Performance of SHOX2 and RASSF1A methylation assay in supernatants and matched cell pellets for the diagnosis of malignant pleural effusion.

BACKGROUND: It is difficult to distinguish between malignant pleural effusion (MPE) and benign pleural effusion (BPE). The purpose of this study was to determine the best specimen type by evaluating the DNA methylation status of SHOX2 and RASSF1A in 3 matched PE components. METHODS: In total, 94 patients were enrolled, including 45 MPE, 35 BPE, and 14 undefined PE (UPE) with malignancies. PE samples were processed into supernatants, fresh-cell pellets, and formalin-fixed and paraffin-embedded (FFPE) cell blocks, respectively. A quantitative real-time PCR was used to detect the methylation status of SHOX2 and RASSF1A. RESULTS: SHOX2 and RASSF1A methylation levels were significantly higher in the 3 MPE sample types than those of BPE (P < 0.05). The area under the curve using cell-free DNA (cf-DNA) was the highest. The detection sensitivity of SHOX2 and RASSF1A in fresh-cell DNA, cf-DNA and FFPE cell-block were 71.1% (32/45), 97.8% (44/45) and 66.7% (28/42), respectively, with specificities of 97.1% (34/35), 94.3% (33/35), and 96.9% (31/32). Notably, a combination of the cytological analysis and cf-DNA methylation assay showed an increase in positivity rate from 75.6% to 100%. CONCLUSIONS: The SHOX2 and RASSF1A methylation assay using cf-DNA, the primary recommended specimen type, can excellently increase the diagnostic sensitivity of MPE. A combination of methylation assay with cytological analysis can be used for auxiliary diagnosis of PE.

Prevalence of osteoporosis in patients awaiting unicompartmental knee arthroplasty: a cross-sectional study.

Objectives: Osteoporosis may contribute to failure of unicompartmental knee arthroplasty (UKA), yet the prevalence of osteoporosis in the population awaiting UKA has not been adequately studied. The objectives of this study were to report the prevalence of osteoporosis in people awaiting UKA and the rate of anti-osteoporosis treatment, and to explore factors associated with osteoporosis prevalence in people awaiting UKA. Methods: Participants awaiting UKA from January 2019 to May 2023 were consecutively enrolled. Participants ' age, gender, BMI, knee K-L score, VAS score, history of previous DXA testing, history of anti-osteoporosis treatment, and possible underlying risk factors were recorded. All participants were given a dual-energy x-ray absorptiometry (DXA) test after the visit. The diagnosis of osteoporosis was made according to the World Health Organization criteria. Compare the prevalence of osteoporosis between people waiting for UKA and the general population. Risk factors associated with osteoporosis were analyzed using multiple linear regression and binary logistic regression models. Results: A total of 340 participants were included in the study, 259 in female and 81 in male, with a mean age of 63.53 years (range: 41-84 years), and all participants completed UKA and had DXA prior to UKA. The prevalence of osteoporosis was 40.88% (44.79% in female and 28.40% in male). The prevalence of osteoporosis was higher in female than in male (p<0.001). The prevalence of osteoporosis in the population waiting for UKA was significantly higher than that in the general population (p < 0.001). DXA testing was performed in 12.06% within 1 year prior to the visit. The percentage of those who had received anti-osteoporosis treatment was 20.59% (20.86% in osteoporosis, 22.39% in Osteopenia and 16.42% in normal bone mass). The correlation between age, gender, body mass index, visual analogue scale score and osteoporosis was statistically significant. Conclusion: Osteoporosis is common in people waiting for UKA, but screening and treatment rates are low. Female patients of advanced age and low weight combined with significant pain should be considered for osteoporosis screening and appropriate treatment before UKA.

Simultaneous diagnosis of tuberculous pleurisy and malignant pleural effusion using metagenomic next-generation sequencing (mNGS).

BACKGROUND: Metagenomic next-generation sequencing (mNGS) has become a powerful tool for pathogen detection, but the value of human sequencing reads generated from it is underestimated. METHODS: A total of 138 patients with pleural effusion (PE) were diagnosed with tuberculous pleurisy (TBP, N = 82), malignant pleural effusion (MPE, N = 35), or non-TB infection (N = 21), whose PE samples all underwent mNGS analysis. Clinical TB tests including culture, Acid-Fast Bacillus (AFB) test, Xpert, and T-SPOT, were performed. To utilize mNGS for MPE identification, 25 non-MPE samples (20 TBP and 5 non-TB infection) were randomly selected to set human chromosome copy number baseline and generalized linear modeling was performed using copy number variant (CNV) features of the rest 113 samples (35 MPE and 78 non-MPE). RESULTS: The performance of TB detection was compared among five methods. T-SPOT demonstrated the highest sensitivity (61% vs. culture 32%, AFB 12%, Xpert 35%, and mNGS 49%) but with the highest false-positive rate (10%) as well. In contrast, mNGS was able to detect TB-genome in nearly half (40/82) of the PE samples from TBP subgroup, with 100% specificity. To evaluate the performance of using CNV features of the human genome for MPE prediction, we performed the leave-one-out cross-validation (LOOCV) in the subcohort excluding the 25 non-MPE samples for setting copy number standards, which demonstrated 54.1% sensitivity, 80.8% specificity, 71.7% accuracy, and an AUC of 0.851. CONCLUSION: In summary, we exploited the value of human and non-human sequencing reads generated from mNGS, which showed promising ability in simultaneously detecting TBP and MPE.

Effect of Hypoglycemic Drugs on Kisspeptin Expression in the Hypothalamic Arcuate Nucleus of PCOS Rats.

Background: To investigate the change in hypothalamic kisspeptin-1 (Kiss1) expression during the development of polycystic ovary syndrome (PCOS) and hypoglycemic drug intervention. Methods: Letrozole lavage was used to construct a polycystic ovary rat model. After successful modeling, we treated PCOS rats with metformin, pioglitazone, and acarbose, and we then observed changes in weight, estrus, glucose tolerance, insulin resistance, sex hormones, and hypothalamic kiss1 expression. Results: PCOS rats exhibited increased body weight, abnormal estrous cycle, impaired glucose tolerance, insulin resistance, increased testosterone level, increased luteinizing hormone level, and increased Kiss1 expression in the hypothalamus. However, intervention with metformin, pioglitazone, and acarbose improved the reproductive and metabolic disorders as well as reduced hypothalamic Kiss1 expression. Conclusion: The expression of hypothalamic Kiss1 may play an important role in the pathogenesis of PCOS. Metformin, pioglitazone, and acarbose may reduce the expression of hypothalamic Kiss1 by improving insulin resistance, thereby improving reproductive and metabolic disorders in PCOS rats.

[Comparison of outcomes of mixed unicompartmental knee arthroplasty and total knee arthroplasty in the treatment of medial osteoarthritis of the knee].

OBJECTIVE: To evaluate outcomes of mixed unicompartmental knee arthroplasty(UKA) and total knee arthroplasty(TKA) in the treatment of medial osteoarthritis(OA) of the knee. METHODS: Retrospective analysis of 156 patients, 44 males and 112 females, aged from 50 to 75 years old with an average of(58.76±4.97) years old, who underwent knee arthroplasty from October 2017 to October 2019. The patients were divided into two groups:81 cases(81 knees) underwent TKA, including 23 males and 58 females, aged from 51 to 75 years old with an average of (58.60±5.01) years old, and 75 case (75 knees) underwent UKA with mixed phase 3 Oxford, including 21 males and 54 females, aged from 50 to 72 years old with an average of (58.92±4.95) years old. The two groups were compared regarding to the clinical outcomes, assessed using surgical information and complications, American Knee Society score(AKSS) clinical score and functional score. Radiographs were assessed using hip-knee-ankle angle(HKA), tibial component valgus/varus angle(TCVA), tibial component posterior slope angle(TCPSA), femoral component valgus/varus angle(FCVA), femoral component posterior slope angle(FCPSA), looking for bearing dislocation, prosthesis loosening, progression of OA in lateral compartment. RESULTS: Intraoperative bleeding, operative time and hospital days were significantly better in the UKA group than in the TKA group (P<0.05), and there were no postoperative complications in either group. Patients in both groups were enrolled with an average follow-up time of (38.01±8.90) months, ranged from 24 to 54 months. AKSS functional, AKSS clinical, HKA in both groups significantly improved at the final follow-up compared with those before operation. At the final follow-up, the UKA group was significantly better than the TKA group in AKSS functional and AKSS clinical, whereas HKA in the TKA group was better. At the final follow-up. TCVA and FCVA between the two groups were not significantly different, while TCPSA and FCPSA in the UKA group were significantly greater than the TKA group. No signs of progression of OA to the lateral compartment were observed. CONCLUSION: Mixed phase 3 Oxford UKA in medial unicompartmental knee osteoarthritis was considerably better than TKA for less blood loss, shorter operation time, shorter hospital stay, rapid postoperative recovery, helping achieve satisfactory function, provided satisfactory outcome.

[A Real-world Study on the Expression Characteristics of PD-L1 in Patients 
with Advanced EGFR Positive NSCLC and Its Relationship with the 
Therapeutic Efficacy of EGFR-TKIs].

BACKGROUND: Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) is the first choice for first-line treatment of advanced patients with EGFR positive non-small cell lung cancer (NSCLC). For advanced NSCLC patients with negative drive gene and positive programmed cell death ligand 1 (PD-L1) or highly expressed NSCLC patients, the first-line treatment recommends immune checkpoint inhibitors (ICIs) monotherapy or ICIs combined chemotherapy. Therefore, the first-line treatment strategy for advanced NSCLC patients with EGFR positive at different PD-L1 expression levels is worth further exploring. Many previous studies have suggested that the expression of PD-L1 is obviously affected by EGFR mutation, and the expression of PD-L1 is likely to be related to the mechanism of EGFR-TKIs resistance. The purpose of this study was to analyze the expression characteristics of PD-L1 in patients with advanced EGFR positive NSCLC and its relationship with the efficacy of EGFR-TKIs. METHODS: 159 patients with newly diagnosed advanced NSCLC with EGFR positive (including 141 patients with EGFR sensitive mutations) were enrolled to analyze the relationship between clinicopathological characteristics and PD-L1 expression. The factors affecting the therapeutic effect of EGFR-TKIs in 141 patients with EGFR sensitive mutations were also explored. RESULTS: The PD-L1 expression of the included patients was classified according to the tumor promotion score (TPS): negative (TPS<1%) accounted for 47.2%, low expression (1%≤TPS<50%) accounted for 32.1%, and high expression (TPS≥50%) accounted for 20.7%. Among them, patients with solid predominant cancer cell pathomorphology classification are more likely to have high expression of PD-L1, accounting for 52.9% (P<0.0001). The median progression-free survival (mPFS) of patients with PD-L1 negative expression, low expression and high expression who received first generation EGFR-TKIs treatment were 12.4 months, 10.5 months and 3.7 months, respectively, with significant statistical difference (P<0.0001). During the EGFR-TKIs treatment, patients with a history of radiotherapy for lung lesions have a longer PFS benefit than those without a history of radiotherapy (mPFS: 17.0 months vs 9.3 months, P<0.0001). CONCLUSIONS: The expression level of PD-L1 in advanced EGFR positive NSCLC patients was significantly correlated with the pathomorphology classification of cancer cells. The efficacy of EGFR-TKIs in patients with PD-L1 overexpression NSCLC was significantly poor. PFS can be significantly prolonged during targeted therapy combined with radiotherapy of lung lesions.