ObjectiveTo detect the effect of adeno-associated-virus induced Kringles5 gene on retinal neovascularization in rats with retinopathy of prematurity (ROP), and to explore the new ways of treatment for ROP.MethodspSNAV-Kringle5-gfp carrier was constructed by subclone and adeno-associated-virus was packed to form rAAV-Kringle5-gfp. ROP model was set up under circumstances of high oxygen in 21 SD rats which were divided into experimental (21 eyes) and control group (21 eyes). Eighteen eyes from each group was used to making the histologic section of retina, and the other 3 eyes in each group was detected by polymerase chain reaction (PCR) and Western blotting. There were 5 rats in the normal control group. AAV-Kringle5-gfp with the dosage of 10 μl and titer of 2.5×1012vg/ml was injected into the eyes in experimental group, while rAAVlacZ with the same dosage and titer of 2.5×1011vg/ml was injected in to the eyes in control group. The expression of target gene in ocular tissues was observed under the fluoroscope. Twelve weeks later, the rats were executed, and the staining of Ⅷ factor related antigens in retinal vascular endothelial cells was performed and number of nucleolus of vascular endothelial cells were counted. ResultsThe plasmid of pSNAV-Kringle5-gfp was correct according to the sequence measurement; the expression of rAAV-Kringle5-gfp was found in vitreous cavity and on retina; the expression of target gene was found on the level of mRNA and protein; the number of nucleolus of vascular endothelial cells on the surface of retina was (19.954 2±3.825 7) in experimental group and (7.335 2±2.731 3) in the control group, which had significant difference between the two groups (P＜0.01）.ConclusionsAdeno-associated-virus induced Kringles5 gene can inhibit the occurrence of retinal neovascularization in patients with ROP.(Chin J Ocul Fundus Dis, 2005,21:288-291)
Perioperative monitoring of blood coagulation is critical to better understand causes of hemorrhage, to guide hemostatic therapies, and to predict the risk of bleeding. Point-of-care (POC) coagulation monitoring devices assessing the viscoelastic properties of whole blood may overcome several limitations of routine coagulation tests in the perioperative setting. The advantage of these techniques is that they have the potential to measure the clotting process, starting with fibrin formation and continue through to clot retraction and fibrinolysis at the bedside, with minimal delays. Furthermore, the coagulation status of patients is assessed in whole blood, allowing the plasmatic coagulation system to interact with platelets and red blood cells, and thereby providing useful additional information on platelet function. Viscoelastic POC coagulation devices are increasingly being used in clinical practice, especially in the management of patients undergoing cardiac and liver surgery, assessment of hypo-and hypercoagulable states, guiding pro- and anticoagulant therapies, monitoring of antiplatelet therapy and procoagulant therapy. To ensure optimal accuracy and performance, standardized procedures for blood sampling and handling, strict quality controls and trained personnel are required.
ObjectiveTo evaluate the correlation of activated partial thromboplastin time (aPTT), activated clotting time (ACT) and the activity of anti-factor Ⅹa activity with the concentration of unfractionated heparin (UFH) during extracorporeal membrane oxygenation (ECMO) in children after cardiac surgery.MethodsThe clinical data of children (aged 6 months to 6 years) who received ECMO support after cardiac surgery in Fuwai Hospital from January 2010 to October 2020 were retrospectively collected. And the aPTT value, ACT value, anti-Ⅹa activity and the corresponding UFH dose measured simultaneously during ECMO were recorded. According to the Extracorporeal Life Support Organization anticoagulation guideline, the bleeding events of children during ECMO support were defined, and the children were divided into a bleeding group and a non-bleeding group according to whether bleeding events occurred. Pearson correlation was used to evaluate the correlation between ACT, aPTT or anti-Ⅹa activity and UFH in the same patient.ResultsA total of 58 children, including 33 males and 25 females, aged 27.31±34.17 months, were enrolled and divided into the bleeding group (n=39) and the non-bleeding group (n=19). Univariate analysis showed that compared with children in the non-bleeding group, children in the bleeding group had lower red blood cell counts (P=0.049), hemoglobin concentration (P=0.010), and hematocrit (P=0.046) on the day of ECMO installation. In addition, the transfusion volume of fresh frozen plasma (P=0.034) and fibrinogen (P=0.033) in the bleeding group was relatively more, and the proportion of exploratory thoracotomy for hemostasis was high (P=0.000); there was a moderate degree of correlation between anti-Ⅹa and UFH (r=0.418, P=0.013) but there was no correlation between ACT or aPTT and UFH.ConclusionThe aPTT value and ACT value are poorly correlated with the concentration of UFH transfused during ECMO in children after cardiac surgery, while the anti-Ⅹa activity is moderately correlated with it.
【Abstract】ObjectiveTo investigate the changes and significances of the activity of blood coagulation and fibrinolysis in hepatectomy patients accompanied with chronic hepatic disease. MethodsThirtyfive patients who were accompanied with cirrhosis undertook surgery in the second affiliated hospital of Chongqing Medicall University from year 2003 to 2004 were divided into two groups: the first group of 18 cases received hepatectomy and the second group received nonhepatectomy surgical treatment. The (prothrombin time PT), (activated partial thromboplatin time APTT), (thrombin time TT), and the content of (Fibrinogen Fbg) and (Ddimer DD) in the blood drawn from peripheral veins were quantitatively measured by a fullyautomatic chromogenic and immunological assay machine (ACLFutura 9000,USA) at the phases of before operation, right after operation and 24hour after operation, respectively. ResultsAPTT in hepatectomy group increased significantly (P＜0.01) and were much higher than the nonhepatectomy group at corresponding phases (P＜0.01). PT in hepatectomy group increased even more significantly compared with that of preoperation and right after the operation (P＜0.01). The differences of TT at varying phases in hepatectomy group were of no significance (Pgt;0.05). There was also no significant difference of PT, APTT, and TT in nonhepatectomy group at varying phases. ConclusionThe function of blood coagulation is relatively poor and the secondary activity of fibrinolysis is overactivated in hepatectomy patients accompanied with chronic hepatic disease, which indicates a high risk of hemorrhage.
Objective To evaluate the predictive value of lactate dehydrogenase (LDH) to albumin (Alb) ratio (LAR) in the prognosis of severe pneumonia patients complicated with DIC. Methods A total of 312 patients with severe pneumonia hospitalized in the intensive care unit (ICU) of the Affiliated Changzhou No.2 People's Hospital with Nanjing Medical University from January 1, 2018 to March 1, 2023 were retrospectively collected. The clinical parameters, such as gender, age, underlying diseases, and lactate dehydrogenase, albumin etc. l of the first test on admission were collected. LAR, sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) within 24 hours were calculated. The firstly endpoint of the study was the incidence of disseminated intravascular coagulation (DIC), the secondary endpoint was the 30-day in-hospital mortality in severe pneumonia patients with DIC. Univariate and multivariate logistic regression were used to analyze the risk factors of severe pneumonia with DIC. The receiver operating characteristic curve (ROC curve) was drawn and the area under the ROC curve (AUC) was calculated to evaluate the predictive value of LAR for the incidence of DIC in patients with severe pneumonia. Results The level of LAR was higher in the severe pneumonia patients with DIC than the severe pneumonia patients without DIC [LAR median ratio 12.72 (8.72, 21.89) vs. 7.23 (5.63, 10.90), P<0.001]. Multiple logistic regression analysis showed that LAR [OR=1.071, 95%CI 1.038 - 1.106, P<0.001] was the independent risk factor of the incidence of DIC in the patients with severe pneumonia. ROC curve analysis showed that the AUC for LAR to predict the incidence of DIC was 0.723, 95%CI 0.650 - 0.796, P<0.001. When the LAR cut-off value was 8.08, the sensitivity was 79.7% and the specificity was 56.1%. Kaplan-Meier survival analysis curve showed that the patients in the above LAR cut-off value group had a significantly lower 30-day survival rate than those in the below LAR cut-off value group (P<0.001). In the subgroup analysis and numerical variable transformed analysis, LAR was still the risk factor of DIC. Conclusion The increased LAR is a high risk factor of the incidence of DIC and mortality in patients with severe pneumonia, which is useful for predicting prognosis of patients with severe pneumonia.
目的：了解左旋門冬酰胺酶（L-ASP）對兒童急性淋巴細胞白血病凝血功能變化的影響。方法：觀察86例患兒在誘導緩解后治療期間，L-ASP使用前后活化部分凝血活酶時間(APTT)、凝血酶原時間(PT)、凝血酶時間(TT)、纖維蛋白原(FIB)、抗凝血酶Ⅲ(AT-Ⅲ)、D-二聚體變化情況。結果：與用藥前比,用藥結束后一天的PT、APTT、TT均顯著延長(P＜0.01);FIB、AT-Ⅲ顯著降低(P＜0.01),D-二聚體顯著升高(P＜0.01);用藥結束后1周時PT、APTT、TT、D-二聚體較用藥前差異無顯著性,FIB、AT-Ⅲ雖有回升,但仍低于正常(P＜0.01)。結論：L-ASP可引起ALL患兒凝血功能異常，尤其對FIB、AT-Ⅲ影響明顯，應引起臨床高度重視。Ｌ-Ａｓｐ主要影響蛋白質的合成而引起蛋白質成份的凝血因子減少,從而引起凝血功能障礙,且對纖維蛋白原的合成影響更為顯著。
ObjectiveTo explore the use of agkistrodon halys antivenin, and the influence of its infusion time on the coagulation function of the patient bitten by agkistrodon halys. MethodsWe retrospectively analyzed the clinical data of patients suffering from pit viper bites and first diagnosed and treated in the emergency department of our hospital between April 1 and November 30, 2013. According to the allergy test results, patients were divided into two groups: negative and positive. Based on the infusion time, the negative patients were divided into ≤1.5 hours and >1.5 hours groups, and the positive patients were divided into ≤3 hours and >3 hours groups. All patients' gender, age, infusion time, and PT, APTT, TT, FIB, D-DIMER before and after infusion of antivenomous serum were recorded, and blood coagulation indicators before and after infusion of antivenomous serum and the impact of infusion time were compared among different groups. ResultsFor both the negative and positive groups, PT, APTT, TT, FIB, and D-DIMER were statistically improved after infusion of antivenomous serum. The blood coagulation indicators of infusion time ≤1.5 hours group and ≤3 hours group were significantly better than those of infusion time >1.5 hours and >3 hours groups. ConclusionAntivenomous serum can correct coagulation and the faster infusion rate, the more obvious the effect is.
ObjectiveTo explore the predictive value of serum prothrombin induced by vitamin K absence-Ⅱ (PIVKA-Ⅱ) detection for the biological characteristics of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC).MethodsThis retrospective study included 394 patients with HBV-related HCC who were newly diagnosed and treated with surgical resection in West China Hospital of Sichuan University between June 2017 and December 2018. Their clinical information such as tumor size, tumor number, tumor cell differentiation, presence of microvascular invasion (MVI), distant metastasis, and portal vein tumor thrombus was collected from the medical record. The laboratory test results of patients during diagnosis and before surgery were collected, including alpha-fetoprotein (AFP), PIVKA-Ⅱ, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (γ-GGT), etc., and the relationships between PIVKA-Ⅱ levels and tumor biological characteristics were analyzed. Non-normal continuous variables were presented as medium (lower quartile, upper quartile).ResultsCompared with the patients with low HCC serum PIVKA-Ⅱ levels (≤40 mAU/mL), patients with high serum PIVKA-Ⅱ levels (>40 mAU/mL) had larger tumor diameters [5.00 (3.00, 9.00) vs. 2.50 (1.63, 4.95) cm, P<0.001], more severe Barcelona Clinic Liver Cancer (BCLC) stage (P<0.001), and higher AFP [186.05 (6.86, 1 210.00) vs. 17.83 (4.33, 231.95) ng/mL, P<0.001], ALT [38.00 (26.00, 66.25) vs. 32.00 (22.00, 51.00) U/L, P=0.018], AST [42.00 (30.00, 76.00) vs. 34.00 (25.50, 48.25) U/L, P<0.001], and γ-GGT [71.00 (39.00, 165.50) vs. 55.50 (25.00, 93.00) U/L, P=0.005], and were more likely to form portal vein tumor thrombi (16.61% vs. 3.75%, P=0.003) and MVI (43.67% vs. 11.11%, P<0.001). In BCLC stage 0 HCC patients, the positive rate of PIVKA-Ⅱ was only 51.35%. Multivariate logistic regression analysis showed that PIVKA-Ⅱ>40 mAU/mL was an independent predictor of MVI [odds ratio=6.588, 95% confidence interval (CI) (1.645, 26.383), P=0.008]. The area under receiver operating characteristic curve of PIVKA-Ⅱ level predicting MVI was 0.761 [95%CI (0.693, 0.830)], with a sensitivity of 66.22% and a specificity of 79.06%.ConclusionIn HBV-related HCC patients, high PIVKA-Ⅱ is associated with the poor biological characteristics of tumor, and is an independent risk factor for tumor MVI.
ObjectiveTo investigate the effects of thrombospondin-1 active fragment (TSP-1) synthetical peptide VR-10 on proliferation and migration of rhesus choroidal-retinal endothelial (RF/6A) cell and the expressions of apoptosis relative genes in RF/6A cell. MethodsThe survival rate of RF/6A cell were detected by methyl thiazolyl tetrazolium, and migration ability was measured by transwell chamber after exposure to 1.0 μg/ml TSP-1 and synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml) for different times (6, 12, 24, 48 hours). Caspase-3 and factor associated suicide (FAS) protein levels were measured by Western blot. The mRNA level of bcl-2 and FAS ligand (FASL) were measured by reverse transcription-polymerase chain reaction (RT-PCR). ResultsThe survival rate of RF/6A cells was determined by the treatment time and concentration of TSP-1(1.0 μg/ml) and the synthetic peptide VR-10 (0.1, 1.0, 10.0 μg/ml). The lowest survival ratio of RF/6A was 78% (P < 0.001) when cells were treated by 10 μg/ml synthetic peptide VR-10 after 48 hours. TSP-1 and synthetic peptide VR-10 could inhibit migration of RF/6A cells in transwell chamber (P < 0.001). 10.0 μg/ml synthetic peptide VR-10 had the strongest effect, 1.0 μg/ml TSP-1 was the next. Migration inhibition rate was increase with the increase of the concentration of VR-10 (P < 0.001). There was no significant differences between 0.1 μg/ml and 1.0 μg/ml VR-10 (P=0.114). Western bolt showed that RF/6A cell in control group mainly expressed the 32×103 procaspase-3 forms. To 10.0 μg/ml VR-10 treated group, it showed decreased expression of procaspase-3 (32×103) and concomitant increased expression of its shorter proapoptotic forms (20×103). Compared with control group, expression of FAS peptides were significantly increased in 10.0 μg/ml VR-10 treated group. Compared with control group, expression of FasL mRNA was significantly increased in 10.0 μg/ml VR-10 treated group(t=39.365, P=0.001), but the expression of bcl-2 mRNA was decreased(t=-67.419, P=0.000). ConclusionTSP-1 and synthetic peptide VR-10 had the ability to inhibit proliferation and migration of endothelial cell, and also induce apoptosis by increasing FAS/FASL expression and repressing bcl-2 expression.
ObjectiveTo investigate the effects of intraoperative intermittent pneumatic compression (IPC) treatment on coagulation and fibrinolysis, lower extremity venous blood flow velocity, and deep venous thrombosis (DVT) in patients with gastric cancer (GC) after radical gastrectomy. MethodsThe patients with GC who underwent radical gastrectomy at Hebei Provincial People’s Hospital from July 2021 to July 2024 were retrospectively enrolled, then the patients were assigned into control group (the patients who did not receive intraoperative IPC) and study group (the patients who received intraoperative IPC), and the propensity score matching (PSM) method was used to conduct 1∶1 matching based on the basic characteristics such as age, gender, body mass index, comorbidities to ensure baseline comparability between the two groups. The incidences of postoperative DVT and lower extremity swelling, and coagulations [prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT)] and fibrinolysis [D-dimer (D-D), fibrinogen (FIB), and fibrin degradation products (FDP)], as well as lower extremity venous blood flow velocity were analyzed after PSM. The locally weighted regression was used to analyze the correlation between the coagulation and fibrinolytic functions indexes and the lower extremity venous blood flow velocity. ResultsA total of 120 patients were matched (60 cases per group). The baseline characteristics of both groups were comparable (P>0.05). The incidences of DVT on day 7 and lower extremity swelling on day 1, 3, and 7 after surgery in the study group were significantly lower than those in the control group (P<0.05). The results of repeated-measures analysis of variance showed that there were statistically significant differences in the inter-group, time-related, and group-by-time interaction effects of coagulation and fibrinolysis indexes as well as lower extrimety venous blood flow velocities (P<0.05). The impact of the time factor on coagulation and fibrinolysis indexes and lower extrimety venous blood flow velocities varied with intraoperative IPC intervention measures. After surgery, the PT, TT, APTT, and lower extrimety venous blood flow velocities in the study group first decreased and then increased as compared with those before surgery, and the decrease degree was smaller and the increase degree was larger than those in the control group. After surgery, the D-D, FIB, and FDP in the study group first increased and then decreased as compared with those before surgery, and the increase degree was smaller and the decrease degree was larger than those in the control group. Both PT and TT were significantly positively correlated with femoral vein blood flow velocity (r=0.21, P=0.042; r=0.22, P=0.040), and both also showed significant positive correlations with popliteal vein blood flow velocity (r=0.25, P＜0.001; r=0.20, P=0.032). APTT was only significantly positively correlated with popliteal vein blood flow velocity (r=0.33, P＜0.001). D-D was negatively correlated with the flow velocities of the femoral vein, external iliac vein, and popliteal vein (r=–0.23, P=0.012; r=–0.22, P=0.047; r=–0.37, P<0.001). Both FIB and FDP were negatively correlated with the flow velocity of the femoral vein (r=–0.23, P=0.036; r=–0.27, P=0.002). FIB was also negatively correlated with the flow velocity of the popliteal vein (r=–0.26, P=0.038), and FDP was negatively correlated with the flow velocity of the external iliac vein (r=–0.31, P<0.001). ConclusionBased on the results of this study, intraoperative IPC treatment could improve coagulation and fibrinolytic functions of patients with GC after surgery, and has a certain preventive effect on occurrence DVT of lower extremity.