Objective To explore the feasibility of high-pressure injection to transfer human thrombomodulin (hTM) gene into arterial wall of rabbits.Methods Eighty-four healthy New Zealand rabbits were randomly divided into three groups: pcDNA3.1/hTM plasmid group (n=28), pcDNA3.1(+)/neo plasmid group (n=28) and untransfected group (n=28). After gene transfection, the model of arterial injury-blocking was established. Then, the expressions of hTM mRNA and protein in arterial wall were examined by RT-PCR and immunohistochemistry at 3 d, 7 d, 14 d and 28 d after operation. Results Seventeen rabbits died accidentally from the day of operation to 3 d after operation. The expressions of hTM mRNA of different time points in pcDNA3.1/hTM plasmid group were significantly higher than that in pcDNA3.1(+)/neo plasmid group and untransfected group (Plt;0.01). For the expressions of hTM mRNA at different time points in pcDNA3.1(+)/neo plasmid group and untransfected group, the difference of inter-group and intra-group was not significant (Pgt;0.05). hTM protein was expressed in every group and mainly localized in the inner lining of arterial wall. The expressions of hTM protein at different time points in pcDNA3.1/hTM plasmid group were significantly higher than that in pcDNA3.1(+)/neo plasmid group and untransfected group (Plt;0.05). The expression of hTM protein at different time points in pcDNA3.1(+)/neo plasmid group and untransfected group kept relative constancy, the difference of inter-group and intra-group was also not significant (Pgt;0.05). Conclusion High-pressure injection is feasible to transfer pcDNA3.1/hTM plasmid into arterial wall of live animals.
ObjectiveTo systematically review clinical value of des-γ-carboxy prothrombin (DCP) in the diagnostic of primary hepatocellular carcinoma (PHC).MethodsDatabases including PubMed, The Cochrane Library, EMbase, Medline (Ovid), CNKI, VIP, WanFang Data and CBM were electronically searched to collect relevant studies on DCP in the diagnosis of PHC from inception to December 31st, 2018. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then, meta-analysis was performed by using Meta-Disc 1.4 software and RevMan 5.3 software.ResultsA total of 50 studies involving 15 099 cases were included. The results of meta-analysis showed that the pooled sensitivity, pooled specificity, pooled positive likelihood ratio, pooled negative likelihood ratio, pooled diagnostic odds ratio and area under the curve of SROC were 0.69 (95%CI 0.67 to 0.70), 0.89 (95%CI 0.89 to 0.90), 7.35 (95%CI 6.08 to 8.90), 0.31 (95%CI 0.27 to 0.35), 26.63 (95%CI 20.42 to 34.73) and 0.909 9, respectively.ConclusionsSerum DCP has higher diagnostic efficacy for PHC, especially with higher specificity of diagnosis. Due to the limited quality and quantity of included studies, the above results should be validated by more studies.
Objective To systematically review the association between prothrombin gene G20210A mutation and the risk of cerebral venous thrombosis (CVT). Methods Databases including PubMed, Springer, Google Scholar, The Cochrane Library (Issue 1, 2016), CNKI, WanFang Data and CBM were searched for case-control studies concerning the association between prothrombin gene G20210A mutation and cerebral venous thrombosis risk from inception to January 2016. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then, meta-analysis was performed using RevMan 5.3 software and Stata 12.0 software. Results A total of 26 case-control studies were included, involving 1 361 CVT cases and 6 323 controls. The results of meta-analysis showed that: there was a significant association between prothrombin gene G20210A mutation and CVT risk (OR=4.56, 95% CI 3.51 to 5.93,P<0.000 01). Sensitivity analysis showed no significant publication bias was detected confirmed the stability of results. Subgroup analysis showed that G20210A mutation increased CVT risk in adults (OR=5.02, 95% CI 3.81 to 6.60,P<0.000 01), but not in children (OR=1.99, 95% CI 0.83 to 4.79,P=0.12). Conclusion Prothrombin gene G20210A mutation can significantly increase the CVT risk. Due to the limited quality and quantity of included studies, the above results are needed to be validated by more high quality studies.
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 effect of Rex surgery (superior mesenteric vein-left portal vein shunt) with internal jugular vein bypass on the anticoagulant factors and portal pressure in children with extrahepatic portal vein obstruction (EHPVO).MethodsFrom January 2014 to December 2018, children with EHPVO in Xi’an Children’s Hospital were retrospectively analyzed. All children underwent Rex surgery. The anticoagulant factors, blood routine indicators, and portal pressure-related indicators of all children were tested before and 1 year after Rex surgery, and the differences were compared. ResultsA total of 32 children were enrolled, and all children were followed up for 1 year after Rex surgery, and no follow-up was lost. Follow-up ultrasound examination 1 year after surgery showed that the portal vein blood flow in all children was unobstructed, and there was no venous thrombosis. The concentration of protein C, protein S and antithrombin Ⅲ activity of the children 1 year after surgery [(5.91±0.67) μg/mL, (2.43±0.34) μg/mL and (59.64±4.54)%, respectively] were all higher than those before surgery [(3.25±0.82) μg/mL, (2.02±0.37) μg/mL and (50.22±3.91)%, respectively], and the differences were statistically significant (P<0.05). There was no statistically significant difference in the concentration of antithrombin Ⅲ 1 year after surgery compared with that before surgery (P>0.05). The red blood cell count, hemoglobin concentration, white blood cell count and platelet count of the children 1 year after surgery [(4.61±0.17)×1012/L, (128.53±6.55) g/L, (6.09±0.72)×109/L and (104.88±5.74)×109/L, respectively] were all higher than those before surgery [(3.78±0.19)×1012/L, (105.53±5.31) g/L, (3.39±0.58)×109/L and (87.42±5.53)×109/L, respectively], and the differences were statistically significant (P<0.05). The diameter of the left portal vein 1 year after surgery was larger than that before surgery [(7.23±0.66) vs. (2.30±0.69) mm], the spleen volume was smaller than that before surgery [(55.74±4.07) vs. (67.21±4.22) cm3], and the portal vein pressure was lower than that before surgery [(23.37±1.27) vs. (35.29±1.36) cm H2O (1 cm H2O=0.098 kPa)], and the differences were statistically significant (P<0.05). ConclusionRex surgery with internal jugular vein bypass is beneficial to improving the level of anticoagulant factors in children with EHPVO, improving portal vein blood flow and pressure, and effectively relieving hypersplenism, which has a certain promotion value.
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 current situation of randomized controlled trials (RCTs) of thrombin like enzyme research and the ability to provide a reliable basis for the clinical practice. MethodsRCTs identified from four Chinese databases up to the year 2012 were assessed according to international standard, including SinoMed (1978-2012), CNKI (1979-2012), Wanfang Data (1986-2012), and VIP (1989-2012). ResultsA total of 2358 articles were searched and 53 RCTs were identified. The results showed that the quality of these articles was not high enough to meet the needs of clinical practice in China. ConclusionIn China, current quantity and quality of RCTs of thrombin like enzyme can not meet the need of clinical practice. In order to improve the prevention and treatment of hemorrhagic diseases, and surgical bleeding, especially for patients lacking clotting factor, more high-quality RCTs are required.