The aim of this research was to investigate the mechanism of chelidonine on H2 O2-in-duced inflammatory injury in porcine intestinal epithelial cells(IPEC-J2)through transcriptome sequencing.IPEC-J2 cells in the logarithmic growth phase were divided into the blank group(K group),H2O2 group(S group)and chelidonine group(L group),with three replicates in each group.Total RNA was isolated from each group for the purpose of constructing a sequencing li-brary.The assembled data underwent functional annotation,differential gene analysis,as well as GO and KEGG enrichment analyses.qPCR was used to confirm the expression of key differentially expressed genes(DEGs),and ELISA was utilized to assess the effect of chelidonine on the permea-bility of IPEC-J2 cells.The results indicated that the sequencing data met the necessary criteria and demonstrated a strong correlation between samples.The GO functional annotation results suggest that the intervention effects of chelidonine involve biological processes such as oxidative stress re-sponse and G2/M phase transition regulation of the mitotic cycle,and are closely associated with molecular functions,such as transmembrane transport activity.The KEGG enrichment analysis indicates that following H2 O2 treatment,DEGs in IPEC-J2 cells are predominantly enriched in the p53 signaling pathway,the coagulation cascade,the FoxO signaling pathway,and various other sig-naling pathways.Following pretreatment with chelidonine,the DEGs exhibit significant enrichment in several signaling pathways related to inflammation,including the TNF signaling pathway,syn-aptic vesicle cycle,and IL-17 signaling pathway.The results of qPCR were consistent with the se-quencing results.Chelidonine has also been found to effectively inhibit LDH release,elevate GLN content,and decrease DOA content.In conclusion,it can be seen that chelidonine can reduce cell permeability and alleviate H2 O2-induced inflammatory injury in IPEC-J2 cells by modulating in-flammation-related pathways such as the TNF signaling pathway.

Objective The mechanism of Huazhuo xingxue decoction(HZXXD)in the treatment of ischemic stroke was explored through network pharmacology,molecular docking and cell validation.Methods TCMSP,TCMID,BATMAN-TCM database and literature search were used to get the chemical components and related target proteins of Huazhuo Xingxue Decoction,and the targets of dementia,stroke and amnesia were obtained from Genecards database and OMIM database.The traditional Chinese medicine-active components-target-network and protein interaction map were constructed by using Cytoscape,and the target was enriched by KEGG pathway by David database.Western blot was used to investigate the effect of HZXXD on inflammation-related core targets expression using oxygen and glucose deprivation/reoxygenation cell model.Finally,Autodock was used for molecular docking of key active ingredients and important targets to evaluate their binding activity.Results 76 active molecules and 33 common targets of herb-disease were screened out.KEGG bioaccumulation results involve multiple inflammatory signal pathways such as TNF,chemical carcinogenesis-reactive oxygen species and HIF-1.TNF-α was found to be the core target of HZXXD by oxygen glucose deprivation/reoxygenation cell experiments.Five compounds with the strongest binding ability to TNF-α,kaempferol,apigenin,aloe-emodin,baicalein and stigasterol,were screened by traditional Chinese medicine-active ingredient-target network map and molecular docking.Conclusion Huazhuo Xingxue Decoction may down regulate the expression of core target TNF-α,kaempferol,apigenin,aloe emodin,baicalein and stigasterol may be the main active substances for TNF-α binding.

Objective The mechanism of Huazhuo xingxue decoction(HZXXD)in the treatment of ischemic stroke was explored through network pharmacology,molecular docking and cell validation.Methods TCMSP,TCMID,BATMAN-TCM database and literature search were used to get the chemical components and related target proteins of Huazhuo Xingxue Decoction,and the targets of dementia,stroke and amnesia were obtained from Genecards database and OMIM database.The traditional Chinese medicine-active components-target-network and protein interaction map were constructed by using Cytoscape,and the target was enriched by KEGG pathway by David database.Western blot was used to investigate the effect of HZXXD on inflammation-related core targets expression using oxygen and glucose deprivation/reoxygenation cell model.Finally,Autodock was used for molecular docking of key active ingredients and important targets to evaluate their binding activity.Results 76 active molecules and 33 common targets of herb-disease were screened out.KEGG bioaccumulation results involve multiple inflammatory signal pathways such as TNF,chemical carcinogenesis-reactive oxygen species and HIF-1.TNF-α was found to be the core target of HZXXD by oxygen glucose deprivation/reoxygenation cell experiments.Five compounds with the strongest binding ability to TNF-α,kaempferol,apigenin,aloe-emodin,baicalein and stigasterol,were screened by traditional Chinese medicine-active ingredient-target network map and molecular docking.Conclusion Huazhuo Xingxue Decoction may down regulate the expression of core target TNF-α,kaempferol,apigenin,aloe emodin,baicalein and stigasterol may be the main active substances for TNF-α binding.

The aim of this research was to investigate the mechanism of chelidonine on H2 O2-in-duced inflammatory injury in porcine intestinal epithelial cells(IPEC-J2)through transcriptome sequencing.IPEC-J2 cells in the logarithmic growth phase were divided into the blank group(K group),H2O2 group(S group)and chelidonine group(L group),with three replicates in each group.Total RNA was isolated from each group for the purpose of constructing a sequencing li-brary.The assembled data underwent functional annotation,differential gene analysis,as well as GO and KEGG enrichment analyses.qPCR was used to confirm the expression of key differentially expressed genes(DEGs),and ELISA was utilized to assess the effect of chelidonine on the permea-bility of IPEC-J2 cells.The results indicated that the sequencing data met the necessary criteria and demonstrated a strong correlation between samples.The GO functional annotation results suggest that the intervention effects of chelidonine involve biological processes such as oxidative stress re-sponse and G2/M phase transition regulation of the mitotic cycle,and are closely associated with molecular functions,such as transmembrane transport activity.The KEGG enrichment analysis indicates that following H2 O2 treatment,DEGs in IPEC-J2 cells are predominantly enriched in the p53 signaling pathway,the coagulation cascade,the FoxO signaling pathway,and various other sig-naling pathways.Following pretreatment with chelidonine,the DEGs exhibit significant enrichment in several signaling pathways related to inflammation,including the TNF signaling pathway,syn-aptic vesicle cycle,and IL-17 signaling pathway.The results of qPCR were consistent with the se-quencing results.Chelidonine has also been found to effectively inhibit LDH release,elevate GLN content,and decrease DOA content.In conclusion,it can be seen that chelidonine can reduce cell permeability and alleviate H2 O2-induced inflammatory injury in IPEC-J2 cells by modulating in-flammation-related pathways such as the TNF signaling pathway.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.

Objective:To investigate the proteins interacting with Mycobacterium tuberculosis Rv1705c in human body. Methods:Rv1705c was prokaryotically expressed and inclusion bodies were collected for further lysis and the purification of Rv1705c. ELISA assay was used to detect the secretion of IFN-γ after stimulating macrophages with Rv1705c protein. Purified and biotin-labeled Rv1705c sample was incubated on the HuProt? human proteome microarray to screen the interacting proteins. GenePix Pro 6.0 software was used to extract all features of the data obtained from the scanned images and further analysis was performed based on bioinformatics databases such as GO and KEGG. GST pull-down was performed to verify the interaction of Rv1705c with PSMA3 and RSAD2.Results:The purification results showed that Rv1705c was expressed in endosomes. The secretion of IFN-γ increased significantly after stimulating macrophages with Rv1705c. A total of 29 potential Rv1705c-interacting proteins were screened, and nine of them showed signal-to-noise ratio (SNR)>1.6, namely PSMA3, NLN, THOP1, UPF3A, RSAD2, OMG, PNKD, STEAP3 and MED8. Further bioinformatics analysis revealed that PSMA3, RSAD2 and C1QBP were involved in innate immune signaling pathway, and there were interactions of PSMA3 and RSAD2 with IFN. GST pull-down assay validated that PSMA3 and RSAD2 interacted with Rv1705c.Conclusions:This study showed that PSMA3 and RSAD2 interacted with Rv1705c, providing reference for further investigation on the mechanism of Mycobacterium tuberculosis infection.

Objective:To summarize and analyze the clinical and genotype features of female-restricted X-linked syndromic mental retardation-99(MRXS99F, OMIM: 300968)caused by USP9 X gene mutation, and to improve the clinicians′ understanding of the disease. Methods:Clinical data and genotypes of 2 children with MRXS99F treated in the Children′s Hospital of Nanjing Medical University in March 2020 (case 1) and June 2020 (case 2) were analyzed, and the relevant databases at home and abroad were reviewed to summarize the clinical characteristics and gene variation characteristics of the disease.Results:The 2 cases were 6 months old (case 1) and 5 years old (case 2), both showed psychomotor retardation.Case 1 presented a short stature, pigment abnormality, characteristic facial features, hypotonia, recurrent respiratory tract infections, laryngeal cartilage hypoplasia, atrial septal defect, feeding difficulty, hearing loss and brain hypoplasia.Case 2 had abnormal electroencephalogram.As confirmed by whole-exome sequencing, two children carried c. 6972+ 1G>A, c.6437C>T of USP9 X, respectively.Neither of the 2 variations was previously reported.Twenty-two cases of MRXS99F caused by USP9 X gene mutation were reported in 4 literatures globally, and 24 cases were combined with this study.The clinical manifestations of 20/22 children had special faces.All of them accompanied mental retardation combined with motor and language retardation, and carried neonatal variation. Conclusions:This is the first case report of MRXS99F induced by USP9 X gene variation in China.MRXS99F caused by functional deletion and variation of USP9 X gene is mainly characterized by psychomotor retardation, language disorder, special face and multiple congenital malformations.For children with unexplained growth retardation, special face and multiple congenital malformations, genetic testing like high-throughput sequencing should be carried out as early as possible to determine the etiology.

Objective　To investigate the efficacy of tirofiban on transient ischemic attack with high risk of recurrence.Methods　156 patients with non-cardiogenic transient ischemic attack with ABCD2 score ≥ 4 points were hospitalized at the Department of Neurology,Sinopharm Tongmei General Hospital from Jan 2019 to Mar 2021.Eight patients were excluded due to incomplete data or drop-off from follow-up.The remaining 148 patients were randomly divided into two groups,78 in the observation group and 70 in the control group.The observation group was given tirofiban treatment (loading tirofiban via a micropump,and continuous intravenous pumping for 48 hours) and intensive lipid-lowering therapy.The control group was given intensive anti-platelet aggregation therapy with Aspirin enteric-coated tablets,hydroclopidogre and intensive lipid-lowering therapy.We made a comparative analysis between two groups about efficacy,safety and short-term stroke incidence.Results　In the observation group,57 cases were cured,14 cases were effective,and 7 cases were ineffective;6 cases (7.69%) had a stroke (cerebral infarction) within 7 days,7 cases (8.97%) had a stroke within 30 days,and 7 cases (8.97%) had a stroke within 90 days.In the control group,39 cases were markedly effective,15 cases were effective,and 16 cases were ineffective;in the control group,13 cases (18.57%) had a stroke (cerebral infarction) within 7 days,15 cases (21.43%) had a stroke within 30 days,16 cases (22.86%) had a stroke within 90 days.No intracerebral hemorrhage occurred in either group within 90 days.The two groups had statistical significance in terms of effective treatment rate (P<0.05).However,there was no significant difference in the number of stroke cases between the observation group and the control group in the short-term 7 days,30 days,and 90 days (P>0.05).There was no significant difference in coagulation,platelet,liver and kidney function between the two groups after 48 hours of treatment (P>0.05).Conclusion　For patients with transient ischemic attack at high risk of recurrence,tirofiban treatment can effectively control TIA attacks,but it cannot reduce the risk of stroke.

Rapid development of high-throughput technologies has permitted the identification of an increasing number of disease-associated genes (DAGs), which are important for understanding disease initiation and developing precision therapeutics. However, DAGs often contain large amounts of redundant or false positive information, leading to difficulties in quantifying and prioritizing potential relationships between these DAGs and human diseases. In this study, a network-oriented gene entropy approach (NOGEA) is proposed for accurately inferring master genes that contribute to specific diseases by quantitatively calculating their perturbation abilities on directed disease-specific gene networks. In addition, we confirmed that the master genes identified by NOGEA have a high reliability for predicting disease-specific initiation events and progression risk. Master genes may also be used to extract the underlying information of different diseases, thus revealing mechanisms of disease comorbidity. More importantly, approved therapeutic targets are topologically localized in a small neighborhood of master genes in the interactome network, which provides a new way for predicting drug-disease associations. Through this method, 11 old drugs were newly identified and predicted to be effective for treating pancreatic cancer and then validated by in vitro experiments. Collectively, the NOGEA was useful for identifying master genes that control disease initiation and co-occurrence, thus providing a valuable strategy for drug efficacy screening and re-positioning. NOGEA codes are publicly available at https://github.com/guozihuaa/NOGEA.

Objective:To investigate the association between levator hiatus area, pelvic organ prolapse quantification (POP-Q) examination and prolapse symptoms.Methods:The prospective multicenter study enrolled 996 female patients between January 2017 and January 2019. All enrolled patients underwent a standard clinical interview, POP-Q examination and transperineal ultrasound examination. Volume data of pelvic floor ultrasound examinations were obtained at rest, during contraction and during maximal Valsalva maneuver. The association between levator hiatus area, POP-Q examination and prolapse symptoms was analyzed. The performance of levator hiatus area on maximal Valsalva for assessing significant POP(POP-Q stage≥2) and prolapse symptoms were also evaluated.Results:There were significant differences of levator hiatus area at rest, during contraction and during maximal Valsalva among patients with different POP-Q stages (all P<0.001). Levator hiatus area during maximal Valsalva showed the highest correlation with abdominal dragging sensation ( r=0.277, P<0.001). The area under the ROC curve (AUC) of levator hiatus area during maximal Valsalva for significant POP (POP-Q stage≥2) was significantly higher than that for prolapse symptoms (AUC: 0.77 vs 0.69, P<0.001). Conclusions:Levator hiatus area on transperineal has moderate correlation with POP-Q examination and their association is stronger than the correlation between ultrasound findings and prolapse symptoms.