1.Cross-sectional investigation and analysis of the early warning threshold for intraoperative bleeding intervention during dilation and curettage in type Ⅱ cesarean scar pregnancy
Changmei SANG ; Qiuling SHI ; Yanjun KANG ; Zhibiao WANG ; Shuping ZHAO
Chinese Journal of Obstetrics and Gynecology 2025;60(10):782-787
Objective:To investigate nationwidely the alert threshold for intraoperative bleeding intervention during dilation and curettage (D&C) for type Ⅱ cesarean scar pregnancy (CSP) in the first trimester.Methods:A retrospective cross-sectional survey was conducted. From March 11 to April 14, 2022, obstetricians and gynecologists affiliated with professional associations or institutions of Family Planning Subgroup, Chinese Society of Obstetrics and Gynecology, Chinese Medical Associaton were surveyed using “Expert Consultation Questionnaire on Alert Threshold for Intraoperative Blood Loss in CSP”. The questionnaire was distributed via WeChat using the Questionnaire Star platform. Observation indicators included professional expertise of participants, preferred treatment modalities for type Ⅱ CSP, and an alert threshold for intraoperative bleeding during D&C.Results:A total of 361 valid questionnaires were collected. Among 361 participants, 73.7% (266/361) held the title of associate chief physician or higher, 61.2% (221/361) had ≥20 years of clinical experience, and 71.8% (178/248) worked in tertiary general hospitals. The most common treatment for type Ⅱ CSP was D&C following pretreatment with medication, uterine artery embolization (UAE), or high intensity focused ultrasound (HIFU), the percentage was 35.73% (129/361). 51.2% (185/361) of participants recommended an alert threshold of 100 ml for intraoperative bleeding during D&C. Cross-analysis revealed that participants managing <30 cases annually preferred ultrasound-guided D&C (32.0%, 56/175), those managing 30-99 cases or 100-199 cases per year favored pretreatment with medication, UAE or HIFU+D&C (39.0%, 55/141; 52.9%, 18/34), those managing ≥200 cases per year preferred hysteroscopic D&C (4/11). The most frequently selected alert threshold was 100 ml. In the subgroup analysis of participants with ≥100 cases annual admissions for CSP, 46.7% (21/45) of participants chose medication, UAE or HIFU pretreatment+D&C, while 53.3% (24/45) supported setting the alert threshold at 100 ml.Conclusions:The preferred treatment for type Ⅱ CSP is D&C following medication, UAE or HIFU pretreatment, with an intraoperative bleeding alert threshold of 100 ml. Timely alerts and proactive interventions could reduce injury severity, improve outcomes, and optimize CSP management strategies.
2.Study on the application effect of personalized osteotomy guide plate in high tibial osteotomy for knee osteoarthritis
Chao QI ; Xiaoming LI ; Donghui GUO ; Qiuling SHI ; Yunchao ZHAO ; Jun DONG ; Zhengxin MENG ; Xingyue WANG
Journal of Clinical Surgery 2025;33(4):360-364
Objective To explore the application effect of personalized osteotomy guide plate in high tibial osteotomy for patients with knee osteoarthritis(KOA).Methods A total of 99 patients with KOA who underwent open wedge high tibial osteotomy(OWHTO)in our hospital from January 2022 to January 2023 were selected and randomly divided into a study group(50 cases)and a control group(49 cases)using a random number table method.The control group received traditional medial OWHTO treatment,and the study group received a combination of medial OWHTO and personalized osteotomy guide plate treatment.The indexes of operation and postoperative rehabilitation,serum inflammatory stress factor[C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),cortisol(Cor),adrenocorticotropin(ACTH)],anatomical structure of knee joint[tibial plateau posterior Angle(PTSA),proximal medial tibial Angle(MPT A),hip knee ankle Angle(HKA)],knee function,ACL shape and function,postoperative complications were compared between the two groups.Results The amount of bleeding,the number of intraoperative fluoroscopy,and the postoperative drainage volume in the study group were(138.69±24.03)ml,(4.83±1.07)times,and(228.95±38.72)ml,respectively,which were all less than those in the control group(154.28±27.16)ml,(7.15±1.14)times,and(271.61±42.19)ml.In the study group,the operation time,incision length,and hospitalization time were(40.96±7.28)min,(8.96±0.85)cm,and(10.73±2.05)d,respectively,which were all shorter than those in the control group[(52.31±10.12)min,(9.51±1.03)cm,and(12.16±2.37)d],with statistically significant differences(P<0.05).The levels of serum CRP,TNF-α,Cor,and ACTH in the study group on the 3rd day after the operation were(31.36±4.68)mg/L,(26.71±3.84)ng/ml,(241.28±27.45)ng/ml,and(18.65±3.01)pmol/L,respectively,which were lower than those in the control group[(35.07±5.16)mg/L,(30.29±4.15)ng/ml,(279.65±30.12)ng/ml,and(21.73±3.28)pmol/L,respectively],and the differences were statistically significant(P<0.05).The Hospital for Special Surgery(HSS)knee score and Knee Society Score(KSS)of the study group at 12 months after surgery were(81.24±6.85)points and(78.26±6.14)points,respectively,which were higher than those of the control group[(78.08±6.42)points and(75.53±5.82)points,respectively],with statistically significant differences(P<0.05);at the 12th month after surgery,the width of the ACL body in the study group was(5.68±0.71)mm,which was greater than that in the control group[(5.12±0.64)mm].The amount of anterior tibial displacement was(5.81±0.43)mm,which was smaller than that in the control group(6.19±0.41)mm,and the differences were statistically significant(P<0.05);the incidence of postoperative complications in the study group was 4.00%,which was lower than that in the control group(18.37%),and the difference was statistically significant(P<0.05).Conclusion The combined treatment of medial OWHTO and personalized osteotomy guide plate can reduce surgical trauma in patients with KOA,lower the incidence of complications,facilitate patient recovery,while maintaining the morphology and function of the ACL,and improving prognosis.
3.Drug resistance and virulence characteristics of carbapenem-resistant Klebsiella pneumoniae carrying the blaKPC-2 gene
Yongshi ZHAO ; Yan ZHANG ; Jian MAO ; Yingxuan CHEN ; Yaozhu YANG ; Qiuling HUANG ; Yan DU ; Na DU
Chinese Journal of Microbiology and Immunology 2025;45(8):671-679
Objective:To analyze the drug resistance and virulence characteristics of KPC-2-producing carbapenem-resistant Klebsiella pneumoniae(CRKP). Methods:A total of 26 non-repeating CRKP strains clinically isolated from a Class Ⅲ hospital in Kunming from August 2021 to March 2022 were collected. Mass spectrometry and the VITEK 2 Compact system were used to identify the bacteria and perform drug susceptibility tests. PCR was used to amplify the drug resistance and virulence genes carried by the strains. These CRKP strains were divided into a hypervirulent CRKP(CR-hvKP) group and a CR-non-hvKP group according to the characteristic virulence genes of hypervirulent Klebsiella pneumoniae. The virulence phenotypes of CRKP were investigated by wire drawing test, serum resistance test and siderophore qualitative and quantitative tests. The whole genomes of CRKP-67 (a CR-hvKP strain) and CRKP-94 (a CR-non-hvKP strain) were sequenced by the Illumina high-throughput sequencing platform, to further analyze the drug resistance genes, virulence genes, and virulence plasmidds carried by the strains. Results:The drug sensitivity results indicated that all 26 strains were resistant to carbapenem, cephalosporins, fluoroquinolones and β-lactam/β-lactam inhibitor complexes. The resistance rates to amicacin, cotrimoxazole and gentamicin were 61.54%(6/26), 57.69%(15/26) and 73.08%(9/26), respectively. Regarding the drug resistance gene amplification results, the carrying rates of blaKPC-2, blaNDM-1, blaOXA-48, blaVIM, blaIMP, blaSME, blaSHV, blaCTX-M and blaTEM were 100.00%(26/26), 0, 0, 0, 0, 100.00%(26/26), 100.00%(26/26), 15.38% (4/26) and 73.08%(19/26), respectively. In the 26 strains, the carrying rates of toxic genes entB, entC, ureA, uge, wabG, ycf, irp1, irp2, mrkD, fimH and ybtS were 100.00%(26/26), while the carrying rates of virulence genes kfuB, iroN, aero, magA and alls were 0. The positive rate of string test was 66.7%(6/9) in the CR-hvKP group and 0 in the CR-non-hvKP group. The serum killing test showed a high sensitivity rate of 77.78%(7/9), a low sensitivity rate of 11.11%(1/9), and a serum resistance rate of 11.11%(1/9) in the CR-hvKP group. In the CR-non-hvKP group, the high sensitivity rate was 29.41%(5/17); the low sensitivity rate was 17.65%(3/17), and the serum resistance rate was 52.94%(9/17). There was no statistical significance between the two groups( P>0.05). The qualitative results of siderophore showed that all strains produced yellow chelating circles with slightly different color depth and size. The quantitative results of siderophore experiment showed that the average siderophore production level of CR-hvKP group was 40.74%, and that of CR-non-hvKP group was 28.21%. The level was higher in the CR-hvKP group than in the CR-non-hvKP group, and the difference was statistically significant( P<0.05). Whole-genome sequencing results showed that CRKP-67 was ST11 type and contained 3 plasmids. Among them, plasmid pCRKP-67-A carried a series of virulence genes, including iucABCD, iutA, rmpA, rmpA2, iroB and peg344, which were highly virulent characteristic genes. Plasmid pCRKP-67-B carried blaKPC-2, blaCTX-M, blaSHV, blaTEM and other drug-resistant genes. Plasmid pCRKP-67-C carried sul2, tetR, tetA and other drug-resistant genes. The CRKP-94 was of ST340 type and contained a drug-resistant plasmid carrying blaKPC-2, blaCTX-M, blaSHV, blaTEM and other drug-resistant genes. Conclusions:CRKP strains are highly resistant, and are only sensitive to a few antibiotics, and carry a variety of drug resistance genes. The main resistance mechanism to carbapenem antibiotics is the presence of the blaKPC-2 gene, which is located on the plasmids, which results in the spread of carbapenem resistance. The types and quantity of virulence genes carried by the CR-hvKP strain are more and greater respectively than those carried by the CR-non-hvKP strain. The co-existence of drug-resistant and virulence plasmids in CR-hvKP strains may lead to the co-transmission of high drug resistance and hypervirulence, which should be highly valued by relevant departments.
4.Causal relationship between non-Hodgkin lymphoma and chronic hepatitis B virus infection: a two-sample bidirectional Mendelian randomization analysis
Liangliang DONG ; Yongjian HUANG ; Jianqiang YE ; Zilin NIAN ; Lin YANG ; Ting CHEN ; Wenbin LIU ; Qiuling ZHAO ; Juming CHEN ; Lijun LAI ; Qin CHEN
Journal of Leukemia & Lymphoma 2025;34(2):85-91
Objective:To investigate the correlation between non-Hodgkin lymphoma (NHL) and chronic hepatitis B virus (HBV) infection by using the method of two-sample bidirectional Mendelian randomization (MR) analysis.Methods:Genetic variation data for NHL came from the Finnish database (FinnGen) Consortium 2021 public genome-wide association study (GWAS) dataset including 1 088 patients with NHL and 299 952 control subjects. The GWAS dataset for chronic HBV infection was derived from GWAS analysis published in 2021, including 145 NHL patients and 351 740 control subjects. NHL was used as an exposure factor, single nucleotide polymorphism (SNP) significantly associated with NHL was used as an instrumental variable (IV), chronic HBV infection was used as an outcome variable. The two-sample MR analysis was performed by using inverse-variance weighted (IVW) method. Chronic HBV infection was taken as an exposure factor, SNP significantly associated with chronic HBV infection was taken as IV, and NHL was taken as outcome variable, and then reverse two-sample MR analysis was performed. The IVW method used the inverse variance of each IV as the weight to fit, and the ratio method was used to measure SNP one by one and make weighted regression analysis, so as to obtain the overall estimate. MR-Egger regression and the weighted median (WME) method were also used to supplement the IVW method. In sensitivity analysis, leave-one-out sensitivity analysis was used to evaluate the impact of a single SNP. Cochran Q test was used to analyze the heterogeneity of the selected IV. MR-Egger regression was used to measure the average horizontal pleiotropy of IV, and the P-value of directivity was calculated. The MR-pleiotropy residual sum and outlier (MR-PRESSO) Global Test was used to exclude possible horizontal pleiotropic outliers and reduce bias. Results:In the leave-one-out sensitivity analysis, SNP with significant effects on causal associations was excluded. In forward MR analysis, IVs were 10 SNPs associated with NHL; the IVW method indicated that there was no causal association between NHL and chronic HBV infection ( OR = 0.979, 95% CI: 0.925-1.036, P = 0.465). MR-Egger regression ( OR = 0.992, 95% CI: 0.926-1.062, P = 0.825) and WME method ( OR = 0.992, 95% CI: 0.934-1.055, P = 0.805) were used as supplementary methods to obtain the consistent results. In sensitivity analysis, Cochran Q test showed no heterogeneity among IVs (IVW method: P = 0.271, MR-Egger regression: P = 0.239). Horizontal pleiotropy was not found in the MR-Egger regression (intercept was -0.01, P = 0.778) and the MR-PRESSO Global Test ( P > 0.05), suggesting robust results. In the reverse MR analysis, IVs were 8 SNPs associated with NHL; the IVW method ( OR = 1.117, 95% CI: 0.942-1.324, P = 0.202) also found no significant causal relationship between chronic HBV infection and NHL; MR-Egger regression ( OR = 0.777, 95% CI: 0.450-1.343, P = 0.401) and WME method ( OR = 1.120, 95% CI: 0.887-1.415, P = 0.351) also showed similar risk estimates. Sensitivity analysis also suggested the consistency and reliability of the results. Cochran Q test showed no heterogeneity among IVs (IVW method: P = 0.775, MR-Egger regression: P = 0.903). Horizontal pleiotropy was not found by MR-Egger regression (intercept was 0.102, P = 0.548) and MR-PRESSO Global Test ( P > 0.05). Conclusions:MR analysis suggests no causal relationship between NHL and chronic HBV infection.
5.Dual activation of GCGR/GLP1R signaling ameliorates intestinal fibrosis via metabolic regulation of histone H3K9 lactylation in epithelial cells.
Han LIU ; Yujie HONG ; Hui CHEN ; Xianggui WANG ; Jiale DONG ; Xiaoqian LI ; Zihan SHI ; Qian ZHAO ; Longyuan ZHOU ; JiaXin WANG ; Qiuling ZENG ; Qinglin TANG ; Qi LIU ; Florian RIEDER ; Baili CHEN ; Minhu CHEN ; Rui WANG ; Yao ZHANG ; Ren MAO ; Xianxing JIANG
Acta Pharmaceutica Sinica B 2025;15(1):278-295
Intestinal fibrosis is a significant clinical challenge in inflammatory bowel diseases, but no effective anti-fibrotic therapy is currently available. Glucagon receptor (GCGR) and glucagon-like peptide 1 receptor (GLP1R) are both peptide hormone receptors involved in energy metabolism of epithelial cells. However, their role in intestinal fibrosis and the underlying mechanisms remain largely unexplored. Herein GCGR and GLP1R were found to be reduced in the stenotic ileum of patients with Crohn's disease as well as in the fibrotic colon of mice with chronic colitis. The downregulation of GCGR and GLP1R led to the accumulation of the metabolic byproduct lactate, resulting in histone H3K9 lactylation and exacerbated intestinal fibrosis through epithelial-to-mesenchymal transition (EMT). Dual activating GCGR and GLP1R by peptide 1907B reduced the H3K9 lactylation in epithelial cells and ameliorated intestinal fibrosis in vivo. We uncovered the role of GCGR/GLP1R in regulating EMT involved in intestinal fibrosis via histone lactylation. Simultaneously activating GCGR/GLP1R with the novel dual agonist peptide 1907B holds promise as a treatment strategy for alleviating intestinal fibrosis.
6.Cross-sectional investigation and analysis of the early warning threshold for intraoperative bleeding intervention during dilation and curettage in type Ⅱ cesarean scar pregnancy
Changmei SANG ; Qiuling SHI ; Yanjun KANG ; Zhibiao WANG ; Shuping ZHAO
Chinese Journal of Obstetrics and Gynecology 2025;60(10):782-787
Objective:To investigate nationwidely the alert threshold for intraoperative bleeding intervention during dilation and curettage (D&C) for type Ⅱ cesarean scar pregnancy (CSP) in the first trimester.Methods:A retrospective cross-sectional survey was conducted. From March 11 to April 14, 2022, obstetricians and gynecologists affiliated with professional associations or institutions of Family Planning Subgroup, Chinese Society of Obstetrics and Gynecology, Chinese Medical Associaton were surveyed using “Expert Consultation Questionnaire on Alert Threshold for Intraoperative Blood Loss in CSP”. The questionnaire was distributed via WeChat using the Questionnaire Star platform. Observation indicators included professional expertise of participants, preferred treatment modalities for type Ⅱ CSP, and an alert threshold for intraoperative bleeding during D&C.Results:A total of 361 valid questionnaires were collected. Among 361 participants, 73.7% (266/361) held the title of associate chief physician or higher, 61.2% (221/361) had ≥20 years of clinical experience, and 71.8% (178/248) worked in tertiary general hospitals. The most common treatment for type Ⅱ CSP was D&C following pretreatment with medication, uterine artery embolization (UAE), or high intensity focused ultrasound (HIFU), the percentage was 35.73% (129/361). 51.2% (185/361) of participants recommended an alert threshold of 100 ml for intraoperative bleeding during D&C. Cross-analysis revealed that participants managing <30 cases annually preferred ultrasound-guided D&C (32.0%, 56/175), those managing 30-99 cases or 100-199 cases per year favored pretreatment with medication, UAE or HIFU+D&C (39.0%, 55/141; 52.9%, 18/34), those managing ≥200 cases per year preferred hysteroscopic D&C (4/11). The most frequently selected alert threshold was 100 ml. In the subgroup analysis of participants with ≥100 cases annual admissions for CSP, 46.7% (21/45) of participants chose medication, UAE or HIFU pretreatment+D&C, while 53.3% (24/45) supported setting the alert threshold at 100 ml.Conclusions:The preferred treatment for type Ⅱ CSP is D&C following medication, UAE or HIFU pretreatment, with an intraoperative bleeding alert threshold of 100 ml. Timely alerts and proactive interventions could reduce injury severity, improve outcomes, and optimize CSP management strategies.
7.Study on the application effect of personalized osteotomy guide plate in high tibial osteotomy for knee osteoarthritis
Chao QI ; Xiaoming LI ; Donghui GUO ; Qiuling SHI ; Yunchao ZHAO ; Jun DONG ; Zhengxin MENG ; Xingyue WANG
Journal of Clinical Surgery 2025;33(4):360-364
Objective To explore the application effect of personalized osteotomy guide plate in high tibial osteotomy for patients with knee osteoarthritis(KOA).Methods A total of 99 patients with KOA who underwent open wedge high tibial osteotomy(OWHTO)in our hospital from January 2022 to January 2023 were selected and randomly divided into a study group(50 cases)and a control group(49 cases)using a random number table method.The control group received traditional medial OWHTO treatment,and the study group received a combination of medial OWHTO and personalized osteotomy guide plate treatment.The indexes of operation and postoperative rehabilitation,serum inflammatory stress factor[C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),cortisol(Cor),adrenocorticotropin(ACTH)],anatomical structure of knee joint[tibial plateau posterior Angle(PTSA),proximal medial tibial Angle(MPT A),hip knee ankle Angle(HKA)],knee function,ACL shape and function,postoperative complications were compared between the two groups.Results The amount of bleeding,the number of intraoperative fluoroscopy,and the postoperative drainage volume in the study group were(138.69±24.03)ml,(4.83±1.07)times,and(228.95±38.72)ml,respectively,which were all less than those in the control group(154.28±27.16)ml,(7.15±1.14)times,and(271.61±42.19)ml.In the study group,the operation time,incision length,and hospitalization time were(40.96±7.28)min,(8.96±0.85)cm,and(10.73±2.05)d,respectively,which were all shorter than those in the control group[(52.31±10.12)min,(9.51±1.03)cm,and(12.16±2.37)d],with statistically significant differences(P<0.05).The levels of serum CRP,TNF-α,Cor,and ACTH in the study group on the 3rd day after the operation were(31.36±4.68)mg/L,(26.71±3.84)ng/ml,(241.28±27.45)ng/ml,and(18.65±3.01)pmol/L,respectively,which were lower than those in the control group[(35.07±5.16)mg/L,(30.29±4.15)ng/ml,(279.65±30.12)ng/ml,and(21.73±3.28)pmol/L,respectively],and the differences were statistically significant(P<0.05).The Hospital for Special Surgery(HSS)knee score and Knee Society Score(KSS)of the study group at 12 months after surgery were(81.24±6.85)points and(78.26±6.14)points,respectively,which were higher than those of the control group[(78.08±6.42)points and(75.53±5.82)points,respectively],with statistically significant differences(P<0.05);at the 12th month after surgery,the width of the ACL body in the study group was(5.68±0.71)mm,which was greater than that in the control group[(5.12±0.64)mm].The amount of anterior tibial displacement was(5.81±0.43)mm,which was smaller than that in the control group(6.19±0.41)mm,and the differences were statistically significant(P<0.05);the incidence of postoperative complications in the study group was 4.00%,which was lower than that in the control group(18.37%),and the difference was statistically significant(P<0.05).Conclusion The combined treatment of medial OWHTO and personalized osteotomy guide plate can reduce surgical trauma in patients with KOA,lower the incidence of complications,facilitate patient recovery,while maintaining the morphology and function of the ACL,and improving prognosis.
8.Investigation of physician countermeasures for mild progression after treatment with EGFR-TKI in lung adenocarcinoma
Wang QIMING ; Wang LU ; Yu HONGFAN ; Shen BO ; Chen CHENG ; Yan ZHAO ; Huang CHENG ; Wang YUMENG ; Li KAI ; Wang JING ; Shi QIULING
Chinese Journal of Clinical Oncology 2025;52(3):109-114
Objective:To investigate the views of doctors on the incidence and treatment tactics of mild progression after epithelial growth factor receptor tyrosine kinase inhibitor(EGFR-TKI)treatment in lung adenocarcinoma and provide suggestions to optimize the counter-measure strategies.Methods:Convenience sampling was used to conduct an online questionnaire survey for doctors specializing in onco-logy and respiratory diseases.Results:584 valid questionnaires were collected,and all the doctors expressed concerns regarding mild tumor progression after EGFR-TKI treatment.The coping strategies included maintaining the original TKI treatment,adding other treatments to the original TKI,changing the regimen,and performing secondary tissue biopsy,among which,most doctors chose to add other treatments to the original TKI.Conclusions:Doctors have noticed the enlargement of target lesions in still stable disease(SD)and most frequently chose to add other treatments to the original TKI as a coping strategy.This finding can provide a reference for framing future guidelines for large sample prospective clinical studies that are needed to find the most effective synergistic treatment options.
9.Investigation of physician countermeasures for mild progression after treatment with EGFR-TKI in lung adenocarcinoma
Wang QIMING ; Wang LU ; Yu HONGFAN ; Shen BO ; Chen CHENG ; Yan ZHAO ; Huang CHENG ; Wang YUMENG ; Li KAI ; Wang JING ; Shi QIULING
Chinese Journal of Clinical Oncology 2025;52(3):109-114
Objective:To investigate the views of doctors on the incidence and treatment tactics of mild progression after epithelial growth factor receptor tyrosine kinase inhibitor(EGFR-TKI)treatment in lung adenocarcinoma and provide suggestions to optimize the counter-measure strategies.Methods:Convenience sampling was used to conduct an online questionnaire survey for doctors specializing in onco-logy and respiratory diseases.Results:584 valid questionnaires were collected,and all the doctors expressed concerns regarding mild tumor progression after EGFR-TKI treatment.The coping strategies included maintaining the original TKI treatment,adding other treatments to the original TKI,changing the regimen,and performing secondary tissue biopsy,among which,most doctors chose to add other treatments to the original TKI.Conclusions:Doctors have noticed the enlargement of target lesions in still stable disease(SD)and most frequently chose to add other treatments to the original TKI as a coping strategy.This finding can provide a reference for framing future guidelines for large sample prospective clinical studies that are needed to find the most effective synergistic treatment options.
10.Drug resistance and virulence characteristics of carbapenem-resistant Klebsiella pneumoniae carrying the blaKPC-2 gene
Yongshi ZHAO ; Yan ZHANG ; Jian MAO ; Yingxuan CHEN ; Yaozhu YANG ; Qiuling HUANG ; Yan DU ; Na DU
Chinese Journal of Microbiology and Immunology 2025;45(8):671-679
Objective:To analyze the drug resistance and virulence characteristics of KPC-2-producing carbapenem-resistant Klebsiella pneumoniae(CRKP). Methods:A total of 26 non-repeating CRKP strains clinically isolated from a Class Ⅲ hospital in Kunming from August 2021 to March 2022 were collected. Mass spectrometry and the VITEK 2 Compact system were used to identify the bacteria and perform drug susceptibility tests. PCR was used to amplify the drug resistance and virulence genes carried by the strains. These CRKP strains were divided into a hypervirulent CRKP(CR-hvKP) group and a CR-non-hvKP group according to the characteristic virulence genes of hypervirulent Klebsiella pneumoniae. The virulence phenotypes of CRKP were investigated by wire drawing test, serum resistance test and siderophore qualitative and quantitative tests. The whole genomes of CRKP-67 (a CR-hvKP strain) and CRKP-94 (a CR-non-hvKP strain) were sequenced by the Illumina high-throughput sequencing platform, to further analyze the drug resistance genes, virulence genes, and virulence plasmidds carried by the strains. Results:The drug sensitivity results indicated that all 26 strains were resistant to carbapenem, cephalosporins, fluoroquinolones and β-lactam/β-lactam inhibitor complexes. The resistance rates to amicacin, cotrimoxazole and gentamicin were 61.54%(6/26), 57.69%(15/26) and 73.08%(9/26), respectively. Regarding the drug resistance gene amplification results, the carrying rates of blaKPC-2, blaNDM-1, blaOXA-48, blaVIM, blaIMP, blaSME, blaSHV, blaCTX-M and blaTEM were 100.00%(26/26), 0, 0, 0, 0, 100.00%(26/26), 100.00%(26/26), 15.38% (4/26) and 73.08%(19/26), respectively. In the 26 strains, the carrying rates of toxic genes entB, entC, ureA, uge, wabG, ycf, irp1, irp2, mrkD, fimH and ybtS were 100.00%(26/26), while the carrying rates of virulence genes kfuB, iroN, aero, magA and alls were 0. The positive rate of string test was 66.7%(6/9) in the CR-hvKP group and 0 in the CR-non-hvKP group. The serum killing test showed a high sensitivity rate of 77.78%(7/9), a low sensitivity rate of 11.11%(1/9), and a serum resistance rate of 11.11%(1/9) in the CR-hvKP group. In the CR-non-hvKP group, the high sensitivity rate was 29.41%(5/17); the low sensitivity rate was 17.65%(3/17), and the serum resistance rate was 52.94%(9/17). There was no statistical significance between the two groups( P>0.05). The qualitative results of siderophore showed that all strains produced yellow chelating circles with slightly different color depth and size. The quantitative results of siderophore experiment showed that the average siderophore production level of CR-hvKP group was 40.74%, and that of CR-non-hvKP group was 28.21%. The level was higher in the CR-hvKP group than in the CR-non-hvKP group, and the difference was statistically significant( P<0.05). Whole-genome sequencing results showed that CRKP-67 was ST11 type and contained 3 plasmids. Among them, plasmid pCRKP-67-A carried a series of virulence genes, including iucABCD, iutA, rmpA, rmpA2, iroB and peg344, which were highly virulent characteristic genes. Plasmid pCRKP-67-B carried blaKPC-2, blaCTX-M, blaSHV, blaTEM and other drug-resistant genes. Plasmid pCRKP-67-C carried sul2, tetR, tetA and other drug-resistant genes. The CRKP-94 was of ST340 type and contained a drug-resistant plasmid carrying blaKPC-2, blaCTX-M, blaSHV, blaTEM and other drug-resistant genes. Conclusions:CRKP strains are highly resistant, and are only sensitive to a few antibiotics, and carry a variety of drug resistance genes. The main resistance mechanism to carbapenem antibiotics is the presence of the blaKPC-2 gene, which is located on the plasmids, which results in the spread of carbapenem resistance. The types and quantity of virulence genes carried by the CR-hvKP strain are more and greater respectively than those carried by the CR-non-hvKP strain. The co-existence of drug-resistant and virulence plasmids in CR-hvKP strains may lead to the co-transmission of high drug resistance and hypervirulence, which should be highly valued by relevant departments.

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