Coagulopathy is very common in patients in intensive care unit (ICU) and often indicates organ dysfunction or underlying diseases.The application of traditional methods assessing the patients' coagulation status in ICU is limited because they can not reflect the whole process of coagulation.Thromboelastography (TEG),a point-of-care (POC) assay of coagulation,fibrinolysis and platelet function,developed in recent years has been widely used in organ transplant and cardiovascular surgery and so on.However,there is no standard for the use of TEG in ICU.The development and application of TEG in sepsis,multiple trauma,guiding blood transfusion,extracorporeal membrane oxygenation (ECMO),and anticoagulation monitoring were addressed in this review,and its value and application prospect in ICU were analyzed.

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Objective To investigate the pulmonary microvascular responsiveness of diabetic animals to sepsis and the potential mechanism of NO system.Methods Sixty-four Wistar rats of clean grade were randomly (random number) divided into 4 groups,namely normal control group (group A,n =16),diabetes group (group B,n =16),sepsis group (group C,n =16),diabetes and sepsis group (group D,n =16).Diabetic mellitus model was made in rats with injection of streptozotocin,STZ (65 mg/kg).Successful model was defined as the blood glucose value≥ 16.67 mmol/L 48 hours after injection of STZ.All animals were fed 4 weeks before initiation of next experiment.The sepsis model was established by intravenous injection of LPS (10 mg/kg) in rats.RT-PCR was used to determine the mRNA expression of Tie-2 in rats'blood.The ratio of dry/wet of lung tissue and the extravasation of Evans blue dye into the lung were detected.Quantitation of NO in lung tissue and serum was measured by using Griess method.RT-PCR was also used for determination of iNOS,eNOS,DDAH2 mRNA expressions in lung tissue.Data were analyzed with ANONA and LSD method for comparison between groups,and P ＜ 0.05 was considered statistically significant.Results Compared with septic group.,the diabetic rats with sepsis group demonstrated higher expression of Tie-2 mRNA in blood (19.72 ± 0.70) vs.(3.99 ± 0.92),P =0.00,lower ratio of dry/wet in lung tissue (0.19 ±0.01) vs.(0.22 ±0.01),P =0.000,higher permeability of Evans blue dye into lung tissue (3.76 ± 0.77) vs.(1.74 ± 0.24),P =0.000.Serum NO level was lower in group D than that in group C (123.13 ±4.24) vs.(188.30 ±5.18),P =0.000,however,NO levels in lung tissue of both group D and group C were higher than that in control group (53.62 ± 6.70),(23.63± 3.92) vs.(10.37 ± 1.29),P =0.00,and NO level in group D was higher in 2 times than that in group C (P =0.00).However,there were no differences in eNOS expression among groups A,B and C,but the difference in eNOS expression was present between group D with lower expression and group A,that lower in group D (0.07 ±0.02) vs.(0.38 ±0.05),P=0.017.Compared with group C,the expression of iNOS was higher in group D (80.23 ±2.49),(32.48±5.37) vs.(1.74±0.23),P=0.00),and the expression of DDAH2 was lower in group D (0.49 ±0.13),(7.26 ±0.50) vs.(11.96 ±0.55).Conclusions Diabetic rats with sepsis enhanced endothelial cell damages.Diabetes deteriorates the regulatory activity of NO system,suggesting the potential mechanism of the worsened damages of EC in diabetic sepsis host.

Objective To investigate whether heparin has a beneficial effect on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats,and to explore the possible underlying mechanisms. Methods Thirty-two adult Sprague-Dawley(SD)rats were randomly assigned into the control,heparin control,model,and heparin treatment groups,with 8 in each group. ALI rat model was reproduced by intratracheal instillation of LPS at a dose of 1 mg/kg. The rats in the control and heparin control groups received an equal volume of normal saline at the same times. The rats in the heparin control and heparin treatment groups were intravenously received 50 U/kg heparin every 1 hour after the induction of ALI. Animals were sacrificed 24 hours after LPS challenge. Bronchoalveolar lavage fluid(BALF) and lung tissue samples were collected. Histopathological evaluation,lung wet/dry(W/D)ratio,malondialdehyde (MDA),nitric oxide(NO)and myeloperoxidase(MPO)were analyzed. Enzyme-linked immunosorbent assay(ELISA) was used to measure the concentration of inflammatory factor in BALF. Expression of inducible nitric oxide synthase (iNOS)mRNA in the lung of rats was measured by reverse transcription-polymerase chain reaction(RT-PCR). Western Blot was used to determine the expression of transforming growth factor-β1(TGF-β1)and phosphorylation of Smad in the lung tissues. The expression of iNOS in lung was determined by immunohistochemistry. Results In the control and heparin control groups,lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group,ALI characters such as extensive thickening of the alveolar wall,significant infiltration of inflammatory cells,demolished structure of pulmonary alveoli,and hemorrhage were found. In the heparin treatment group,heparin treatment markedly alleviated LPS-induced these pathological changes in lung. Compared with control and heparin control groups,lung W/D ratio,lung MDA,NO and MPO levels,and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6)in BALF in the model group were increased significantly. Compared with the model group, lung W/D ratio,lung MDA,NO and MPO levels,and TNF-αand IL-6 in BALF in the heparin treatment group were significantly decreased〔W/D ratio:7.54±0.17 vs. 10.69±0.15,MDA(mmol/mg):2.01±0.30 vs. 2.51±0.25,NO (μmol/L):3.07±0.21 vs. 3.89±0.14,MPO(U/g):1.94±0.09 vs. 2.74±0.20,TNF-α(μg/L):201.80±0.27 vs. 297.53±0.34,IL-6(μg/L):38.41±0.25 vs. 46.31±0.31,all P<0.05〕. RT-PCR showed that the expression of iNOS mRNA in the heparin treatment group was significantly lower than that in the model group(2-ΔΔCt:3.04±0.18 vs. 4.37±0.15,P<0.05). Western Blot showed that compared with control group,the protein expressions of iNOS and TGF-β1,and phosphorylation of Smad2 and Smad3 were significantly increased,and the heparin could inhibit the protein expressions compared with model group. Immunohistochemistry showed that positive expressions of iNOS in alveolar epithelial cell and capillary endothelial cell in the heparin treatment group were significantly lower than those in the model group. Conclusion Heparin significantly ameliorated the lung injury induced by LPS in rats via the inhibition of nitric oxide synthase expression and the TGF-β/Smad pathway.

OBJECTIVE: To explore the correlation between thromboelastography (TEG) parameters and the risk of venous thromboembolism (VTE) and bleeding in patients receiving anticoagulant therapy in surgical intensive care unit (SICU). METHODS: 205 patients received low molecular weight heparin (LMWH) anticoagulant therapy admitted to SICU of Tianjin Hospital from December 2016 to December 2017 were consecutively enrolled. TEG detection was performed in all patients at 1 day after anticoagulation therapy, and coagulation reaction time (R value), blood clot generation time (K value), blood clot generation rate (α angle) and maximum width value (MA value) were recorded. At the same time, the traditional coagulation function test was carried out, and prothrombin time (PT), activated partial thromboplastin time (APTT) and D-dimer levels were also recorded. The incidence of deep venous thrombosis (DVT), pulmonary embolism (PE) and bleeding during hospitalization were observed. Multivariate Logistic regression analysis was used to analyze the risk factors for VTE and bleeding in patients receiving anticoagulant therapy. RESULTS: Of 205 patients, during the anticoagulant treatment, 14 patients developed DVT, and 4 patients with PE (2 of them were combined with DVT) with an incidence of 7.8% (16/205). There were 2 patients suffering from cerebral hemorrhage, 2 patients with gastric bleeding, and 1 patient with intra-tracheal hemorrhage with an incidence of 2.4% (5/205). Compared with the patients without VTE or bleeding, the R value of TEG in patients with VTE was significantly lowered (minutes: 4.6±2.2 vs. 7.4±1.4, P < 0.01), which was significantly increased in patients with hemorrhagic complications (minutes: 12.1±1.1 vs. 7.4±1.4, P < 0.01). There was no significant difference in the K value, α angle, MA value of TEG, or PT, APTT, D-dimer between the patients with and without VTE or bleeding. Multivariate Logistic regression analysis revealed that the R value of TEG was independent risk factor for incidence of VTE and hemorrhagic complication in SICU patients who receiving anticoagulation therapy [VTE: β = 0.386, odds ratio (OR) = 1.096, 95% confidence interval (95%CI) = 1.021-2.361, P = 0.006; hemorrhagic complication: β = -1.213, OR = 1.051, 95%CI = 1.017-3.458, P = 0.045]. CONCLUSIONS The R value of TEG is associated with the occurrence of VTE and hemorrhagic complications in patients receiving anticoagulant therapy in SICU.
Anticoagulants ; Critical Care ; Heparin, Low-Molecular-Weight ; Humans ; Pulmonary Embolism ; Risk Factors ; Thrombelastography ; Venous Thromboembolism

A detailed three-dimensional nonlinear finite element model of lumbar segment L3-L5 was developed to investigate the influence of vibration on the components of human spine. The results show that the vibration effects of different spinal components are not exactly the same, and the stress near the posterior region of L4-L5 annulus is higher than that of its anterior region. The vibration exerts a great influence on the facet joint of L4-L5 segment. The changing amplitudes of stress and deformation of spine reduce by 50% on the condition that the damping ratio is 0.08.
Biomechanical Phenomena ; Cadaver ; Computer Simulation ; Finite Element Analysis ; Humans ; Lumbar Vertebrae ; physiology ; Models, Biological ; Nonlinear Dynamics ; Stress, Mechanical ; Vibration ; Weight-Bearing ; physiology

Objective: To explore the correlation between thromboelastography (TEG) parameters and the risk of venous thromboembolism (VTE) and bleeding in patients receiving anticoagulant therapy in surgical intensive care unit (SICU). Methods: 205 patients received low molecular weight heparin (LMWH) anticoagulant therapy admitted to SICU of Tianjin Hospital from December 2016 to December 2017 were consecutively enrolled. TEG detection was performed in all patients at 1 day after anticoagulation therapy, and coagulation reaction time (R value), blood clot generation time (K value), blood clot generation rate (α angle) and maximum width value (MA value) were recorded. At the same time, the traditional coagulation function test was carried out, and prothrombin time (PT), activated partial thromboplastin time (APTT) and D-dimer levels were also recorded. The incidence of deep venous thrombosis (DVT), pulmonary embolism (PE) and bleeding during hospitalization were observed. Multivariate Logistic regression analysis was used to analyze the risk factors for VTE and bleeding in patients receiving anticoagulant therapy. Results: Of 205 patients, during the anticoagulant treatment, 14 patients developed DVT, and 4 patients with PE (2 of them were combined with DVT) with an incidence of 7.8% (16/205). There were 2 patients suffering from cerebral hemorrhage, 2 patients with gastric bleeding, and 1 patient with intra-tracheal hemorrhage with an incidence of 2.4% (5/205). Compared with the patients without VTE or bleeding, the R value of TEG in patients with VTE was significantly lowered (minutes: 4.6±2.2 vs. 7.4±1.4, P < 0.01), which was significantly increased in patients with hemorrhagic complications (minutes: 12.1±1.1 vs. 7.4±1.4, P < 0.01). There was no significant difference in the K value, α angle, MA value of TEG, or PT, APTT, D-dimer between the patients with and without VTE or bleeding. Multivariate Logistic regression analysis revealed that the R value of TEG was independent risk factor for incidence of VTE and hemorrhagic complication in SICU patients who receiving anticoagulation therapy [VTE: β = 0.386, odds ratio (OR) = 1.096, 95% confidence interval (95%CI) = 1.021-2.361, P = 0.006; hemorrhagic complication: β = -1.213, OR = 1.051, 95%CI = 1.017-3.458, P = 0.045]. Conclusion The R value of TEG is associated with the occurrence of VTE and hemorrhagic complications in patients receiving anticoagulant therapy in SICU.

It was expected that recombinant Aspergillus niger glucose oxidase could be expressed in Trichoderma reesei with stable activity. T. reesei CBHI promoter--CBHI ss. gene--A. niger glucose oxidase gene--T. reesei CBHI terminator--A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator--pUC19 (pCBHGOD) vector was constructed in E. coli DH5alpha by PCR application and gene cloning methods. T. reesei QM9414 protoplast was transformed by T. reesei CBHI promoter-CBHI ss. Gene--A. niger glucose oxidase gene--T. reesei CBHI terminator-A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator linear DNA fragment (CBHGOD fragment) that was made by digestion of pCBHGOD with Kpn I. T. reesei mutant clone with homologous recombinant A. niger glucose oxidase gene was selected by PCR method. Recombinant glucose oxidase was produced by mutant T. reesei strain under induction of wheat straw for 5 days. Recombinant glucose oxidase molecular mass was showed the same as native A. niger glucose oxidase standard from Sigma company by Western blot analysis. Recombinant glucose oxidase activity was 25u/mL in medium. The yield was 0.5 g/L in comparison with Sigma company glucose oxidase standard. There was no recombinant GOD degradation during Trichoderma reesei cultivation that was showed in Western blot analysis. Trichoderma reesei has capability to be a new recombinant host for Aspergillus niger GOD production.
Aspergillus niger ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Glucose Oxidase ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Trichoderma ; genetics ; metabolism

Objective: To observe the effects of herb-partitioned moxibustion and ginger-partitioned moxibustion on the growth of colon tumors in rats with colitis-associated colon cancer (CACC), and explore the mechanism of moxibustion intervening CACC through the purinergic receptor P2X ligand-gated ion channel 7 (P2X7R)/signal transducer and activator of transcription 3 (STAT3)/vascular endothelial growth factor (VEGF) pathway. Methods: A total of 26 male Sprague-Dawley rats were selected. According to the random number table method, 6 rats were selected as the normal group. The remaining 20 rats were injected intraperitoneally with azoxymethane (AOM) combined with oral dextran sodium sulfate (DSS) to prepare the CACC model. After the model was successfully established, 2 rats were randomly selected for model identification. The remaining 18 rats which were successfully modeled were randomly divided into a model group, a herb-partitioned moxibustion group and a ginger-partitioned moxibustion group, with 6 rats in each group. Moxibustion intervention was performed in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group at Qihai (CV 6) and bilateral Tianshu (ST 25). Moxibustion was performed twice at each point each time, once a day, at a 1-day interval after 6 consecutive interventions, for a total of 30 interventions. After intervention, the colon tumor load, pathological change and histopathological score were observed. Immunohistochemistry was used to detect the expressions of VEGF, P2X7R, phospho-STAT3 (p-STAT3), and nuclear factor-kappa B p65 (NF-κB p65) proteins in rat colon tissue. Western blot was used to detect the levels of p-STAT3 and NF-κB p65 proteins in rat colon tissue. Results: Compared with the normal group, the colon tumor load and histopathological score in the model group were significantly increased (both P<0.001), and different grades of dysplasia were observed in colon tissue from the model group, reaching the degree of adenocarcinoma; the expression level of P2X7R protein in colon tissue was significantly decreased (P<0.001), and the expression levels of p-STAT3, NF-κB p65 and VEGF proteins were significantly increased (all P<0.001) in the model group. Compared with the model group, the colon tumor load, colon histopathological score and the levels of p-STAT3, NF-κB p65 and VEGF proteins in colon tissue were significantly decreased (all P<0.05) in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group while the expression levels of P2X7R protein in colon tissue were significantly increased (both P<0.05). Conclusion: Both herb-partitioned moxibustion and ginger-partitioned moxibustion can reduce the colon tumor load in CACC rats and delay the progression of colon adenomas. The mechanism may be mediated by the P2X7R/STAT3 pathway to inhibit STAT3 phosphorylation, thereby reducing VEGF protein expression.

OBJECTIVE: The purpose of this study was to evaluate the clinical usefulness of CD44v6 in patients with histologically confirmed endometrial cancer. METHODS: Immunohistochemistry was used to determine the expression of CD44v6 in 39 hysterectomy specimens with the diagnosis of endometrial cancer. RESULTS: CD44v6 expression was detected in 8 specimens with no myometrial invasion of endometrial cancer, one specimen with <50% myometrial invasion, and none with >50% myometrial invasion. There was a significant association between CD44v6 expression and no myometrial invasion of endometrial cancer. There was a statistically significant association between CD44v6 expression and lymph node metastasis, lymphovascular involvement. And all cases of CD44v6 expression were FIGO stage I with histological grade 1 or 2. CD44v6 expression was statistically related with estrogen receptor expression. Although statistical significance was not revealed, it was likely that CD44v6 expression was related with progesterone receptor expression with positive predictive value of 6/9 (66.7%). CONCLUSION: Significant relationships between myometrial invasion, lymph node metastasis, lymphovascular involvement, surgical stage and expression of estrogen receptor and CD44v6 expression suggest that CD44v6 expression could be a good prognostic factor. CD44v6 expression may have potential clinical usefulness if this expression is demonstrated in a further study with prehysterectomy sampling specimens containing endometrial cancer.
Diagnosis ; Endometrial Neoplasms* ; Estrogens ; Female ; Humans ; Hysterectomy ; Immunohistochemistry ; Lymph Nodes ; Neoplasm Metastasis ; Receptors, Progesterone