Objective To analyze the multi-target mechanism of Paeoniflorin in the intervention of oral submucosal fibrosis(OSF)systematically,based on molecular dynamics simulation and network pharmacology.Methods Identify potential targets of Paeoniflorin were predicted by using database.OSF-related disease targets and identified drug-disease intersecting targets were screened.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were conducted to validate the molecular binding capabilities between Paeoniflorin and core targets.Finally,molecular dynamics simulations were performed to verify binding stability.Results A total of 20 overlapping targets were identified,including key genes such as transforming growth factor(TGF)-β1,interleukin(IL)-6,and tumor necrosis factor(TNF)-α.TGF-β1,IL-6,and TNF formed the core hub.The enrichment analysis revealed that the target molecules were significantly enriched in the TGF-β1,phosphatidylinositol 3-kinease-actin(PI3K-Akt),and nuclear factor κB(NF-κB)signaling pathways.Molecular docking confirmed high affinity binding of Paeoniflorin to targets including TGF-β1,while molecular dynamics simulations verified stable interactions between Paeoniflorin and both TGF-β1 and B-cell lymphoma-2 targets.Conclusion This study revealed that Paeoniflorin inhibits the inflammatory-fibrotic cascade of OSF through synergistic regulation of TGF-β1/Smad,PI3K-Akt and NF-κB pathways.

Objective To analyze the multi-target mechanism of Paeoniflorin in the intervention of oral submucosal fibrosis(OSF)systematically,based on molecular dynamics simulation and network pharmacology.Methods Identify potential targets of Paeoniflorin were predicted by using database.OSF-related disease targets and identified drug-disease intersecting targets were screened.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were conducted to validate the molecular binding capabilities between Paeoniflorin and core targets.Finally,molecular dynamics simulations were performed to verify binding stability.Results A total of 20 overlapping targets were identified,including key genes such as transforming growth factor(TGF)-β1,interleukin(IL)-6,and tumor necrosis factor(TNF)-α.TGF-β1,IL-6,and TNF formed the core hub.The enrichment analysis revealed that the target molecules were significantly enriched in the TGF-β1,phosphatidylinositol 3-kinease-actin(PI3K-Akt),and nuclear factor κB(NF-κB)signaling pathways.Molecular docking confirmed high affinity binding of Paeoniflorin to targets including TGF-β1,while molecular dynamics simulations verified stable interactions between Paeoniflorin and both TGF-β1 and B-cell lymphoma-2 targets.Conclusion This study revealed that Paeoniflorin inhibits the inflammatory-fibrotic cascade of OSF through synergistic regulation of TGF-β1/Smad,PI3K-Akt and NF-κB pathways.

ObjectiveTo explore new targets and herbal medicines of total ginsenosides in ameliorating alcoholic hepatitis （AH） by data mining and experimental validation and to provide new directions for the clinical treatment of AH. MethodGSE28619 was selected as the test set from the GEO database and GSE83148 and GSE103580 were selected as the validation sets. The limma package and weighted gene co-expression network analysis （WGCNA） were employed to identify the AH-related differentially expressed genes and modular genes， and Venny was used to extract the common genes. The protein-protein interaction （PPI） network was constructed and the enrichment analysis was carried out. The hub genes were further screened and evaluated for their diagnostic value. After validation with the datasets， new potential targets of AH and traditional Chinese medicine were predicted. Molecular docking between the targets and active ingredients of traditional Chinese medicine was performed， and the results were validated by experiments. Eight out of 48 SD rats were randomly selected into a blank group and received an equal amount of normal saline. The rest rats were subjected to modeling with ethanol by gavage and then randomized into low- （10 mg·kg^-1）， medium- （20 mg·kg^-1）， and high-dose （40 mg·kg^-1） total ginsenosides， model， and positive control （metadoxine， 117 mg·kg^-1） groups. After 3 weeks of gavage， serum samples were collected for the measurement of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） levels， and liver samples were collected for hematoxylin-eosin （HE） staining. Western blot and Real-time PCR were employed to determine the protein and mRNA levels， respectively， of potential targets in the liver tissue. ResultData mining predicted the potential genes： Proto-oncogene FOS and collagen type Ⅰ alpha 2 （COL1A2）. Experimental validation showed that the liver injury was alleviated after drug administration compared with that after modeling. The serum AST and ALT levels were reduced after drug administration. The protein and mRNA levels of FOS were significantly up-regulated， while those of COL1A2 were down-regulated after drug administration. ConclusionTotal ginsenosides ameliorate HA via FOS and COL1A2.

ObjectiveTo construct a macro-evaluation tool for dampness syndrome （DS） animal model， which will provide a basis for experimental research on dampness syndrome in traditional Chinese medicine （TCM）. MethodsConceptual framework of this study was clarified through discussions within the core working group， and dimensions of the evaluation of the animal model of DS were identified according to TCM principles. We searched CNKI， Wanfang， VIP and SinoMed databases from the inception to June 30th， 2023， on experiments involving dampness syndrome animals to create a pool of items about DS animal models. The core items were selected and extracted for factor analysis and cluster analysis. An expert importance rating questionnaire was developed based on the results of the literature review， analyzing the distribution of item scores， importance averages， and coefficient of variation. Through a comprehensive analysis of literature， expert importance scoring， and specific expert opinions， items that did not meet anyone of the criteria of average importance rating ≥2.04， coefficient of variation ≤30%， or literature eva-luation frequency ≥2% were removed， thereafter， the macro-evaluation tool for DS animal model was preliminarily constructed. ResultsSpirit and body state， autonomic activity state， body surface characteristics， diet， urination and defecation， tongue manifestation， and motor behavior assessment were constructed as the seven dimensions in the evaluation of DS animal model. A total of 348 papers about animal experiments were included and analyzed， resulting in a saturated pool of 72 items， which was refined to 38 core items of DS animal models. Factor analysis obtained 16 common factors， which were further clustered into two categories， named dampness transforming from heat syndrome and dampness transforming from cold syndrome. The expert importance scoring showed that the Kendall harmony coefficient was 0.359 （P＜0.05） indicating a high level of agreement， coordination and reliability among the experts. Notably， 50% or more of the experts considered the items thick and greasy tongue coating， unclean perianal area， loose stools， lethargy， unformed stools， and listless expression as very important. The median scores for all items were 2.04（1.73， 2.37）， with a coefficient of variation ranging from 19.73% to 53.38%. After expert evaluation， the macro-evaluation tool for DS animal model in TCM with 33 items and corresponding criteria for assessing the formation of DS models was finally contructed. ConclusionThe Macro-evaluation tool for DS animal model in TCM is highly scientific， credible， and operable， and can be utilized in DS animal experiments after its characteristics are actually evaluated.

Objective:To explore the influencing factors for hypoglycemia in elderly patients during peri-colonoscopy, construct and validate a risk prediction model.Methods:The factors influencing hypoglycemia in elderly patients during the peri-colonoscopy period were identified through a literature review and semi-structured interviews. After two rounds of Delphi expert consultation, the survey questionnaire was determined. From January to September 2023, convenience sampling was used to select elderly patients who underwent colonoscopy in the Department of Gastroenterology at the Affiliated Hospital of Qingdao University as participants for a questionnaire survey. Univariate and multivariate Logistic regression was used to explore the influencing factors of hypoglycemia in elderly patients during the peri-colonoscopy period, and a nomogram model of hypoglycemia risk in elderly patients during the peri-colonoscopy period was drawn. The area under the receiver operating characteristic curve ( AUC) of the subjects and the Hosmer-Lemeshow goodness of fit test were used to evaluate the model's predictive performance. The clinical decision curve of DCA was implemented to evaluate the model's clinical benefit ability. Results:A total of 558 questionnaires were distributed (392 for the modeling group and 166 for the validation group) and 558 valid questionnaires were collected, with a valid response rate of 100.00%. Among 558 elderly patients, a total of 130 cases (89 in the modeling group and 41 in the validation group) experienced hypoglycemia during the peri-colonoscopy period, with an incidence of 23.30%. Multivariate Logistic regression analysis showed that serum albumin, age, previous hypoglycemia frequency, insulin use, fasting time, and nutritional risk were independent influencing factors (all P<0.05). The AUCs of the modeling and validation groups were 0.933 and 0.899, respectively. Hosmer Lemeshow test showed that the model had good calibration accuracy, and the DCA curve indicated that the model had good clinical effectiveness. Conclusions:The nomogram model has good predictive performance and can intuitively and concisely predict the risk of hypoglycemia in elderly patients during the peri-colonoscopy period, providing reference for medical and nursing staff.

Background: Excessive growth of keratinocytes is the critical event in the etiology of psoriasis.However, the underlying molecular mechanism of psoriatic keratinocyte hyperproliferation is still unclear. Objective: This study aimed to figure out the potential contributory role of S-phase kinase-associated protein 2 (SKP2) in promoting the hyperproliferation of keratinocytes in psoriasis. Methods: We analyzed microarray data (GSE41662) to investigate the gene expression of SKP2in psoriatic lesion skins compared with their adjacent non-lesional skin. Then, we further confirmed the mRNA and protein expression of SKP2 in human psoriatic skin tissues, imiquimod (IMQ)-induced psoriatic mice back skins and tumor necrosis factor α (TNF-α), interleukin (IL)-17A and IL-6-stimulated keratinocytes by using real-time quantitative polymerase chain reaction and western blot (WB). Furthermore, we explored the potential pathogenic role and its underlying cellular mechanism of SKP2 in promoting keratinocytes hyperproliferation through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell cycle detection, 5-ethynyl-2′-deoxyuridine staining and WB. Finally, we determined whether inhibition of SKP2 can effectively alleviate the keratinocytes hyperproliferation in vivo. Results: We identified that SKP2 is aberrantly upregulated in the psoriatic lesion skin and cytokines-stimulated keratinocytes. Moreover, upregulated SKP2 augments cytokines-induced keratinocytes hyperproliferation. Mechanistically, enhanced SKP2 increased the S phase ratio through inhibiting Cyclin-Dependent Kinase Inhibitor p27 (P27 Kip1) expression. Correspondingly, suppression of SKP2 with SMIP004 can significantly ease the epidermis hyperplasia in vivo. Conclusion Our results suggest that elevated SKP2 can empower keratinocytes proliferation and psoriasis-like epidermis hyperplasia via downregulation of P27 Kip1. Therefore, targeting SKP2-P27 Kip1 axis might be a promising therapeutic strategy for the treatment of psoriasis in future.

Objective:To investigate the clinical outcomes of open reduction and cannulated screw fixation directly via the anterior approach for femoral neck fracture of Garden type Ⅳ which is difficultly reducible in young and middle-aged patients.Methods:Forty patients aged younger than 65 years old were treated by open reduction and cannulated screw fixation directly via the anterior approach for femoral neck fractures which were difficulty reducible at Department of Orthopaedics, Mindong Hospital from March 2015 to October 2017. They were 28 males and 12 females, aged from 22 to 62 years (mean, 49.2 years). All the fractures were type Ⅳ by the Garden classification. The time from injury to operation averaged 3.2 d (range, from 1 to 6 d). Their postoperative fracture nonunion and femoral head necrosis were recorded. The reduction quality and hip function were evaluated using Garden reduction index and Harris hip scores.Results:All the 40 patients were followed up for 11 to 44 months (mean, 22.3 months). Nonunion occurred in one patient who had to accept secondary artificial hip replacement. No avascular necrosis of the femoral head occurred in any patients. The anteroposterior Garden reduction indexes at 2 d after surgery and at the last follow-up for the 40 patients were respectively 156.97°±1.40° and 157.07°±1.47°, showing no significant difference( t=-0.302, P=0.765); so were the lateral Garden reduction indexes (171.82°±4.48° and 172.10°±4.27°) ( t=-2.333, P=0.817). The Harris hip scores at the final follow-up for the hip ranged from 77 to 98 (average, 92); 31 cases were rated as excellent, 6 as good, 2 as fair and one as poor, giving an excellent to good rate of 92.5%. Conclusion:In the treatment of femoral neck fracture of Garden type Ⅳ which is difficultly reducible in young and middle-aged patients, if closed reduction has failed, open reduction and cannulated screw fixation directly via the anterior approach can achieve satisfactory short-term outcomes because it facilitates fracture reduction and fixation under direct vision, promotes fracture union and reduces avascular necrosis of the femoral head.

Molecular beacon probe was designed based on a specific DNA sequence of Nocardia to PCR detection of thisbacterium.The strains of Nocardia、Gordina and Rhodococcus were inoculated in Brain Heart Infusion Agar medium separately,then the growth condition was observed,DNA was extracted as a template;the molecular beacon probe was designed based on the partial secA 1 gene sequences of Nocardia strains,and the probe was added into the reaction system of real time fluorescence quantitative PCR (RT-PCR),and the fluorescence signal was tested at the end of PCR.Showed that the amplified secA1 gene of Nocardia could produce positive fluorescence signal in RT-PCR,but those of Gordonia and Rhodococcus with control groups showed negative results because of no fluorescence signal.In conclusion as a housekeeping gene,secA1 is an ideal target molecule to identify the actinomycetes strains on the species level in the systematic evolution research,and the technique of fluorescence molecular beacon probe is accurate,rapid and sensitive for detecting the Nocardia strains with secA1 gene.

Objective To analyze the genomic sequences of Cryptococcus neoformans var grubii strains of two genotypes with different virulence and to screen out the virulence-associated genes. Methods A clinical strain (IFM56800) with the strongest virulence and an environmental strain (IFM56731) with the weakest virulence were screened out for whole genome sequencing analysis. The results of sequencing analy-sis were comprehensively analyzed by using the method of comparative genomics. Genetic variations were ex-tensively screened by using the strategies of non-synonymous single nucleotide polymorphisms ( nsSNPs), nonsense SNPs and the insertions or deletions ( InDels) causing frameshift mutations. The filtered genes were sequenced in 20 experimental strains. The whole RNAs were extracted and then the full-length cDNAs were sequenced by using the rapid amplification of 5′ and 3′ cDNA ends (RACE) method. Results By whole genome sequencing, valid data with high coverage (127 times and 111 times) was obtained in both the environmental strain IFM56731 and the clinical strain IFM56800. The data of InDels and SNPs were statisti-cally analyzed, respectively. Six genes were chosen for further analysis based on the strategies of nonsense SNPs and the InDels causing frameshift mutations. The six genes were amplified and sequenced in all of the experimental strains, three of which were further analyzed with cDNA sequencing. Ultimately, the location and structure of CNAG_01032 gene were determined. The predicted nonsense mutation locus was verified to present in the actual mRNA. Conclusion The strategies of nonsense SNPs and the InDels causing frame-shift mutations showed high-efficiency in screening potential virulence-associated genes. The CNAG_01032 gene was screened out as a novel virulence-associated gene.