Objective To establish a reference interval for the pepsinogen(PG)Ⅰ,PGⅡ and PGⅠ/PGⅡ in healthy adult subjects in Zhejiang Province.Methods The data of 45 504 healthy adult subjects were collected,and the levels of serum PGⅠ and PG Ⅱ were detected by chemiluminescence microparticle immunoassay.The reference range of PGs were determined according to the CLSI-C28-A3 file.Results The median serum PGⅠ concentration in male was 132.62μg/L,PGⅡ concentration was 8.10μg/L,and PGⅠ/PGⅡ was 15.9.For females,they were 107.44μg/L,6.96μg/L and 15.0,respectively.PGI,PGⅡ concentration and PGⅠ/PGⅡ ratio were significantly higher in males than in females(P<0.001).The levels of PGⅠ and PGⅡ increased with age(P<0.001).Serum PGⅠ reference interval for male:59.79-234.97μg/L for 19-39 years old,63.33-294.62μg/L for 40-59 years old,64.25-333.61μg/L for≥60 years old,PGⅡ reference interval:3.33-22.60μg/L for 19-39 years of age,3.79-33.89μg/L for 40-59 years of age,4.15-42.08μg/L for≥60 years of age,PGⅠ/PGⅡ reference interval:those aged 19-39 years ranged from 7.3 to 31.4,those aged 40-59 years ranged from 5.8 to 30.9,and those aged≥60 years ranged from 3.9 to 30.7.The reference intervals of female PGⅠ were 48.79-215.68μg/L,52.10-276.01μg/L,and 64.34-317.20μg/L,respectively.The reference intervals of female PgⅡwere:2.87-23.93μg/L,3.41-33.31μg/L and 3.88-39.16μg/L,respectively.The reference intervals of of female PGⅠ/PGⅡ are 6.6-28.1,5.2-27.9 and 3.6-26.2.Conclusion This study determined the reference range of serum PGs deficiency in healthy subjects of different sex and age in Zhejiang Province.

Objective     To evaluate the early clinical outcomes of the Renatus® balloon-expandable valve in the treatment of severe aortic stenosis. Methods    From November 2021 to April 2022, a total of 38 patients who received Renatus® balloon-expandable valve for severe aortic stenosis in Guangdong Provincial People's Hospital were included. There were 22 males and 16 females, with an average age of 73.7±5.3 years. Mean aortic gradient and peak aortic jet velocity at baseline, post-procedure, and follow-up were compared. Clinical outcomes including all-cause mortality, perivalvular leakage, serious adverse cardiovascular events and the occurrence of permanent pacemaker implantation were assessed. Results    All patients completed the procedure successfully without conversion to thoracotomy or perioperative death. The post-implant mean aortic pressure gradient was decreased from 41.5 (27.8, 58.8) mm Hg to 6.0 (3.0, 8.0) mm Hg, and the peak aortic jet velocity was also decreased from 4.1±0.9 m/s to 1.7±0.4 m/s (P<0.001). Pacemakers were required in 2 (5.3%) patients. The median follow-up time was 27.5 (23.0, 87.5) d, with a follow-up rate of 100.0%. The mean aortic gradient was 8.0 (7.0, 10.8) mm Hg and peak aortic jet velocity was 2.0±0.3 m/s, showing significant improvement compared with those in the preoperative period (P<0.001). No severe aortic regurgitation or paravalvular leak was observed. There was no serious cardiovascular adverse event or reoperative event during the study period. Conclusion    Transcatheter aortic valve replacement with the domestic Renatus® balloon-expandable valve system is a safe and effective procedure for selected patients with severe aortic stenosis who are at high risk or not candidates for surgical aortic valve replacement.

An 81-year-old male patient was admitted to Guangdong Provincial People's Hospital due to chest distress and shortness of breath after activity for half a year. Examination after admission revealed severe aortic insufficiency, tricuspid aortic valve and extremely horizontal aorta with an aortic root angulation of 99°. The Society of Thoracic Surgeons score was 7%. And taking the strong demand of the patient and his family into consideration, we decided to perform transapical transcatheter aortic valve replacement after multidisciplinary evaluation. The procedure was successfully performed by means of low deployment land zone and traction of pre-exist Prolene suture. Three-month follow-up confirmed the normal function of aortic prosthetic valve without residual regurgitation. This case provides a reference for the interventional treatment in patients with extremely horizontal aorta.

Objective    To determine the clinical efficacy of transapical transcatheter mitral valve-in-valve treatment for patients with deteriorated mitral bioprosthesis after aortic-mitral double valve replacement. Methods    The clinical data of 9 patients who underwent transapical transcatheter mitral valve-in-valve implantation after aortic-mitral double valve replacement due to mitral bioprosthesis deterioration from May 2020 to January 2021 in our hospital were retrospectively analyzed, including 4 males and 5 females with a mean age of 72.44±7.57 years. Results    Surgeries were performed successfully in all patients with no conversion to median sternotomy. The mean procedural time was 101.33±48.49 min, the mechanical ventilation time was 23.11±26.54 h, the ICU stay was 1.89±1.05 d and the postoperative hospital stay was 6.11±2.02 d. Residual mild mitral regurgitation was only observed in 1 patient. Only 1 patient needed postoperative blood transfusion. No major complications were observed in all patients. There was no death in postoperative 90 days. Conclusion    For patients with deteriorated mitral bioprosthesis after aortic-mitral double valve replacement, transapical transcatheter mitral valve-in-valve implantation achieves good clinical results and effectively  improves the hemodynamics without increasing the risk of postoperative left ventricular outflow tract obstruction. The surgery is feasible and effective.

Objective:To investigate the role of peptidyl arginine deiminase 4 (PAD4) in anti-β 2GP1/β 2GP1 complex-induced formation of neutrophil extracellular trapping networks (NETs). Methods:Peripheral blood neutrophils were isolated from healthy humans by density gradient centrifugation. PAD4 expression was detected by Western blot after the neutrophils were incubated with anti-β 2GP1/β 2GP1 complex (100 μg/ml) for a certain period of time. PAD4 inhibitor Cl-amidine (10 μmol/L) was used to pretreat neutrophils. Changes in the expression of citrullinated histone 3 (CitH3) at protein level and the relative content of myeloperoxidase (MPO)-DNA were detected by Western blot and ELISA, respectively. A mouse thrombus model of antiphospholipid syndrome (APS) was established by inferior vena cava stenosis. Intervention experiments were performed by intraperitoneal injection of Cl-amidine (50 mg/kg). The expression of CitH3 at protein level in plasma was detected by Western blot. The concentration of circulating free DNA (cf-DNA) in plasma was measured with fluorescent staining. Thrombus in inferior vena cava was collected and weighted to evaluate whether inhibiting the activity of PAD4 would suppress the APS-IgG-induced formation of NETs and thrombosis. Differences among groups were analyzed by t test or one-way analysis of variance (ANOVA). Results:The expression of PAD4 induced by anti-β 2GP1/β 2GP1 complex was significantly down-regulated in the cytoplasm, but increased in the nucleus [(3.67±0.32) vs (1.47±0.19), t=10.22, P<0.05; (0.57±0.19) vs (2.97±0.31), t=11.49, P<0.05]. Cl-amidine significantly inhibited the anti-β 2GP1/β 2GP1 complex-induced expression of CitH3 protein by neutrophils [(2.46±0.47) vs (0.46±0.13), t=12.24, P<0.01], and reduced the MPO-DNA content in the culture supernatants [(4.09±0.94) vs (2.80±0.57), t=4.23, P<0.05]. In vivo, Cl-amidine significantly inhibited the expression of CitH3 protein [(3.97±0.56) vs (1.09±0.45), t=11.83, P<0.01] and decreased the content of cf-DNA [(2 685.0±735.8) vs (1 784.0±577.0), t=3.93, P<0.05] in plasma of APS mice. Compared with the experimental APS mice in the control group, the weight of thrombus in the APS mice pretreated with Cl-amidine was significantly reduced [(8.22±3.06) vs (4.89±1.90), t=2.27, P<0.05]. Conclusions:PAD4 was involved in the formation of NETs induced by anti-β 2GP1/β 2GP1 complex, which might play an important role in APS thrombosis.

Objective: To analyze the plague monitoring results in Ulanqab City of Inner Mongolia in 2018, to master the changes in rat density and the prevalence of plague in rats, and provide a basis for scientific prevention and control of plague. Methods: According to "The Plague Monitoring Scheme of Inner Mongolia", we surveyed Siziwang Banner, Chahar Right Back Banner, Huade County, and Shangdu County of Ulanqab City from April to November 2018 to monitor the plague. Rat density was surveyed using a one-day bow clamp method; small rodent was surveyed using a 5 m clamping method. Rodents were obtained by sample method, 5 m clamping method, daily method, collecting dead animals and the like, and fleas were picked up from the captured rats and rat nest. The rodents and fleas were carried out pathogen detection, the serum of rodents was tested by indirect hemagglutination test. Laboratory test results were analyzed based on the "Diagnostic Criteria for Plague" (WS 279-2008). Results: Totally 1 463 mice were captured overlapping a monitored area of 416 hm², the average rat density was 3.52 per hectare; the number of Meriones unguiculatus was 1 235, and the rat density was 2.97 per hectare. Totally 1 603 mice were grooming, 404 mice with fleas, the flea infected rate was 25.20%, the number of fleas were 1 348, and the flea index was 0.84. A total of 22 mouse nests were dug, 17 nests with fleas, the flea infected rate was 77.27%, the number of fleas were 131, and the flea index was 5.95. Totally 1 603 rodents were checked by etiology, the results showed that 7 plague rats were all Meriones unguiculatus. Totally 1 479 fleas of 581 groups were cultured, 14 fleas of 5 groups were detected, 3 fleas of 1 group in Siziwang Banner, and 11 fleas of 4 groups in Huade County. Totally 243 samples of murine animal serum were tested and the results were all negative. Conclusions The epidemic of plague in Ulanqab City is in an active state, so monitoring should be strengthened in this area to prevent the prevalence of human plague.

Objective To detect the levels of neutrophil extracellular traps (NETs) in patients with antiphospholipid syndrome (APS) and to preliminarily explore its formation mechanism.Methods Plasma samples from 27 APS patients and 30 healthy controls were collected.The circulating free DNA (cf-DNA) in plasma was detected by the PicoGreen nucleic acid quantitative assay kit,and the concentration of citrulline histone 3 (CitH3) was analyzed by enzyme-linked immuno sorbent assay (ELISA).The association of cf-DNA/NETs with thrombotic events in APS patients was further analyzed.The neutrophils in healthy controls were separated by density gradient centrifugation and stimulated with anti-β2GPl/β2GPI complex (100 μg/mL) for 4 h,and the cf-DNA/NETs in the culture supernatant was determined.TLR-4 inhibitor-TAK242 (5 μmol/L) was further used to observe whether the stimulation of the anti-β2GPI/β2GPI complex on cells could be intervened.The differences between groups were analyzed by analysis of variance (ANOVA) or rank sum test,Sidak or Dunnett's test were used to compare the mean of multiple samples and the correlation between variables was analyzed by Spearman's correlation test.Results The concentration of cf-DNA/NETs and CitH3 were significantly increased in plasma of APS patients compared with that in healthy controls [175.7(70.6,205.7) ng/ml vs 29.8(7.6,115.7) ng/ml,Z=-3.654,P＜0.05;19.5(7.8,26.4) ng/ml vs 3.3(0.84,10.3) ng/ml,Z=-3.932,P＜0.05],and there was a significant positive correlation between the cf-DNA/NETs and CitH3 (r=0.447,P=0.019).In the APS group,there was no significant difference in cf-DNA/NETs between patients with arterial thrombosis and those with venous thrombosis [177.1(67.8,297.2) ng/ml vs 184.7(82.4,233.9) ng/ml,Z=-0.301,P=0.786],whereas cf-DNA/NETs in the patients who experienced a new thrombotic event in 1 month was significantly higher than those with a history of thrombosis [192.1(83.6,328.8) ng/ml vs 90.0(42.8,184.7) ng/ml,Z=-2.006,P=0.046].In vitro,anti-β2GPI/β2GPI complex (100 μg/ml) stimulated the release of cf-DNA/NETs from neutrophils,which was significantly increased compared with the control group (t=10.39,P＜0.05),while TAK242 significantly inhibited the stimulating effects of anti-β2GPI/β2GPI complex on cells (t=4.22,P＜0.05).Conclusion The level of cf-DNA/NETs in peripheral blood of APS patients is significantly increased,which may play an important role in APS thrombosis.Anti-β2GPI/β2GPI complex induces the formation of cf-DNA/NETs through TLR4 and participates in the pathological process of APS.

Objective To establish and assess the new method of APTT assay based on the combinations of Mg2+and Ca2+for lupus anticoagulants(LA)measurements.Methods This prospective study included 309 trisodium citrate anticoagulated plasma samples from 244 random patients and 65 patients with different autoimmune diseases(AID)to establish and assess the method of LA measurement, respectively.Final concentrations of 0,2.0, 4.0, 8.0,16.0 mmol/L Mg2+were added into 25 mmol/L Ca2+solution, and Actin reagent was used to measured plasma APTT of 94 patients.The applied concentration of Mg2+-Ca2+solution was confirmed through the special and significant alteration of APTT from LA-positive and -negative plasma observed in the presence of Mg 2+(test solution).Based on Actin reagent use,the test solution and 25 mmol/L Ca2+solution were applied to measure APTT of patients and normal individuals, respectively, and the ratio of Mg2+-Ca2+-APTT to Ca2+-APTT(Mg2+-Ca2+-APTT indices)and normalized Mg2+-Ca2+-APTT indices(NAR)were calculated, respectively.Mixed plasma NAR was measured,and CV%was calculated to evaluate the repeatability and stability of Mg 2+-Ca2+-APTT method.APTT of 150 patients was measured with the test solution and Actin reagent to calculate Mg 2+-Ca2+-APTT indices, and normalized LA ratio was determined with dRVVT method.The applicability of Mg2+-Ca2+-APTT assay was assessed through comparisons of the results from the two methods.Finally, NAR and NLR of 65 patients with AID(including 26 SLE patients)were measured with Mg2+-Ca2+-APTT assay and dRVVT method, respectively, and ROC curve was also used to assess the efficacy of the two methods for LA measurements.Results In all LA-negative plasma,APTT increased from 28.1 ±4.5 s to 61.2 ±7.9 s in normal APTT group,47.2 ±8.9 s to 97.5 ±10.3 s in increased APTT group,and 27.6 ± 5.1 s to 61.2 ±7.9 s in ACA-positive group when Mg2+increased from 0 to 8 mmol/L in Mg2+-Ca2+solution(F=34.12, 38.9 and 28.35,P<0.01).Following increased Mg2+concentration, APTT shortened from 0 to 4.0 mmol/L, but simultaneously prolonged from 4.0 to 16.0 mmol/L in LA-positive plasma with prolonged or normal APTT(F=31.55 and 39.51, P<0.01), and APTT was significantly higher in 8.0 mmol/L than that in 4.0 mmol/L(P<0.001).The test concentration of Mg 2+/Ca2+solution was 4.0 mmol/L.The within, inter-day CV% of NAR was 1.39%,2.30%, and 3.44%, respectively. According to the judging criteria of <0.966 and >1.034 of Mg2+/Ca2+indices, there was 141 patients with increased indices and NLR <1.20, and 9 patients with decreased ones and NLR≥1.20 in all 150 patients.The area under ROC curve of NAR and NLR for LA detection was 0.913(95%CI:0.848-0.978) and 0.892(95%CI:0.817-0.966), respectively, and the cut-off value was 0.87 and 1.13, respectively. The sensitivity and specificity of NAR(85% and 77%)was higher than that of NLR(81% and 74%), respectively.The accordant rate of positive,negative,and total results between NAR and NLR was 94.4%, 98.5%,and 98%,respectively.Conclusion The method of APTT assay based on Mg2+combining Ca2+for LA measurements is feasible,and can be used to detect plasma LA of patients.

Objective To study the value of deferred stent implantation in patients with high thrombus burden of acute ST-segment elevation myocardial infarction (STEMI). Methods Select 106 cases with a high thrombus burden within 12 hours of onset in patients with STEMI ,the infarct-related artery thrombus aspiration after antegrade flow of TIMI 2-3 and≤2 points of the thrombus aspiration(TS)patients were randomly divided into immediate stenting group(n = 43)and deferred stenting group(n = 40). Two groups of patients were compared with the myocardial blush grade(MBG),the incidence of slow-/no-reflow ,the incidence of compound endpoints in 6 months and the cardiac function after PCI for 6 months. Results After stenting,the MBG of deferred group was significantly higher than that of immediate group ,the incidence of slow-/no-reflow and the compound endpoints events within 6 months in deferred group was significantly lower than that in the immediate group. After PCI for 6 months,the improvement of LVEF in the deferred group was significantly higher than that in the immediate group, the left ventricular end diastolic dimension(LVEDD)in deferred group was significantly lower than that in immedi-ate group,and the differences were statistically significant(P < 0.05). Conclusions The high thrombus burden in patients with acute STEMI after thrombus aspiration ,deferred stent implantation can significantly reduce the rate of slow-/no-reflow ,improve myocardial perfusion ,reduce the incidence of compound endpoints events ,im-prove cardiac function in patients.

Objective To investigate the correlation between serum total bile acid level and coronary atherosclerosis. Methods The clinical data of 1408 patients who had underwent coronary angiography were retrospectively analyzed. The patients were divided into coronary atherosclerosis group (stenosis ≥ 50%, 681 cases) and coronary normal group (stenosis < 50%, 727 cases) according to the results of coronary angiography. The general clinical data, serum total bile acid, serum creatinine, fasting plasma glucose, low-density lipoprotein cholesterol (LDL-C) and so on were compared between 2 groups, and the indexes analyzed by Spearman correlation analysis and multivariate Logistic regression analysis. Results There were no significant differences between 2 groups in the sex constitution, the family history of hyperlipidemia and the history of lipid-lowering therapy (P>0.05). The rate of smoking, rate of hypertension, rate of diabetes, age, body mass index (BMI), creatinine, fasting plasma glucose, total bile acid and low density lipoprotein cholesterol in coronary atherosclerosis group were significantly higher than those in coronary normal group:18.6%(127/681) vs. 14.2%(103/727), 64.6%(440/681) vs. 45.8%(333/727), 48.5%(330/681) vs. 22.7%(165/727), (58.9 ± 12.2) years vs. (56.7 ± 13.1) years, (25.6 ± 4.3) kg/m2 vs. (24.9 ± 4.5) kg/m2, (70.28 ± 15.94)μmol/L vs. (52.79 ± 12.75)μmol/L, (6.82 ± 2.73) mmol/L vs. (5.57 ± 2.35) mmol/L, (7.86 ± 4.38)μmol/L vs. (5.63 ± 3.71)μmol/L and (3.32 ± 0.69) mmol/L vs. (2.28 ± 0.57) mmol/L, and there were statistical differences (P<0.05 or <0.01). The Spearman correlation analysis result showed that coronary atherosclerosis was positively correlated with men, age, diabetes, hypertension, BMI, serum creatinine and total bile acid (r=0.084, 0.068, 0.322, 0.263, 0.073, 0.248 and 0.176; P < 0.05 or < 0.01). The multivariate Logistic regression analysis result showed that men, diabetes, hypertension, serum creatinine, BMI ( >24 kg/m2) and total bile acid levels were risk factors of coronary atherosclerosis (P<0.05 or<0.01). Conclusions The serum total bile acid level is positively correlated with the severity of coronary atherosclerosis, which may be one of the independent risk factors for coronary atherosclerosis.