This article reports a rare case of autoinflammatory disease presenting initially with skin induration and swelling after trauma as the initial manifestation, followed by progressive limb weakness. The patient was a middle-aged female who developed skin induration and swelling after trauma, which gradually progressed to limb weakness, dysarthria and bilateral facial paralysis, accompanied by livedo reticularis of the lower extremities, diffuse skin induration of the limbs, and beaded subcutaneous nodules in the right upper limb. The patient had a susceptibility to infection since childhood and a history of chronic livedo reticularis. Skin pathological examination revealed panniculitis. A comprehensive etiological screening for special infections and autoimmune diseases was completed with an unremarkable results, and whole-exome sequencing showed no abnormal findings. Following a multidisciplinary discussion combined with RNA sequencing results, the patient was diagnosed with an autoinflammatory disease, with a suspected type Ⅰ interferonopathy. Treatment with tofacitinib resulted in gradual improvement of clinical symptoms. This case highlights the importance of detailed medical history collection, systematic physical examination and multidisciplinary collaborative diagnosis and treatment, and underscores the pivotal role of molecular diagnosis in the confirmation of rare diseases. It can provide a reference for the clinical diagnosis and management of similar rare cases.

BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

ObjectiveTo investigate the therapeutic effect and mechanism of Xueshisanjia San against liver fibrosis by regulating PTEN-induced putative kinase （PINK1）/Parkin signaling pathway-mediated mitophagy. MethodsForty specific pathogen free （SPF）-grade male C57BL/6 mice were randomized into the control， model， silibinin （100 mg·kg^-1）， high-dose （15.16 g·kg^-1） Xueshisanjia San， and low-dose （7.58 g·kg^-1） Xueshisanjia San groups. The mouse model of liver fibrosis was constructed by intraperitoneal injection of 20% carbon tetrachloride solution. The treatment lasted for 6 weeks. Blood was collected from the abdominal aorta after intraperitoneal anesthesia， and the liver was separated. Liver pathology was examined by hematoxylin-eosin （HE） staining， Masson staining， and Sirius Red staining. Transmission electron microscopy （TEM） was employed to observe the mitochondrial morphology in the liver tissue. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， C-reactive protein （CRP）， total bilirubin （TBil）， transforming growth factor-β₁ （TGF-β₁）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） in the serum of mice were measured by enzyme-linked immunosorbent assay. Immunohistochemistry， immunofluorescence assay， and Western blot were employed to determine the protein levels of liver fibrosis markers α-smooth muscle actin （α-SMA） and collagen Ⅰ， as well as mitophagy markers microtubule-associated protein 1 light chain 3 （LC3）， p62， Beclin-1， PINK1， Parkin， and translocase of outer mitochondrial membrane 20 （TOM20）. ResultsCompared with the control group， the model group exhibited elevated levels of ALT， AST， CRP， TBil， IL-6， TGF-β₁， and TNF-α in the serum （P<0.05）， pathological changes such destroyed structure of hepatic lobules， disarrangement of hepatic cells， and collagen accumulation， swollen， vacuolated， and fragment mitochondria， down-regulated expression of p62 and TOM20， and up-regulated expression of LC3， Beclin-1， PINK1， and Parkin （P<0.05）. Compared with the model group， all the treatment groups exhibited declined levels of ALT， AST， CRP， TBil， IL-6， TGF-β₁， and TNF-α in the serum （P<0.05）， alleviated pathological damage of liver tissue and mitochondrial damage， up-regulated expression of p62 and TOM20， and down-regulated expression of α-SMA， COL1A1， LC3， Beclin1， PINK1， and Parkin （P<0.05）. ConclusionXueshisanjia San may prevent excessive mitophagy and improve mitochondrial quality by inhibiting PINK1/Parkin signaling pathway， thereby alleviating liver fibrosis.

Objective:To establish an ion chromatography-integrated amperometric detection method for iodide in water.Methods:After the water sample was filtered through a filter membrane, the AS 11-HC anion chromatography column of ion chromatography method was used to separate iodide ions under the conditions of 70 mmol/L sodium hydroxide solution as the eluent, injection volume of 100 μl, column temperature of 30 ℃, and flow rate of 1.0 ml/min. The results were determined by silver working electrode integral amperometric detection method. Under the optimized experimental conditions, methodological evaluations such as method calibration curves, detection limits, quantification limits, precision, and accuracy were conducted.Results:Iodide followed a square correction curve within the concentration range of 0 - 100 μg/L, with a correlation coefficient ( r) > 0.999 9. The detection limit of the method was 0.30 μg/L, and the quantification limit was 1.00 μg/L. The determination results of the national standard substances GBW09113f and GBW09114f for iodine composition analysis in water were within the reference range [(8.4 ± 1.2), (55 ± 6) μg/L]. The recovery rates of low, medium, and high concentration spiked samples with low background values ranged from 91.7% to 97.2%, and the relative standard deviation ranged from 0.40% to 1.60%. Conclusion:This method has the characteristics of simple water sample pretreatment, high sensitivity, and good accuracy, which can meet the determination of trace iodides in bulk water samples for iodine deficiency disorders monitoring.

Objective:To explore the morphological characteristics of tumor cells and the key diagnostic points of immunocytochemistry in neoplastic serous effusions.Methods:A retrospective case series study was conducted. A total of 522 samples which were ultimately diagnosed as neoplastic serous effusions by immunocytochemistry were collected in Shanxi Province Cancer Hospital from January 2019 to December 2023. The microscopic morphological characteristics of tumor cells in the samples were analyzed, and the diagnostic points of immunocytochemistry and the differential diagnostic points between rare tumors and adenocarcinoma were summarized.Results:Among the 522 samples of neoplastic serous effusion, there were 305 cases of pleural effusion, 178 cases of abdominal effusion, and 39 cases of pericardial effusion. Immunocytochemical diagnosis revealed 380 cases of adenocarcinoma [198 cases (52.1%) of pleural effusion, 155 cases (40.8%) of peritoneal effusion, and 27 cases (7.1%) of pericardial effusion], 55 cases of small cell carcinoma [47 cases (85.5%) of pleural effusion, 1 case (1.8%) of peritoneal effusion, and 7 cases (12.7%) of pericardial effusion], 23 cases of squamous cell carcinoma [18 cases (78.3%) of pleural effusion, 4 cases (17.4%) of peritoneal effusion, and 1 case (4.3%) of pericardial effusion], 5 cases of large cell neuroendocrine carcinoma (4 cases of pleural effusion and 1 case of pericardial effusion), and 31 cases of lymphoma [21 cases (67.7%) of pleural effusion, 7 cases (22.6%) of peritoneal effusion, and 3 cases (9.7%) of pericardial effusion], 20 cases of malignant mesothelioma [17 cases (85.0%) of pleural effusion and 3 cases (15.0%) of peritoneal effusion], 6 cases of ovarian borderline tumors (all of which were peritoneal effusions), and 2 cases of yolk sac tumors (both of which were peritoneal effusions). Under the microscope, the morphological characteristics of tumor cells in squamous cell carcinoma, large cell neuroendocrine carcinoma, lymphoma, malignant mesothelioma, ovarian borderline tumor, and yolk sac tumor were similar to those of adenocarcinoma cells, making them prone to misdiagnosis as adenocarcinoma. Immunocytochemical examination was necessary for the clear diagnosis.Conclusions:Adenocarcinoma accounts for the vast majority of neoplastic serous effusions, while the cell morphologies of other rare tumors overlap with adenocarcinoma. Careful observation and combination with clinical data and immunocytochemical examination results of patients are necessary for making the correct diagnosis.

AIM:This study aims to investigate the synergistic cytotoxic effects of chrysin and venetoclax on acute myeloid leukemia(AML)cells and to elucidate the underlying mechanisms.METHODS:Human AML cell lines MV411 and MOLM13 were cultured in vitro and treated with chrysin in combination with venetoclax.Cell viability was as-sessed using the CCK8 assay,while flow cytometry was employed to measure cell cycle distribution and apoptosis rates.Western blot was used to detect the expression of apoptosis-related proteins and protein kinase B(PKB/Akt)/nuclear factor-κB(NF-κB)signaling pathway-related proteins.RESULTS:The results from the CCK8 assay and flow cytometry demon-strated that treatment with 16 and 32 μmol/L chrysin significantly inhibited the viability of AML cells and increased the proportion of cells in G1 phase,as well as the apoptosis rate.Notably,the cells in combination treatment group exhibited a marked reduction in proliferation and an elevated apoptosis rate compared with either chrysin or venetoclax group alone.Western blot analysis indicated that increasing concentrations of chrysin led to an elevation in cleaved poly(ADP-ribose)polymerase(PARP)level,alongside a down-regulation of proteins associated with the Akt/NF-κB signaling pathway.Fur-thermore,the combination treatment significantly up-regulated cleaved PARP level and down-regulated Akt/NF-κB path-way-related proteins compared with the treatment with chrysin or venetoclax alone.CONCLUSION:Chrysin and veneto-clax synergistically inhibit the proliferation of AML cells and promote apoptosis by modulating the Akt/NF-κB signaling pathway.

Objective:To analyze the application value of ultrasound Thyroid Imaging Reporting and Data System (TI-RADS) classification in the differential diagnosis of thyroid nodules and its correlation with patients' midkine, thymidine kinase 1, and thyroid function indicators.Methods:This study was a prospective study. A total of 108 patients with thyroid nodules (120 nodules) who visited Jinan 2 nd People's Hospital from December 2022 to February 2024 were included. Ultrasound examinations were conducted to complete the TI-RADS classification, and serum levels of midkine (MK) and thymidine kinase 1 (TK1), as well as thyroid function indicators, were measured. Based on the patients' pathological results, the consistency between the TI-RADS diagnostic results and the pathological findings was analyzed. The diagnostic efficacy of TI-RADS classification was calculated. The incidence of malignant nodules across various TI-RADS categories was assessed, and the differences in serum levels of MK, TK1, and thyroid function indicators between patients with benign and malignant nodules were compared. Additionally, a correlation analysis was performed to analyze the correlation between the TI-RADS classification of thyroid nodules and the serum levels of MK, TK1, and thyroid function indicators. Results:Pathological examination results revealed 38 malignant nodules and 82 benign nodules. The consistency between ultrasound TI-RADS classification diagnoses and pathological results was good ( Kappa = 0.90, P < 0.001), with diagnostic sensitivity, specificity, and accuracy of 86.84% (33/38), 97.56% (80/82), and 94.17% (113/120), respectively. Among the ultrasound TI-RADS classifications, the malignant rates for category Ⅳb and category Ⅴ thyroid nodules were the highest, at 92.00% (23/25) and 100.00% (10/10), respectively. Serum levels of MK, TK1, and thyroid-stimulating hormone (TSH) were significantly higher in patients with malignant thyroid nodules than in those with benign thyroid nodules (all P < 0.05). Correlation analysis indicated that TI-RADS classification was positively correlated with serum levels of MK, TK1, and TSH ( r = 0.56, 0.60, 0.52, all P < 0.05). Conclusions:Ultrasound TI-RADS classification can be used to differentiate the nature of thyroid nodules, demonstrating high accuracy with low risks of missed or misdiagnoses. It is also closely related to indicators such as MK, TK1, and TSH, providing a basis for the detection of thyroid cancer.

Objective:To summarize the clinical and genetic characteristics of 23 children with pathogenic ACAN gene variants, enhance the understanding of this disorder, and explore possible regulatory mechanisms. Methods:A retrospective case series summary.The clinical characteristics and genetic analysis results of 23 children with ACAN gene variants treated in the Department of Endocrinology, Genetics and Metabolism, Children′s Hospital of Soochow University from January 2016 and September 2024 were analyzed retrospectively.Transcriptome sequencing was performed on peripheral blood samples from 3 of affected children and 3 age-matched healthy children as controls.Differentially expressed genes (DEGs) in the peripheral blood transcriptome profiles were identified.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted to explore the potential signaling pathways involved. Results:Among the 23 cases, there were 13 males and 10 females, aged from 2 years and 8 months to 12 years old, with 11 cases presenting advanced bone age.Thirteen cases were treated with growth hormone (GH), including 6 cases who received concomitant gonadotropin-releasing hormone analogue therapy.The treatment duration ranged from 3 to 70 months, resulting in varying degrees of height improvement in all treated patients.Transcriptomic analysis identified 811 DEGs, with 516 up-regulated and 295 down-regulated.GO and KEGG enrichment analyses revealed that the heterozygous ACAN variants were significantly associated with FcγR-mediated phagocytosis, nuclear factor-κB signaling pathway, the intestinal immune network for IgA production, rheumatoid arthritis, and systemic lupus erythematosus signaling pathways. Conclusions:The predominant clinical manifestations of patients with ACAN gene variants are short stature and advanced bone age.Although GH provocation tests may indicate normal GH levels, GH therapy can be effective in improving height.Immune-related factors may play a role in the pathogenesis of this disorder.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.