ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

Objective To genetically investigate the protective effects of statins on breast cancer. Methods Instrumental variables for the statin target gene HMGCR and five other cholesterol-regulated genes (LDLR, PCSK9, ABCG8, APOB, and NPC1L1) were obtained from previous expression quantitative trait locus (eQTL) studies. Cholesterol-regulated genes predicted by these instrumental variables served as the exposure factors. Mendelian randomization based on pooled data (SMR) was conducted to explore the genetic effects of exposure factors on the incidence risk of all breast cancers, ER+ breast cancer, and ER-breast cancer. Instrumental variables for total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) were derived from a previous human genome-wide association study and restricted to be chromosomally located within 100 kb of the above cholesterol regulatory genes; the instrumental variables could predict TC, LDL-C, or non-HDL-C levels under the regulation of the abovementioned cholesterol-associated genes which were used as exposure factors. Two-sample Mendelian randomization (IVW, MR-PRESSO, and MR-Egger) was used to explore the genetic effects of exposure factors on the risk of all breast cancers, ER+ breast cancer, and ER− breast cancer. Results SMR analysis reported that elevated HMGCR expression was significantly associated with the increased incidence risk of all breast cancers and ER+ breast cancer (P=0.044 and P=0.039, respectively) but not with the change in incidence risk of ER− breast cancer (P=0.190); the other five regulatory genes were not significantly correlated with the change in incidence risk of all breast cancers, ER+ breast cancer, and ER− breast cancer (all P>0.05). IVW analysis reported that under the regulation of HMGCR, elevated levels of peripheral TC, LDL-C, and non-HDL-C significantly increased the incidence risk of all breast cancers (P=1.160e-05, P=1.248e-05, and P=1.869e-05) and the incidence risk of ER+ breast cancer (P=3.181e-04, P=2.231e-04, and P=3.520e-04), but they were not associated with a change in the incidence risk of ER− breast cancer (P=0.062, P=0.133, and P=0.055). The results of MR-PRESSO and MR-Egger analyses supported the IVW results. Conclusion Statins could reduce the incidence risk of ER+ breast cancer at the genetic level, but there is no such protective effects on ER− breast cancer.

Objective·To evaluate and compare the clinical efficacy and safety of rituximab(RTX)monotherapy and RTX combined with glucocorticoids in the treatment of high-risk primary membranous nephropathy(PMN).Methods·A retrospective cohort study was conducted to include 87 high-risk PMN patients who received RTX treatment in Xin Hua Hospital,Shanghai Jiao Tong University School of Medicine,between December 2018 and February 2024.Patients were divided into RTX monotherapy group and RTX combined with glucocorticoid group(combination therapy group)according to their treatment regimens.After adjusting for confounding factors using propensity score matching(PSM),intention-to-treat(ITT)and per-protocol(PP)analyses were performed to evaluate the primary efficacy outcomes.Results·A total of 58 patients were included after PSM,with 29 in each group.At 12 months after treatment,ITT analysis showed that the overall remission and complete remission(CR)of RTX monotherapy were 72.41%and 20.69%respectively,compared to 79.31%and 55.17%in the combination therapy group.The difference in CR rates between the two groups was statistically significant(P=0.007).The differences in serum creatinine,blood urea nitrogen,and estimated glomerular filtration rate(eGFR)were not statistically significant.However,the 24-hour urinary protein levels in the combination therapy group were significantly lower than those in the RTX monotherapy group(P=0.024).PP analysis showed that the CR rate of the combination therapy group was also significantly higher than that of the RTX monotherapy group(P=0.026).During follow-up,serum creatinine levels and eGFR remained stable in both groups.The incidence of adverse events was comparable between the two groups,with no statistically significant differences.Conclusion·Compared with RTX monotherapy,RTX combined with glucocorticoids significantly improves the CR rate in high-risk PMN patients without increasing the incidence of adverse events.This combination regimen appears to be an effective treatment strategy,though its long-term efficacy and safety warrant further confirmation through large-scale prospective studies.

Objective:To investigate the causal relationship and pathogenesis between gastroesophageal reflux disease(GERD)and rheumatoid arthritis(RA).Methods:Genetic correlation between GERD and RA was evaluated using linkage disequi-librium score regression(LDSC),and potential causal relationship between GERD and RA was analyzed by Mendelian randomization(MR)method.GWAS data for GERD were obtained from the European Bioinformatics Institute,while GWAS data for RA were obtained from the FinnGen consortium and large-scale genome-wide association studies.Transcriptional analysis based on Genecard,DrugBank and GEO databases was then conducted to explore mechanisms of action for GERD and RA,and to identify potential biomarkers.Results:LDSC analysis result showed a genetic correlation between GERD and RA(rg=0.34,P=1.07E-20).Further MR analysis revealed a positive association between GERD and RA risk(OR=1.44,95%CI:1.27～1.63,P=1.17E-08),which was supported by validation data(OR=1.61,95%CI:1.34～1.94,P=3.56E-07).Conversely,there was no evidence to suggest that RA increases the risk of GERD.CXCL10 and EDNRB were identified as potential biomarkers for diagnosing RA in GERD patients.Conclusion:There is a positive causal relationship between GERD and RA,while not vice versa.Transcriptional data analysis indicated that CXCL10 and EDNRB can serve as novel biomarkers for diagnosing RA in GERD patients.

The clustered regularly interspaced short palindromic repeat-CRISPR-associated protein (CRISPR-Cas) system is renowned for its exceptional gene-editing capabilities. In recent years, the discovery of the trans-cleavage activity of Cas12 and Cas13 proteins has shown great potential in the field of molecular diagnostics. The trans-cleavage activity of the CRISPR-Cas system has been widely applied in point-of-care testing (POCT). This article will provide a detailed discussion on the principles, advantages, and challenges of combining the CRISPR-Cas system with lateral flow assays, colorimetric methods, microfluidic technologies, smartphone applications, and portable detection sets. The CRISPR-Cas system demonstrates versatility in the POCT field with its ability to detect nucleic acids, pathogenic microorganisms, and non-nucleic acid targets. As CRISPR-Cas-based POCT continues to advance, these developments provide strong support for the formulation of personalized medical and public health strategies, further promoting the realization of precision medicine.

OBJECTIVE To analyze the clinical distribution characteristics and drug resistance of Enterobacter cloa-cae in Gansu Province from 2019 to 2023,providing reference for the prevention and control of E.cloacae infec-tions in this region.METHODS Data on the distribution and drug resistance of E.cloacae from hospitals in mem-ber units of the Gansu Antimicrobial Surveillance Network between 2019 and 2023 were collected.In vitro drug susceptibility testing was performed by the Kirby-Bauer disk diffusion(K-B)method,minimum inhibitory con-centration method,and fully automated instrumentation,followed by analysis of the susceptibility results.RESULTS From 2019 to 2023,a total of 402 490 bacterial strains were isolated and cultured in Gansu Province,including 17 417 strains of E.cloacae,with a detection rate of 4.33％.The bacteria were primarily iso-lated from sputum specimens(62.81％),followed by urine(7.37％)and wound pus specimens(6.07％).The de-partmental distribution was dominated by internal medicine(44.96％)and surgery(28.50％).The highest detec-tion rate was observed in the adult group(15-65 years,45.79％).E.cloacae exhibited varying degrees of resist-ance to over 20 antibacterial drugs,but the overall drug resistance rate showed a declining trend(P＜0.05).The highest drug resistance rate was observed for cefazolin(96.40％-98.38％),while the lowest was for tigecycline(0.44％—2.92％).Carbapenem-resistant E.cloacae demonstrated an increasing trend in drug resistance rates,with imipenem resistance ranging from 2.79％to 3.71％(P=0.044)and meropenem resistance ranging from 1.29％to 3.41％(P＜0.05).CONCLUSIONS The isolation rate of E.cloacae in Gansu Province remains stable,with a declining trend in drug resistance to multiple antibacterial drugs.However,the increasing drug resistance to carbapenems warrants attention.

In order to effectively prevent and control the occurrence of catheter-associated urinary tract infection and ensure the safety of both patients and medical personnel,the National Health Commission of the People's Re-public of China officially released the recommended health industry standard"Standard for prevention and control of catheter-associated urinary tract infection"(WS/T862-2025)in Aug.2025.This paper provides an interpreta-tion of the standard,covering its drafting background,basis and content,to assist relevant medical personnel in healthcare institutions in enhancing their understanding and recognition of the standard,and to further promote its implementation and enforcement.

Hepatitis virus is the main pathogen causing liver inflammation and damage. Early detection is crucial for the effective treatment of hepatitis. The detection technology of hepatitis virus has gone through multiple stages, including immunological detection technology and nucleic acid detection technology. The emergence of emerging molecular detection technologies makes its detection more sensitive and convenient, including nanotechnology, Raman spectroscopy technology, microfluidic technology and biosensor technology. The development of these technologies has promoted the early diagnosis of hepatitis, but their clinic applications are still facing challenges. In the future, the development of hepatitis virus detection technology is expected to transform in the form of multidimensional and interdisciplinary innovation process, with its core objectives being the realization of more precise, convenient, and accessible detection methods, thereby comprehensively advancing the progress of hepatitis prevention and control efforts.

Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.