This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

PURPOSE: Deep vein thrombosis (DVT) of the left and right lower extremities was treated in the same way, but the left and right extremities received different levels of attention. This study aimed to investigate the differences between the right and left lower extremity deep vein thrombosis (LEDVT). METHODS: Clinical characteristics of LEDVT patients from July 2020 to June 2022 were retrospectively analyzed to compare the incidence of LEDVT on different limbs, demographics, predisposing factors, and anatomical characteristics. The exclusion criteria were bilateral LEDVT and recurrent thrombosis. Measured data was analyzed using independent samples t-test or Mann-Whitney test. Count data were analyzed by Chi-square test. A p < 0.05 was considered a statistically significant difference. RESULTS: There were 478 patients included in this study and the ratio of left to right LEDVT on the left and right limbs was 3.16:1 (363:115). Left LEDVT predominantly affected female, with the major aged > 50 years (50 - 60 years: 16.80%; > 60 years: 57.30%). The primary predisposing factor was iliac vein compression syndrome, with iliofemoral thrombosis being the main type. Male patients with LEDVT on the right limb were predominant and the age of onset was usually ≤ 60 years (52.17%). The main predisposing factor was recent surgery or trauma (< 30 days) and femoropopliteal thrombosis was the main type. In more detail, the left iliac vein was compressed mainly in the proximal segment, and the right iliac vein was compressed mainly in the intermediate and distal segments. Recent surgery or trauma to the locomotor system and genitourinary system often induced LEDVT. CONCLUSION The incidence of LEDVT on the left is significantly higher than that on the right. LEDVT on different sides has different characteristics, which is crucial for prevention and diagnosis in the relevant population so there are also differences in treatment of the affected limbs.
Humans ; Venous Thrombosis/etiology* ; Male ; Female ; Middle Aged ; Lower Extremity/blood supply* ; Retrospective Studies ; Adult ; Aged ; Age Factors ; Sex Factors ; Risk Factors ; Acute Disease ; Incidence ; Aged, 80 and over ; Young Adult ; Adolescent

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

Objective To investigate the effect of astragaloside Ⅳ on high glucose-induced pyroptosis and invasive migration of human chorionic trophoblast cells(HTR-8/SVneo).Methods HTR-8/SVneo cells were divided into 4 groups:control group(untreated),high glucose group(high glucose stimulation)and astragaloside Ⅳ 50 and 100 μmol/L group(high glucose stimulation + astragaloside).Cell activity was detected by Cell Counting Kit 8(CCK-8),cell invasion and migration abilities were determined by Transwell assay and scratch assay,respectively,cell pyroptosis was assessed by Hoechst 33342/propidium iodide(PI)dual fluorescence staining.The protein expression levels of NOD-like receptor thermoprotein structural domain(NLRP3),cleaved-Caspase-1,GSDMD-NT,and IL-18 were detected by Western Blot.Results Compared with the control group,HTR-8/SVneo cell viability was significantly reduced in the high glucose group,the rate of cell migration was significantly reduced,the number of invasive cells was significantly reduced,the percentage of PI-positive cells was significantly increased,and the levels of NLRP3,cleaved-Caspase-1,GSDMD-NT and IL-18 protein expression levels were significantly increased(P<0.05 or P<0.01);compared with the high glucose group,cell viability was significantly higher in the astragaloside Ⅳ treated group,the rate of cell migration was significantly increased,the number of invasive cells was significantly increased,the percentage of PI-positive cells was significantly decreased,and the protein expressions of number of NLRP3,cleaved-Caspase-1,GSDMD-NT,IL-18 were significantly decreased(P<0.05 or P<0.01).Conclusion AstragalosideⅣcan inhibit high glucose-induced HTR-8/SVneo cell pyrolysis and improve cell invasion and migration ability.

AIM To establish the quality standard of the zhujieshen Formula Granules based on standard decoction.METHODS The contents and transfer rates of ginsenoside Ro and chikusetsu saponin Ⅳa in standard decoction and formula granules were determined by HPLC,after which the transfer rates were calculated.The HPLC characteristic chromatograms of standard decoctions were established,after which cluster analysis and principal component analysis were adopted.Then the HPLC characteristic chromatograms of formula granules were established.RESULTS Nine common peaks were found in the HPLC characteristic chromatograms of seventeen batches of standard decoctions with the similarities of more than 0.9(except for S6,S12),which were clustered into two categories,and the accumulative variance contribution rate of three principal components reached 86.7％.The contents of ginsenoside Ro in three batches of formula granules were 83.1-88.6 mg/g,and the transfer rates were 53.1％-55.5％.The contents of chikusetsusaponin Ⅳa were 14.8-15.0 mg/g,and the transfer rates were 47.4％-48.1％.Nine common peaks were found in the HPLC characteristic chromatograms of three batches of formula granules with the similarities of 0.998,0.998 and 0.999,respectively.CONCLUSION This reasonable and reliable method can comprehensively evaluate the quality of Zhujieshen Formula Granules,and provide a reference for the quality control.

Objective To investigate the effect of varying blood pressure stratification on renal function in the diabetic population.Methods A prospective cohort study was conducted,enrolling 9 489 diabetic patients from a total of 101 510 Kailuan Group employees who underwent health examinations between July 2006 and October 2007.The follow-up period was(8.6±4.0)years.Participants were categorized into four groups based on their baseline blood pressure levels:normal blood pressure(systolic blood pressure<120 mmHg and diastolic blood pressure<80 mmHg),elevated blood pressure(systolic blood pressure 120-130 mmHg and diastolic blood pressure<80 mmHg),stage 1 hypertension(systolic blood pressure 130-140 mmHg and/or diastolic blood pressure 80-90 mmHg),and stage 2 hypertension(systolic blood pressure≥140 mmHg and/or diastolic blood pressure≥90 mmHg).The incidence density of chronic kidney disease(CKD)was compared among these groups.A multivariate Cox proportional hazards regression model was employed to assess the effects of different blood pressure levels on renal function in diabetic patients,with the stability of the results confirmed using a multivariate time-dependent Cox proportional hazards model.Sensitivity analysis was conducted after excluding cases of cardiovascular disease(CVD)during follow-up,and cases using antihypertensive and antidiabetic medications at baseline.Results(1)At baseline,stage 1 hypertension patients demonstrated statistically significant higher differences with age and body mass index(BMI)compared to normal blood pressure group(P<0.05).(2)By the end of the follow-up,2 294 cases of CKD were identified,including 1 117 cases of estimated glomerular filtration rate(eGFR)decline and 1 575 cases of urinary protein.The incidences density of CKD,eGFR decline and urinary protein for stage 1 hypertension group were 39.4,16.3 and 25.5 per thousand person-years,respectively,all of which were statistically significant different from normal blood pressure group(log-rank test,P<0.01).(3)Multivariate Cox regression analysis revealed that,compared to the normal blood pressure group,stage 1 hypertension was associated with a 29%increased risk of CKD(HR=1.29,95%CI 1.09-1.52)and a 40%increased risk of eGFR decline(HR=1.40,95%CI 1.08-1.80)in diabetic individuals.Conclusion Stage 1 hypertension significantly increases the risk of CKD and eGFR decline in diabetic individuals,with a particularly notable effect on the risk of eGFR decline.

Objective To explore the efficacy of high tibial osteotomy(HTO)combined with medial meniscus centraliza-tion in knee osteoarthritis.Methods A total of 26 patients who underwent surgery from October 2018 to October 2020 were re-viewed.Among them,14 patients underwent high tibial osteotomy combined with arthroscopic meniscus centralization surgery were centralized group,including 8 males and 6 females,with an average age of(50.2±1.4)years old and follow-up time of(16.8±4.0)months.Twelve patients with high tibial osteotomy were in the control group,including 6 males and 6 females,with an average age of(50.9±1.8)years and follow-up time of(19.0±4.8)months.Operation time,the knee Lysholm score,knee 2000 IKDC score,MRI,femoral tibial angle(FTA),hip knee ankle angle(HKA),and intraoperative and postoperative compli-cations were recorded.Results All the incisions healed without any complication.The operation time in the centralized group was longer than that in the control group[(65.0±2.1)min vs(52.0±2.1)min,P＜0.05].The medial meniscus extrusion reduction value in the centralized group was significantly reduced compared with the control group[(2.8±1.4)mm vs(1.1±2.2)mm,P＜0.05].The FTA,HKA,knee Lyshlom score,and 2000 IKDC score between two groups were no significantly(P＞0.05).Postop-erative knee Lyshlom score and knee 2000 IKDC score improved in both groups(P＜0.05).Conclusion HTO combined with centralization of medial meniscus can improve the reduction of medial meniscus and improve knee function.The medium and long-term curative effect still needs long-term follow-up of more cases.

Objective To compare clinical efficacy of robot-assisted(RA)and remote sensing navigation alignment(RSNA)system-assisted total knee arthroplasty(TKA).Methods From March 2023 to June 2023,60 patients who underwent the first unilateral TKA due to severe knee osteoarthritis(KOA)were admitted and divided into RSNA group and RA group according to different treatment methods,with 30 patients in each group.There were 5 males and 25 females in RSNA group,aged from 56 to 81 years old with an average of(66.33±7.16)years old;body mass index(BM1)ranged from 19.87 to 38.54 kg·m-2 with an average of(28.40±6.18)kg·m-2;the courses of disease ranged from 5 to 36 months with an average of(18.20±8.98)months;RSNA system was used to assist the positioning of osteotomy.There were 7 males and 23 females in RA group,aged from 55 to 82 years old with an average of(67.83±8.61)years old;BMI ranged from 19.67 to 37.25 kg·m-2 with an aver-age of(28.01±4.89)kg·m-2;the courses of disease ranged from 3 to 33 months with an average of(17.93±9.20)months;RA was performed.Operation time,incision length,latent blood loss at 2 weeks after operation and incidence of lower extremity thrombosis were compared between two groups.Hip-knee ankle angle(HKAA),HKAA deviation,lateral distal femoral angle(LDFA),medial proximal tibial angle(MPTA)and posterior tibial slope(PTS)were compared between two groups;Western Ontario McMaster Universities Osteoarthritis Index(WOMAC)and Knee Society score(KSS)were used to evaluate functional recovery before operation,3 and 6 months after operation.Results The operation was performed successfully in both groups,and there were no serious complications such as vascular and nerve injury during operation.The wound healed well at stage Ⅰafter operation,and the follow-up time was 6 months.The operation time,latent blood loss at 2 weeks after operation and inci-sion length in RSNA group were(94.35±5.75)min,(130.54±17.53)mland(14.73±2.14)cm,respectively;while(102.57±6.88)min,(146.33±19.47)ml and(16.78±2.32)cm in RA group,respectively.RSNA group was better than RA group(P＜0.05).No deep vein thrombosis occurred in both groups at 2 weeks after operation,5 patients occurred intermuscular vein throm-bosisin in RSNA group and 8 patients in RA group,the difference was not statistically significant(P＞0.05).In RSNA group,HKAA,LDFA and MPTA were(173.00±5.54)°,(86.96±3.45)°,(82.79±3.35)° before operation,and(178.34±1.85)°,(89.92±0.42)°,(89.84±0.73)° at 1 week after operation,respectively.In RA group,HKAA,LDFA and MPTA were(173.31±6.48)°,(87.15±3.40)° and(82.99±3.05)° before operation,and(178.52±1.79)°,(90.03±0.39)° and(90.15±0.47)° at 1 week after operation,respectively.HKAA,LDFA and MPTA were significantly improved in both groups at 1 week after oper-ation(P＜0.05).There were no significant difference in HKAA,LDFA,MPTA and PTS between two groups before operation and 1 week after operation(P＞0.05).There was no significant difference in deviation distribution of HKAA at 1 week after op-eration(x2=2.61 1,P=0.456).There were no significant difference in WOMAC and KSS between two groups before operation,3 and 6 months after operation(P＞0.05),and postoperative WOMAC and KSS at 3 and 6 months between two groups were im-proved compared with those before operation(P＜0.05).Conclusion Both RA and RSNA system assisted TKA could obtain ac-curate osteotomy,RA has higher surgical accuracy,RSNA system assisted operation has less trauma,and operation is simpler.

Objective To explore clinical accuracy of remote sensing navigation alignment(RSNA)system in total knee arthroplasty(TKA)and its influence on postoperative clinical efficacy.Methods From May 2021 to May 2022,60 knee os-teoarthritis(KOA)patients with Kellgren-Lawrence(K-L)grade Ⅲ to Ⅳ treated by unilateral primary TKA were selected and divided into RSNA group and traditional operation group according to treatment methods,and 30 patients in each group.There were 6 males and 24 females in RSNA group,aged from 55 to 86 years old with an average of(68.06±8.23)years old;body mass index(BMI)ranged from 22.15 to 34.58 kg·m-2 with an average of(28.20±3.01)kg·m-2;the courses of disease ranged from 2 to 60 months with an average of(18.80±14.80)months;13 patients with grade Ⅲ and 17 patients with grade Ⅳaccording to K-L grading.In traditional operation group,there were 8 males and 22 females,aged from 57 to 85 years old with an average of(67.26±6.32)years old;BMI ranged from 23.94 to 34.55 kg·m-2 with an average of(27.49±2.32)kg·m-2;the courses of disease ranged from 3 to 60 months with an average of(21.30±16.44)months;14 patients with grade Ⅲ and 16 pa-tients with grade Ⅳ according to K-L grading.Western Ontario and McMaster Universities(WOMAC)osteoarthritis index and Knee Society score(KSS)were used to evaluate functional recovery of patients.Hip-knee-ankle angle(HKAA),distal femoral valgus angle(FVA)and distal fermoral flexion angle(DFFA)were measured before operation.HKAA and HKAA deviation angle were measured at 1 week after operation,and defective rate of lower limb force line,femur prosthesis valgus angle(FP-VA)and femoral prosthesis flexion angle(FPFA),respectively,were calculated.Results There were no serious complications such as vascular and nerve injury during operation,and wound healed at stage Ⅰ.Both groups were followed up for 6 months.There were no significant difference in WOMAC index,KSS,HKAA,FVA and DFFA between two groups before operation(P＞0.05).The force line defect rate,HKAA,HKAA deviation angle,FPVA deviation angle and FPFA of RSNA group were 6.7％,(178.74±1.56)°,(1.25±1.56)°,(1.84±0.16)° and(4.85±2.46)°,respectively;while in traditional operation group were 20％,(176.73±3.46)°,(3.27±3.46)°,(2.44±0.26)°,(6.60±1.86)°;the difference between two groups were statistically sig-nificant(P＜0.05).There were no significant difference in WOMAC index and KSS between two groups at 3 and 6 months after operation(P＞0.05).Conclusion RSNA system could reduce defective rate of lower limb force line,FPVA deviation angle and FPFA after TKA,which is more accurate and easy to operate than traditional intramedullary localization surgery while ensuring postoperative efficacy.

Objective:Of all spontaneous bleeding complications in patients with acute ST-elevation myocardial infarction (STEMI), upper gastrointestinal bleeding (UGIB) is the most common and of specific interest, because it can be prevented by several prophylactic measures. The purpose of this study was to investigate the in-hospital incidence, associated outcomes, and predictors of UGIB after STEMI.Methods:In this retrospective study, we analyzed the records of 2 791 patients with acute STEMI admitted to Beijing Anzhen Hospital Affiliated to Capital Medical University between January 2018 and January 2022. The patients were divided into the UGIB group ( n=61) and non-UGIB group ( n=2 730) according to the presence or absence of upper gastrointestinal hemorrhage, respectively. Baseline clinical conditions, coronary lesions, in-hospital deaths, and in-hospital adverse events were compared between the two groups. Logistic regression analysis was also performed for risk factors that could lead to UGIB. Results:The in-hospital incidence of UGIB after STEMI was 2.2% (61/2 791). Hospital stay was significantly longer in the UGIB group [8(6, 12) days vs. 5 (4, 7) days, Z=3.28, P<0.001] and in-hospital mortality was significantly higher in the UGIB group than in the non-UGIB group (9.8% vs. 0.8%, χ2=0.63, P=0.001). Patients with UGIB were significantly older than those without UGIB (63±11 years vs. 58±11 years, t=-3.75, P<0.001). The serum creatinine level of UGIB patients was significantly higher than that of non-UGIB patients [(80(62, 98) mmol/L vs. 73(64, 84) mmol/L, Z=1.68, P=0.007], the red blood cell count of UGIB patients was significantly lower than that of non-UGIB patients [4.1(3.8, 4.6)×10 12/L vs. 4.6(4.2, 4.9)×10 12/L, Z=2.61, P<0.001], and the hemoglobin concentration of UGIB patients was significantly lower than that of non-UGIB patients [129(109, 141) g/L vs. 141(130, 152) g/L, Z=2.52, P<0.001]. Brain natriuretic peptide levels were significantly higher in UGIB patients than in non-UGIB patients [331(165, 644) ng/L vs. 181(89, 333) ng/L, Z=2.42, P<0.001]. Logistic regression analysis showed that age ( OR=1.045, 95% CI 1.009-1.082, P=0.013); hemoglobin ( OR=1.594, 95% CI 1.150-2.210, P=0.005); hematocrit ( OR=0.181, 95% CI 0.060-0.546, P=0.002); and mean hemoglobin concentration ( OR=0.845, 95% CI 0.752-0.951, P=0.005) were independent risk factors for UGIB in patients with STEMI. Logistic regression analysis of risk factors for in-hospital death revealed that concurrent UGIB was an independent risk factor for in-hospital death in patients with STEMI ( OR=2.954, 95% CI 0.635-13.751, P=0.024). Conclusions:The incidence of in-hospital UGIB in STEMI patients was 2.2%, and the in-hospital mortality rate of STEMI complicated with UGIB increased to 9.8%. Concurrent UGIB was an independent risk factor for in-hospital death in patients with STEMI. The most important predictors of in-hospital UGIB in patients with STEMI were age, hemoglobin, hematocrit, and mean hemoglobin concentration.