ObjectiveTo determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of granulocyte colony-stimulating factor (G-CSF), and the role of Toll-like receptor 4 (TLR4) signaling pathway in this process.Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. ExperimentⅠ: the cells were divided into four groups as follows: control group, LPS stimulation group (LPS 10μg/mL), LPS+ 0.1 U/mL UFH group, and LPS+ 1 U/mL UFH group. HPMECs in UFH groups were treated with 0.1 U/mL or 1 U/mL UFH 15 minutes before LPS stimulation, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The concentrations of interleukin-6 (IL-6) and G-CSF in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 24 hours after LPS challenge to detect the effect of UFH on HPMECs. ExperimentⅡ: HPMECs were treated with 5μg/mL of rhodobacter sphaeroides LPS (LPS-RS, antagonist for TLR4) 4 hours before the addition of PBS or LPS. The concentrations of IL-6 and G-CSF in cell culture supernatants were determined 24 hours after LPS stimulation to detect the effect of TLR4 on LPS-induced HPMEC injury. ExperimentⅢ: HPMECs were divided into four groups as before: control group, LPS stimulation group, LPS+ 0.1 U/mL UFH group, LPS+ 1 U/mL UFH group. Treatments to cells were the same as experimentⅠ. The protein expression of TLR4 in HPMECs was determined by Western Blot 1 hour after LPS stimulation to detect the effect of UFH on TLR4.Results① Compared with control group, the levels of IL-6 and G-CSF in LPS stimulation group were increased [IL-6 (ng/L): 655.9±58.3 vs. 75.5±18.2, G-CSF (ng/L): 388.7±36.2 vs. 35.3±12.6, both P< 0.05]. Compared with those of LPS stimulation group, in LPS+ 0.1 U/mL UFH group and LPS+ 1 U/mL UFH group, the levels of IL-6 and G-CSF were significantly decreased [IL-6 (ng/L): 518.2±64.6, 489.1±75.6 vs. 655.9±58.3, G-CSF (ng/L): 298.8±41.0, 273.4±33.2 vs. 388.7±36.2, allP< 0.05]. The results indicated that 1 U/mL UFH had better results, though there was no statistical significance between the results of two UFH groups.② LPS-induced up-regulation of IL-6 and G-CSF levels was prevented by LPS-RS [IL-6 (ng/L): 139.1±37.6 vs. 655.9±58.3, G-CSF (ng/L): 73.7±19.7 vs. 388.7±36.2, bothP< 0.05]. LPS-RS alone had no effect on cytokines [IL-6 (ng/L):118.2±42.1 vs. 75.5±18.2, G-CSF (ng/L): 48.4±26.8 vs. 35.3±12.6, bothP> 0.05].③ Compared with control group, the protein expression of TLR4 (grey value) in LPS stimulation group was significantly upregulated after 1 hour (0.87±0.23 vs. 0.36±0.12,P< 0.05). UFH with 0.1 U/mL and 1 U/mL lowered TLR-4 protein expression induced by LPS (0.68±0.18, 0.62±0.26 vs. 0.87±0.23, bothP< 0.05).ConclusionsThe expressions of IL-6 and G-CSF were increased obviously in LPS treated HPMECs. UFH might take its therapeutic effect through TLR4-dependent pathway.

Objective To systemically review the efficacy and safety of heparin for treatment of sepsis.Methods Database search of IM/MEDLINE,Cochrane Library,SCIE,CBM,CNKI,VIP Data,WanFang Data (from January 2000 to June 2012) was conducted.The quality of included randomized controlled trials (RCTs) about heparin for treatment of sepsis was assessed,and relevant data were extracted according to the inclusion and exclusion criteria.Then meta analysis was performed using RevMan 5.1.Results 17 trials with 1 167 participants were included.The results of meta-analysis showed:compared with the control group,heparin significantly decreased 28-day mortality in patients with sepsis [odds ratio (OR) =0.59,95％ confidence interval (95％CI) 0.45-0.77,P=0.0001] ; heparin did not deteriorate coagulation disorders,but corrected sepsis-induced platelet (PLT) count reduction [mean difference (MD) =13.94,95％ CI 10.15 to 17.72,P＜0.000 01],while it had no significant effect on the activated partial thromboplastin time (APTT) and prothrombin time (PT,APTT:MD=-3.18,95％CI-6.88 to 0.53,P=0.09; PT:MD=-0.68,95％ CI-1.48 to 0.12,P=0.09).There was no significant difference between the two groups in the incidence of bleeding either.Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of heparin group was significantly lower than that of the control group (MD=-2.58,95％CI-3.29 to-1.87,P＜0.000 01),and the incidence of multiple organ dysfunction syndrome (MODS) was significantly lower than that of the control group (OR=0.32,95％CI 0.17 to 0.61,P=0.000 6).In addition,heparin could shorten intensive care unit (ICU) stay (MD=-4.43,95％CI-6.79 to-2.07,P=0.000 2),whereas it showed no significant effect on the total length of hospital stay.Conclusions Heparin can ameliorate sepsis,and has high degree of safety and lower hospital expense.Due to limitation of the quality of included studies,larger sample and well-designed RCTs are needed to further support this conclusion.

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.

Objective To investigate the influence of liraglutide combined with metformin and simple metformin on insulin re‐sistance in the patients with type 2 diabetes mellitus(T2DM ) .Methods Thirty patients with T2DM treated by liraglutide combined with metformin were collected as the experimental group ,while other 30 patients with T2DM treated by single metformin were col‐lected as the control group .The changes of HbA1c ,HOMA‐IR ,adiponectin ,NO and ET before treatment and at 3 months after treatment were compared between the two groups .Results The decrease degree of HbA1c ,HOMA‐IR ,NO and ET and the in‐crease degree of adipodectin in the experimental group were more significant than those in the control group with statistical differ‐ences between the two groups(P<0 .05) .Conclusion Liraglutide has more significant effect than metformin for improving the in‐sulin resistance in the patients with T2DM .

Objective To investigate the expression of ?-methylacyl CoA racemase (AMACR,P504S) and p63 in prostate carcinoma tissues and the clinicopathologic significance in the diagnosis of the disease. Methods The expression of AMACR and p63 in 39 cases of prostate carcinoma and 24 cases of benign prostate lesions were examined using immunohistochemical method. The association of the AMACR and p63 expression in prostate carcinoma tissues with the differentiation and clinical staging was evaluated. Results The positive rates of AMACR and p63 were 85%(33/39) and 10%(4/39),respectively,in prostate carcinoma tissues; and those were 21%(5/24) and 88%(21/24),respectively,in benign prostatic hyperplasia. There were significant differences of these values between prostate carcinoma and benign prostatic hyperplasia (P0.05). Conclusions Over-expression of AMACR and under-expression of p63 may be the preliminary incident of the formation of prostate carcinoma,which may induce carcinoma into occurrence and progression.Thus it may be an early diagnostic parameter and a novel target for prostate carcinoma treatment.

Neutrophil extracellular traps (NETs) are net-like structure composed of DNA and nuclear proteins, which are produced by activated neutrophils under the circumstances of a variety of pathogens or drugs. As part of defensive mechanism, NETs have been proved to restrict the spread of pathogens and release of antimicrobial molecules. NETs can not only strengthen the adhesion between neutrophils and platelets, promote platelet mediated procoagulant reaction, but also lead to endothelial cell damage and coagulopathy in sepsis. In addition, NETs also plays an important role in pathophysiological processes of venous thrombosis. Therefore, NETs may become the biomarkers of evaluating coagulation dysfunction and potential therapy target in sepsis.

Objective To investigate the changes of neutrophil extracellular traps (NETs) level in plasma of sepsis patients and judge its clinical value for early diagnosing of sepsis.Methods A prospective observational study was conducted. The patients after surgery aged > 18 years and expected to stay in the ICU > 24 hours admitted to intensive care unit (ICU) of the First Affiliated Hospital of China Medical University from November 2014 to February 2015 were enrolled. According to the criteria of sepsis diagnosis in 1991, patients were divided into non-sepsis group and sepsis group. The healthy people who taken a physical examination were enrolled in the healthy control group. 3 mL peripheral venous blood was collected at 1 hour after admission to ICU. A fasting blood was collected in the healthy control group in the morning. The plasma free DNA (cf-DNA/NETs) was determined by using the fluorescence microplate reader, white blood cell (WBC), neutrophil ratio (NEU), procalcitonin (PCT), C-reactiveprotein (CRP) in peripheral blood of the patients were detected, and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) scores were calculated. The correlation between plasma NETs and the risk factors in sepsis patients was analyzed by Spearman correlation analysis. The value of cf-DNA/NETs and WBC level in the diagnosis of sepsis was analyzed by using the receiver operating characteristic curve (ROC).Results Twenty-three sepsis patients, 20 non-sepsis patients, and 22 healthy persons were enrolled. There were no differences in baseline variables including gender and age among three groups, which indicated baseline data equalization. The plasma concentration of cf-DNA/NETs in sepsis group was significantly higher than that in non-sepsis group and healthy control group (μg/L: 453.44±185.37 vs. 188.35±29.66, 203.83±43.25, bothP < 0.05),and there was no significant difference between last two groups (P > 0.05). WBC, NEU, PCT, CRP, APACHE Ⅱ and SOFA score in sepsis group were significantly higher than those of non-sepsis group [WBC (×109/L): 9.52±5.51 vs. 5.97±2.28, NEU: 0.787±0.110 vs. 0.655±0.067, PCT (mg/L): 7.14 (3.60, 13.29) vs. 6.07 (3.57, 7.91), CRP (mg/L): 64.44±13.14 vs. 27.00±19.47, APACHE Ⅱ: 10.25±4.92 vs. 6.00±1.22, SOFA: 6.0±5.1 vs. 5.0±1.2, allP < 0.05]. Correlation analysis showed that the level of NETs had no obvious correlation with gender, age, WBC, NEU, PCT, CRP, APACHE Ⅱ and SOFA scores (r value was 0.322, 0.262, 0.194, 0.312, 0.227, 0.454, 0.433, 0.333, respectively, allP > 0.05). The area under the ROC curve (AUC) of plasma cf-DNA/NETs for the diagnosis of sepsis was 0.981. When the cut-off valueof plasma cf-DNA/NETs was > 257.96μg/L, the sensitivity was 91.3%, specialty was 95.2%, and Youden index was 0.865. AUC of WBC in the diagnosis of sepsis was 0.663. When the cut-off value of WBC was > 6.0×109/L, the sensitivity was 78.3% and specificity was 25.0%.Conclusion The plasma cf-DNA/NETs levels increased significantly in sepsis patients. In the diagnosis of sepsis, plasma NETs levels had better advantages over WBC. NETs can be used as a biomarker for early diagnosis of sepsis.

Objective To determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of chemokines and nuclear factor-κB (NF-κB) signaling pathway. Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. The cells were divided into control group, LPS challenge group, 1 kU/L or 10 kU/L UFH+LPS group, and NF-κB inhibitor N-tosyl-L-lysyl chloromethyl-ketone (TLCK) group (TLCK+LPS group). HPMECs in LPS challenge group were treated with 10 mg/L LPS. UFH pretreatment with different dosages groups were treated with 1 kU/L or 10 kU/L UFH 15 minutes before LPS challenge. Cells in the TLCK+LPS group were treated with 10 μmol/L of TLCK 30 minutes before the addition of LPS, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The cells were harvested 1 hour after LPS challenge, and the nuclear translocation of NF-κB was determined by immunofluorescence assay to detect the effect of UFH on NF-κB activation. The levels of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 3 hours and 6 hours after LPS challenge to detect the effect of UFH on LPS induced expression of chemokines and its mechanism of effect on NF-κB signaling pathway in HPMECs. Results ① In the control group, NF-κB was mostly located in the cytosol as shown by immunofluorescence. Treatment of HPMECs with LPS significantly increased the translocation of NF-κB from the cytosol to nucleus. UFH suppressed LPS-induced NF-κB activation both in 1 kU/L and 10 kU/L dosages, and 10 kU/L UFH gave even better results. ② Compared with control group, the levels of IL-8 and MCP-1 in the supernatants in LPS challenge group were significantly increased at 3 hours and 6 hours after LPS challenge [IL-8 (ng/L): 387.1±26.4 vs. 23.8±8.1 at 3 hours, 645.5±69.6 vs. 125.7±18.7 at 6 hours; MCP-1 (ng/L): 3 654.9±467.9 vs. 721.6±61.3 at 3 hours, 8 178.5±792.6 vs. 1 324.7±148.7 at 6 hours, all P < 0.05]. Compared with that of LPS challenge group, in 1 kU/L and 10 kU/L UFH pretreatment groups, the levels of IL-8 and MCP-1 were significantly decreased [IL-8 (ng/L): 315.3±24.8, 275.8±31.1 vs. 387.1±26.4 at 3 hours, 557.8±43.3, 496.9±38.7 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 2 924.1±267.9, 2 668.3±522.6 vs. 3 654.9±467.9 at 3 hours, 7 121.7±557.2, 6 563.9±576.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. The results indicated that 10 kU/L UFH yielded better results. However, inhibition study using the known NF-κB inhibitor TLCK could decrease LPS-induced increase in IL-8 and MCP-1 levels [IL-8 (ng/L): 162.4±21.3 vs. 387.1±26.4 at 3 hours, 274.1±22.6 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 1 478.2±138.5 vs. 3 654.9±467.9 at 3 hours; 3 667.6±259.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. Conclusions The levels of IL-8 and MCP-1 were increased obviously in LPS treated HPMECs. UFH might suppress LPS-activated NF-κB signaling pathway, contributing to the inhibitory effects of chemokines in HPMECs.

Objective To investigate the effects on knee function between patella plasty and patellar replacement in total knee arthroplasty.Methods From August 2010 to November 2010,48 patients (69 knees) of osteoarthritis performed TKA were covered in this study.All the patients were randomly assigned to the following two groups:one group contained 24 patients (34 knees) performed patella plasty,and the other group contained other 24 patients (35 knees) performed patellar replacement.There was no significant difference between the two groups in age,weight,height,BMI,patellar score and the American Knee Society Score (KSS).Every patient was followed up for 6 weeks,3 months,6 months,one year,and two years.All the results,which were used to compare the difference between the two groups,in KSS knee score,knee function score,patellar score,the incidence of anterior knee pain,and imaging findings.Results In this clinical study,20 patients (30 knees) in patellar replacement group and 20 patients (29 knees) in patella plasty group were followed up.No significant difference was found in the postoperative KSS knee score between 2 groups at each time point.The KSS knee function score in replacement group was significantly higher than that in arthroplasty group at 6,12 and 24 months after operation.The incidence of anterior knee pain after the surgery in replacement group was significant different from the plasty group at every time point after operation.There was no significant difference in tibiofemoral angle,patellar ligament ratio,patellar tilt angle,congruence angle and lateral patellar displacement between two groups at last follow-up.Conclusion Total knee arthroplasty with patella replacement can improve both knee function and patella function,and reduce the incidence of postoperative anterior knee pain.

Objective To explore the clinical effect of the elderly far lateral lumbar disc herniation via Quadrant channel system under the Wiltse approach. Methods Thirty-three elderly patients suffering from far lateral lumbar disc herniation were treated by surgery, among whom 18 patients were treated by Quadrant minimally invasive channel system (Quadrant group), and 13 patients were treated by percutaneous transforaminal endoscopic discectomy (PTED group). The patients were followed up for 12 months, and the operation time, blood loss and length of incision were investigated. The JOA scores were compared preoperatively and postoperatively. Results The patients in two groups were followed up for 24 months, except 1 patient withdrew in Quadrant group. The incision length and blood loss in PTED group were superior to those in the Quadrant group:2.0-2.5 cm vs. 0.7 cm, (46.50± 15.20) ml vs.(23.87±6.58) ml, P<0.01, but the operation time in PTED group was longer than that in Quadrant group: (77.17 ± 8.71) min vs. (63.20 ±10.87) min, P<0.01.The two groups had achieved good curative effect, while JOA score in PTED group was superior to the Quadrant group 1 week after operation ( P<0.05), but in 3 month and 12 month two groups had no significant difference ( P>0.05). Conclusion Surgical treatment on elderly far lateral lumbar disc herniation via quadrant channel under wiltse approach is an effective method, and can achieve similar effect with PTED.