High-risk human papillomavirus (HR-HPV), especially types 16 and 18, is strongly linked to cervical cancer (CC) incidence, accounting for approximately 70 % of cases globally. The current HPV detection methods are complex, requiring stringent preservation and transportation conditions, and contamination poses a risk. We thus developed a stable, simple, and efficient multiplex polymerase chain reaction (PCR)-based HPV detection method allowing for the simultaneous detection and differentiation of 14 high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) in a single closed-tube reaction. Lyophilized microspheres were prepared via vacuum freeze-drying and integrated into the PCR system. The sensitivity, specificity, precision, stability, morphology, and interference resistance of the system were evaluated. The results obtained with the developed system and with fluorescence PCR melting curves and flow cytometric hybridization were comparable; Sanger sequencing was used to verify discordant results. The lyophilized microspheres showed excellent inter-batch consistency, with coefficients of variation ranging narrowly from 0.14 % to 0.36 %. The lyophilized micro-spheres were mechanically stable, maintained their morphological and structural integrity, and furthermore demonstrated stability at room temperature for up to 1 year. The assay exhibited a limit of detection of 350-500 copies/mL, and demonstrated high specificity with no observed cross-reactivity and high resistance to challenging levels of common clinical interferents. The results using 1569 clinical samples were consistent with those of flow fluorescence hybridization and fluorescence PCR melting curves, with kappa values of 0.883 to 1.000. The sensitivity of the HR-HPV assay used for primary CC screening was 83.76 % and the negative predictive value was 98.17 %. The method shows promise for CC screening owing to its ambient stability, sensitivity, and operational simplicity.

This review aims to synthesize recent evidence to clarify the causal and mechanistic roles of lipid metabolism disorders as core drivers of type 2 diabetes mellitus (T2DM), and to summarize emerging therapeutic advances targeting key nodes within pathophysiological cascade, thereby providing an integrated, translational framework for the precision management of T2DM. Normoglycemic obese individuals with insulin resistance (IR) frequently exhibit elevated plasma free fatty acids (FFAs), triglycerides (TGs), and low-density lipoprotein cholesterol (LDL-C), alongside reduced high-density lipoprotein cholesterol (HDL-C). These observations indicate that dyslipidemia is not merely a comorbidity but an important early driver of T2DM progression. Lipid metabolism disorders initiate a unifying pathological cascade: ectopic lipid spillover induces systemic IR in the liver and skeletal muscle; lipotoxic stress directly compromises pancreatic β-cell function; and progressive disruption of inter-organ axes, including the gut–adipose–liver axis and brain–periphery axis, further exacerbates systemic metabolic deterioration. Recent evidence supports emerging therapeutic strategies, including traditional medicine-based approaches and interventions targeting lipotoxicity and the gut–brain–liver–fat axis to restore metabolic health. Lipid metabolism disorders are not merely complications of T2DM, but critical drivers of disease progression. Adipose dysfunction acts as the initiating event, triggering a pathological cascade that induces systemic IR, impairs pancreatic β-cell function via lipotoxic intermediates, and disrupts inter-organ communication networks. Targeted interventions along this axis therefore represent a promising strategy for precision therapy.

OBJECTIVE:To evaluate the impact of varying lipid turbidity (LT) levels on D-dimer measurements after routine centrifugation (RC) and high-speed centrifugation (HC) in lipemic samples. METHODS:Lipemic samples (triglyceride > 1.7 mmol/L) were classified into four LT grades via HIL testing and visual inspection. Coagulation parameters (APTT, TT, FIB, PT-INR, and D-dimer) were compared between RC (2600 × g/10 min) and HC (10 000 × g/10 min) in 104 lipemic and 30 non-lipemic control samples. RESULTS:Significant differences (p < 0.05) were observed in all coagulation indices between RC and HC for lipemic samples. LT level was positively correlated with TG and total cholesterol (TC). Deviation rates for APTT, TT, FIB, and PT-INR were below 15%, while D-dimer deviation rates exceeded 50% in severe LT (grades 3-4). D-dimer levels in the lipid layer were significantly higher than those in the plasma layer (p < 0.05). HC failed to resolve interference in severely turbid samples, where D-dimer adhered to chylomicron-rich lipid fractions. CONCLUSION:Contrary to CLSI recommendations, high-speed centrifugation (HC) significantly reduced D-dimer concentrations in the lower plasma layer of severely turbid specimens (e.g., type II hyperlipidemia), failing to mitigate turbidity interference. This discrepancy may lead to misdiagnosis of high-risk thrombotic conditions, such as pancreatitis. The upper lipid layer, predominantly composed of chylomicrons, exhibits a strong binding affinity with D-dimer, further complicating accurate measurement. Direct dilution, however, may resolve measurement failures in severe LT samples by maintaining analyte integrity while eliminating lipid interference.

The distribution of peripheral blood lymphocytes may change due to the influence of diseases. This study aims to investigate the impact of Hepatitis B virus (HBV) infection on the distribution of peripheral blood lymphocyte subsets in patients with Rheumatoid Arthritis (RA). Two hundred ninety-eight patients were recruited from a retrospective cohort of patients with RA. Patients with RA (n = 43) who had hepatitis B surface antigen (HBsAg) positivity in the serum were categorized into the HBV group (HBV-RA group), while 255 RA patients without HBsAg positivity were assigned to the control group. The patients in the HBV-RA group were further divided into two subgroups based on their levels of HBV DNA: those with levels below the lower limit of quantification (< 20 IU/ml) formed the HBV DNAlow group, while those with levels above or equal to this limit (≥ 20 IU/ml) constituted the HBV DNAhigh group. Demographic, clinical and laboratory data were also collected. Compared with those of the control group, a higher proportion of CD19+ B cells and CD8+ T cells and a lower CD4+/CD8+ ratio were observed in the HBV-RA group (all P < 0.05). The same trend was observed in the HBV DNAhigh group compared to the HBV DNAlow group (all P < 0.05). Additionally, based on multivariable logistic regression analysis, the male gender, DAS-28 ≥ 2.6, a high proportion of CD19 + B and CD8 + T cells, and the elevated levels of rheumatoid factor (RF) were found to be significantly associated with RA concurrent with HBV infection (all P < 0.05). In the HBV-RA group, a correlation analysis was conducted revealing a positive association between CD19 + B cells and DAS-28 score, as well as CD8 + T cells and DAS-28 score. There was no statistically significant difference in CD4/CD8 ratio between different DAS-28 groups, however the study revealed a significant negative association between the ratio of CD4 +/CD8 + and the DAS-28 score. The high proportion of CD19 + B and CD8 + T cells were closely associated with RA concurrent with HBV infection. • The high proportion of CD19+ B cells and CD8+ T cells were closely associated with RA concurrent with HBV infection. • CD19+ B cells and CD8+ T cells demonstrated a positive correlation with the DAS-28 score. • CD4+/CD8+ ratio demonstrated a negative correlation with the DAS-28 score. • Male gender, DAS-28 ≥ 2.6 and the elevated level of RF were closely associated with RA concurrent with HBV infection.

To evaluate the ovarian reserve (OR) in women with antiphospholipid syndrome (APS), systemic lupus erythematosus (SLE), especially SLE-associated APS, and to determine the association between OR and clinical and laboratory parameters.

Rheumatoid arthritis (RA) is a chronic inflammatory disorder characterized by pain, swelling, stiffness, and impaired function. Attenuating inflammation is a crucial objective in RA management. Diet and nutrition are believed to influence RA symptomatology, with a low-protein diet being one potential nutritional strategy, although its underlying mechanisms remain to be fully elucidated. In this research, serum derived from arthritic transgenic K/BxN mice was administered to naive mice to establish a K/BxN rheumatoid arthritis model. Physiological assessments and histological staining were performed to evaluate joint pathology. (Enzyme-linked immunosorbent assay) ELISA was used to measure inflammatory cytokines. Flow cytometry and immunofluorescence were applied to characterize macrophage phenotypes. Transcriptomic analysis elucidated molecular pathways under the effect of a low-protein diet and verified by immunoblotting. Mitochondrial reactive oxygen species (ROS) was detected by Mito-SOX. Protein expression was silenced through the application of siRNA transfection. Our results indicate that a low-protein diet significantly alleviates disease symptoms and decreases pro-inflammatory cytokine levels in synovial fluid. Furthermore, this dietary intervention inhibits M1 macrophage polarization while promoting a shift towards the M2 phenotype. Transcriptomic analysis revealed that the beneficial effects of the low-protein diet in alleviating rheumatoid arthritis are closely linked to the NRF2 pathway. In vitro, low protein treatment can promote the activity of NRF2 via inhibiting the ubiquitin mediated proteolysis and activate the NRF2/SIRT3/SOD2 pathway to inhibit the production of ROS, which will further inhibit the M1 macrophage polarization. NRF2 knockdown can abolish the effects of low-protein treatment, indicating that the inhibition of M1 polarization and the anti-inflammatory response induced by low-protein treatment are dependent on NRF2. In summary, our findings propose that low-protein diet can inhibit synovial macrophage M1 polarization via activating NRF2/SIRT3/SOD2 pathway to reduce mitochondrial ROS production. This mechanism effectively decreases synovial inflammation and alleviates RA symptoms.

Studies have indicated that low high-density lipoprotein cholesterol (HDL-C) level is an important risk factor for diabetic kidney disease (DKD) in patients with type 2 diabetes (T2D). However, whether higher HDL-C levels decrease the risk of developing DKD remains unclear. This study aimed to clarify the relationship between HDL-C levels and DKD risk in individuals with T2D in China. In total, 936 patients with T2D were divided into DKD and non-DKD groups. The association between HDL-C levels and DKD risk was evaluated using logistic regression analysis and restricted cubic spline curves adjusted for potential confounders. Threshold effect analysis of HDL-C for DKD risk was also performed. Higher HDL-C levels did not consistently decrease the DKD risk. Furthermore, a nonlinear association with threshold interval effects between HDL-C levels and the incidence of DKD was observed. Patients with HDL-C ≤ 0.94 mmol/L or HDL-C > 1.54 mmol/L had significantly higher DKD risk after adjusting for confounding factors. Interestingly, the association between high HDL-C levels and increased DKD risk was more significant in women. A U-shaped association between HDL-C levels and DKD risk was observed; therefore, low and high HDL-C levels may increase the DKD risk in patients with T2D.

Objectives: To evaluate the ovarian reserve (OR) in women with antiphospholipid syndrome (APS), systemic lupus erythematosus (SLE), especially SLE-associated APS, and to determine the association between OR and clinical and laboratory parameters. Methods: We compared the antral follicle count (AFC), anticardiolipin antibody, and anti-Mullerian hormone (AMH), inhibin B (INHB), antiphospholipid (aPL) antibody, follicle-stimulating hormone (FSH), progesterone (P), testosterone (T), and estradiol (E2) among patients with primary APS (PAPS), SLE-APS, and SLE who were treated at Jinhua Central Hospital between 2017 and 2020. We conducted correlations and logistic regression analyses to identify the risk factors of OR failure in women with APS. Results: Serum AMH were positively correlated with AFC and INHB in APS patients, and low AMH was independent risk factor for OR decline in APS patients. The ROC curve showed a high accuracy for AMH in the prediction of OR failure. Compared to healthy subjects (HS), patients with PAPS, SLE-APS, and SLE exhibited lower serum AMH, AFC, INHB, and E2 levels and higher FSH and levels (p<0.05). Of all the patients, those with SLE-APS manifested the lowest serum AMH, AFC, INHB, and E2 levels and the highest FSH levels (p<0.05). Conclusions: APS and SLE patients showed lower indications of OR, including AFC and AMH, compared to HS. SLE-APS patients also appeared to have a lower OR than either SLE or PAPS patients.

Abstract Objectives To evaluate the ovarian reserve (OR) in women with systemic lupus erythematosus (SLE)-antiphospholipid syndrome (APS), and to determine the association between OR and clinical and laboratory parameters.Methods We compared the antral follicle count (AFC) and blood parameters of patients who were diagnosed with APS or SLE but had not yet been treated. We compared the presence of anticardiolipin antibody, and concentrations of anti-Müllerian hormone (AMH), inhibin B (INHB), follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), testosterone (T), and estradiol (E2) among patients with primary APS(PAPS), SLE-associated APS, and SLE who were treated at Jinhua Central Hospital between 2017 and 2020. We also conducted correlations and logistic regression analyses to identify the risk factors of OR failure in women with APS.Results Serum AMH levels were positively correlated with AFC and INHB levels in APS patients, and low AMH and high LH were independent risk factor for OR decline in APS patients. The receiver operating characteristic (ROC) curve also showed a high accuracy for AMH in the prediction of OR failure. Compared to healthy subjects (HS), patients with PAPS, SLE-APS, and SLE exhibited lower serum AMH, AFC, INHB, and E2 levels and higher FSH and LH levels (P < 0.05). Of all the patients, those with SLE-APS manifested the lowest serum AMH, AFC, INHB, and E2 levels and the highest FSH and LH levels (P < 0.05).Conclusions APS and SLE patients showed lower indications of OR, including AFC, AMH and LH, compared to HS. SLE-APS patients also appeared to have a lower OR than either SLE or PAPS patients. We noted that the low E2 levels in APS and SLE patients on day 3 of the menstrual cycle were not able to inhibit FSH release.

Introduction Bone marrow-derived mesenchymal stem cells (BMSCs)-derived exosomes are involved in the modulation of tissue repair and regeneration. CircRNAs play important roles in BMSCs exosomes. The current study sought to explore the role of circRNAs in exosomes derived from BMSCs of postmenopausal osteoporosis (PMOP) patients and the underlying mechanisms. Methods RNA was extracted from BMSCs exosomes of PMOP and a control group. RNA microarray and bioinformatics analyses were used to explore the expression profile and functions circRNAs. Differentially expressed circRNAs from 20 PMOP and 20 controls were analyzed using RT-qPCR. Results A total of 237 upregulated and 279 downregulated circRNAs were identified in the current study. The top-10 most upregulated circRNAs in the PMOP group were hsa_circ_0069691, hsa_circ_0005678, hsa_circ_0006464, hsa_circ_0015813, hsa_circ_0000511, hsa_circ_0076527, hsa_circ_0009127, hsa_circ_0047285, hsa_circ_0027741, and hsa_circ_0090949. The top-10 most downregulated circRNAs were hsa_circ_0048669, hsa_circ_0090247, hsa_circ_0070899, hsa_circ_0087557, hsa_circ_0045963, hsa_circ_0090180, hsa_circ_0058392, hsa_circ_0040751, hsa_circ_0067910, and hsa_circ_0049484. RT-PCR verified dysregulation of 5 circRNAs including hsa_circ_0009127, hsa_circ_0090759, hsa_circ_0058392, hsa_circ_0090247, and hsa_circ_0049484. Moreover, a circRNA-microRNA-mRNA interaction network was developed based on differentially expressed circRNAs. Functional analysis showed that pathways involved in the regulation of autophagy, PI3K-Akt signaling, FoxO signaling, and MAPK signaling were associated with the differentially expressed circRNAs in PMOP patients. Conclusion The findings of this study show dysregulated circRNAs in BMSCs exosomes of PMOP patients, which may affect the progression of PMOP. These circRNAs can be used as predictive biomarkers and as therapeutic targets for the treatment of PMOP.

Epidemiological evidence linking metals bound to ambient particulate matters with aerodynamic diameter less than 2.5 μm (PM2.5) and maternal thyroid function is limited. In this study, we investigated the association of PM2.5-bound metals with maternal thyroid hormones (TH) during the first trimester. We retrospectively reviewed data for 2528 pregnant women attending prenatal care in Jinhua Maternal and Child Health Care Hospital, Jinhua, China, from January to December 2018. Information including thyroid hormone levels and demographics was retrieved from existing medical records. We analyzed the concentration of 10 metals for collected particulate samples, and estimated their exposure levels during the first trimester for each woman. We employed multivariate linear regression models to estimate the association of exposure to individual PM2.5-bound metals with serum levels of maternal TH, and weighted quantile sum (WQS) to estimate the overall association of exposure to PM2.5-bound metals within a mixture. Higher exposures to most of the PM2.5-bound metals were associated with lower levels of maternal free thyroxine (FT4) and free triiodothyronine (FT3). The thyroid peroxidase antibody (TPOAb) or thyroglobulin antibody (TgAb) status had no effect modification on the observed associations. WQS analyses further suggested that Be, Ni, Tl and Ba contributed the most to the associations. These findings highlight the associations of exposure to PM2.5-bound metals with maternal thyroid function, and emphasize the public health significance of commitments to improve air quality.

[目的]基于高通量测序技术,探究绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)肾阴虚证的潜在机制.[方法]分离培养PMOP肾阴虚证和骨量正常对照组骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs),从BMSCs外泌体中提取RNA,利用芯片测序技术和生物信息学分析mRNA的表达谱和功能.使用定量逆转录聚合酶链式反应(quantitative reverse transcription polymerase chain reaction,RT-qPCR)分析20例PMOP肾阴虚证患者和20例骨量正常对照组受试者差异表达的mRNA.[结果]与骨量正常对照组比较,PMOP肾阴虚证BMSCs外泌体检测出174个差异表达的mRNA,其中92个上调,82个下调;RT-qPCR鉴定出9个关键mRNA(ATM、COL8A1、GABARAPL1、HLA-DQA1、HSPA1L、MAX、PPP2R2D、PTHLH和WWTR1),与基因芯片数据相一致,两组之间差异有统计学意义(P<0.001).功能分析表明,PMOP肾阴虚证患者差异表达的mRNA可能靶向参与丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、自噬调节、磷脂酰肌醇3激酶-蛋白激酶B(phosphatidylinositol 3 kinase-protein kinase B,PI3K-AKT)和叉头框转录因子O(forkhead box transcription factor O,FoxO)亚族等信号通路.[结论]PMOP肾阴虚证患者BMSCs外泌体中差异表达的基因可能作为新型的生物标志物,为PMOP肾阴虚证的靶向治疗提供有价值的信息.

Abstract Objectives The clinical implications of different blood indices in patients with coronavirus disease-2019 (COVID-19) were analyzed at different stages. Methods We compared blood test results of 17 COVID-19 patients treated in Jinhua Central Hospital between January 1 and March 5, 2020 at different stages. We also compared the initial blood results of 17 COVID-19 patients with 115 influenza virus A/B (Flu A/B)-positive patients, 19 Mycoplasma pneumonia (MP)-positive patients and 50 healthy subjects (HSs). Results (1) The white blood cell count (WBC) and absolute neutrophil count (NEU#) were lower in the SARS-CoV-2 group than in the MP and Flu A/B groups; the eosinophil percentage (EO%) and absolute eosinophil count (EO#) were lower in the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) group than in the MP group (p<0.05). (2) Aspartate aminotransferase (AST) levels were significantly lower when patients were discharged from the hospital (p<0.05), EO% and EO# recovered at discharge, and returned to normal levels during follow-up (p<0.05). (3) When the throat swab was nucleic acid-negative but the stool was still positive, lymphocyte percentage (LY%) and absolute lymphocyte count (LY#) decreased (p<0.05). (4) As the cycle threshold (Ct) value of the nucleic acid increased or decreased, EO# showed a consistent trend. Conclusions Blood cell count indices upon hospital admission could be helpful to give some tips of diagnosis of SARS-CoV-2-infection, Flu A/B-infection and MP-infection; AST and EO# could be used to predict the outcome of patients. Feces turned negative for nucleic acid more slowly than throat swabs; LY# was lower during the fecal-positive period and low Ct values of fecal nucleic acid were negatively associated with the patient’s recovery level.

目的 比较4种自身抗体抗核周因子(APF)、抗角蛋白抗体(AKA)、抗环瓜氨酸多肽抗体(CCP)和类风湿因子(RF)在类风湿性关节炎(RA)诊断中的价值.方法 回顾2019年3至12月金华市中心医院检查的257例RA患者,采用间接免疫荧光法(IIF)检测抗APF、抗AKA,化学发光法检测抗CCP,速率散射法检测RF,计算4种自身抗体的阳性率,并采用ROC曲线分析其诊断价值.结果 抗APF、抗AKA、抗CCP、RF在257例RA患者中的阳性率分别为42.80％、31.13％、79.77％、83.66％.4项联合抗APF/抗CCP/RF/抗AKA和3项联合抗APF/抗CCP/RF的阳性率均为94.16％,高于抗CCP的79.77％,差异有统计学意义(P<0.05).抗APF与抗CCP、RF呈负相关(r=-0.470、-0.231,均P<0.05),与抗AKA、RA病程、晨僵均呈正相关(r=0.488、0.132、0.130,均P<0.05).抗APF和抗CCP与RA关节外表现有统计学差异(P<0.05).当抗APF最佳临界值为2.5时,AUC为0.886,灵敏度87.2％.结论 抗APF在RA中有更高的灵敏度,并且抗APF与RA病程、晨僵以及关节外器官受累相关,抗APF在RA诊断中有一定的诊断价值.

Dynamic monitoring of the concentration variation of IgM and IgG in patients with SARS‐CoV‐2 infections and exploring their diagnostic value for coronavirus disease‐19 (COVID‐19).