: (1) control, (2) saline, (3) ultrasound, (4) vortex vibration (VV), (5) low-pulse lavage (LPL), (6) high-pulse lavage (HPL), and (7) high-speed centrifugation (HSC). OCAs were evaluated using weight measurement, micro-computed tomography (micro-CT), macroscopic and histological evaluation, DNA quantification, and chondrocyte activity testing. Additionally, the subchondral bone was zoned to assess the bone marrow and nucleated cell contents. One-way ANOVA and paired two-tailed Student's t-test are used for statistical analysis. RESULTS: Histological evaluation and DNA quantification showed no significant reduction in marrow elements compared to the control group after the OCAs were treated with saline, ultrasound, or VV treatments; however, there was a significant reduction in marrow elements after LPL, HPL, and HSC treatments. Furthermore, HSC more effectively reduced the marrow elements of OCAs in the middle and deep zones compared with LPL (p < 0.0001) and HPL (p < 0.0001). Macroscopic evaluation revealed a significant reduction in blood, lipid, and marrow elements in the subchondral bone after HSC. Micro-CT, histological analyses, and chondrocyte viability results showed that HSC did not damage the subchondral bone and cartilage; however, LPL and HPL may damage the subchondral bone. CONCLUSION: HSC may play an important role in decreasing immunogenicity and therefore potentially increasing the success of OCA transplantation.

TMT quantitative proteomics reveals key proteins relevant to microRNA-1-mediated regulation in osteoarthritis.

Osteoarthritis (OA) is the second-commonest arthritis, but pathogenic and regulatory mechanisms underlying OA remain incompletely understood. Here, we aimed to identify the mechanisms associated with microRNA-1 (miR-1) treatment of OA in rodent OA models using a proteomic approach. First, N = 18 Sprague Dawley (SD) rats underwent sham surgery (n = 6) or ACL transection (n = 12), followed at an interval of one week by randomization of the ACL transection group to intra-articular administration of either 50 µL placebo (control group) or miR-1 agomir, a mimic of endogenous miR-1 (experimental group). After allowing for eight weeks of remodeling, articular cartilage tissue was harvested and immunohistochemically stained for the presence of MMP-13. Second, N = 30 Col2a1-cre-ERT2 /GFPf1/fl -RFP-miR-1 transgenic mice were randomized to intra-articular administration of either placebo (control group, N = 15) or tamoxifen, an inducer of miR-1 expression (experimental group, N = 15), before undergoing surgical disruption of the medial meniscus (DMM) after an interval of five days. After allowing for eight weeks of remodeling, articular cartilage tissue was harvested and underwent differential proteomic analysis. Specifically, tandem mass tagging (TMT) quantitative proteomic analysis was employed to identify inter-group differentially-expressed proteins (DEP), and selected DEPs were validated using real-time quantitative polymerase chain reaction (RT-qPCR) technology. Immunohistochemically-detected MMP-13 expression was significantly lower in the experimental rat group, and proteomic analyses of mouse tissue homogenate demonstrated that of 3526 identified proteins, 345 were differentially expressed (relative up- and down-regulation) in the experimental group. Proteins Fn1, P4ha1, P4ha2, Acan, F2, Col3a1, Fga, Rps29, Rpl34, and Fgg were the *top ten most-connected proteins, implying that miR-1 may regulate an expression network involving these proteins. Of these ten proteins, three were selected for further validation by RT-qPCR: the transcript of Fn1, known to be associated with OA, exhibited relative upregulation in the experimental group, whereas the transcripts of P4ha1 and Acan exhibited relative downregulation. These proteins may thus represent key miR-1 targets during OA-regulatory mechanisms, and may provide additional insights regarding therapeutic mechanisms of miR-1 in context of OA.

The influence of prostate volume on clinical parameters in prostate cancer screening.

PURPOSE: The purpose of the study was to evaluate the diagnostic significance of two new and a few clinical markers for prostate cancer (PCa) at various prostate volumes (PV). METHODS: The study subjects were divided into two groups. Among them, there were 70 cases in the PV ≤30 ml group (benign prostatic hyperplasia [BPH]: 32 cases, PCa: 38 cases) and 372 cases in the PV > 30 ml group (BPH: 277 cases, PCa: 95 cases). SPSS 26.0 and GraphPad Prism 8.0 were used to construct their receiver operating characteristic (ROC) curves for diagnosing PCa and calculating their area under the ROC curve (AUC). RESULTS: In the PV ≤30 ml group, the diagnostic parameters based on prostate-specific antigen (PSA) had a decreased diagnostic significance for PCa. In the PV > 30 ml group, PSAD (AUC = 0.709), AVR (AVR = Age/PV, AUC = 0.742), and A-PSAD (A-PSAD = Age×PSA/PV, AUC = 0.736) exhibited moderate diagnostic significance for PCa, which was better than PSA-AV (AUC = 0.672), free PSA (FPSA, AUC = 0.509), total PSA (TPSA, AUC = 0.563), (F/T) PSA (AUC = 0.540), and (F/T)/PSAD (AUC = 0.663). Compared with AVR, A-PSAD exhibited similar diagnostic significance for PCa, but higher than PSA density (PSAD). CONCLUSIONS: Choosing appropriate indicators for different PVs could contribute to the early screening and diagnosis of PCa. The difference in the diagnostic value of two new indicators (A-PSAD and AVR), and PSAD for PCa may require further validation by increasing the sample size.

Clinical research analysis based on prostate cancer screening diagnosis.

This study aimed to analyse the clinical characteristics and risk factors of patients with positive prostate biopsy at 4-20 ng/mL of prostate-specific antigen (PSA), construct a new parameter based on this characteristics and assess its diagnostic value for prostate cancer (PCa). Logistic regression analysis was used to clarify the risk factors of PCa, and a new parameter based on the results was constructed. Compare the diagnostic value of various diagnostic parameters for PCa. Logistic multivariate regression analysis revealed that age (OR, 5.269; 95%CI, 2.762-10.050), comorbid diabetes (OR, 2.437; 95%CI, 1.162-5.111), PSA (OR, 2.462; 95%CI, 1.198-5.059) and prostate volume (PV) (OR, 0.227; 95%CI, 0.100-0.516) are risk factors for PCa. The age, PSA and PV of patients were combined to construct a new parameter, that is A-PSAD = (age × total PSA [TPSA])/PV]. The area under the receiver-operating characteristic curve(AUC) of A-PSAD (0.728) for PCa diagnosis was higher than the AUCs of TPSA (0.581), free prostate-specific antigen (0.514), (F/T)PSA (0.535) and PSAD (0.696), with significant differences. Age, history of diabetes, TPSA and PV are risk factors for PCa(PSA:4-20ng/mL); in addition, A-PSAD has a moderate diagnostic value for PCa and may become a new indicator for PCa screening.

Impact of Surgical Wait Time on Survival in Patients With Upper Urinary Tract Urothelial Carcinoma With Hydronephrosis.

BACKGROUND AND OBJECTIVES: Due to the inevitability of waiting time for surgery, this problem seems to have become more pronounced since the outbreak of COVID-19, and due to the high incidence of preoperative hydronephrosis in upper urinary tract urothelial carcinoma (UTUC) patients, it is particularly important to explore the impact of preoperative waiting time and hydronephrosis on upper urinary urothelial carcinoma. METHODS: 316 patients with UTUC who underwent radical surgery at a high-volume center in China between January 2008 and December 2019 were included in this study. We retrospectively collected the clinicopathologic data from the medical records, including age, sex, smoking history, ECOG performance status (ECOG PS), body mass index (BMI), tumor location and size, number of lesions, T stage, N stage, surgical approach and occurrence of hydronephrosis, lymph node invasion, lymph node dissection, surgical margin, tumor necrosis, infiltrative tumor architecture, lymphovascular invasion and concomitant bladder cancer. Surgical wait time was defined as the interval between initial imaging diagnosis and radical surgery of UTUC. Hydronephrosis was defined as abnormal dilation of the renal pelvis and calyces due to obstruction of the urinary system. Firstly, all patients were divided into short-wait (<31 days), intermediate-wait (31-90 days) and long-wait (>90 days) groups according to the surgical wait time. The clinicopathological characteristics of each group were evaluated and the survival was compared. For patients with hydronephrosis, we subsequently divided them into two groups: short-wait (≤60 days) and long-wait (>60 days) groups according to the surgical wait time. Univariate and multivariate COX regression analysis were performed to evaluate the prognostic risk factor for patients with hydronephrosis. RESULTS: A total of 316 patients with UTUC were included in this study with a median surgical wait time of 22 days (IQR 11-71 days). Of the 316 patients, 173 were classified into the short-wait group (54.7%), 69 into the intermediate-wait group (21.8%) and 74 into the long-wait group (23.5%). The median follow-up time for all patients was 43 months (IQR 28-67months). The median surgical wait times of the short-wait, intermediate-wait and long-wait group were12 days (IQR 8-17days), 42days (IQR 37-65days) and 191days (IQR 129-372days), respectively. The 5-year overall survival (OS) of all patients was 54.3%. The 5-year OS of short-wait, intermediate-wait and long-wait groups were 56.4%, 59.3% and 35.1%, respectively (P=0.045). The 5-year cancer-specific survival (CSS) of short-wait, intermediate-wait and long-wait groups were 65.8%, 70.9% and 39.6%, respectively (P=0.032). In the subgroup analysis, we divided 158 UTUC patients with hydronephrosis into short-wait group (≤60 days) and long-wait group (> 60 days), 120 patients were included in the short-wait group and 38 patients in the long-wait group. The median surgical wait times of the short-wait and long-wait group were 14days (IQR 8-28days) and 174days (IQR 100-369days), respectively. The 5-year OS of long-wait group was significantly lower than the OS of short-wait group (44.2% vs. 55.1%, P =0.023). The 5-year CSS of long-wait and short-wait group were 49.1% and 61.7%, respectively (P=0.041). In multivariate Cox regression analysis of UTUC patients with hydronephrosis, surgical wait time, tumor grade, pathological T stage, and tumor size were independent risk factors for OS and CSS. Lymph node involvement was also a prognostic factor for CSS. CONCLUSION: For patients with UTUC, the surgical wait time should be limited to less than 3 months. For UTUC patients with hydronephrosis, the OS and CSS of patients with surgical wait time of more than 60 days were relatively shorted than those of patients with surgical wait time of less than 60 days.

Effects of captopril and valsartan on ventricular remodeling and inflammatory cytokines after interventional therapy for AMI.

The effects of captopril and valsartan on ventricular remodeling and inflammatory cytokines after interventional therapy for acute myocardial infarction (AMI) were investigated. A total of 94 patients with AMI admitted to Honggang Hospital of Dongying from July 2016 to June 2017 were selected as study subjects. The patients were treated with interventional therapy and randomly divided into the observation group (n=47) and the control group (n=47). The control group received aspirin after operation, while the observation group received captopril and valsartan after operation. Three-dimensional ultrasonography was performed to evaluate ventricular remodeling. The related parameters included left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF), end-systolic sphericity index/end-diastolic sphericity index (ESSI/EDSI), systolic dyssynchrony index (SDI), diastolic dyssynchrony index (DDI), dispersion end systole (DISPES), DDI-late and DISPED-late. The levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay (ELISA). The incidence of adverse reactions after treatment was compared. After treatment, LVEF in the control group was significantly lower than that in the observation group, while LVEDV, LVESV and the ratio of early diastolic (E) and late diastolic (A) (E/A) in the control group were significantly higher than those in the observation group (p<0.05). EDSI, DDI-late and DISPED-late in the control group were significantly higher than those in the observation group (p<0.05). ESSI, SDI and DISPES in the control group were significantly higher than those in the observation group (p<0.05). The levels of interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP) and tumor necrosis factor-α (TNF-α) in the observation group were significantly lower than those in the control group at 1, 4 and 8 weeks after treatment (p<0.05). The administration of captopril and valsartan after interventional therapy for AMI can effectively improve the cardiac function of patients, improve the synchronism of left ventricular diastole and contraction, and reduce the level of inflammation. It is safe and reliable, and has important clinical significance.