ObjectiveTo explore the possible mechanism of Yigan Fupi Prescription （抑肝扶脾方， YFP） in treating diarrhea-type irritable bowel syndrome （IBS-D） by investigating the AKT/mTOR signaling pathway. MethodsSixty SD rats were randomly divided into control group， model group， YFP low-， medium-， and high-dose group， and pinaverium bromide group， with 10 rats in each group. All groups but the control group， were subjected to 21 days of tail-clamping stimulation and 14 days of senna leaf gavage to establish a liver stagnation and spleen deficiency-type IBS-D rat model. After successful modeling， the YFP low-， medium-， and high-dose group were administered 0.96， 1.93， and 3.87 g/（kg·d） of the prescription， respectively. The pinaverium bromide group was given 13.5 mg/（kg·d）， while the control and model groups were given 10 ml/（kg·d） distilled water. All groups were administered once daily for 14 consecutive days. General conditions of the rats were recorded during the experiment， and after modeling and drug administration， body weight， Bristol stool score， abdominal withdrawal reflex （AWR） score， and histo pathology of colon tissue were observed under HE staining. ELISA was used to detect serum levels of tumor necrosis factor α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Immunofluorescence was employed to detect the levels of AKT/mTOR pathway-related proteins including phosphorylated AKT （p-AKT）/AKT and phosphorylated mTOR （p-mTOR）/mTOR in the colon tissue. Western Blotting was used to detect the levels of autophagy-related proteins， including UNC-51-like kinase 1 （ULK1）， Beclin1 and LC3， and tight junction proteins including Occludin and ZO-1 in the colon tissue. ResultsAfter modeling， compared to the control group， the body weight of rats in the other groups decreased， and Bristol stool scores， as well as AWR scores under 20， 40， 60， and 80 mmHg increased （P＜0.05 or P＜0.01）. After drug administration， compared to the control group， the model group showed reduced body weight， decreased ULK1， Beclin1， LC3Ⅱ/LC3Ⅰ， Occludin， and ZO-1 protein levels in the colon tissue （P＜0.05 or P＜0.01）， and increased Bristol stool scores， AWR scores， serum TNF-α， IL-1β， and IL-6 levels， as well as p-AKT/AKT and p-mTOR/mTOR protein relative expression levels （P＜0.05 or P＜0.01）. Pathological results showed a significant reduction in goblet cells in the upper part of the glandular layer of the colon， with mild inflammatory cell infiltration. The submucosal collagen fibers were dissolved， with unclear boundaries， pale staining， and microvascular congestion and dilation. Compared with the model group， the YFP low-， medium-， and high-dose group and the pinaverium bromide group showed increased body weight， Beclin1， Occludin， and LC3Ⅱ/LC3Ⅰ protein levels （P＜0.05 or P＜0.01）， and decreased Bristol stool scores， AWR scores under 40， 60， and 80 mmHg， serum IL-1β， IL-6， TNF-α levels， and p-AKT/AKT， p-mTOR/mTOR protein relative expression levels （P＜0.05 or P＜0.01）. The pathological morphology of the rats in the YFP groups and pinaverium bromide group showed varying degrees of improvement. Compared with the pinaverium bromide group， the YFP low- and medium-dose group showed increased AWR scores under 20， 40， and 60 mmHg （P＜0.05）. The YFP low-dose group had reduced TNF-α， IL-1β， and IL-6 levels， and increased p-mTOR/mTOR protein relative expression levels occured in all YFP groups （P＜0.05）. Compared with the YFP low-dose group， the YFP high-dose group and pinaverium bromide group showed decreased AWR scores under different pressure levels and reduced p-AKT/AKT protein relative expression levels， while the YFP medium- and high-dose group had elevated serum TNF-α， IL-1β levels and reduced p-mTOR/mTOR protein relative expression levels （P＜0.05）. ConclusionYFP can effectively improve the pathological injury of colon tissue in IBS-D model rats with liver stagnation and spleen deficiency， reduce Bristol stool and AWR scores， and its mechanism may be related to reducing level of inflammatory factors and inhibiting AKT/mTOR pathway-related proteins in colon tissue， thereby enhancing the expression of autophagy-related proteins in the colon tissue.

Idiopathic pulmonary fibrosis (IPF), a chronic interstitial lung disease, is characterized by aberrant wound healing, excessive scarring and the formation of myofibroblastic foci. Although the role of alternative splicing (AS) in the pathogenesis of organ fibrosis has garnered increasing attention, its specific contribution to pulmonary fibrosis remains incompletely understood. In this study, we identified an up-regulation of serine/arginine-rich splicing factor 7 (SRSF7) in lung fibroblasts derived from IPF patients and a bleomycin (BLM)-induced mouse model, and further characterized its functional role in both human fetal lung fibroblasts and mice. We demonstrated that enhanced expression of Srsf7 in mice spontaneously induced alveolar collagen accumulation. Mechanistically, we investigated alternative splicing events and revealed that SRSF7 modulates the alternative splicing of pyruvate kinase (PKM), leading to metabolic dysregulation and fibroblast activation. In vivo studies showed that fibroblast-specific knockout of Srsf7 in conditional knockout mice conferred resistance to bleomycin-induced pulmonary fibrosis. Importantly, through drug screening, we identified lomitapide as a novel modulator of SRSF7, which effectively mitigated experimental pulmonary fibrosis. Collectively, our findings elucidate a molecular pathway by which SRSF7 drives fibroblast metabolic dysregulation and propose a potential therapeutic strategy for pulmonary fibrosis.

Objective To explore the mechanism of the demethylase fat mass and obesity associatal(FTO)gene on the proliferation of human liver cancer cell line HepG2.Methods HepG2 cells were divided into the control group,FTO group(transfected with FTO over-expression plasmid),si-FTO group(transfected with si-FTO)and si-FTO+si-FoxO1 group(simultaneously transfected with si-FTO and si-FoxO1).The expression of FTO in cells was detected by RT-qPCR.Cell proliferation,invasion and apoptosis were examined using CCK-8 assay,Transwell chamber assay and flow cytometry,respectively.Dot blot assay was performed to measure m6 A methylation,and protein expression of FoxO1 in cells was detected by Western blot.Results Analysis of overall survival in liver cancer patients using The Cancer Genome Atlas(TCGA)showed higher expression of FTO associated with shorter survival(P＜0.05).Compared with normal liver cells HL7702,FTO relative expression was significantly increased in human liver cancer cells HepG2(P＜0.05).The relative expression of FTO in si-FTO group cells was lower than that in the control group,while the relative expression of FTO in FTO group was higher than that in the control group(P＜0.05).The relative expression of FTO in the si-FTO+si-FoxO1 group was higher than that in the si-FTO group(P＜0.05).Compared with the Control group,cell viability,count of invasive cells,relative level of m6 A were significantly decreased,while apoptosis rate and protein expression of FoxO1 were significantly increased in the si-FTO group(P＜0.05).Cell viability,count of invasive cells,and relative expression level of m6 A were sig-nificantly higher,while apoptosis rate and protein expression of FoxO1 were significantly lower in the FTO group compared to the Control group(P＜0.05).Compared with the si-FTO group,cell viability,invasive cells and rela-tive level of m6 A were significantly increased,while apoptosis rate and protein expression of FoxO1 were significant-ly decreased in the si-FTO+si-FoxO1 group(P＜0.05).Conclusion High expression of FTO is associated with poor clinical prognosis.Knockdown of the demethylase FTO gene inhibits proliferation and invasion of liver cancer cells and induces their apoptosis.The mechanism is potentially related to the regulation of FoxO1.

Objective To investigate the clinical manifestations,molecular genetics and gonadal pathol-ogy characteristics of patients with disorders of sex development(DSD),and to summarize the clinical experi-ence of identifying rare diseases from common symptoms.Methods The clinical data of 416 patients with DSD diagnosed and treated in the multidisciplinary center of DSD of Guangzhou Women and Children's Medical Cen-ter from May 2018 to August 2023 were retrospectively analyzed,summarized and discussed.Results Accord-ing to chromosome karyotype,416 cases of DSD were classified into three types:92 cases(22.1％)of abnormal sex chromosome karyotype,285 cases(68.5％)of 46,XY karyotype and 39 cases(9.4％)of 46,XX karyotype.Among the 92 patients with abnormal sex chromosome karyotype,59 cases were raised as males,18 cases(30.5％)complained of short penis with hypospadias and cryptorchidism.The most common karyotype was 45,X/46,XY(58 cases,63.0％).Among the 285 patients with 46,XY karyotype,238 cases were raised as males,and 63 cases(26.5％)complained of short penis and hypospadias;47 cases were raised as females,and 13 ca-ses(27.7％)complained of inguinal mass.A total of 216 patients with 46,XY karyotype were subjected to whole exome gene detection,and 155 cases(71.8％)were found to have molecular pathogenesis with the clinical phe-notype.Among the 39 patients with 46,XX karyotype,19 cases were raised as males,and 8 cases(42.1％)com-plained of short penis and hypospadias.In the 18 cases of gonad biopsy,17 cases showed testicular tissue in go-nads.Whole exome sequencing was performed in 14 cases.NR5A1 gene heterozygous mutation,SRY gene muta-tion and SOX3 gene mutation were found in 2 cases,respectively(14.3％).Twenty cases were raised as females,and 14 cases(70.0％)complained of clitoral hypertrophy.Gonad biopsy was performed in 8 cases,with 7 cases of ovotestis(87.5％)and 1 case of NR5A1 gene heterozygous mutation(14.3％).Conclusions The etiologies of DSD are complex and diverse,and the clinical manifestations are various,which can be manifested as hypospa-dias,micropenis,cryptorchidism and other common symptoms of the urinary system.Different etiologies have dif-ferent treatment options.Therefore,chromosome karyotype,molecular genetic testing and gonadal pathology can be used to clarify the cause of disease,especially for rare diseases,improve the detection rate,reduce the rate of missed diagnosis,and ensure reasonable treatment,especially sex selection.

Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.

Bladder perfusion is one of the main methods for the treatment of bladder cancer. In order to further improve the standardization of bladder cancer bladder perfusion operation for nursing staff, this paper, guided by evidence-based methods, formed the expert consensus on the whole process management of bladder perfusion for bladder cancer through Delphi expert consultation and expert demonstration meeting, and provided guidance for the standardization of clinical nursing practice and management institutionalization of bladder cancer bladder perfusion from seven aspects, namely, perfusion environment, operators, drug allocation, operation process, adverse reactions, health education and continuous nursing.

OBJECTIVE To systematically evaluate the safety of paroxetine in the treatment of pregnant patients with depression in the second and third trimesters of pregnancy， and provide reference for rational clinical use of it. METHODS Retrieved from Cochrane Library， PubMed， Embase， VIP， CNKI， Wanfang database and SinoMed database， by manual search， randomized controlled studies or observational studies were collected on depression patients who were given paroxetine vs. selective serotonin reuptake inhibitor （SSRI） in the second and third trimesters of pregnancy during the inception to Aug. 2022. Methodological qualities of the included studies were assessed by Cochrane Handbook 5.1.0 or Newcastle-Ottawa Scale （NOS）. Meta-analysis was performed with RevMan 5.4.1 software. RESULTS Finally， 9 observational studies were included， and all included studies were of high quality in NOS scale. Meta-analysis was performed on 8 cohort studies. Meta-analysis showed that the total incidence of adverse pregnancy outcomes of mothers and infants [RR＝0.99， 95%CI（0.89，1.10），P＝0.87]， total incidence of maternal adverse pregnancy outcomes [RR＝0.98， 95%CI （0.87，1.10）， P＝0.69] and premature birth [RR＝0.89， 95%CI （0.43， 1.83）， P＝0.75] in the second and third trimesters of pregnancy were lower than that with other SSRI， without statistical significance. The incidence of neonatal complications with paroxetine in the second and third trimesters of pregnancy was higher than that with other SSRI， but the difference was not statistically significant [RR＝1.02， 95%CI （0.82，1.29）， P＝0.84]. One study reported that the incidence of neonatal pulmonary hypertension in paroxetine group was higher than that in other SSRI group （0.4% vs. 0.3%）. CONCLUSIONS The safety of peroxetine in the second and third trimesters of pregnancy is comparable with that of other SSRI， but it is necessary to be alert to the occurrence of neonatal pulmonary hypertension.

Objective:To explore the value of the Wilcoxon-Mann-Whitney generalized dominance ratio (T max-weighted ratio) based on residual tissue time to peak (T max) delayed severity weighting in predicting the moderate to severe edema after acute anterior circulation ischemic stroke. Methods:The clinical and imaging features of patients with acute anterior circulation ischemic stroke from January 2019 to April 2022 in Yidu Central Hospital of Weifang were retrospectively analyzed. A total of 85 patients were enrolled, including 60 males and 25 females, with the age from 34 to 93 (67±11) years old. Patients underwent non-contrast CT, CT angiography of the head and neck, and CT perfusion imaging of the head, and ischemic core volume and the ratio of T max 4-6 s volume, T max 6-8 s volume, T max 8-10 s volume, and T max >10 s volume relative to the entire hypo-perfused area (T max>4 s volume) was measured, and the T max-weighted ratio was calculated, the collateral circulation were assessed. Patients were divided into mild edema group and moderate to severe edema group according to whether local swelling exceeded 1/3 of the unilateral cerebral hemisphere on non-contrast CT at 24-48 h. The indicators were compared between the two groups by independant t test, Mann-Whitney U and χ 2 test. The performance to predict moderate to severe edema was assessed using the receiver operating characteristic (ROC) curve. The univariate and multivariate logistic regression was used to analyze the risk factors for the moderate to severe edema. Differences in baseline National Institutes of Health Stroke Scale (NIHSS) score and infarct core volume were equalized by 1∶1 propensity score matching (PSM) and the differences of T max-weighted ratio between the two groups were further compared. Results:There were 52 cases in the mild edema group and 33 cases in the moderate to severe edema group. Baseline NIHSS score, T max>10 s volume, ischemic core volume, T max-weighted ratio and proportion of poor collateral circulation were higher in the moderate-severe edema group than those in the mild edema group ( P<0.001), T max 4-6 s volume was lower than in the mild edema group ( P<0.001). ROC analysis showed that the area under the curve (AUC) of T max-weighted ratio for predicting the incidence of moderate to severe edema was 0.885 (95%CI 0.798-0.944), with an optimal cut-off value of 1.17, sensitivity of 84.85% and specificity of 82.69% before PSM. The predictive ability based on T max-weighted ratio was similar to ischemic core volume( Z=0.64, P=0.520), T max 4-6 s volume ( Z=1.48, P=0.140) and superior to T max 6-8 s volume( Z=5.65, P<0.001), T max 8-10 s volume( Z=4.46, P<0.001), T max >10 s volume ( Z=2.91, P=0.004). Multivariate logistic regression analysis showed that T max-weighted ratio>1.17 was an independent predictor of the development of moderate to severe edema (OR=10.40,95%CI 2.65-40.83, P=0.001) through adjusted for baseline NIHSS score and ischemic core volume. After PSM, 14 patients in each group were included; the T max-weighted ratio was higher in the moderate-to-severe edema group than that in the mild edema group ( P<0.001), and the differences in other factors were not statistically significant (all P>0.05); ROC analysis showed that the AUC of T max-weighted ratio to predict the occurrence of moderate-to-severe edema was 0.852 (95%CI 0.667-0.957). Conclusion:The T max-weighted ratio can predict the occurrence of moderate-to-severe edema in brain tissue after acute anterior circulation ischemic stroke.

H 1-antihistamines are widely used in the treatment of various allergic diseases, but there are still many challenges in the safe and rational use of H 1-antihistamines in pediatrics, and there is a lack of guidance on the prescription review of H 1-antihistamines for children.In this paper, suggestions are put forward from the indications, dosage, route of administration, pathophysiological characteristics of children with individual difference and drug interactions, so as to provide reference for clinicians and pharmacists.

Objective To investigate the clinical and imaging features of testicular interstitial lesions in children.Methods The clinical and imaging data of 8 children with testicular interstitial lesions confirmed by pathology were analyzed retrospectively.Results There were 8 cases of precocious puberty,5 cases of elevated sex hormones,and 2 cases of congenital adrenal hyperplasia(CAH).The lesions were located in the unilateral testis in 6 cases,bilateral in 2 cases,single in 4 cases,and multiple in 4 cases,with diameters 3-14 mm,all of them with clear boundaries.4 cases of leydig cell hyperplasia(LCH)showed heterogeneous hypoechoic lesions,with slightly significant blood flow signal on color Doppler flow imaging(CDFI);CT showed iso-density lesions with mild enhanced after enhancement;MRI showed iso-signal on T1WI,and T2WI fat suppression were iso-signal mainly with mild enhanced after enhancement.2 cases of leydig cell tumor(LCT)showed heterogeneous hypoechoic lesions with significant blood flow signals inside and around on CDFI,CT showed iso and slightly low density with low-density necrosis,which were inhomogeneously and significantly enhanced after enhancement.2 cases of testicular adrenal rest tumor(TART)showed heterogeneous hypoechoic lesions with significant blood flow signals inside and around on CDFI;CT showed iso and slightly low density,with inhomogeneouly and significantly enhanced after enhancement;MRI showed iso-signal mainly on T1WI,and T2WI fat suppression were slightly low signal mainly with mixed signal,which were inhomogeneously and significantly enhanced after enhancement,complicated with adrenal hyperplasia.Conclusion The imaging findings of testicular interstitial lesions in children have certain characteristics,and the combination of clinical manifestations,basic diseases,laboratory and a variety of imaging examinations are helpful in improving diagnostic efficiency.