ObjectiveTo investigate the mechanism of Qinggan Jianpi Houxue prescription against hepatic fibrosis in mice using metabolomics. MethodsA hepatic fibrosis mouse model was established by intraperitoneal injection of 20% carbon tetrachloride （CCl₄， diluted with olive oil at a ratio of 4∶1） at a dose of 2.5 mL·kg^-1 for 4 weeks. A total of 54 C57BL/6J mice were randomly divided into the blank control group， model group， Qinggan Jianpi Houxue prescription high-dose group （47.32 g·kg^-1）， medium-dose group （23.66 g·kg^-1）， low-dose group （11.83 g·kg^-1）， and Biejiajian pill group （1.365 g·kg^-1）， with 9 mice in each group. Concurrently with modeling， each administration group received the corresponding dose continuously for 4 weeks， once daily. After 4 weeks， liver tissues and blood samples were collected for liver function and hepatic pathological examinations. Non-targeted metabolomics was employed to detect hepatic metabolites. Principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were performed to explore the metabolites and their involved metabolic pathways in the liver tissues of mice in the blank control group， model group and Qinggan Jianpi Houxue prescription high-dose group. Western blot validation was conducted at both the cellular and animal levels. ResultsCompared with the blank control group， the model group showed significantly increased levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）， along with increased inflammatory cell infiltration， and formation of fibrous septa. The predominantly enriched pathway was the forkhead box O （FoxO） signaling pathway. Compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group exhibited significantly decreased ALT and AST levels （P<0.01）. In all administration groups of Qinggan Jianpi Huoxue prescription， inflammatory cell infiltration was reduced， fibrous septa were narrowed and diminished， collagen fibers were reduced， and the collagen area was significantly decreased. Non-targeted metabolomics identified 79 differential metabolites， among which 49 were up-regulated and 30 down-regulated， primarily enriched in the tricarboxylic acid （TCA） cycle and pyruvate metabolism pathways. Western blot results demonstrated that， compared with the blank control group， the model group exhibited significantly upregulated protein expressions of α-smooth muscle actin （α-SMA）， lactate dehydrogenase A （LDHA）， FoxO1， pyruvate kinase M2 （PKM2）， hexokinase 2 （HK2）， and phosphofructokinase 1 （PFK1） in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration increased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， glucose transporter 1 （GLUT1）， and PFK1 were significantly up-regulated （P<0.05， P<0.01）. Conversely， compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group showed significantly downregulated protein expression of α-SMA， LDHA， FoxO1， PKM2， HK2， and PFK1 in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration decreased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， GLUT1， and PFK1 were significantly down-regulated （P<0.05， P<0.01）. ConclusionQinggan Jianpi Huoxue prescription can alleviate the pathological progression of hepatic fibrosis in mice， potentially by regulating the TCA cycle and pyruvate metabolism through FoxO1， and by modulating glycolytic metabolic reprogramming in hepatic stellate cells， thereby promoting liver tissue repair.

Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Solitary fibrous tumors（SFT）is rare Mesenchymal tissue tumor，originally identified and usually found in the pleura，but the incidence of SFT in the spermatic cord is particularly rare.This article reported a case of a 37-year-old patient who was admitted for a right inguinal mass over the past 2 years. Preoperative MR examination showed a right spermatic cord-derived mass. Under spinal anesthesia，excision of the right spermatic cord mass was performed. Postoperative pathological diagnosis was spermatic cord solitary fibrous tumor. Twelve months of follow-up revealed no metastasis and recurrence on imaging examination. Solitary fibrous tumors of the spermatic cord are rare and require a combination of pathological examination and immunohistochemistry to confirm the diagnosis. Once the diagnosis is confirmed，tumor resection should be performed as soon as possible，and benign lesions are also at risk of recurrence，and long-term follow-up is required after surgery.

Objective:To explore the feasibility of joint screening of the two primary immunodeficiency diseases [severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia(XLA)] and spinal muscular atrophy(SMA) in newborns by multiplex real-time quantitative PCR technology, and to provide evidence for early screening, diagnosis and treatment of children.Methods:Cross-sectional study. From July 2021 to January 2023, a total of 103 240 dry blood spots samples of newborns were collected which were delivered to Neonatal Disease Screening Center of Zhejiang by cold chain transportation. The concentrations of the T cell receptor excision ring (TREC), Kappa deletion of the recombinant excision loop (KREC), and exon 7 deletion of Survival Motor Neuron 1 (SMN1) gene in dry blood spots were simultaneously detected by multiplex real-time fluorescence quantitative PCR, taken ribonuclease P/MRP 30 000 subunits (RPP30) as an internal reference gene. The positive newborns were further diagnosed by other laboratory tests and gene sequencing was taken as gold standard. Children samples from 1 case of SCID, 3 cases of XLA and 2 cases of SMA were used for positive verification. The correlation between detected concentration of TREC/KREC and basic information in newborns were analyzed. The differences among groups for each factor were analyzed.Results:One case of SCID, 2 cases of XLA, 9 cases of SMA and 7 cases of other genetic diseases (4 cases of DiGeorge syndrome, 1 case of trisomy 21 syndrome, 1 case of Noonan syndrome and 1 case of super male syndrome) were identified by multiplex real-time fluorescence quantitative PCR. The positive predictive values of screening neonatal SCID, XLA and SMA were 2.44% (1/41), 2.78% (2/72) and 9/9 respectively. Taking the samples from clinically diagnosed 1 case of SCID, 3 cases of XLA and 2 cases of SMA as positive validation samples, which were all identified. The detected results of TREC/KREC correlated with time of blood collection, sex, weight, gestational age and delivery mode of newborns, whose r values were 0.162/0.187, 0.066/0.032, 0.045/0.042, ?0.015/?0.088 and 0.014/0.068 respectively (all P<0.05). Conclusions:Relying on current neonatal screening platform in Zhejiang, it is feasible to screen jointly two kinds of primary immunodeficiency diseases and spinal muscular atrophy in newborns by multiple real-time fluorescence quantitative PCR technology.

This article reports a case of a 55-year-old patient who was admitted to the hospital due to an 8-year history of a right epididymal mass and intermittent right scrotal heaviness for 1 month. Preoperative ultrasound examination revealed a right epididymal cyst and right testicular hydrocele. Under general anesthesia, a right epididymal mass resection and right testicular tunica vaginalis inversion procedure were performed. Postoperative pathological diagnosis was epididymal papillary cystadenoma. The patient has been followed up for 1 year with no recurrence and malignancy. Epididymal papillary cystadenoma, a rare benign tumor, lacks specific clinical and imaging features, often leading to misdiagnosis as seminal vesicle cysts or malignant tumors. If associated with von Hippel-Lindau syndrome, bilateral occurrence should be considered to prevent missed diagnosis and treatment delay.

Lung transplantation is an important means to treat end-stage lung disease and improve the survival rate and quality of life of patients. However, many postoperative complications seriously affect the prognosis of recipients. Accurate identification of key prognostic factors and construction of individualized and accurate prediction models are of great significance for postoperative prognosis evaluation, treatment strategy formulation and clinical decision-making. In recent years, the clinical prediction model of lung transplantation has gradually changed from traditional statistical methods to machine learning-driven. Compared with traditional models such as Cox regression and Logistic regression, machine learning models such as random forest, support vector machine and artificial neural network have certain advantages in postoperative survival rate prediction, early warning of complications and pulmonary function evaluation. However, their application is also affected by insufficient sample size and poor interpretability of models. Under the condition of small samples, the traditional model still has important value in prediction accuracy. The appropriate prediction model should be selected according to the clinical status of lung transplantation in China, considering the factors such as sample size, variable complexity and model interpretability. In the future, a multi-center, large-sample lung transplantation database should be constructed to further optimize and tap the potential of machine learning algorithms to improve the robustness and clinical applicability of the model.

This article reports a case of a 55-year-old patient who was admitted to the hospital due to an 8-year history of a right epididymal mass and intermittent right scrotal heaviness for 1 month. Preoperative ultrasound examination revealed a right epididymal cyst and right testicular hydrocele. Under general anesthesia, a right epididymal mass resection and right testicular tunica vaginalis inversion procedure were performed. Postoperative pathological diagnosis was epididymal papillary cystadenoma. The patient has been followed up for 1 year with no recurrence and malignancy. Epididymal papillary cystadenoma, a rare benign tumor, lacks specific clinical and imaging features, often leading to misdiagnosis as seminal vesicle cysts or malignant tumors. If associated with von Hippel-Lindau syndrome, bilateral occurrence should be considered to prevent missed diagnosis and treatment delay.

Solitary fibrous tumors（SFT）is rare Mesenchymal tissue tumor，originally identified and usually found in the pleura，but the incidence of SFT in the spermatic cord is particularly rare.This article reported a case of a 37-year-old patient who was admitted for a right inguinal mass over the past 2 years. Preoperative MR examination showed a right spermatic cord-derived mass. Under spinal anesthesia，excision of the right spermatic cord mass was performed. Postoperative pathological diagnosis was spermatic cord solitary fibrous tumor. Twelve months of follow-up revealed no metastasis and recurrence on imaging examination. Solitary fibrous tumors of the spermatic cord are rare and require a combination of pathological examination and immunohistochemistry to confirm the diagnosis. Once the diagnosis is confirmed，tumor resection should be performed as soon as possible，and benign lesions are also at risk of recurrence，and long-term follow-up is required after surgery.

Objective:To explore the feasibility of joint screening of the two primary immunodeficiency diseases [severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia(XLA)] and spinal muscular atrophy(SMA) in newborns by multiplex real-time quantitative PCR technology, and to provide evidence for early screening, diagnosis and treatment of children.Methods:Cross-sectional study. From July 2021 to January 2023, a total of 103 240 dry blood spots samples of newborns were collected which were delivered to Neonatal Disease Screening Center of Zhejiang by cold chain transportation. The concentrations of the T cell receptor excision ring (TREC), Kappa deletion of the recombinant excision loop (KREC), and exon 7 deletion of Survival Motor Neuron 1 (SMN1) gene in dry blood spots were simultaneously detected by multiplex real-time fluorescence quantitative PCR, taken ribonuclease P/MRP 30 000 subunits (RPP30) as an internal reference gene. The positive newborns were further diagnosed by other laboratory tests and gene sequencing was taken as gold standard. Children samples from 1 case of SCID, 3 cases of XLA and 2 cases of SMA were used for positive verification. The correlation between detected concentration of TREC/KREC and basic information in newborns were analyzed. The differences among groups for each factor were analyzed.Results:One case of SCID, 2 cases of XLA, 9 cases of SMA and 7 cases of other genetic diseases (4 cases of DiGeorge syndrome, 1 case of trisomy 21 syndrome, 1 case of Noonan syndrome and 1 case of super male syndrome) were identified by multiplex real-time fluorescence quantitative PCR. The positive predictive values of screening neonatal SCID, XLA and SMA were 2.44% (1/41), 2.78% (2/72) and 9/9 respectively. Taking the samples from clinically diagnosed 1 case of SCID, 3 cases of XLA and 2 cases of SMA as positive validation samples, which were all identified. The detected results of TREC/KREC correlated with time of blood collection, sex, weight, gestational age and delivery mode of newborns, whose r values were 0.162/0.187, 0.066/0.032, 0.045/0.042, ?0.015/?0.088 and 0.014/0.068 respectively (all P<0.05). Conclusions:Relying on current neonatal screening platform in Zhejiang, it is feasible to screen jointly two kinds of primary immunodeficiency diseases and spinal muscular atrophy in newborns by multiple real-time fluorescence quantitative PCR technology.

Lung transplantation is an important means to treat end-stage lung disease and improve the survival rate and quality of life of patients. However, many postoperative complications seriously affect the prognosis of recipients. Accurate identification of key prognostic factors and construction of individualized and accurate prediction models are of great significance for postoperative prognosis evaluation, treatment strategy formulation and clinical decision-making. In recent years, the clinical prediction model of lung transplantation has gradually changed from traditional statistical methods to machine learning-driven. Compared with traditional models such as Cox regression and Logistic regression, machine learning models such as random forest, support vector machine and artificial neural network have certain advantages in postoperative survival rate prediction, early warning of complications and pulmonary function evaluation. However, their application is also affected by insufficient sample size and poor interpretability of models. Under the condition of small samples, the traditional model still has important value in prediction accuracy. The appropriate prediction model should be selected according to the clinical status of lung transplantation in China, considering the factors such as sample size, variable complexity and model interpretability. In the future, a multi-center, large-sample lung transplantation database should be constructed to further optimize and tap the potential of machine learning algorithms to improve the robustness and clinical applicability of the model.