1.Detection and genetic evolution analysis of pathogens borne by Pulex irritans in selected areas of Xinjiang
Xinxin HAN ; Jing ZHAO ; Xuefeng LIU ; Yitao LI ; Tingting WU ; Junang DAI ; Mengyang YAN ; Zhihua SUN ; Hui ZHANG
Chinese Journal of Zoonoses 2025;41(8):852-858
This study identified the types and pathogen carrying status of fleas on the surface of sheep in some areas of southern Xinjiang,and analyzed the genetic evolution differences with respect to related pathogens.The aim was to provide a reference for the local prevention and control of fleas and insect borne infectious diseases.A total of 1 586 fleas were collected from agricultural and pas-toral areas of Tumushuke City and Hotan Prefecture.Flea species were identified on the basis of morphology and the Pulex irritans mi-tochondrial COII gene.Flea DNA was extracted,and PCR was conducted to amplify the Bartonella gltA gene;Arsenophonus,Ana-plasma,Ehrlichia,and Wolbachia 16S rRNA genes;RickettsiaOmpA,17kDa,16S rRNA genes,and Yersinia pestis 16S rDNA gene.The amplified products were sequenced,and the homology of the genes of the three detected pathogens(gltA gene of Bartonella,16S rRNA gene of Wolbachia,and Anaplasma phagocytophilum)with respect to known corresponding genes of the same pathogen in Gen-Bank was analyzed.Phylogenetic trees were constructed with the adjacency method in MEGA 11.0.According to morphological and mo-lecular biology identification results,all fleas collected in this study were Pulex irritans.PCR indicated that the target gene fragments had been added to the mitochondrial COII,BartonellagltA,Wolbachia,and autophagosomal 16S rRNA genes of human fleas,all of which were consistent with the expected fragment sizes.Target bands were not amplified from Ehrlichia,Arsenophonus,spotted fever group Rickettsia,and Yersinia pestis.According to homology and genetic evolution analysis of human flea mitochondrial COII and the corresponding genes of the above-described pathogens,the COX2 gene(ON455234.1)of human fleas in Tumushuke city and Iran ob-tained in this study showed the highest homology(99.84%).The COII gene(NC_063709.1)of human fleas in Hetan City and Hunan region showed the highest homology(100%).Our findings further confirmed that the flea species was Pulex irritans.The PCR amplifi-cation results indicated that the collected Pulex irritans carried multiple pathogens,among which Bartonella and Wolbachia had the highest infection rates,and the infection rate with Anaplasma phagocytophilum was relatively low.This study is the first to discover flea species on the surface of sheep in some areas of southern Xinjiang.Our findings preliminarily confirmed that Bartonella,Wolba-chia,and Anaplasma phagocytophilum are the main Pulex irritans pathogens.
2.The Study on Application Timing of Intra-aortic Balloon Pump in ST-egment-elevation Myocardial Infarction Patients Complicated with Ventricular Septal Rupture
Jie FANG ; Xuefeng GUANG ; Xiaofeng LIN ; Hailong DAI ; Tongxi ZUO ; Qi LAI
Journal of Kunming Medical University 2025;46(5):96-100
Objective To study the effect of intra-aortic balloon pump(IABP)in the therapy of ST-segment-elevation myocardial infarction(STEMI)patients complicated with ventricular septal rupture(VSR).Methods A retrospective analysis was performed on 35 STEMI patients complicated with VSR.Those patients were admitted in Yan'an Hospital Affiliated to Kunming Medical University from January 2019 to June 2023.Patients were divided into the combine-treated group(20 cases)and the drug-treated group(15 cases)according to the therapeutic strategies.The combine-treated group received IABP implantation and drug therapy,and the drug-treated group only received drug therapy.The clinical characteristics,hemodynamic and cardiac function improvement and mortality were evaluated.Hemodynamic and cardiac function improvement were compared between the two groups.Results There were no statistically significant differences in age,male proportion and size of VSR between the two groups(P>0.05).In the combine-treated group,the average heart rate,the average arterial blood pressure,central venous pressure,the LVEF value of cardiac ultrasound,pleural effusion,B-Line of lung,the level of B-type natriuretic peptide(BNP),the level of serum creatinine and the level of serum lactic acid were improved at 72 h after IABP use(all P<0.05).All these indicators got worse in the drug-treated group.The mortality rate of the combine-treated group was markedly lower than that of the drug-treated group(P<0.05).The mortality rate of patients who received IABP implantation within 72 h after VSR was lower than that of patients who received IABP implantation beyond 72 h after VSR.Conclusion For patients with AMI complicated with VSR,implantation of IABP can significantly improve hemodynamics,cardiac function and reduce mortality.
3.Laboratory testing for non-factor therapy in hemophilia: progress and challenges from traditional to innovative approaches
Jing DAI ; Xiaoli MA ; Yu LIU ; Xuefeng WANG
Chinese Journal of Laboratory Medicine 2025;48(8):953-961
Hemophilia A and hemophilia B are hereditary coagulation bleeding disorders. Traditional treatment primarily relies on factor replacement therapy using coagulation factor Ⅷ(FⅧ) or coagulation factor Ⅸ (FⅨ)products. Although conventional therapies can alleviate bleeding symptoms to some extent, they also have certain limitations, such as the need for frequent infusions, the risk of infection, and the potential development of inhibitors in some patients. Currently, treatment strategies for hemophilia are gradually shifting toward non-factor therapies, providing the appearence of novel non-factor drugs. However, these novel therapies not only interfere with traditional coagulation assays but may also fail to comprehensively evaluate their efficacy and safety through conventional coagulation tests. Therefore, there is an urgent need to develop and optimize new laboratory testing methods to ensure accurate assessment of patients′ responses to non-factor therapies and the hemostatic capacity of the drugs themselves. Although some studies have explored the coagulation factor equivalence of non-factor agents, such equivalence cannot fully reflect the actual hemostatic effect in patients after treatment and is therefore unsuitable as a prognostic indicator. Compared with assessing coagulation factor equivalence, total coagulation assays, such as the thrombin generation assay (TGA), can more accurately evaluate efficacy and safety. TGA can take into account multiple factors in the coagulation process, providing a more comprehensive assessment of coagulation function. Furthermore, combining TGA with patient symptoms for comprehensive analysis can enhance its prognostic predictive ability, offering more reliable support for clinical decision-making.
4.The value of thrombin generation and activated protein C resistance in thrombotic risk assessment for patients with lupus anticoagulant
Guiting ZHANG ; Aili JUERAITI ; Yu LIU ; Guanqun XU ; Han SHEN ; Jing DAI ; Xuefeng WANG ; Yeling LU
Chinese Journal of Laboratory Medicine 2025;48(8):979-984
Objective:To analyze the levels of thrombin generation and activated protein C resistance (APC-R) in lupus anticoagulant (LA)-positive patients, and to assess their effectiveness in predicting thrombotic risk in these patients.Methods:Retrospective case-control study. A total of 185 patients with positve LA [91 males, 94 females; age (47.59±19.14) years] in Ruijin Hospital of Shanghai Jiaotong University School of Medicine from November 1st, 2024 to March 31st, 2025 were included. Patients were stratified into thrombotic ( n=91) and non-thrombotic groups ( n=94) based on clinical diagnosis and imaging evidence of thrombosis. The basic characteristics and routine laboratory coagulation levels of LA-positive patients were analyzed. Post-test plasma samples were collected from 43 cases with positive or strongly positive LA, categorized into thrombotic ( n=23) and non-thrombotic ( n=20) groups. Additionally, plasma was collected from 80 healthy controls [40 males and 40 females, age (38.37±15.74) years]. Using simple random sampling method, plasma samples from 10 selected males and 10 selected females were mixed to make 1 group of healthy control, thus accordingly resulted in a total of 4 healthy control groups. Thrombin generation assays (TGA) were then employed to measure prothrombin generation and activated protein C resistance (APC-R) levels in the healthy control, non-thrombotic, and thrombotic groups. One-way analysis of variance was utilized to compare thrombin generation and APC-R levels across these groups. Results:Among the routine laboratory coagulation indexes, the median levels of activated partial thromboplastin time (APTT), fibrin degradation product (FDP) and protein C (PC) in thrombotic group were 30.9 (28.8, 35.5) s, 2.5 (1.3, 2.8) mg/L, and 107.0 (93.0, 127.0)%, respectively, which were significantly higher compared with the non-thrombosis group (all P<0.05). However, between the thrombotic and non-thrombotic group, no statistically significant differences were observed for the levels of prothrombin time (PT), thrombin time (TT), fibrinogen (Fg), or D-dimer (D-D) ( P>0.05). The TGA results showed that the total thrombin generation, the maximal thrombin generation and APC-R levels of patients in the thrombotic group were (1 118.72±387.34) nmol/L·min, (106.01±59.00) nmol/L and (0.33±0.22), respectively, which were significantly higher compared with those in the non-thrombotic group (all P<0.05). Conclusion:Significantly increased thrombin generation and enhanced APC-R were present in the LA-positive patients with thrombosis, indicating the important values of thrombin generation and APC-R in assessing thrombosis risk among this population.
5.Study on discrepancies in measured values of an extended half-life recombinant FⅧ product across different assay systems
Yanyan SHAO ; Yu LIU ; Guanqun XU ; Jing DAI ; Xuefeng WANG
Chinese Journal of Laboratory Medicine 2025;48(8):1015-1021
Objective:To compare the accuracy of one-stage clotting assay and chromogenic substrate assay for testing an extended half-life recombinant FⅧ and to explore standardized conversion models between methods.Methods:Observational study. FⅧ activity (FⅧ:C) in plasma samples with theoretical values of 1 000, 800, 600, 500, 400, and 300 IU/L was measured using both one-stage clotting assay (employing Siemens Actin FSL reagent, Werfen SynthASil reagent, Stago PTT-A reagent) and the chromogenic substrate assay from Hyphen Biomed. Differences in FⅧ:C measured by the various methods were compared using the SNK test. Recovery rates were calculated to evaluate the accuracy of each assay. Sample activity was verified using the thrombin generation assay (TGA). Correlations between activities determined by the different assay systems were assessed using linear regression analysis.Results:Observational study. FⅧ activity (FⅧ:C) in diluted plasma samples with theoretical values of 1 000, 800, 600, 500, 400, and 300 IU/L was measured using both one-stage clotting assay (employing Siemens Actin FSL reagent, Werfen SynthASil reagent, Stago PTT-A reagent) and the chromogenic substrate assay from Hyphen Biomed. Differences in FⅧ:C measured by the various methods were compared using the SNK test. Recovery rates were calculated to evaluate the accuracy of each assay. Sample activity was verified using the thrombin generation assay (TGA). Correlations between activities determined by the different assay systems were assessed using linear regression analysis.Conclusion:Some marked one-stage clotting assay system has limitations in the clinical detection of extended half-life recombinant FⅧ. While the chromogenic substrate assay provides more accurate results. The one-stage clotting assay values can undergo cross-assay correction for FⅧ:C using a standardized conversion coefficient, which can further elevate the accuracy of monitoring hemophilia treatment efficacy.
6.Laboratory diagnosis of congenital coagulation factor Ⅶ deficiency combined with antiphospholipid syndrome and literature review
Zhijun MENG ; Guanqun XU ; Yu LIU ; Chi MA ; Qiulan DING ; Jing DAI ; Wenman WU ; Xuefeng WANG
Chinese Journal of Laboratory Medicine 2025;48(8):1091-1094
Congenital coagulation factor Ⅶ deficiency is a rare autosomal incomplete recessive disorder caused by a defect in the coagulation factor Ⅶ (FⅦ) gene, with an incidence of approximately 1 in 500 000. Antiphospholipid antibody syndrome is relatively common and is a common cause of acquired thrombosis. However, the combination of the latter and the former is extremely rare in clinical practice, which brings difficulties to diagnosis and treatment. This article reported the laboratory examination, diagnosis and treatment of a patient with congenital coagulation factor Ⅶ deficiency and antiphospholipid syndrome after portal vein thrombosis, and reviewed the relevant literature.
7.Perioperative management of a patient with hemophilia a and high-titer inhibitors and literature review
Zhijun MENG ; Jie XIANG ; Guanqun XU ; Yu LIU ; Qiulan DING ; Jing DAI ; Wenman WU ; Xuefeng WANG
Chinese Journal of Hematology 2025;46(8):766-768
A 64-year-old male patient with hemophilia A was scheduled for the surgical removal of a pulmonary mass. Preoperative evaluation revealed that the coagulation factor Ⅷ (FⅧ) activity was 0.5%, with an F Ⅷ inhibitor level of 32 BU/ml; the R value could not be detected on the thromboelastogram. Thoracoscopic lobectomy was successfully completed. On the day of the operation and the first day after the operation, 6 mg of recombinant activated coagulation factor Ⅶ (rFⅦa) was intravenously administered every 6 h. On postoperative day 1, the patient’s blood pressure dropped and the HGB gradually declined from 102 g/L to 65 g/L. Chest X-ray revealed a large amount of pleural effusion on the left side, and urgent thoracoscopic thoracic exploration was performed. A total of 3200 mL fresh blood was cleared, and a thoracic drainage tube was placed. On postoperative day 2, the rFⅦa dose was increased to 6 mg, which was intravenously administered every 4 h, and concentrated red cells were intermittently infused to correct anemia. Four days later, due to the inability to obtain rFⅦa, PCC (50 IU/kg every 8 hours) was administered. Additionally, treatment with methylprednisolone (40 mg/d) and cyclophosphamide (200 mg, every 2 weeks) was initiated to remove FⅧ inhibitors. The thoracic drainage tube was removed on postoperative day 9, and the patient was successfully discharged 3 weeks later.
8.Detection and genetic evolution analysis of pathogens borne by Pulex irritans in selected areas of Xinjiang
Xinxin HAN ; Jing ZHAO ; Xuefeng LIU ; Yitao LI ; Tingting WU ; Junang DAI ; Mengyang YAN ; Zhihua SUN ; Hui ZHANG
Chinese Journal of Zoonoses 2025;41(8):852-858
This study identified the types and pathogen carrying status of fleas on the surface of sheep in some areas of southern Xinjiang,and analyzed the genetic evolution differences with respect to related pathogens.The aim was to provide a reference for the local prevention and control of fleas and insect borne infectious diseases.A total of 1 586 fleas were collected from agricultural and pas-toral areas of Tumushuke City and Hotan Prefecture.Flea species were identified on the basis of morphology and the Pulex irritans mi-tochondrial COII gene.Flea DNA was extracted,and PCR was conducted to amplify the Bartonella gltA gene;Arsenophonus,Ana-plasma,Ehrlichia,and Wolbachia 16S rRNA genes;RickettsiaOmpA,17kDa,16S rRNA genes,and Yersinia pestis 16S rDNA gene.The amplified products were sequenced,and the homology of the genes of the three detected pathogens(gltA gene of Bartonella,16S rRNA gene of Wolbachia,and Anaplasma phagocytophilum)with respect to known corresponding genes of the same pathogen in Gen-Bank was analyzed.Phylogenetic trees were constructed with the adjacency method in MEGA 11.0.According to morphological and mo-lecular biology identification results,all fleas collected in this study were Pulex irritans.PCR indicated that the target gene fragments had been added to the mitochondrial COII,BartonellagltA,Wolbachia,and autophagosomal 16S rRNA genes of human fleas,all of which were consistent with the expected fragment sizes.Target bands were not amplified from Ehrlichia,Arsenophonus,spotted fever group Rickettsia,and Yersinia pestis.According to homology and genetic evolution analysis of human flea mitochondrial COII and the corresponding genes of the above-described pathogens,the COX2 gene(ON455234.1)of human fleas in Tumushuke city and Iran ob-tained in this study showed the highest homology(99.84%).The COII gene(NC_063709.1)of human fleas in Hetan City and Hunan region showed the highest homology(100%).Our findings further confirmed that the flea species was Pulex irritans.The PCR amplifi-cation results indicated that the collected Pulex irritans carried multiple pathogens,among which Bartonella and Wolbachia had the highest infection rates,and the infection rate with Anaplasma phagocytophilum was relatively low.This study is the first to discover flea species on the surface of sheep in some areas of southern Xinjiang.Our findings preliminarily confirmed that Bartonella,Wolba-chia,and Anaplasma phagocytophilum are the main Pulex irritans pathogens.
9.Perioperative management of a patient with hemophilia a and high-titer inhibitors and literature review
Zhijun MENG ; Jie XIANG ; Guanqun XU ; Yu LIU ; Qiulan DING ; Jing DAI ; Wenman WU ; Xuefeng WANG
Chinese Journal of Hematology 2025;46(8):766-768
A 64-year-old male patient with hemophilia A was scheduled for the surgical removal of a pulmonary mass. Preoperative evaluation revealed that the coagulation factor Ⅷ (FⅧ) activity was 0.5%, with an F Ⅷ inhibitor level of 32 BU/ml; the R value could not be detected on the thromboelastogram. Thoracoscopic lobectomy was successfully completed. On the day of the operation and the first day after the operation, 6 mg of recombinant activated coagulation factor Ⅶ (rFⅦa) was intravenously administered every 6 h. On postoperative day 1, the patient’s blood pressure dropped and the HGB gradually declined from 102 g/L to 65 g/L. Chest X-ray revealed a large amount of pleural effusion on the left side, and urgent thoracoscopic thoracic exploration was performed. A total of 3200 mL fresh blood was cleared, and a thoracic drainage tube was placed. On postoperative day 2, the rFⅦa dose was increased to 6 mg, which was intravenously administered every 4 h, and concentrated red cells were intermittently infused to correct anemia. Four days later, due to the inability to obtain rFⅦa, PCC (50 IU/kg every 8 hours) was administered. Additionally, treatment with methylprednisolone (40 mg/d) and cyclophosphamide (200 mg, every 2 weeks) was initiated to remove FⅧ inhibitors. The thoracic drainage tube was removed on postoperative day 9, and the patient was successfully discharged 3 weeks later.
10.Laboratory testing for non-factor therapy in hemophilia: progress and challenges from traditional to innovative approaches
Jing DAI ; Xiaoli MA ; Yu LIU ; Xuefeng WANG
Chinese Journal of Laboratory Medicine 2025;48(8):953-961
Hemophilia A and hemophilia B are hereditary coagulation bleeding disorders. Traditional treatment primarily relies on factor replacement therapy using coagulation factor Ⅷ(FⅧ) or coagulation factor Ⅸ (FⅨ)products. Although conventional therapies can alleviate bleeding symptoms to some extent, they also have certain limitations, such as the need for frequent infusions, the risk of infection, and the potential development of inhibitors in some patients. Currently, treatment strategies for hemophilia are gradually shifting toward non-factor therapies, providing the appearence of novel non-factor drugs. However, these novel therapies not only interfere with traditional coagulation assays but may also fail to comprehensively evaluate their efficacy and safety through conventional coagulation tests. Therefore, there is an urgent need to develop and optimize new laboratory testing methods to ensure accurate assessment of patients′ responses to non-factor therapies and the hemostatic capacity of the drugs themselves. Although some studies have explored the coagulation factor equivalence of non-factor agents, such equivalence cannot fully reflect the actual hemostatic effect in patients after treatment and is therefore unsuitable as a prognostic indicator. Compared with assessing coagulation factor equivalence, total coagulation assays, such as the thrombin generation assay (TGA), can more accurately evaluate efficacy and safety. TGA can take into account multiple factors in the coagulation process, providing a more comprehensive assessment of coagulation function. Furthermore, combining TGA with patient symptoms for comprehensive analysis can enhance its prognostic predictive ability, offering more reliable support for clinical decision-making.

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