BACKGROUND:Fibromyalgia syndrome,as a common rheumatic disease,is related to central sensitization and immune abnormalities.However,the specific mechanism has not been elucidated,and there is a lack of specific diagnostic markers.Exploring the possible pathogenesis of this disease has important clinical significance. OBJECTIVE:To screen the potential diagnostic marker genes of fibromyalgia syndrome and analyze the possible immune infiltration characteristics based on bioinformatics methods,such as weighted gene co-expression network analysis(WGCNA),and machine learning. METHODS:Gene expression profiles in peripheral serum of fibromyalgia syndrome patients and healthy controls were obtained from the gene expression omnibus(GEO)database.The differentially co-expressed genes were screened in the expression profile by differential analysis and WGCNA analysis.Least absolute shrinkage and selection operator(LASSO)and support vector machine-recursive feature elimination(SVM-RFE)machine learning algorithm were further used to identify hub biomarkers,and draw receiver operating characteristic curve(ROC)to evaluate the accuracy of diagnosing fibromyalgia syndrome.Finally,single sample gene set enrichment analysis(ssGSEA)and gene set enrichment analysis(GSEA)were used to evaluate the immune cell infiltration and pathway enrichment in patients with fibromyalgia syndrome. RESULTS AND CONCLUSION:Eight down-regulated differentially expressed genes(DEGs)were obtained after differential analysis of the GSE67311 dataset according to the conditions of log2|(FC)|>0 and P<0.05.After WGCNA analysis,497 genes were included in the module(MEdarkviolet)with the highest positive correlation(r=0.22,P=0.04),and 19 genes were included in the module(MEsalmon2)with the highest negative correlation(r=-0.41,P=6×10-5).After intersecting DEGs and the module genes of WGCNA,seven genes were obtained.Four genes were screened out by LASSO regression algorithm and five genes were screened out by SVM-RFE machine learning algorithm.After the intersection of the two,three core genes were identified,which were germinal center associated signaling and motility like,integrin beta-8,and carboxypeptidase A3.The areas under the ROC curve of the three core genes were 0.744,0.739,and 0.734,respectively,indicating that they have good diagnostic value and can be used as biomarkers for fibromyalgia syndrome.The results of immune infiltration analysis showed that memory B cells,CD56 bright NK cells,and mast cells were significantly down-regulated in patients with fibromyalgia syndrome compared with the control group(P<0.05),and were significantly positively correlated with the above three biomarkers(P<0.05).The enrichment analysis suggested that there were nine fibromyalgia syndrome enrichment pathways,mainly related to olfactory transduction pathway,neuroactive ligand-receptor interaction,and infection pathway.The above results showed that the occurrence and development of fibromyalgia syndrome are related to the involvement of multiple genes,abnormal immune regulation,and multiple pathways imbalance.However,the interactions between these genes and immune cells,as well as their relationships with various pathways need to be further investigated.

BACKGROUND:The repair process of myocardial injury involves complex cellular and molecular mechanisms,especially mitochondrial calcium homeostasis,macrophage autophagy and pyroptosis pathways.Traditional Chinese medicine(TCM)has shown significant clinical efficacy in improving myocardial injury,but its mechanism of action needs to be thoroughly investigated. OBJECTIVE:To investigate the role of mitochondrial calcium homeostasis-mediated macrophage autophagy and pyroptosis pathways in myocardial injury,and to summarize the progress of TCM in this field. METHODS:A computerized search was performed for relevant literature from the database inception to March 2024 in the Web of Science,PubMed and CNKI.The search terms were"mitochondrial calcium homeostasis,macrophage autophagy,macrophage pyroptosis,traditional Chinese medicine,myocardial injury,myocardial injury reperfusion"in Chinese and English.Through literature review,we analyzed the relationship between mitochondrial calcium homeostasis and macrophage autophagy and pyroptosis,explored the mechanism of their roles in myocardial injury,and summarized the pathways of multi-targeted,multi-pathway effects of TCM. RESULTS AND CONCLUSION:The maintenance of mitochondrial calcium homeostasis has been found to be closely related to the normal function of cardiomyocytes.Macrophages can participate in the repair process of myocardial injury through autophagy and pyroptosis pathways.Autophagy contributes to cell clearance and regulation of inflammatory response,while pyroptosis affects myocardial repair by releasing inflammatory factors.TCM regulates mitochondrial calcium homeostasis and macrophage function through multiple mechanisms.For example,astragalosid regulates calcium homeostasis by lowering mitochondrial membrane potential and inhibiting cytochrome C,and epimedium glycoside plays a role in reducing β-amyloid deposition.In addition,herbal compounds and single drugs promote myocardial repair by activating or inhibiting specific signaling pathways,such as PI3K/AKT and nuclear factor-κB signaling pathways.Future studies should focus on the interactions between mitochondrial calcium homeostasis,autophagy and pyroptosis pathways,as well as how TCM can exert therapeutic effects through these pathways to provide new strategies and drugs for the treatment of myocardial injury.

Background Obstructive sleep apnea (OSA) is a sleep disorder characterized by recurrent episodes of obstruction of the upper airway during sleep. Given the substantial number of OSA patients, it is urgently in need to address the burden on society. Current available evidence linking outdoor light at night (LAN) to OSA is scarce. Objective To explore the relationships regarding outdoor LAN and OSA among residents in Southern China. Methods A total of 3925 hospitalized patients in Guangdong Provincial People's Hospital Sleep Center were recruited between January 1, 2005 and December 31, 2015. The severity of OSA was determined by apnea-hypopnea index (AHI) using polysomnography or home sleep test. The annual Suomi National Polar-orbiting Partnership Visible Infrared Imaging Radiometer Suite (NPP-VIIRS)-like LAN data with a 500 m spatial resolution were used to evaluate outdoor LAN levels of the participants 1, 3, and 5 years before undergoing sleep monitoring. Generalized linear regression models were utilized to examine the associations of outdoor LAN with OSA. Results The ORs (95%CIs) of OSA, mild OSA, and moderate OSA were 1.175 (1.021, 1.351), 1.215 (1.038, 1.421), and 1.195 (1.003, 1.425) respectively for per interquartile range (IQR) increment in three-year average outdoor LAN. The results of stratified analyses showed a higher OR of 1.933 (95%CI: 1.424, 2.637) in female than male participants (P _interaction < 0.001). Additionally, the ORs of OSA did not substantially change using one-year/ five-year instead of three-year average outdoor LAN. Conclusion Our findings demonstrate that an elevated outdoor LAN is significantly associated with higher odds of OSA (especially mild and moderate OSA) in Southern China, especially in females. An effective outdoor LAN management and policy-making framework has been suggested as potential preventive measures to reduce burden of OSA.

AIM:To investigate the relationship between the level of monocyte chemoattractant protein-1(MCP-1)in the aqueous humor and macular microcirculation in patients with macular edema secondary to retinal vein occlusion(RVO-ME).METHODS:A total of 327 patients(327 eyes)with RVO-ME treated in our hospital from July 2022 to July 2024 were selected as the research objects. According to the recurrence or not, they were divided into non-recurrence group(291 cases, 291 eyes)and recurrence group(36 cases, 36 eyes). The clinical data, macular microcirculation index and MCP-1 level were collected. Unordered multinomial Logistic regression was used to analyze the effect of MCP-1 on RVO-ME recurrence after excluding the influence of other factors. Cubic spline was used to analyze the dose-response relationship between macular microcirculation indexes and MCP-1 expression level and RVO-ME recurrence. Multiple linear regression was used to analyze the relationship between macular microcirculation parameters and MCP-1 expression level, and the difference of MCP-1 expression level in recurrence under different macular microcirculation parameters was analyzed. Bootstrap method was used to test the mediating effect of macular microcirculation indexes on MCP-1 expression level and RVO-ME recurrence.RESULTS: The course of ME, the incidence of vitreomacular traction(VMT), the incidence of fibrous membrane epiretinal membrane(ERM), best corrected visual acuity(BCVA), hyperreflective foci in the inner and outer retinal layers(HRF), the frequency and dose of anti-vascular endothelial growth factor(VEGF)injection in the recurrence group were significantly higher than those in the non-recurrence group(all P<0.05). The external limiting membrane(ELM)integrity and ellipsoid zone(EZ)integrity in the recurrence group were significantly worse than those in the non-recurrence group(all P<0.05). The central foveal thickness(CFT), central macular thickness(CMT), superficial capillary plexus(SCP)vascular density, deep capillary plexus(DCP)vascular density and MCP-1 in the recurrence group were higher than those in the non-recurrence group(all P<0.05), and the foveal avascular zone(FAZ)area was lower than that in the non-recurrence group(P<0.05). Logistic analysis showed that MCP-1 was a risk factor for RVO-ME recurrence before and after adjusting for confounding factors. There was a significant non-linear dose-response relationship between macular microcirculation indexes and MCP-1 expression and the risk of RVO-ME recurrence(non-linear test, all P<0.001). The vascular density of CFT, CMT, SCP and DCP was positively correlated with the expression level of MCP-1(all P<0.05), while the FAZ was negatively correlated with MCP-1 expression level(P<0.05). With the increase of vessel density in CFT, CMT, SCP and DCP, and the decrease of FAZ, the expression level of MCP-1 increased, and the risk of RVO-ME recurrence showed an upward trend. The proportion of MCP-1 in Q3(>28.47 pg/mL)group was the highest(P<0.05). Macular microcirculation parameters play a mediating effect between MCP-1 expression level and RVO-ME recurrence.CONCLUSION: The level of MCP-1 in aqueous humor is positively correlated with RVO-ME recurrence, and it is closely related to macular microcirculation. Macular microcirculation has a mediating effect between MCP-1 level and RVO-ME recurrence.

Objective: To explore the relationship between parenting styles and depressive symptoms in adolescents with nightmare disorder, so as to provide a scientific basis for formulating effective family intervention measures and psychological counseling. Methods: From January 2023 to August 2024, 90 adolescents diagnosed with nightmare disorder and admitted to Hangzhou Seventh Peoples Hospital, along with 176 healthy controls from the urban areas of Hangzhou, were recruited as participants in the study. All participants were assessed using the Nightmare Experience Questionnaire (NEQ), Family Relationship Questionnaire (FRQ), and Plutchik-van Praag Selfreport Depression Scale (PVP). The ttest and Chisquare test were conducted to compare two groups. Pearson correlation and stepwise multiple linear regression were employed to explore the correlations between PVP and NEQ or FRQ. The Process model was used to testing the mediating effects among NEQ/FRQ/PVP. Results: The nightmare disorder group had higher scores in nightmare frequency, the four factors of NEQ (physical effect, negative emotion, meaning interpretation, horrible stimulation), and PVP than the healthy control group (24.86±18.89, 10.12±3.67, 19.01±3.51, 17.02±3.31, 15.14±3.26, 14.02±4.38; 2.34±1.04, 6.49±2.18, 17.63±4.76, 13.91±4.24, 12.40±4.49, 9.39±3.28)(t=15.79, 10.11, 2.43, 6.09, 5.14, 27.46, P<0.05). The nightmare disorder group reported significantly lower scores in FRQ general attachment and maternal encouragement than the healthy control group (7.22±2.81, 16.39±3.28) (t=-5.53, -4.95). In contrast, they exhibited significantly higher scores in maternal abuse, maternal dominance, paternal freedom release, and paternal dominance than the healthy control group (8.23±1.80, 13.11±3.73, 18.36±3.37, 12.04±3.29; 6.07±1.85, 8.48±3.80, 15.15±2.51, 9.47±3.03) (t=6.70, 8.96, 5.90, 7.04, P<0.01). The results of Pearson correlation analysis showed that, in the nightmare disorder group, the PVP score was positively correlated with negative emotion, nightmare frequency, maternal abuse, and maternal dominance score (r=0.14, 0.63, 0.26, 0.51, P<0.05). The results of multiple linear regression analysis showed that when using FRQ score to predict NEQ score, the adjusted R2 in the nightmare disorder group was 0.01-0.59. Mother abuse could prediced physical effect (β=0.33); maternal dominance significantly predicted negative emotion, horrible stimulation, and nightmare frequency (β=0.29, 0.30, 0.79); paternal freedom release could predict negative emotion (β=0.26), paternal dominance predicted both negative emotion and nightmare frequency (β=0.22, 0.45) (P<0.05). Mediation analysis further revealed that, in the nightmare disorder group, PVP scores served as a mediating variable between FRQ and NEQ. Conclusion Abusive, controlling, and neglectful family upbringing styles as well as depression maybe are key factors that may contribute to the development of nightmare disorder among adolescents.

Objective: To analyze the changes in homologous immunity after RhCE-matched transfusion in positive patients with RhCE blood group antibodies, and to provide precise transfusion strategies for chronic anemia patients. Methods: Patients with chronic anemia in our hospital from January 2020 to March 2024 (continuously receiving blood transfusions for more than 6 months) were enrolled, and 63 cases of unexpected antibody screening positive and identified as RhCE blood group antibodies were selected as the research subjects. The changes in unexpected antibody yield rate after ABO and RhCcDEe isotype blood transfusion were observed. Patients with MNS, Kidd, or Lewis blood group antibodies were screened for corresponding negative donors using monoclonal antibodies for extended typing transfusion based on RhCcEe typing, and the changes in unexpected antibody yield rate after transfusion were observed. Blood group genotyping was performed when serological techniques failed to resolve discrepancies or detect abnormal antigen expression. Results: After RhCcDEe-matched transfusions, RhCE antibodies disappeared in 62 patients, while 1 patient developed anti-Ce. The latter did not develop blood type isotype immunity after receiving RhccEE donor blood. Among the 62 patients, 9 developed unexpected antibodies against other systems: anti-M (4 cases), anti-Mur (2), anti-S (1), anti-Jka (1), and anti-Lea (1). No additional alloimmunization occurred after extended antigen-matched transfusions. A patient with serologically weak e phenotype was genotyped as DCe/DcE, with gene sequencing revealing an 827C>A mutation in exon 6 of the RHCE gene, forming the RHCE 01.31 allele. Conclusion: Precise transfusion strategies incorporating RhCE, MNS, Kidd, and Lewis blood group antigen typing can reduce the probability of blood group homologous immunity. RhCE complex antibodies and RhCE variants pose difficulties for clinical RhCE typing transfusion, which can be addressed through cross-matching and genetic analysis.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

Background: The immune system plays a critical role in the development and progression of osteoporosis and fractures. However, the causal relationships between antibody immune responses, immune cells, and these bone conditions remain unclear. This study aimed to explore these relationships using Mendelian randomization (MR) analysis. Methods: We collected complete blood count data from patients with fractures and healthy individuals and analyzed their differences. Then, we conducted a 2-sample, 2-step MR analysis to investigate the causal effects of antibody immune responses on osteoporosis and fractures, using inverse-variance weighted (IVW) as the primary method. We also explored whether immune cells mediate the pathway between antibodies and osteoporosis or fractures. Finally, we analyzed the functions and expression levels of key genes involved. Results: Overall, the fracture group exhibited increased white blood cell count, absolute neutrophil count, absolute monocyte count, platelet count, and their respective proportions, while absolute lymphocyte count, absolute eosinophil count, absolute basophil count, red blood cell count, and their proportions were decreased. We identified 44 causal relationships between antibodies and osteoporosis or fractures, with 7 supported by multiple MR methods, and 5 showing odds ratios significantly deviating from 1 in the IVW analysis. Epstein-Barr virus-related antibodies had a notable impact on osteoporosis and fractures. The human leukocyte antigen (HLA) gene family, particularly HLA-DPB1, emerged as a significant risk factor. However, immune cells were not found to mediate these effects. Conclusions This study elucidated the causal relationships between antibody immune responses, immune cells, and osteoporosis or fractures. The HLA gene family plays a crucial role in the interaction between antibodies and these bone conditions, with HLA-DPB1 identified as a key risk gene. Immune cells do not serve as mediators in this process. These findings provide valuable insights for future research.

Background: The immune system plays a critical role in the development and progression of osteoporosis and fractures. However, the causal relationships between antibody immune responses, immune cells, and these bone conditions remain unclear. This study aimed to explore these relationships using Mendelian randomization (MR) analysis. Methods: We collected complete blood count data from patients with fractures and healthy individuals and analyzed their differences. Then, we conducted a 2-sample, 2-step MR analysis to investigate the causal effects of antibody immune responses on osteoporosis and fractures, using inverse-variance weighted (IVW) as the primary method. We also explored whether immune cells mediate the pathway between antibodies and osteoporosis or fractures. Finally, we analyzed the functions and expression levels of key genes involved. Results: Overall, the fracture group exhibited increased white blood cell count, absolute neutrophil count, absolute monocyte count, platelet count, and their respective proportions, while absolute lymphocyte count, absolute eosinophil count, absolute basophil count, red blood cell count, and their proportions were decreased. We identified 44 causal relationships between antibodies and osteoporosis or fractures, with 7 supported by multiple MR methods, and 5 showing odds ratios significantly deviating from 1 in the IVW analysis. Epstein-Barr virus-related antibodies had a notable impact on osteoporosis and fractures. The human leukocyte antigen (HLA) gene family, particularly HLA-DPB1, emerged as a significant risk factor. However, immune cells were not found to mediate these effects. Conclusions This study elucidated the causal relationships between antibody immune responses, immune cells, and osteoporosis or fractures. The HLA gene family plays a crucial role in the interaction between antibodies and these bone conditions, with HLA-DPB1 identified as a key risk gene. Immune cells do not serve as mediators in this process. These findings provide valuable insights for future research.