Objective:To establish a modified one-stage clotting assay (mOSA) based on the STA-R Evolution coagulation analyzer for quantifying emicizumab (EMI) concentration and to preliminarily evaluate its analytical performance; meanwhile to explore the clinical utility of the standard one-stage clotting assay (sOSA) in indirectly predicting EMI levels through surrogate factor Ⅷ (FⅧ) activity.Methods:A total of 30 pediatric patients with hemophilia A (HA) treated with EMI in the Hemophilia Treatment Center of Shenzhen Children′s Hospital from January 2023 to March 2025 were enrolled, and 48 post-treatment plasma samples were collected. EMI standards (2.5~100 μg/ml) were prepared using FⅧ-deficient plasma to establish the mOSA detection system. The linearity, accuracy, and precision of the method were evaluated. Surrogate FⅧ activity was measured by sOSA to estimate EMI concentrations, and its correlation with mOSA-derived EMI concentrations was analyzed using Spearman correlation analysis. The equivalent FⅧ activity in patient plasma samples was measured using a human chromogenic substrate assay-based FⅧ activity detection reagent, and Spearman correlation analysis was employed to evaluate its correlations with both the EMI concentrations measured by the mOSA method and estimated by the sOSA method respectively.Results:The established mOSA method for EMI detection showed excellent linearity in the range of 2.5?100 μg/ml ( Y=1.047 X?1.033, R 2=0.995, P<0.001). Average spike recovery rates at 25, 50, and 75 μg/ml were 101.55%(25.39/25.00), 105.31%(52.66/50.00), and 98.20%(73.65/75.00), respectively. Coefficients of variations of within-and inter-batch were 3.47%?4.80% and 6.30%?8.96%, respectively. A prediction model for EMI concentration was established as follows: estimated EMI concentration (μg/ml)=0.095×[alternative FⅧ activity (%) measured by sOSA]+2.652 ( R2=0.999, P<0.001). Validation demonstrated a strong correlation between the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method ( r=0.989, P<0.001), with good consistency ( Y=1.014 X+0.684, R2=0.972, P<0.001). Both the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method showed extremely strong correlations with the equivalent FⅧ activity ( r=0.986 and 0.987, respectively; P<0.001 for both). Conclusions:The mOSA system established on the STA-R Evolution analyzer demonstrates robust linearity, accuracy, and reproducibility, fulfilling clinical requirements for therapeutic drug monitoring of EMI. The sOSA method provides reliable indirect estimation of EMI concentrations through surrogate FⅧ activity, offering critical support for emergency decision-making.

Objective:To establish a modified one-stage clotting assay (mOSA) based on the STA-R Evolution coagulation analyzer for quantifying emicizumab (EMI) concentration and to preliminarily evaluate its analytical performance; meanwhile to explore the clinical utility of the standard one-stage clotting assay (sOSA) in indirectly predicting EMI levels through surrogate factor Ⅷ (FⅧ) activity.Methods:A total of 30 pediatric patients with hemophilia A (HA) treated with EMI in the Hemophilia Treatment Center of Shenzhen Children′s Hospital from January 2023 to March 2025 were enrolled, and 48 post-treatment plasma samples were collected. EMI standards (2.5~100 μg/ml) were prepared using FⅧ-deficient plasma to establish the mOSA detection system. The linearity, accuracy, and precision of the method were evaluated. Surrogate FⅧ activity was measured by sOSA to estimate EMI concentrations, and its correlation with mOSA-derived EMI concentrations was analyzed using Spearman correlation analysis. The equivalent FⅧ activity in patient plasma samples was measured using a human chromogenic substrate assay-based FⅧ activity detection reagent, and Spearman correlation analysis was employed to evaluate its correlations with both the EMI concentrations measured by the mOSA method and estimated by the sOSA method respectively.Results:The established mOSA method for EMI detection showed excellent linearity in the range of 2.5?100 μg/ml ( Y=1.047 X?1.033, R 2=0.995, P<0.001). Average spike recovery rates at 25, 50, and 75 μg/ml were 101.55%(25.39/25.00), 105.31%(52.66/50.00), and 98.20%(73.65/75.00), respectively. Coefficients of variations of within-and inter-batch were 3.47%?4.80% and 6.30%?8.96%, respectively. A prediction model for EMI concentration was established as follows: estimated EMI concentration (μg/ml)=0.095×[alternative FⅧ activity (%) measured by sOSA]+2.652 ( R2=0.999, P<0.001). Validation demonstrated a strong correlation between the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method ( r=0.989, P<0.001), with good consistency ( Y=1.014 X+0.684, R2=0.972, P<0.001). Both the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method showed extremely strong correlations with the equivalent FⅧ activity ( r=0.986 and 0.987, respectively; P<0.001 for both). Conclusions:The mOSA system established on the STA-R Evolution analyzer demonstrates robust linearity, accuracy, and reproducibility, fulfilling clinical requirements for therapeutic drug monitoring of EMI. The sOSA method provides reliable indirect estimation of EMI concentrations through surrogate FⅧ activity, offering critical support for emergency decision-making.

Objective:To observe the clinical efficacy of manual therapy based on the Discovery of Posture Secret (DPS) in treating adolescent idiopathic scoliosis (AIS).Methods:Thirty-six AIS patients were randomly divided into an observation group of 17 and a control group of 16. In addition to 10 minutes of side-shift training each week, the control group was given 20 minutes of sling exercise training (SET), while the observation group underwent weekly 20-minute manual therapy sessions based on DPS on the days without SET. Before and after the 4 weeks of treatment spine curvature (Cobb angle), clavicular angle (CA), angle of trunk rotation (ATR) and vertebral rotation (VR) were measured, and the Scoliosis Research Society′s patient questionnaire 22 (SRS-22) was administered.Results:After the treatment the average Cobb angle, CA, ATR and SRS-22 score of the observation group had improved significantly compared with before the treatment. The average Cobb angle, ATR and SRS-22 score of the control group were also significantly higher, but the improvements were not as great as in the observation group. There was no significant difference in VR improvement between the two groups.Conclusion:Manual therapy based on the DPS can effectively ameliorate the scoliosis and shoulder imbalance of AIS patients.

Objective:To evaluate the relationship between blood transfusion factors (massive transfusion, transfusion of stored red blood cells and anhepatic transfusion) and early postoperative outcomes in the patients undergoing orthotopic liver transplantation.Methods:Patients with end-stage liver disease who underwent orthotopic liver transplantation and intraoperative blood transfusion from January 2021 to March 2022 in our hospital were retrospectively selected, and their clinical data were collected. The patients were divided into massive transfusion group (M group, total red blood cell transfusion ≥10 U) and non-massive transfusion group (NM group) according to the amount of intraoperative blood transfused. The patients were divided into fresh blood group (NS group) and stored blood group (S group, red blood cell storage time > 2 weeks) according to the storage time of red blood cells transfused. The patients were divided into transfusion in anhepatic stage group (group T) and nontransfusion in anhepatic stage group (group NT) according to whether blood was transfused in the anhepatic stage. Multivariate logistic regression, generalized linear model and generalized linear mixed model were used to analyze the relationship between blood transfusion factors and primary outcome measures (postoperative pulmonary complications, circulation overload, acute kidney injury, abdominal infection, thrombosis) and between blood transfusion factors and secondary outcome measures (duration of ICU stay, postoperative hospitalization, postoperative temperature ≥38.5 ℃ at any time point after surgery, postoperative liver and kidney function index, coagulation function index, platelet count and preoperative difference).Results:One hundred and six patients were enrolled in this study. The results of multivariate logistic regression analysis: Massive blood transfusion and infusion of stored blood were risk factors for postoperative pulmonary complications, massive blood transfusion was a risk factor for abdominal infection, Child-Turcotte-Pugh score for liver function and time of anhepatic phase were risk factors for postoperative acute kidney injury, Child-Turcotte-Pugh score was a risk factor for circulatory overload, and age and massive blood transfusion were risk factors for thrombosis ( P<0.05). The results of generalized linear model analysis: Intraoperative stored blood transfusion and blood transfusion during the anhepatic phase were risk factors for prolonged duration of ICU stay, massive blood transfusion, intraoperative stored blood transfusion and blood transfusion during the anhepatic phase were risk factors for prolonged hospitalization ( P<0.05), and there was no correlation between massive blood transfusion, intraoperative stored blood transfusion and blood transfusion during the anhepatic phase and temperature ≥38.5 ℃ at any time point after surgery ( P>0.05). The results of generalized linear mixed model analysis: There was statistically significant difference in the difference between preoperative and postoperative liver and kidney function indexes, coagulation function indexes and platelet counts in the patients with or without massive transfusion, stored blood transfusion or blood transfusion during the anhepatic phase ( P<0.05). Conclusions:Massive blood transfusion and infusion of stored blood are the main risk factors for early adverse outcomes after orthotopic liver transplantation in the patients.

Animals ; Female ; Mice ; Mosaicism

Preoperative neoadjuvant chemoradiotherapy NCR) combined with total mesorectal excision (TME) is the standard treatment mode for locally advanced rectal cancer.Compared with postoperative NCR,preoperative NCR increases the tumor down-staging,sphincter-preserving rate and local control rate.Patients who attain pathological complete response (pCR) after preoperative NCR have better prognosis compared with their counterparts.This article reviews the research progress on preoperative NCR in recent years.

To evaluate effects of reinfusion of the remaining blood filtered by leukocyte depletion filter on postoperative cellular immune function after cardiopulmonary bypass (CPB). Methods Forty patients who underwent selective cardiac valve replacement surgery with CPB in department of anesthesiology of Haikou Municipal Hospital from January to June in 2018 were enrolled. All the patients were divided into the control group and experimental group according to the random number table method, with 20 patients in each group. In the experimental group, patients received residual pump blood transfusion which had been filtered by leukocyte depletion filter and stored in sterile blood collection bags. In the control group, patients received residual pump blood transfusion which was stored in sterile blood collection bags without being filtered. The remaining blood was reinfused after CPB in two groups. Blood samples were taken before CPB (T1), 2 hours following CPB (T2), and 1, 3, 5 days after reinfusion of the remaining blood (T3, T4, T5), the levels of T lymphocyte subsets CD3+, CD4+, CD8+ and natural killer cells (NK cells) were detected by flow cytometer, and CD4+/CD8+ ratio was calculated. The levels of plasma tumor necrosis factor-α (TNF-α), interleukins (IL-2, IL-6, IL-8) were measured by enzyme linked immunosorbent essay (ELISA). The duration of mechanical ventilation, the length of intensive care unit (ICU) stay, the length of hospital stay, and incidence of wound and pulmonary infection after surgery were compared between two groups. Results Among 40 patients, there were 22 males and 18 females; with an age of (47.88±12.29) years old; and with 25 cases of American Society of Anesthesiologists (ASA) physical status Ⅱ, and 15 cases of ASAⅢ. There was no statistical difference in the volume of the remaining blood between the two groups (mL: 959.00±116.84 vs. 971.50±115.68, P > 0.05). Compared with T1, the levels of T lymphocyte subsets CD3+, CD4+, CD8+, NK cells and plasma levels of IL-2 were significantly decreased from T2, the CD4+/CD8+ ratio was significantly decreased from T3 in two groups, but there was no statistical difference in CD3+, CD4+, CD8+, NK cells, CD4+/CD8+ ratio or plasma level of IL-2 at each time between the two groups. Compared with T1, the plasma levels of TNF-α, IL-6 and IL-8 were significantly increased at T2 in two groups and then decreased gradually. The plasma levels of TNF-α, IL-6 and IL-8 from T3 in experimental group were lower than those in control group [TNF-α (ng/L): 28.49±4.66 vs. 33.82±4.30, IL-6 (ng/L): 25.98±4.51 vs. 31.38±5.42, IL-8 (ng/L):38.98±4.67 vs. 45.76±5.33, all P < 0.05], they restored to the level of T1 at T5. In addition, compared with control group, the duration of mechanical ventilation, the length of ICU stay in experimental group were significantly decreased (hours: 8.07±1.30 vs. 9.16±1.52, 28.22±2.78 vs. 31.25±3.18, both P < 0.05), and there was no statistical difference in the length of hospital stay (days: 20.65±2.76 vs. 22.45±3.22), incidence of wound and pulmonary infection (25.0% vs. 15.0%, 5.0% vs. 15.0%) between the two groups (all P > 0.05). Conclusion Reinfusion of the remaining blood filtered by leukocyte depletion filtercan inhibit inflammatory responses and don't affect the function of cellular immunity, and don't increase the incidence of infection.

Objective To evaluate the lung protection of residual pump blood processed by microaggregate blood filter-reinfusion in patients undergoing cardiac surgery under cardiopulmonary bypass (CPB).Methods A total of 40 patients,aged 28-55 yr,weighing 46-66 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,with New York Heart Association of Ⅱ or Ⅲ,with left ventricular ejection fraction＞50％,scheduled for elective cardiac valve replacement under general anesthesia,were divided into 2 groups (n =20 each) using a random number table method:residual pump blood reinfusion group (group C) and residual pump blood processed by microaggregate blood filter-reinfusion group (group M).Residual pump blood was collected immediately after the end of CPB.The residual pump blood was stored in sterile blood collection bags without being filtered in group C.The residual pump blood was stored in sterile blood collection bags after being processed by microaggregte blood filter in group M.Residual pump blood was intravenously reinfused after the CPB pipe was removed.At 10 min before CPB (T1),immediately after the end ofCPB (T2),immediately after processing (T3) and at 12 and 24 h after residual pump blood reinfusion (T4,5),blood samples were collected to measure blood components and serum tumor necrosis factor-alpha and intedeukin-6 concentrations (by enzyme-linked immunosorbent assay).Airway plateau pressure was recorded,arterial blood samples were collected for blood gas analysis,and static lung compliance,oxygenation index and respiratory index were calculated at T1.5.The postoperative mechanical ventilation time,duration of intensive care unit stay,length of hospital stay,and incidence of hypoxemia and pulmonary infection were recorded.Results Compared with group C,white blood cell count was significantly decreased at T3-5,and static lung compliance and oxygenation index were increased,respiratory index was decreased,serum tumor necrosis factor-alpha and interleukin-6 concentrations were decreased,postoperative mechanical ventilation time and duration of intensive care unit stay were shortened,and the incidence of hypoxemia was decreased at T4.5 in group M (P＜0.05).Conclusion Residual pump blood proccessed by microaggregate blood filter-reinfusion can reduce systemic inflammatory responses and exerts lung protection to some extent in the patients undergoing cardiac surgery under CPB.

Objective To observe the changes of subfoveal choroidal thickness (SFCT) in diabetic patients with non-proliferative diabetic retinopathy (NPDR) and clinically significant macular edema(CSME),and then investigate diabetic SFCT and the relationship of diabetic CT with diabetic retinopathy (DR).Methods The patients were divided into 2 groups according to clinical guidelines of DR in China in 2014,including NPDR CSME + group(21 eyes) and NPDR CSME-group(36 eyes).All patients were underwent best corrected visual acuity (BCVA),intraocular pressure,axial length,slit lamp microscope,indirect ophthalmoscope,EDI-OCT examination,and mean arterial blood pressure measurement.The differences of BCVA,central retinal thickness (CRT) and SFCT between NPDR CSME + group and NPDR CSME-group were studied as well by means of using SPSS 18.0 for data statistics.Results There was no significant difference in gender,age,eye axis and intraocular pressure between NPDR CSME + group and NPDR CSME-group (all P ＞ 0.05).There was significant difference in BCVA between the two groups (P =0.001).The mean SFCT were (328.24 ± 101.92) μm in the NPDR CSME + group and (235.31 ± 66.98) μm in the NPDR CSME-group,and the difference was statistically significant (t =4.156,P=0.000).And plotting changes in CRT against changes in SFCT in patients with CSME revealed a positive correlation(r =0.473,P =0.000).Conclusion SFCT in NPDR CSME + patient is thicker than that in NPDR CSME-patient.And plotting changes in CRT against changes in SFCT in patients with CSME reveals a positive correlation.