This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

This paper introduces a real-world cohort research model for community-acquired pneumonia （CAP） based on the Jiangsu Traditional Chinese Medicine （TCM） Dominant Diseases Diagnosis and Treatment Data Platform. Firstly， data cleaning is performed by standardizing diagnosis， symptoms， treatment and imaging， intelligently extracting unstructured information， and cleaning and constructing a standardized database. Secondly， for cohort establishment， CAP patients across the province are screened in accordance with CAP diagnostic criteria to build a high-quality disease-specific cohort. Lastly， in terms of protocol design， the characteristics of TCM research and the CAP disease profile are considered to determine appropriate inclusion and exclusion criteria， estimate sample size， define interventions， outcomes and economic evaluations， providing a reference for real-world TCM research on CAP.

OBJECTIVE To investigate the mechanism of cordycepin in delaying the transformation from acute kidney injury （AKI） to chronic kidney disease （CKD） （short for “AKI-CKD”）. METHODS Network pharmacology and bioinformatics were used to analyze and predict the signaling pathways of cordycepin in delaying AKI-CKD progression and its relationship with the ferroptosis pathway. Cell experiments were performed to verify the predicted results. Human renal tubular epithelial HK-2 cells were divided into blank group， cordycepin group， model group， and H 2 O 2 +cordycepin group. Except for the blank group and cordycepin group， all other groups were treated with 150 μmol/L H 2 O 2 for 72 h to induce persistent oxidative stress injury in cells. After 72 h of cordycepin （40 μmol/L） intervention， the protein and mRNA expression levels of glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4），nuclear factor-erythroid 2-related factor 2（Nrf2）， and heme oxygenase-1 （HO-1） in cells were detected. Furthermore， the Nrf2 inhibitor ML385 was added on the basis of H 2 O 2 +cordycepin to verify the role of the Nrf2/HO-1 pathway. RESULTS Network pharmacology and bioinformatics analysis showed that there were 42 overlapping genes related to AKI-CKD and ferroptosis that interact with cordycepin ferroptosis， among which 38 were annotated as ferroptosis-related genes. HO-1 and Nrf2 might be important targets for cordycepin to inhibit ferroptosis. The binding energies of cordycepin with Nrf2 and HO-1 proteins were －8.5 and －6.7 kcal/mol， respectively. Cell experiments showed that compared with the model group， the protein and mRNA expression levels of GPX4， Nrf2 and HO-1 were significantly increased （ P ＜0.05），while the protein and mRNA expression levels of ACSL4 were significantly decrease d （ P ＜0.05） in the H 2 O 2 +cordycepin group. After the addition of the Nrf2 inhibitor ML385， the effects of cordycepin on the above proteins and mRNA were significantly reversed （ P ＜0.05）. CONCLUSIONS Cordycepin can inhibit ferroptosis by activating the Nrf2/HO-1 pathway， reduce persistent oxidative stress injury of renal tubular epithelial cells， and delay the progression of AKI-CKD.

Autosomal dominant polycystic kidney disease （ADPKD） is a systemic hereditary renal disorder and can affect multiple organs， and congenital hepatic fibrosis is one of the manifestations of liver involvement and is an important complication of ADPKD. Symptomatic management is currently the main treatment method for this disease， and disease-specific drugs such as tolvaptan have limited indications and cannot correct the underlying genetic defect. This article reports a case of ADPKD with congenital hepatic fibrosis， and sirolimus was identified as the individualized treatment regimen based on peripheral blood functional genomic analysis and drug sensitivity prediction platform. The patient achieved significant improvements in symptoms and quality of life after treatment， with a stable kidney volume. This case shows that functional genomics has a potential value in guiding individualized treatment of rare genetic disorders， which provides new treatment ideas and practice paths for similar patients.

The abnormal accumulation of neutrophil extracellular traps （NETs） can promote the initiation and progression of pancreatic cancer， which is considered a potential therapeutic target for this disease. The Miraculous Pivot·Inquiry About Statement （《灵枢·口问》） have recorded the concept of "defensive qi stagnation". Based on the recognition that the function of defensive qi is similar to the immune function of neutrophils， and combining traditional Chinese medicine theory with clinical practice， it is proposed that the abnormal accumulation of NETs may be a pathological product of "defensive qi stagnation"， with the spleen being the critical site of pathology. Further exploring the application strategy of cleaning spleen and restoring defensive qi method in pancreatic cancer treatment， it is proposed to employ three approaches such as dredging method to eliminate spleen stagnation and inhibit pancreatic cancer proliferation， cleaning method to remove spleen dampness and suppress the inflammatory micro-environment， and tonifying method to strengthen Weiqi and to improve the immune microenvironment， which aims to provide new insights for the clinical treatment of pancreatic cancer with traditional Chinese medicine.

BACKGROUND:Cannabidiol is effective in ameliorating the body's inflammatory response,but no clear mechanistic studies have been conducted to ameliorate skeletal muscle inflammation induced by exhaustive exercise. OBJECTIVE:To explore the mechanism by which cannabidiol improves skeletal muscle inflammation during exhaustive exercise by using transcriptome sequencing technology. METHODS:Thirty-six Sprague-Dawley rats were randomly divided into six groups:blank control group,exercise coconut oil group,exercise control group,50 mg/kg cannabidiol group,60 mg/kg cannabidiol group,and 70 mg/kg cannabidiol group,with six rats in each group.Except for rats in the blank control group,rats in each group were subjected to swimming exercise for 9 days to produce the exhaustive exercise model.At the end of each swimming exercise,rats in the cannabidiol groups were given 2 mL of fat-soluble cannabidiol at different concentrations(50,60,and 70 mg/kg)by gavage;rats in the exercise coconut oil group were given the same volume of coconut oil by gavage until the end of the exercise on the 9th day;and rats in the blank control group and the exercise control group were not given any special treatment.The levels of inflammatory factors and differentially expressed genes in the skeletal muscle of rats in each group were determined using ELISA and transcriptome sequencing techniques.Differentially expressed genes obtained were subjected to KEGG analysis,and the accuracy of the sequencing data was verified by fluorescence quantitative PCR. RESULTS AND CONCLUSION:The results of ELISA showed that the contents of interleukin-6(P<0.05),tumor necrosis factor-α(P<0.01),interleukin-10 and other inflammatory factors in the exercise group increased significantly compared with the blank control group and the coconut oil group.After cannabidiol intervention,the mass concentrations of interleukin-6 and tumor necrosis factor-α showed a sequential decrease with increasing cannabidiol concentration.By comparing GO and KEGG databases,the functional properties of differentially expressed genes were analyzed,and the results showed that the differentially expressed genes were mainly involved in the tumor necrosis factor signaling pathway and the Toll-like receptor signaling pathway.RT-qPCR results showed that the trends of five randomly selected differentially expressed genes were in agreement with the transcriptome sequencing results.To conclude,cannabidiol can improve skeletal muscle inflammation caused by exhaustive exercise.

Sexual dimorphism in the brain underlies behavioral differences between sexes. The bed nucleus of the stria terminalis (BNST) is a complex nucleus that differs between males and females, but the sexual dimorphism in cytoarchitecture and the connectome of its oval subdivision (ovBNST) remains largely unexplored. By combining snRNA-seq and transgenic labeling, we found a higher density of ovBNST proenkephalin (ovBNST^PENK) neurons in male than female mice. Anatomically, we virally mapped the efferents and afferents of ovBNST^PENK neurons, finding reciprocally dimorphic connections with the hypothalamus and striatum. Gene enrichment analysis suggests that ovBNST^PENK neurons are modulated by the upstream dopamine pathway. Functionally, by applying caspase-3-mediated depletion of ovBNST^PENK neurons, we found that loss of these neurons enhanced locomotor activity in male but not female mice, without altering the anxiety-like phenotypes in either sex. Our study may pave the way for a better understanding of the anatomical and functional profiles of ovBNST^PENK neurons from a sexually dimorphic perspective.
Animals ; Male ; Female ; Septal Nuclei/physiology* ; Sex Characteristics ; Neurons/physiology* ; Enkephalins/metabolism* ; Mice ; Mice, Transgenic ; Protein Precursors/metabolism* ; Mice, Inbred C57BL ; Neural Pathways/physiology*

Objective:To screen differentially expressed circular RNAs(circRNA)associated with lymph node metastasis in cervical cancer and investigate their molecular mechanism and biological function.Methods:Cancer tissue samples were collected from 40 cervical cancer patients(20 with lymph node metastasis and 20 without lymph node metastasis).Three specimens were randomly selected from each group,and the circRNA expression profile was analyzed using high-throughput sequencing technology,and perform KEGG and GO enrichment analy-sis was performed on the differentially expressed circRNA derived mother genes.Real-time fluorescence quantita-tive PCR(qRT-PCR)technology was used to validate four significantly upregulated circRNAs with high differential multiples,and a ceRNA network was constructed for database prediction.The relationship between circRNA with the most significant differential expression and clinical pathological features was analyzed.Transfect circRNA small interfering RNA(siRNA)into cervical cancer cell lines.Cell lines were divided into circRNA knockdown group and negative control(si-NC)group.The proliferation of SiHa cells and HeLa cells in cervical cancer was detected by Cell Counting Kit-8(CCK-8).Results:①A total of 2039 differentially expressed circRNAs were detec-ted.The differentially expressed circRNA derived mother genes are mainly enriched in ubiquitin mediated protein hydrolysis,MAPK signaling pathway,endocytosis,apoptosis and other signaling pathways.②Four significantly up-regulated circRNAs with high differential multiples in the transfer group were selected,which are hsa_circ_0067492,hsa_circ_0004904,hsa_circ_0085465,and hsa_circ_0007718.qRT-PCR showed that hsa_circ_0004904,hsa_circ_0085465,and hsa_circ_0007718 were all highly expressed in cervical cancer tissues with lymph node metas-tasis(P<0.01).Hsa_circ_0004904 could interact with microRNAs(miRNAs)such as hsa-miR-3916.Hsa_circ_0085465 could interact with miRNAs such as hsa-miR-4659b-3p,and hsa_circ_0007718 could interact with miRNAs such as hsa-miR-127-5p.③The high and low expression groups of hsa_circ_0004904 showed statistically signigi-cant differences in lymph node metastasis and lymphovascular invasion status(P<0.05).Compared with the si-NC group,the proliferation ability of SiHa cells and HeLa cells was significantly reduced after transfection with hsa_circ_0004904 at 24,48,and 72 hours(P<0.05).Conclusions:Differentially expressed circRNAs were identi-fied in cervical cancer patients with lymph node metastasis.Silencing these circRNAs significantly inhibited the proliferation of cervical cancer cells,suggesting their potential involvement in the progression of lymph node me-tastasis.These circRNAs may serve as specific biomarkers for predicting lymph node metastasis.

Objective:To explore the effect of Bian-stone warming and ironing therapy on gastrointestinal blood flow in patients with severe cerebrovascular disease undergoing therapeutic hypothermia.Methods:From January 2023 to October 2024, 92 patients with severe cerebrovascular disease in the Neurosurgical Intensive Care Unit of Xuanwu Hospital of Capital Medical University were selected for the study using convenience sampling method. The patients were divided into a control group and an experimental group of 46 cases each according to the randomized numerical table method. Control group was given routine nursing, and experimental group carried out Bian-stone warming and ironing therapy on the basis of control group. Peak systolic velocity (PSV), end-diastolic velocity (EDV), and time-averaged mean velocity (TAMV) of the superior mesenteric artery of the two groups of patients were monitored before and after five days of intervention, respectively, and the situation of gastric retention and bowel sounds were recorded and compared between the groups.Results:PSV was higher in experimental group than in control group after intervention, and the difference was statistically significant ( P<0.05). The difference in EDV between the two groups was not statistically significant ( P>0.05). TAMV of experimental group was higher than that of control group after intervention, and the difference in TAVM before and after intervention of experimental group was smaller than that of control group, and the differences were all statistically significant ( P<0.05). The number of cases of gastric retention and weakened or disappeared bowel sounds in experimental group was less than that in control group after intervention, and the differences were all statistically significant ( P<0.05) . Conclusions:Bian-stone warming and ironing therapy improves gastrointestinal blood flow rate and reduces feeding intolerance in patients with severe cerebrovascular disease undergoing therapeutic hypothermia.

This study investigated the expression of the soluble nontoxic mutant CRM197 of diphtheria toxin,to prepare a safe and effective recombinant CRM197 protein.Codon-optimized CRM197 gene sequences were cloned into the pCold Ⅱ and pET-28a(+)prokaryotic expression vectors.The successfully cloned recombinant plasmids were screened and transformed into Escherichia coli BL21(DE3)competent cells.After induction of expression,the protein expression type was determined through SDS-PAGE analysis.The conditions for expression of the solublerecombinant protein were then optimized.The recombinant CRM197 protein was purified through two rounds of Ni-NTA affinity chromatography.The purified protein was used to immunize mice,and antibody levels in the se-rum were measured with ELISA.The codon adaptation index(CAI)of the optimized sequence increased from 0.72 to 0.93.The recom-binant plasmids pET-28a(+)-CRM197 and pCold Ⅱ-CRM197 were successfully constructed,as confirmed through colony PCR and double digestion.Expression analysis revealed that pET-28a(+)-CRM197 was expressed primarily as inclusion bodies,whereas pColdⅡ-CRM197 was expressed predominantly in soluble form.The conditions for soluble protein expression via pCold Ⅱ-CRM197 were optimized.When the inoculum was 3%and the IPTG concentration was 0.4 mmol/L,induction at 20 ℃ for 24 h significantly increased the expression of the soluble target protein.The pCold Ⅱ-CRM197 recombinant protein was purified from the supernatant with Ni-NTA affinity chromatography,thus resulting in a target protein with a purity greater than 98%.ELISA after three rounds of immuniza-tion indicated that the levels of IgG,IgM,IgG1,and IgG2a antibodies in the serum in immunized mice were significantly higher than those in the control group.In summary,the CRM197 recombinant protein was successfully prepared with the pCold Ⅱvector and exhib-ited high soluble expression,high purity,and favorable immunogenicity.