The clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein 9 (CRISPR /Cas9) immune system is an adaptive immune system widely distributed in bacteria and archaea. It precisely defends against invasion by exogenous phages, viruses, and plasmids through sequence-specific endogenous immune response mechanisms. As the most prominent member of this family, the CRISPR/Cas9 system has evolved into the most widely applied, flexible, and efficient technical platform in the field of genome engineering due to its exceptional genome modification capabilities. Within the CRISPR/Cas9 system, the Cas9 protein, precisely guided by a single-stranded guide RNA (gRNA), can specifically recognize target DNA sequences and induce double-strand breaks. This activates the cell’s DNA repair mechanisms, enabling gene knockout, knock-in, or modification. Demonstrating significant advantages in specificity, flexibility, and operability, CRISPR/Cas9 technology has shown immense potential in the medical field, opening new avenues for modernizing traditional Chinese medicine (TCM) research. On one hand, this technology can be used to construct precise disease models and tailor personalized treatment plans. It enables in-depth elucidation of the molecular mechanisms underlying the action targets and signaling pathways of TCM formulas and active components, thereby unraveling the scientific secrets of their complex mechanisms of action. On the other hand, it demonstrates powerful tool value in improving TCM germplasm resources, identifying and screening superior varieties, evaluating the controllability of TCM quality, and producing innovative drugs, providing technical support for the standardization and precision of TCM. Simultaneously, the high-throughput omics data generated by CRISPR technology is driving artificial intelligence (AI) to construct virtual disease models and drug prediction systems. This empowers the intelligent screening of effective TCM components, the precise prediction of potential targets, and the exploration of “reducing toxicity while enhancing efficacy” through formula combinations. This synergistic innovation between CRISPR and AI aligns perfectly with precision medicine’s urgent demand for personalized, efficient drug development, injecting new momentum into the modernization and transformation of TCM. This paper first systematically reviews and explains the developmental trajectory, structural basis, and action mechanisms of the CRISPR/Cas9 system, tracing its scientific evolution from a bacterial immune system to a gene-editing tool. It then comprehensively outlines the current state of convergence between precision medicine concepts and modernization research in TCM, analyzing the synergistic points and potential spaces for their integration. Against the backdrop of rapid precision medicine advancement, this paper emphasizes how CRISPR/Cas9 gene editing technology empowers in-depth analysis of TCM mechanisms—including specific applications in disease model construction, therapeutic target validation, and multi-target network regulation studies. It further elaborates on its multidimensional practical contributions to modernizing TCM, spanning key domains such as germplasm resource innovation, bioactive compound biosynthesis, quality standardization control, and novel TCM drug development. Finally, this paper envisions the future landscape of deep integration between CRISPR technology and AI: from data-driven intelligent drug screening to high-throughput precision discovery of effective TCM components, and further to intelligent model construction based on “reducing toxicity while enhancing efficacy” mechanisms. The synergistic convergence of these multidimensional technologies will pioneer new scientific paradigms and translational pathways for TCM modernization, propelling TCM toward leapfrogging development in the era of precision medicine.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

AIM To investigate the clinical effects of Yifei Tongluo Decoction combined with azithromycin on patients with childern severe Mycoplasma pneumoniae pneumonia due to Toxic Heat Blocking Lung.METHODS One hundred and fifty-six patients were randomly assigned into control group(78 cases)for 7-day administration of azithromycin,and observation group(78 cases)for 7-day administration of both Yifei Tongluo Decoction and azithromycin.The changes in clinical effects,disappearance time of local symptoms,mycoplasmas,pulmonary imaging score,Toxic Heat Blocking Lung score,pulmonary function indices(PEF,VPTEF,MMF,TPTEF),inflammatory factors(sB7-H3,Cgp-39,sICAM-1,CCL5),immune function indices(RBC-C3bR,RBC-ICR,CD3+,CD4+)and safety indices were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05),along with shorter disappearance time of local symptoms(P＜0.05).After the treatment,the two groups displayed decreased mycoplasmas,pulmonary imaging score,Toxic Heat Blocking Lung score,inflammatory factors,RBC-ICR(P＜0.05),and increased pulmonary function indices,RBC-C3bR,CD3+,CD4+(P＜0.05),especially for the observation group(P＜0.05).No obvious difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with childern severe Mycoplasma pneumoniae pneumonia due to Toxic Heat Blocking Lung,Yifei Tongluo Decoction combined with azithromycin can safely and effectively alleviate clinical symptoms,improve pulmonary functions,and reduce body inflammatory responses.

Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.

Objective To investigate the distribution and antimicrobial resistance profiles of clinical isolates in Xi'an No.3 Hospital from 2019 to 2023.Methods Clinical isolates were collected from January 1,2019 to December 31,2023.Antimicrobial susceptibility testing was carried out according to a unified protocol of China Antimicrobial Resistance Surveillance Network using Kirby-Bauer method or automated systems.The data were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2023.Results A total of 6 621 clinical isolates were collected from 2019 to 2023,including 1 569(23.7％)strains of Gram-positive bacteria and 5 052(76.3％)strains of Gram-negative bacteria.The prevalence of methicillin-resistant S.aureus,S.epidermidis and other Staphylococcus species(except SS.pseudintermedius and S.schleiferi)was 39.0％,62.3％,and 74.4％,respectively.Methicillin-resistant strains showed much higher resistance rates to most of other antimicrobial agents than methicillin-sensitive strains.No Staphylococcus strains were found resistant to vancomycin or linezolid.E.faecium strains demonstrated much higher resistance rates to most antimicrobial agents tested than E.faecalis.The prevalence of linezolid-resistant E.faecalis and vancomycin-resistant E.faecium was 0.9％and 0.4％,respectively.The prevalence of penicillin-nonsusceptible strains(PISP+PRSP)was 5.8％in nonmeningitis S.pneumoniae isolates.The prevalence of ESBL-producing E.coli,K.pneumoniae,and P.mirabilis in Enterobacterales was 48.5％,37.8％,and 47.2％,respectively.Among Enterobacterales strains,K.pneumoniae had the highest resistance rate to imipenem(18.2％)and meropenem(17.9％).Other Enterobacterales were highly sensitive to carbapenems.The resistance rates of P.aeruginosa to imipenem and meropenem were 22.5％and 19.5％,respectively.The resistance rates of A.baumannii to imipenem and meropenem were 65.0％and 71.6％,respectively.Conclusions Antibiotic resistance is still serious in this hospital.Nearly half of the strains of E.coli,K.pneumoniae and P.mirabilis produced ESBLs.K.pneumoniae and A.baumannii showed high resistance rates to carbapenems.Antimicrobial resistance surveillance should be performed appropriately.Relevant departments need to strengthen cooperation to curb the spread of drug-resistant bacteria.

Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.

Objective To investigate the distribution and antimicrobial resistance profiles of clinical isolates in Xi'an No.3 Hospital from 2019 to 2023.Methods Clinical isolates were collected from January 1,2019 to December 31,2023.Antimicrobial susceptibility testing was carried out according to a unified protocol of China Antimicrobial Resistance Surveillance Network using Kirby-Bauer method or automated systems.The data were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2023.Results A total of 6 621 clinical isolates were collected from 2019 to 2023,including 1 569(23.7％)strains of Gram-positive bacteria and 5 052(76.3％)strains of Gram-negative bacteria.The prevalence of methicillin-resistant S.aureus,S.epidermidis and other Staphylococcus species(except SS.pseudintermedius and S.schleiferi)was 39.0％,62.3％,and 74.4％,respectively.Methicillin-resistant strains showed much higher resistance rates to most of other antimicrobial agents than methicillin-sensitive strains.No Staphylococcus strains were found resistant to vancomycin or linezolid.E.faecium strains demonstrated much higher resistance rates to most antimicrobial agents tested than E.faecalis.The prevalence of linezolid-resistant E.faecalis and vancomycin-resistant E.faecium was 0.9％and 0.4％,respectively.The prevalence of penicillin-nonsusceptible strains(PISP+PRSP)was 5.8％in nonmeningitis S.pneumoniae isolates.The prevalence of ESBL-producing E.coli,K.pneumoniae,and P.mirabilis in Enterobacterales was 48.5％,37.8％,and 47.2％,respectively.Among Enterobacterales strains,K.pneumoniae had the highest resistance rate to imipenem(18.2％)and meropenem(17.9％).Other Enterobacterales were highly sensitive to carbapenems.The resistance rates of P.aeruginosa to imipenem and meropenem were 22.5％and 19.5％,respectively.The resistance rates of A.baumannii to imipenem and meropenem were 65.0％and 71.6％,respectively.Conclusions Antibiotic resistance is still serious in this hospital.Nearly half of the strains of E.coli,K.pneumoniae and P.mirabilis produced ESBLs.K.pneumoniae and A.baumannii showed high resistance rates to carbapenems.Antimicrobial resistance surveillance should be performed appropriately.Relevant departments need to strengthen cooperation to curb the spread of drug-resistant bacteria.

Objective To investigate the effect of two treatment regimens combining Tacrolimus(TAC)with different Rituximab(RTX)dosages,and to provide clinical reference for treatment strategies.Methods A retrospective analysis was conducted on patients diagnosed with idiopathic membranous nephropathy(IMN)and treated with RTX combined with TAC regimen(RTX+TAC group and low-dose RTX+TAC group)in The First Affiliated Hospital of Xinxiang Medical University.Propensity score matching(PSM)was performed at a 1:1 ratio,and a total of 60 patients were enrolled,with 30 in each group.In low-dose RTX(375 mg/m2 at the first and fifteenth day respectively)+TAC group,if circulating B cells(CD19?)exceeded 5 cells/μL after 3 months,a 200 mg RTX infusion was administered.In RTX(1g at the first and fifteenth day respectively)+TAC group,if complete remission(CR)was not achieved by 6 months,an additional 1000 mg RTX infusion was administered.The incidence of CR,partial remission,and adverse events were followed up for 12 months after medication in both groups.Results(1)Both groups showed significant reductions in 24-hour proteinuria,with the RTX+TAC group demonstrating a notably higher decrease compared to the low-dose RTX+TAC group.Statistical differences were observed between the two groups at the 1st and 3rd months of treatment(P<0.05).Albumin levels gradually increased,and there were differ-ences between the two groups at both the 1st and 3rd months(P<0.05).The anti-phospholipase A2 antibody levels decreased significantly after one month of treatment[3.45(1.90,22.10)vs.3.28(8.30,23.08)RU/mL],P>0.05.At 3 months of treatment,the overall clinical remission rate was 63.3％for the RTX+TAC group compared to 36.7％for the low-dose RTX+TAC group(P<0.05).At 12 months,the RTX+TAC group achieved an overall remission rate of 86.7％,while the low-dose RTX+TAC group reached 83.3％,showing no statistical significance(P>0.05).After one month of treatment,the RTX+TAC group achieved a complete serological immunological remission rate of 33.3％,significantly higher than the 3.3％in the low-dose RTX+TAC group(P<0.05).(2)The cumulative remission rate of the RTX+TAC group was higher than that of the low-dose RTX+TAC group during the first 6 months of follow-up.The remission rate in the low-dose RTX+TAC group increased significantly after 6 months.Log-rank test showed no statistical difference between the survival curves of the two groups(P=0.37).(3)Based on a multifactorial COX regression analysis of factors related to remission in patients with IMN,for every unit increase in serum immunological remission time,the risk of patients achieving remission decreased by 13.5％(HR=0.87,P=0.016).The risk of remission for patients with high titers of anti-PLA2R antibodies decreased by 60.2％(HR=0.39,P=0.018).Conclusions Different RTX dosages yielded comparable overall clinical remission rates without significantly increasing adverse events.RTX+TAC regimen achieves higher early CR rate.Serological remission time and high titer anti-PLA2R antibodies are associated with clinical outcomes.

Objective To investigate the effect of two treatment regimens combining Tacrolimus(TAC)with different Rituximab(RTX)dosages,and to provide clinical reference for treatment strategies.Methods A retrospective analysis was conducted on patients diagnosed with idiopathic membranous nephropathy(IMN)and treated with RTX combined with TAC regimen(RTX+TAC group and low-dose RTX+TAC group)in The First Affiliated Hospital of Xinxiang Medical University.Propensity score matching(PSM)was performed at a 1:1 ratio,and a total of 60 patients were enrolled,with 30 in each group.In low-dose RTX(375 mg/m2 at the first and fifteenth day respectively)+TAC group,if circulating B cells(CD19?)exceeded 5 cells/μL after 3 months,a 200 mg RTX infusion was administered.In RTX(1g at the first and fifteenth day respectively)+TAC group,if complete remission(CR)was not achieved by 6 months,an additional 1000 mg RTX infusion was administered.The incidence of CR,partial remission,and adverse events were followed up for 12 months after medication in both groups.Results(1)Both groups showed significant reductions in 24-hour proteinuria,with the RTX+TAC group demonstrating a notably higher decrease compared to the low-dose RTX+TAC group.Statistical differences were observed between the two groups at the 1st and 3rd months of treatment(P<0.05).Albumin levels gradually increased,and there were differ-ences between the two groups at both the 1st and 3rd months(P<0.05).The anti-phospholipase A2 antibody levels decreased significantly after one month of treatment[3.45(1.90,22.10)vs.3.28(8.30,23.08)RU/mL],P>0.05.At 3 months of treatment,the overall clinical remission rate was 63.3％for the RTX+TAC group compared to 36.7％for the low-dose RTX+TAC group(P<0.05).At 12 months,the RTX+TAC group achieved an overall remission rate of 86.7％,while the low-dose RTX+TAC group reached 83.3％,showing no statistical significance(P>0.05).After one month of treatment,the RTX+TAC group achieved a complete serological immunological remission rate of 33.3％,significantly higher than the 3.3％in the low-dose RTX+TAC group(P<0.05).(2)The cumulative remission rate of the RTX+TAC group was higher than that of the low-dose RTX+TAC group during the first 6 months of follow-up.The remission rate in the low-dose RTX+TAC group increased significantly after 6 months.Log-rank test showed no statistical difference between the survival curves of the two groups(P=0.37).(3)Based on a multifactorial COX regression analysis of factors related to remission in patients with IMN,for every unit increase in serum immunological remission time,the risk of patients achieving remission decreased by 13.5％(HR=0.87,P=0.016).The risk of remission for patients with high titers of anti-PLA2R antibodies decreased by 60.2％(HR=0.39,P=0.018).Conclusions Different RTX dosages yielded comparable overall clinical remission rates without significantly increasing adverse events.RTX+TAC regimen achieves higher early CR rate.Serological remission time and high titer anti-PLA2R antibodies are associated with clinical outcomes.

Objective:To investigate the expression characteristics of the SLFN11 gene in pancreatic cancer and its relationship with cis-diamminedichloroplatinum(DDP)resistance.Methods:This study analyzed multiple publicly available datasets, including GSE207611, GSE186205, GSE73978, GSE217845, and GSE238163 from the gene expression omnibus(GEO)datasets.These datasets comprise gene expression data and single-cell sequencing data from pancreatic cancer cell lines and patients, which were utilized to examine the expression characteristics of the SLFN11 gene and its association with cisplatin resistance.Receiver operating characteristic (ROC) curves were generated, and the area under the curve (AUC) value along with the confidence interval ( CI) were calculated.Additionally, Kaplan-Meier survival analysis and single-cell RNA sequencing(scRNA-seq)data analysis were conducted. Results:The differential expression of the SLFN11 gene between tumor and adjacent normal tissues was statistically significant( P<0.05). The AUC value for SLFN11 in diagnosing pancreatic cancer was 0.761.SLFN11, along with its associated genes PTN and SLC15A3, collectively influenced the survival of pancreatic cancer patients.Results from single-cell sequencing demonstrated high expression of SLFN11 in mesenchymal stem cells. Conclusions:The SLFN11 gene demonstrates significant diagnostic and prognostic value in pancreatic cancer and may play a role in the mechanism of cisplatin resistance.