OBJECTIVE To systematically evaluate the quality of the Heat-clearing and symptom-relieving formula and screen potential marker components that influence the quality of the formula. METHODS The contents of 11 components （calycosin-7- O - β -D-glucoside， ononin， hyperoside， isoquercitrin， baicalin， baicalein， cryptotanshinone， tanshinone Ⅱ A ， tanshinone Ⅰ， senkyunolide A， ferulic acid） in the Heat-clearing and symptom-relieving formula were determined by high-performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）. Using the contents of the aforementioned components as variables， cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were conducted using OriginPro 2024 software and SIMCA 14.1 software； marker components affecting the quality of the Heat-clearing and symptom-relieving formula were then screened based on the criteria of variable importance in the projection （VIP） value＞1 and P ＜0.05. The comprehensive evaluation of 20 batches of samples was carried out using the entropy weight-technique for order preference by similarity to ideal solution（TOPSIS） and grey correlation analysis （GCA） methods. RESULTS The contents of the above 11 components were 7.993-72.866， 4.542-31.228， 727.666-1 901.884， 496.846-1 293.279， 1 995.501-6 779.150， 54.500-241.280， 150.302-304.339， 79.698-189.206， 257.118-682.418， 5.498-21.687， 7.524-26.935 μg/g. CA， PCA and OPLS-DA results showed that 20 batches of samples were grouped into 2 categories. Q1， Q3， Q4， Q7-Q9， Q12， Q15， Q16 were grouped into one category， and the rest were grouped into another category； VIP values of ferulic acid， tanshinone Ⅱ A ， baicalin， cryptotanshinone， calycosin-7- O - β -D-glucoside and ononin were all greater than 1 （ P ＜0.05）. Both the entropy weight-TOPSIS and GCA methods showed that the samples ranked in the top 11 according to the euclidean distance and relative correlation degree were Q2， Q5， Q6， Q10， Q11， Q13， Q14， Q17-Q20. CONCLUSIONS The established HPLC-MS/MS method is rapid， accurate and highly sens itive. Combined with chemical pattern recognition analysis， entropy weight-TOPSIS and GCA methods， this method can be used to evaluate the quality of the Heat-clearing and symptom-relieving formula. Ferulic acid， tanshinone Ⅱ A ， baicalin， cryptotanshinone， calycosin-7- O - β -D-glucoside and ononin may be the marker components that affect the quality of this formula. The overall quality of 11 batches of the Heat-clearing and symptom-relieving formula， including Q17， is relatively superior.

ObjectiveTo explore the effects of Jingui Shenqiwan （JSW） and Liuwei Dihuangwan （LDW） on the reproductive ability and brain function of normal mice and compare the actions of the two medications. MethodsSeven groups of female and male mice were divided at a ratio of 2∶1. Except for the control group， the other six groups were as follows： a group of both males and females receiving JSW （3.0 g·kg^-1）， a group of both males and females receiving LDW （4.5 g·kg^-1）， a group of males receiving water and females receiving JSW， a group of males receiving water while females receiving LDW， a group of females receiving water while males receiving JSW， and a group of females receiving water while males receiving LDW. Each group was administered the drug for 14 days and then caged together at a 2∶1 （female∶male） ratio to detect the number of pregnant mice and calculate the pregnancy rate. Pregnant mice continued receiving the drug until they naturally gave birth， which was followed by the observation of newborn mice， calculation of their average number， and the measurement of the offspring's preference for sugar water and neonatal recognition index. At the end of the experiment， the weights of the thymus and spleen were measured to calculate the organ coefficients， and mRNA or protein expression was analyzed in the brain and testes or ovaries. A 1% sucrose solution was used to examine the euphoria of their brain reward systems， while novel object recognition test （NOR） was applied to assess their memory capabilities. mRNA expression was detected using real-time quantitative polymerase chain reaction （Real-time PCR） assay， and protein expression was analyzed with Western blot. ResultsCompared with the control group， oral administration of JSW to both male and female mice for 14 days significantly increased the pregnancy rate of female mice on day 2 after being caged together （P<0.05）， while LDW showed a trend but no statistical significance. Additionally， compared with the control group， JSW could upregulate the gene expression of gonadotropin-releasing hormone （GnRH） in the thalamus， as well as reproductive stem cell factor （SCF） and tyrosine kinase receptor （c-Kit） in the testes and reproductive stem cell marker mouse vasa homologue （MVH） in the ovaries， upregulate the expression of proteins influencing neuronal functional activity， such as brain-derived neurotrophic factor （BDNF）， in hippocampal neurons （P<0.05）， and enhance sucrose preference in male mice （P<0.05）. Compared with the control group， JSW significantly increased sucrose preference and novel object recognition index in offspring mice （P<0.05）， which was related to the upregulation of hippocampal dopamine D1 receptor （D1R） and N-methyl-D-aspartate receptor （Nmdar） gene expression. Compared with the control group， both JSW and LDW could upregulate the protein expression of glucocorticoid receptor （GR）， BDNF， and tyrosine kinase receptor B （TrkB） in the hippocampus of offspring mice （P<0.05）. ConclusionJSW significantly enhances the reproductive ability of normal mice， which is not only related to the release of gonadotropin but also associated with its regulation of brain function. Additionally， JSW has a certain regulatory effect on the brain function of the offspring mice.

ObjectiveTo explore the effect of Shenge Bushen Capsules （SBC） on sexual development in normal 3-week-old mice. MethodsThe experiment consisted of two parts. In the first part， mice were divided into four groups： The control group and the low， medium， and high-dose SBC groups （234.7， 469.4， 938.7 mg·kg^-1， respectively）. In the second part， mice were divided into four groups： Control group， Pseudostellariae Radix polysaccharide （PRP） group， total flavonoids group， and SBC group， all receiving a dose of 469.4 mg·kg^-1. After 7 days of administration， the vaginal opening of female mice and the descent of testes and scrotum in male mice， as well as the ovarian and testicular organ indices， were observed. After 4 weeks of administration， female and male mice were housed together for 2 days， and the pregnancy rate of females was monitored. After delivery， the pregnant female mice continued receiving the treatment for 4 weeks， and the sexual development of their offspring， including vaginal opening， testicular descent， and organ indices of ovaries and testes， was observed. Serum sex hormones were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of gonadotropin-releasing hormone （GnRH） and growth hormone （GH） proteins in the hypothalamus was assessed by Western blot. ResultsCompared with the control group， there was no significant effect on the vaginal opening of female mice or the descent of testes in male mice after 7 days of SBC administration. After 4 weeks of administration， the pregnancy rate in the low-dose group was significantly reduced （P<0.05）， but no significant effects were observed in the other groups. The three doses of SBC did not significantly affect the ovarian or testicular organ indices， and there was no significant upregulation in the expression of GnRH or GH in the hypothalamus. The primary component of SBC， Pseudostellariae Radix polysaccharide， significantly reduced the vaginal opening in female mice after 7 days of administration （P<0.05）. After 4 weeks， the serum estradiol levels of non-pregnant female mice were decreased （P<0.05）， but there was no significant effect on the expression of GnRH or GH proteins in the hypothalamus of either male or female mice. Additionally， there were no significant effects on precocious puberty indicators， such as vaginal opening and testicular descent， in the offspring mice. ConclusionSBC does not significantly promote precocious puberty in young mice， and it does not have any noticeable effects on the pregnancy rate of adult mice or the sexual development of their offspring.

Objective:To develop a camelid single-domain antibody (SdAb) recognizing linear B-cell epitopes in glutamate dehydrogenase of Clostridium difficile(CD-GDH), and to apply it in Western blot and ELISA. Methods:Purified recombinant CD-GDH was used as bait to screen phage-displayed camelid SdAb library and obtain positive clones. Then those clones were confirmed by Western blot, and their variable domain of heavy chain of heavy chain antibody(VHH) nucleotide sequence were determined. The VHH sequence was synthesized after codon optimization and cloned into the expression vector pET28a. The SdAb was then expressed and purified, and its ability to detect CD-GDH protein in multiple assays was further explored.Results:Six positive clones were obtained, among which clone GA4 was chosen for recombinant expression in Escherichia coli and further purification. The purified GA4 binded well with CD-GDH with a Kd value of 3 nmol/L. In Western blot and ELISA, GA4 was proven to be able to selectively detect both recombinant and endogenous CD-GDH. Conclusions:A camelid SdAb targeting a linear B-cell epitope in CD-GDH is successfully developed, which provides a very useful tool for detecting CD-GDH.

Objective To study the risk factors for in-hospital mortality in patients with acute myocardial infarction(AMI)undergoing emergency percutaneous coronary intervention(EPCI).Methods Nine hundred and eight patients with acute myocardial infarction(AMI)who underwent EPCI at Beijing Jingmei Group General Hospital between September 2016 and December 2022 were enrolled and divided into an in-hospital death group(36 cases)and a survival group(872 cases)based on in-hospital outcomes.The basic information,clinical indicators,results of coronary angiography,outcomes of EPCI,and complications were compared between the two groups.Multiple logistic regression analysis was conducted to identify the risk factors for in-hospital mortality during EPCI.Results The age and serum creatinine levels of the death group were higher than those of the survival group(P＜0.05),and the estimated glomerular filtration rate(eGFR)and hemoglobin(Hb)levels were lower than those of the survival group(P＜0.05).The proportion of diabetes mellitus,cerebral infarction,cardiogenic shock,left main culprit lesion,rapid ventricular arrhythmia,TIMI flow grade＜3 after EPCI for culprit vessel,and intra-aortic balloon pump(IABP)implantation in the death group were higher than those of the survival group(P＜0.05).The results of multiple logistic regression analysis showed that age,cardiogenic shock,TIMI flow grade＜3 after culprit vessel intervention,and rapid ventricular arrhythmia had a statistically significant impact on in-hospital mortality in EPCI(P＜0.05).Conclusion Age,cardiogenic shock,TIMI flow grade＜3 after PCI for culprit vessels,and rapid ventricular arrhythmia are independent risk factors for in-hospital mortality in EPCI patients with AMI.

Objective To evaluate the role and potential mechanism of apolipoprotein E(APOE)in drug resistance of colon cancer by bioinformatic tools and cellular experiments.Methods After downloading the microarray dataset GSE196900 from the GEO database,the online tool GEO2R was used to identify genes that were expressed differently in the drug-resistant and control groups.The differently expressed genes were then examined for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.The STRING database and Cytoscape software were used to build protein-protein interaction(PPI)networks and find hub genes.Hub genes'predictive significance in colon cancer was further assessed.Western blod and qRT-PCR were used to identify changes in APOE expression,whereas Transwell was used to identify changes in the colon cancer cells'capacity for invasion and migration.Results The analysis of GO and KEGG enrichment revealed that the differential genes derived from the GSE196900 dataset were primarily focused on receptor-ligand activity and cytokine-cytokine receptor interaction pathways.Using the CytoNCA plug-in in Cytoscape software,ten hub genes were obtained through PPI construction.Of these,the prognosis of the patients with colon cancer was negatively correlated with the expression of the APOE gene(P<0.05)and the overexpression of the APOE gene might significantly increase the migration and nvasivenessability of colon cancer cells(P<0.05).Conclusion The increased expression of APOE significantly promotes the migration and invasion ability of colon cancer cells,which may be one of the mechanisms by which APOE gene promotes tumor progression in the patients with colon cancer.

Early diagnosis and therapy of the periodontitis are crucial for the prognosis.Although the latest international classification of periodontal diseases and peri-implant diseases in 2018 can improve the accuracy of diagnosis and reduce the misdiagnosis and missed diagnoses,it is extremely complex in practice.At present,the application of artificial intelligence in the field of dentistry is becoming more and more extensive,which has good performance in detecting tooth types and periodontal bone loss.The use of artificial intelli-gence to detect periodontal lesions and calculate periodontitis staging and grading,and guide clinical personalized and precise treatment has become a hot topic of current research.This paper summarizes and analyzes the current research status of artificial intelligence in periodontal radiographic diagnosis,and provides ideas for further research.

Objective To understand the changing profiles of antimicrobial susceptibility of the bacterial strains isolated from patients at Liuzhou Workers'Hospital in Guangxi from 2020 to 2022.Methods The bacteria were isolated,identified,and underwent antimicrobial susceptibility testing using VITEK 2 Compact,disk diffusion method,or E-test.The results were interpreted according to the breakpoints recommended by CLSI M100 32nd Edition in 2022.The data were analyzed using WHONET 5.6 software.Results A total of 26 254 nonduplicate strains were collected from 2020 to 2022,including Gram-positive bacteria(27.9％)and gram-negative bacteria(72.1％).The prevalence of methicillin-resistant strains was 20.0％in SS.aureus(MRSA),and 72.2％in coagulase-negative Staphylococcus(MRCNS).Methicillin-resistant staphylococcal strains were more resistant to most antimicrobial agents than methicillin-susceptible strains(MSSA and MSCNS).None of the staphylococcal strains was resistant to vancomycin,linezolid or tigecycline.Enterococcus faecium strains showed higher resistance rates to most antimicrobial agents than Enterococcus faecalis.None of enterococcal strains was resistant to vancomycin.A few enterococcal strains were resistant to linezolid.Overall,691 strains of the non-meningitis Streptococcus pneumoniae were isolated from children and 123 strains were isolated from adults.The prevalence of penicillin-resistant SS.pneumoniae(PRSP)was 0.4％in the strains from children and 1.6％in the strains from adults.None of S.pneumoniae strains was intermediate to penicillin.The prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKpn)was 1.2％,1.2％,and 13.8％in 2020,2021,and 2022,respectively.The prevalence of carbapenem-resistant P.aeruginosa(CRPae)and carbapenem-resistant Acinetobacter baumannii(CRAba)was 10.7％and 68.4％in 2020,17.5％and 75.2％in 2021,14.3％and 77.3％in 2022,respectively.About 84.6％of the 1 269 strains of Haemophilus influenzae were isolated from children and 15.4％isolated from adults.The prevalence of beta-lactamase-producing strains was 39.4％in the isolates from children and 46.8％in the isolates from adults.The β-lactamase-producing H.influenzae was resistant to ampicillin.Furthermore,some β-lactamase-nonproducing ampicillin-resistant(BLNAR)H.influenzae strains(27.0％)were also identified.Conclusions Antimicrobial resistance is still serious in this hospital,especially high prevalence of carbapenem-resistant organisms(CRO).Hospital infection prevention and control measures,antibiotic stewardship,and proactive CRO screening should be strengthened.More clinical specimens should be collected for suspected infections.Antimicrobial treatment should be prescribed empirically in time and adjusted when the results of antimicrobial susceptibility testing are available.

Objective To evaluate the efficacy of combining surgical treatment with biological agents for perianal fistulizing Crohn's disease(pfCD).Methods Sixty patients with perianal fistulizing Crohn's disease(pfCD)admitted to our hospital from May 2021 to December 2023 were randomly allocated into two groups:Treatment Group A(n=30)and Treatment Group B(n=30).Treatment Group A received pfCD surgery combined with inflix-imab(IFX)and azathioprine(AZA),while Treatment Group B underwent pfCD surgery along with ustekinumab(UST)and AZA.The CDAI score,PDAI score,and Assche score were monitored for both groups,and postoperative MRI examinations were conducted to evaluate the healing of pfCD.Results There were statistically significant differences in CDAI,PDAI,and Assche scores between pre-treatment and post-treatment comparisons within treat-ment groups A and B(P<0.05),as well as in the magnitude of change at each time point.Comparisons of CDAI,PDAI,and Assche scores at 8,16,24,and 32 weeks,and PDAI scores at 40 weeks between groups A and B using independent samples t-tests did not yield statistically significant results(P>0.05).However,significant differences were observed for CDAI and Assche scores at 40 weeks(P<0.05).Significant changes in CDAI,PDAI,and Assche scores were noted at 8,16,24,32,and 40 weeks post-treatment within both groups A and B(P<0.05).Multiple comparisons using the LSD method revealed that the changes in these scores at each time point were statistically significant(P<0.05).The data indicate a temporal trend in the changes of CDAI,PDAI,and Assche scores,with group B showing a more rapid decline compared to group A.In terms of fistula response rates,both groups A and B achieved 100%(30/30).However,the clinical healing rate of fistulas was higher in group B at 86.7%(26/30)compared to 76.7%(23/30)in group A.Conclusion The combination of surgical treatment with IFX/UST plus AZA is safe and effective for treating pfCD.However,the long-term efficacy of combining surgical treatment with UST appears to be superior.

Objective:To explore the clinical features and risk factors for pediatric severe Mycoplasma pneumoniae pneumonia (SMPP) complicated with pulmonary embolism. Methods:SMPP patients admitted to Department of Pediatrics, Jiaxing First Hospital and Pediatric Intensive Care Unit, Shanghai Children′s Hospital from December 2019 to December 2023 were included in this retrospective case-control study.According to whether they were complicated with pulmonary embolism, SMPP patients were divided into a pulmonary embolism group and a non-pulmonary embolism group.Clinical characteristics of the two groups, including general data, laboratory examination and imaging data were compared and analyzed.The t-test and Mann-Whitney rank-sum test were used to compare the measurement data, and the χ2 test was used to compare the count data.The risk factors of SMPP patients developing pulmonary embolism were analyzed by the univariate method. Results:There were 10 out of 62 SMPP children developing pulmonary embolism, showing an incidence of 16.13%.In the pulmonary embolism group, there were 5 boys and 5 girls, with a median age of 7.50 (5.75, 9.25) years.There were 52 children in the non-pulmonary embolism group, including 29 boys and 23 girls, with a median age of 6.50(5.00, 8.00)years.The hospitalization time, body temperature, total white blood cell count, neutrophil count, C-reactive protein levels, lactate dehydrogenase levels, prothrombin time, activated partial thromboplastin time, D-dimer (D-D) levels, fibrinogen degradation product levels, pleural effusion and atelectasis rates in the pulmonary embolism group were significantly higher than those in the non-pulmonary embolism group (all P<0.05). Fibrinogen levels in the pulmonary embolism group were significantly lower than those in the non-pulmonary embolism group ( P<0.05). Univariate Logistic regression analysis showed that the D-D level was a risk factor for SMPP patient developing pulmonary embolism.The receiver operating characteristic curve analysis revealed that the D-D level had the largest area under the curve for predicting pulmonary embolism of 0.990(95% CI: 0.972-1.000, P<0.001), with a sensitivity of 100%, a specificity of 92%, and a cut-off value of 4.63 mg/L. Conclusions:SMPP children complicated with pulmonary embolism are prone to high inflammation and impaired coagulation function.The increase of D-D levels is a risk factor for the development of pulmonary embolism in SMPP.