Background Polystyrene microplastics (PS-MPs) attract widespread public attention due to their adverse effects on mammalian reproductive systems. However, it is currently unclear whether ferroptosis is related to testicular damage and decreased sperm quality in mice exposed to PS-MPs. Objective To clarify the reproductive damage in male mice exposed to PS-MPs and investigate the mechanism of ferroptotic effects. Methods Five-week-old male BALB/c mice were randomly divided into four experimental groups, including one control group and three PS-MPs groups at low dose (0.5 mg·kg⁻¹), medium dose (5 mg·kg⁻¹), and high dose (50 mg·kg⁻¹), respectively, with 6 mice in each group. The treatment was delivered by gavage for 35 consecutive days (one time per day). After the mice were neutralized, the wet weights of testis and epididymis were measured, and organ coefficients were then calculated. Sperm was counted by hematimetry, and sperm motility and adenosine triphosphate (ATP) level were evaluated using CCK-8 and CellTiter Glo ® Kit 2.0 Assay respectively. In addition, serum testosterone, follicle-stimulating hormone, and luteinizing hormone were determined using ELISA kit, total testicular iron content was measured using tissue iron kit, and pathological changes in testicular tissue were observed after hematoxylin-eosin (HE) staining. We also used glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) assays to examine their changes to better understand the physiological status of testicular tissue. Finally, the expression levels of ferroptosis-associated proteins glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) were detected by Western blotting. Results Compared with the control group, the testicular index in the high dose group decreased, and the epididymal index decreased in all dose groups (P<0.05). The results of sperm quality analysis showed that the sperm count in each dose group was lower than that of the control group; the sperm motility decreased, sperm malformation rate increased, and ATP level in sperm decreased in the medium and high dose groups. The results of HE staining showed that the spermatogenic epithelium was disordered and the arrangement of spermatogenic cells were loose in the low dose group, the spermatogenic gap was enlarged in the middle dose group, and the cells in the high dose group were vacuolated and even azoospermic. The results of serum sex hormone levels showed that the serum testosterone levels decreased in each dose group, the serum follicle-stimulating hormone levels decreased in the medium and high dose groups, and the serum luteinizing hormone levels decreased in the high dose group (P<0.05). The iron content in the testicular tissue homogenate of the high dose group increased (P<0.05). The levels of GSH and SOD in the homogenate of testicular tissue decreased in the medium and high dose groups, while the levels of MDA increased (P<0.05). The results of Western blotting showed that the protein expression level of GPX4 in the testis in the high dose group was lower than that in the control group. The protein expression levels of SLC7A11 in the medium and high dose groups were lower than that in the control group. The results of correlation analysis showed that the expression level of GPX4 was positively correlated with sperm count, and negatively correlated with MDA level (P<0.05). SLC7A11 expression level was positively correlated with sperm count, and negatively correlated with sperm malformation rate and MDA level (P<0.05). Conclusion PS-MPs exposure leads to decreased sperm quality, testicular damage, and decreased serum sex hormone levels in male mice, and its mechanism of action may involve ferroptosis.

Objective:To investigate the diagnostic value of serum Aspergillus fumigatus immunoglobulin G (IgG), bronchoalveolar lavage fluid (BALF) galactomannan antigen test (GM test) and combined detection of the two methods in chronic pulmonary aspergillosis (CPA). Methods:A total of 310 patients with suspected CPA who were hospitalized in Wuhan Pulmonary Hospital from January 2020 to December 2022 were screened. BALF-GM test and serum Aspergillus fumigatus IgG were detected. According to the inclusion and exclusion criteria, 151 confirmed or clinically diagnosed CPA patients were enrolled and 60 non-CPA cases were included as controls. The underlying diseases, clinical symptoms and imaging findings of the two groups were analyzed using case-control study. Statistical comparison was performed by chi-square test. The efficacies of BALF-GM test, serum Aspergillus fumigatus IgG test, and their combined detection for CPA were analyzed by receiver operating characteristic curve. Results:There were 130 confirmed cases and 21 clinically diagnosed cases among 151 patients with CPA, including 106 males (70.2%) with age of (54.6±15.5) years. The age of 60 non-CPA patients was (53.6±17.8) years, including 42 males (70.0%). The proportions of pulmonary tuberculosis, bronchiectasis and chronic obstructive pulmonary disease, and the incidence of hemoptysis in CPA group were 79.47%(120/151), 86.09%(130/151), 26.49%(40/151) and 43.71%(66/151), respectively, which were all higher than those in non-CPA group (10.00%(6/60), 25.00%(15/60), 8.33%(5/60) and 11.67%(7/60), respectively), and the differences were statistically significant ( χ2=86.14, 74.56, 8.44 and 19.48, respectively, all P<0.05). The proportion of interstitial lung disease in non-CPA group was 28.33%(17/60), which was higher than that in CPA group (2.65%, 4/151), and the difference was statistically significant ( χ2=31.61, P<0.001). The common imaging findings of CPA patients were pulmonary cavity (70.86%, 107/151), cavity inclusions (41.72%, 63/151) and lung damage (33.77%, 51/151). The specificity and sensitivity of serum Aspergillus fumigatus IgG for CPA diagnosis were 81.7% and 68.9%, respectively, and the area under the curve (AUC) was 0.753 (95% confidence interval (95% CI) 0.681 to 0.825, P< 0.001). The specificity and sensitivity of BALF-GM test were 76.7% and 67.5%, respectively, and the AUC was 0.724 (95% CI 0.649 to 0.800, P<0.001). The specificity and sensitivity of the two methods in series combined detection were 95.0% and 44.4%, respectively, and those of the two methods in parallel detection were 60.3% and 94.5%, respectively. The AUC of the combined diagnosis of CPA by the two methods was 0.843 (95% CI 0.783 to 0.903, P<0.001). Conclusions:Serum Aspergillus fumigatus IgG combined with BALF-GM test has a good diagnostic efficacy for CPA. The detection of serum Aspergillus fumigatus IgG is non-invasive and highly specific, which is beneficial to the early diagnosis and treatment of CPA.

Objective To comprehensively analyze the circulatory RNA-long non-coding RNA-microRNA-messenger RNA(circRNA-lncRNA-miRNA-mRNA)network in cervical cancer and construct a prognostic model.Methods Differential and key genes were ana-lyzed using bioinformatics based on data from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.Prognostic mRNA models were constructed based on TCGA database using univariate,Least Absolute Shrinkage and Selection Operator(LASSO),and multivariate Cox regression analyses and validated using the GEO database.The R package and Cystoscape software were used to construct a nomogram model and competing endogenous(ceRNA)network of circRNA-lncRNA-miRNA-mRNA in cervical cancer.Results A prognostic model including five mRNAs was constructed using univariate,LASSO,and multivariate Cox regression analyses,which had area under the receiver operating characteristic curve AUC values of 0.71,0.71,and 0.70 at 1,2,and 3 years,respec-tively,indicating its sensitivity and specificity in cervical cancer prognosis.Predictive results were validated using the GSE44001 dataset.The C-index of the nomogram model for this prognostic model was 0.707.In this study,a ceRNA network comprising 39 circRNAs,27 lncRNAs,12 miRNAs,and five mRNAs was constructed.Conclusion The network constructed in this study can help comprehensively elucidate the mechanism of ceRNAs in cervical cancer,and the construction of prognostic and Nomogram models can predict patient prog-nosis.

Purpose: Molecular residual disease (MRD) is a promising biomarker in colorectal cancer (CRC) for prognosis and guiding treatment, while the whole-exome sequencing (WES) based tumor-informed assay is standard for evaluating MRD based on circulating tumor DNA (ctDNA). In this study, we assessed the feasibility of a fixed-panel for evaluating MRD in CRC. Materials and Methods: Seventy-five patients with resectable stage I-III CRC were enrolled. Tumor tissues obtained by surgery, and preoperative and postoperative day 7 blood samples were collected. The ctDNA was evaluated using the tumor-agnostic and tumor-informed fixed assays, as well as the WES-based and panel-based personalized assays in randomly selected patients. Results: The tumor-informed fixed assay had a higher preoperative positive rate than the tumor-agnostic assay (73.3% vs. 57.3%). The preoperative ctDNA status failed to predict disease-free survival (DFS) in either of the fixed assays, while the tumor-informed fixed assay-determined postoperative ctDNA positivity was significantly associated with worse DFS (hazard ratio [HR], 20.74; 95% confidence interval [CI], 7.19 to 59.83; p < 0.001), which was an independent predictor by multivariable analysis (HR, 28.57; 95% CI, 7.10 to 114.9; p < 0.001). Sub-cohort analysis indicated the WES-based personalized assay had the highest preoperative positive rate (95.1%). The two personalized assays and the tumor-informed fixed assay demonstrated same results in postoperative landmark (HR, 26.34; 95% CI, 6.01 to 115.57; p < 0.001), outperforming the tumor-agnostic fixed panel (HR, 3.04; 95% CI, 0.94 to 9.89; p=0.052). Conclusion Our study confirmed the prognostic value of the ctDNA positivity at postoperative day 7 by the tumor-informed fixed panel. The tumor-informed fixed panel may be a cost-effective method to evaluate MRD, which warrants further studies in future.

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.

OBJECTIVE: To explore the genetic basis for a child with mental and motor retardation, language impairment, facial dysmorphism and epilepsy. METHODS: Whole exome sequencing was carried out to detect pathogenic variant in the proband, and candidate variant was selected based on his phenotype. Sanger sequencing was used to verify the variant in the proband, his parents and other family members. RESULTS: The proband was found to carry a frameshifting mutation of MBD5 gene, namely c.2217delT (p.F739Lfs*6), which was inherited from his mother and unreported previously. Sanger sequencing confirmed that his brother carried the same mutation with a similar phenotype. His mother also had poor language expression when she was young, in addition with poor academic performance, though she could do some housework and had no history of convulsion. CONCLUSION A novel pathogenic variant of the MBD5 gene was discovered, which has enriched the mutational spectrum of the MBD5 gene. Above discovery has enabled genetic counseling and prenatal diagnosis for the family.
Child ; DNA-Binding Proteins/genetics* ; Female ; Humans ; Intellectual Disability/genetics* ; Male ; Mutation ; Pedigree ; Phenotype ; Pregnancy ; Whole Exome Sequencing

Objective:To investigate the clinical characteristics and gene variation of asparagine synthase deficiency that is caused by ASNS gene variation. Methods:In Department of Neuroendocrine Pediatrics, Affiliated Hospital of Qingdao University from October 2018 to February 2020, the clinical data of a family of asparagine synthase deficiency were analyzed retrospectively.The pathogenic mutation of the proband was screened by the full exon analysis technique.The pathogenic sites of candidate genes were determined by combining the phenotype of the proband.In the heterotopic spot of the proband, his parents and other family members were verified by Sanger sequencing.Meanwhile, the relevant literature database was consulted, and the reported ASNS mutation related cases were collected and reviewed. Results:The female with proband visited the hospital at the age of 4 months, and she had recurrent convulsions at the age of about 3 months.Physical examination showed that the child suffered from microcephaly, and mental and motor retardation.Meanwhile, video electroencephalogram examination displayed extensive moderate high amplitude spiny slow wave and sharp slow wave.Exon sequencing illustrated that the compound heterozygous variants of ASNS gene were c. 1211G>A (p.R404H) and c. 1643C>T (p.S548F), respectively.c.1211G>A was a known pathogenic variant, and c. 1643C>T was a new variant.The proband′s younger brother visited the hospital at the age of 2 months, developed convulsions at the age of 1 month, and developed mental and motor retardation.Electroencephalogram displayed that bilateral posterior head was dominant, multiple foci and extensive spike wave, and spike slow wave and fast wave were distributed.Sanger sequencing revealed the same ASNS compound heterozygous variants as the proband.Both of them died of status convulsion at the age of 7 months and 6 months, respectively. Conclusions:This study is helpful to further understand the clinical features of the disease and reveal a new pathogenic mutation of ASNS gene, so as to enrich the mutation spectrum of ASNS gene, thus providing important basis for clinical treatment and genetic counseling.

Objective:To investigate the diagnostic value of neutrophil CD64 index in sepsis patients in intensive care unit (ICU).Methods:A prospective case-control study was conducted, the patients admitted to ICU of Jiangbei People's Hospital Affiliated to Nantong University from December 2016 to June 2020 were enrolled. According to the criteria of Sepsis 3, 107 patients diagnosed with sepsis were classified as the sepsis group, 112 patients without infection were classified as control group. Peripheral venous blood samples were collected within 24 hours after ICU admission, neutrophil CD64 index, C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC) were detected. Receiver operating characteristic curve (ROC curve) was used to evaluate the diagnostic value of neutrophil CD64 index, CRP, PCT and WBC for sepsis.Results:The neutrophil CD64 index, CRP and PCT in sepsis group were significantly higher than those in control group [neutrophil CD64 index: 9.03±5.59 vs. 3.18±1.50, CRP (mg/L): 146.9±68.3 vs. 46.5±35.8, PCT (ng/L): 31.82±14.71 vs. 1.87±1.42, all P < 0.05]. ROC curve analysis showed that neutrophil CD64 index, CRP and PCT had certain diagnostic value for sepsis, the area under ROC curve (AUC) were 0.924, 0.915 and 0.879, respectively, the 95% confidence intervals (95% CI) were 0.871-0.978, 0.855-0.975, 0.807-0.951, respectively, P values were 0.016, 0.017 and 0.026, respectively. Among the three indicators, the diagnostic value of neutrophil CD64 index was much higher. When the optimal cut-off value was 4.32, the sensitivity and specificity were 83.6% and 88.7%, respectively, which were higher than the sensitivity (75.1%, 76.3%) and specificity (87.2%, 82.5%) of CRP and PCT. Conclusion:Neutrophil CD64 index is a valuable biomarker for the diagnosis of sepsis in ICU.

Objective:To analyze the clinical characteristics of intestinal tuberculosis improving the diagnosis rate of intestinal tuberculosis.Methods:From January 2014 to June 2018, at Wuhan Pulmonary Hospital, the data of clinical symptoms, laboratory examination, imaging, endoscopy, surgery and pathological examination of 205 patients with intestinal tuberculosis were retrospectively analyzed. Descriptive analysis was performed for statistical analysis.Results:Among 205 patients with intestinal tuberculosis, 145 cases were male and 60 cases were female, aged 14 to 85 years old. A total of 189 cases (92.2%) were complicated with lung tuberculosis, of which 151 cases (79.9%) were positive for sputum acid fast staining. A total of 126 cases were tested for feces acid fast staining, of which 83 cases (65.9%) were positive. A total of 60 cases (29.3%) were tested for GeneXpert Mycobacterium tuberculosis/rifampicintablet (GeneXpert MTB/RIP), of which 49 cases (81.7%) were positive. A total of 44 cases of intestinal tuberculosis were diagnosed by biopsy under electronic enteroscopy, and 21 cases were pathologically diagnosed with intestinal tuberculosis after surgical resection. The 21 patients were tested for GeneXpert MTB/RIP, of which 19 cases (90.5%) were positive and 10 cases (47.6%) were positive for tuberculin test. Six patients were clinically diagnosed with intestinal tuberculosis after effective treatment of antituberculosis drugs. Conclusions:Combination of clinical symptoms and laboratory, imaging, endoscopic and pathological examination, as well as the therapeutic effect of diagnostic antituberculosis treatment could make comprehensive diagnosis of intestinal tuberculosis. The GeneXpert MTB/RIP examination is of great value in the diagnosis of intestinal tuberculosis.

BACKGROUND:Steroid-induced osteonecrosis of the femoral head (SONFH) is a common bone disease characterized as high morbidity and poor prognosis,but the pathogenesis is unclear.Oxidative stress treatment is closely related to the occurrence and development of SONFH,and has tremendous potential in the treatment of SONFH,which can be realized by Nano-Se.OBJECTIVE:To observe the protective effect of porous Se@SiO2 nanocomposite on chondrocytes by antioxidant stress,and to further explore its mechanism of protection and treatment of SONFH.METHODS:(1) In vitro experiment:The rat chondrocytes were isolated,cultured and identified.Then,the chondrocytes were cultured with porous Se@SiO2 nanocomposite to suppress the production of reactive oxygen species (ROS).(2) In vivo experiment:A total of 36 rats were randomly divided into three groups.Steroid-induced group and experimental group were treated with intraperitoneal injection of lipopolysaccharide and intramuscular injection of methylprednisolone to induce SON FH models.Seven days after modeling,the experimental group was intraperitoneally injected with porous Se@SiO2 nanocomposite.No intervention was done in control group (blank control).At 8 weeks after modeling,rat bilateral femoral heads were taken for hematoxylin-eosin staining and Micro-CT scanning.RESULTS AND CONCLUSION:Results from the ROS detection and TUNEL apoptosis tests showed that the level of ROS in the chondrocytes was significantly reduced after intervention with Se@SiO2 (P ＜ 0.05).Micro-CT scanning findings showed that the bone mineral density,bone volume,bone area/bone volume,trabecular number,trabecular thickness,and trabecular separation in the steroid-induced and experimental groups were significantly different from those in the control group (P ＜ 0.05).Hematoxylin-eosin staining results showed smooth femoral head,normal bone cells,chondrocytes and trabecular bone,as well as few empty bone lacunae and fat cells in the control group,while in the steroid-induced group,there was bone trabecular fracture,fat cell hypertrophy fusion,a large number of empty bone lacunae and obvious osteonecrosis.These manifestations were significantly improved in the experimental group.To conclude,the porous Se@SiO2 nanocomposite has good antioxidative stress ability,suppresses the ROS production and exerts therapeutic effects on SONFH.