Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer.

BACKGROUND: The development of venous thromboembolism (VTE) is associated with high mortality among gastric cancer (GC) patients. Neutrophil extracellular traps (NETs) have been reported to correlate with the prothrombotic state in some diseases, but are rarely reported in GC patients. AIM: To investigate the effect of NETs on the development of cancer-associated thrombosis in GC patients. METHODS: The levels of NETs in blood and tissue samples of patients were analyzed by ELISA, flow cytometry, and immunofluorescence staining. NET generation and hypercoagulation of platelets and endothelial cells (ECs) in vitro were observed by immunofluorescence staining. NET procoagulant activity (PCA) was determined by fibrin formation and thrombin-antithrombin complex (TAT) assays. Thrombosis in vivo was measured in a murine model induced by flow stenosis in the inferior vena cava (IVC). RESULTS: NETs were likely to form in blood and tissue samples of GC patients compared with healthy individuals. In vitro studies showed that GC cells and their conditioned medium, but not gastric mucosal epithelial cells, stimulated NET release from neutrophils. In addition, NETs induced a hypercoagulable state of platelets by upregulating the expression of phosphatidylserine and P-selectin on the cells. Furthermore, NETs stimulated the adhesion of normal platelets on glass surfaces. Similarly, NETs triggered the conversion of ECs to hypercoagulable phenotypes by downregulating the expression of their intercellular tight junctions but upregulating that of tissue factor. Treatment of normal platelets or ECs with NETs augmented the level of plasma fibrin formation and the TAT complex. In the models of IVC stenosis, tumor-bearing mice showed a stronger ability to form thrombi, and NETs abundantly accumulated in the thrombi of tumor-bearing mice compared with control mice. Notably, the combination of deoxyribonuclease I, activated protein C, and sivelestat markedly abolished the PCA of NETs. CONCLUSION: GC-induced NETs strongly increased the risk of VTE development both in vitro and in vivo. NETs are potential therapeutic targets in the prevention and treatment of VTE in GC patients.

Effect of trichostatin a on SGC-7901 gastric cancer cells.

AIM: To explore Trichostatin A (TSA) effect on SGC-7901 gastric cancer cells. METHODS: MTT, fluorescence microscopy, and flow cytometry were used to assess TSA effect on cell growth and apoptosis in SGC-7901. Immunocytochemistry was used to evaluate the expression of acetylated histone H4 in SGC-7901 cells.Gene expression profile was determined by microarray assays. Glycoprotein hormones alpha subunit (CGA) gene and protein expressions in SGC-7901 cells were evaluated by Real-time PCR and Western blot, respectively. In addition, CGA protein levels in gastric adenocarcinoma and normal adjacent tissues were assessed by immunohistochemistry. RESULTS: TSA inhibited SGC-7901 cell growth. In addition, cell proliferation was significantly decreased (P = 0.02) in TSA treatment groups (0.93 ± 0.07) compared with controls (1.15 ± 0.07). Apoptosis related morphological changes, including nuclear chromatin condensation and fluorescence strength, were observed by fluorescence microscopy. These findings corroborated the increased expression of acetylated histone H4 observed in TSA treated cells compared to controls, as determined by immunocytochemistry. Interestingly, treatment of SGC-7901 cells with TSA (75 ng/ml) resulted in CGA gene down-regulation (P = 0.0381). Accordingly, CGA protein levels were decreased in TSA treated SGC-7901 cells. Finally, immunohistochemistry analysis showed that CGA expression was significantly higher in gastric adenocarcinoma tissues than normal adjacent tissues (P = 0.001). CONCLUSION: TSA induces cell apoptosis and increases the levels of acetylated histone H4 in SGC-7901 cells. In addition, TSA treatment decreases the expression in gastric cancer cells of the CGA gene, which is upregulated in gastric adenocarcinoma tissues.

Differential expression of glycoprotein non-metastatic melanoma protein B (GPNMB) involved in trichostatin A-induced apoptosis in gastric cancer.

In this study, we investigated the effect of trichostatin A (TSA) on the gastric cancer cell line BGC-823. The effect of TSA on growth inhibition and apoptosis of BGC-823 cells was examined. The gene expression profile was determined by microarray. Western blotting was used to study the levels of acetylated histone H4 and Glycoprotein non-metastatic melanoma protein B (GPNMB) proteins. GPNMB gene expression was measured by real-time PCR. GPNMB protein levels in gastric adenocarcinoma tissues and adjoining normal tissues were detected by immunohistochemistry. The results showed that a significant decrease in cell population following treatment with 75 ng/mL TSA for 48 h (0.87 ± 0.04) as compared to control (1.14 ± 0.06) (P = 0.02). Apoptotic cells were increased in TSA (75 ng/mL for 48 h) treated group as compared to the control group (from 2.02% to 19.74%) by flow cytometry. The expression of acetylated histone H4 was increased in TSA treated (75 ng/mL for 48 h) group (from 1.00 ± 0.26 to 1.87 ± 0.33, F = 5.862, P = 0.0038) as compared to the control group by Western blotting. After 48 h TSA treatment (75 ng/mL), BGC-823 cells showed decrease in GPNMB gene expression (from 1.00 ± 0.21 to 0.59 ± 0.11, F = 6.214, P = 0.0018). Immunohistochemistry showed that GPNMB expression in gastric adenocarcinoma was significantly higher than the adjoining normal tissues (P = 0.000). To conclusion, our results support that TSA can induce apoptosis, and increase acetylated histone H4 in BGC-823 cells. GPNMB expression is decreased in BGC-823 cells after TSA treatment. GPNMB is overexpressed in gastric adenocarcinoma tissue. GPNMB involved in TSA-induced apoptosis might participate in gastric cancer.

[Analysis of clinical characteristics of gastrointestinal cancer in Heilongjiang province, China 1998 to 2007].

OBJECTIVE: To provide basic information for epidemiological research of gastrointestinal (GI) malignant tumors. METHODS: Data of GI cancer diagnosed in 15 hospitals of Heilongjiang province between January 1998 and December 2007 were analyzed retrospectively. The data mainly involved the age of onset, initial symptoms, pathological types, clinical staging and types of surgical procedure. RESULTS: Gastric cancer was the most common type (45.8%) among the 33,540 GI cancer cases, then were rectal cancer (27.3%) and colon cancer (26.8%). Right colon cancer cases were more common than the left ones (1.3:1.0), particularly in people over 80 (2.1:1.0). Only 1.3% of colorectal cancer could be found in age under 30 years old. In patients aged 50 to 70, advanced gastric cancer accounted for 70.6%, advanced colon cancer 73.4% and advanced rectal cancer 72.4%. Well-moderately differentiated adenocarcinoma in early gastric cancer was 49.7%, early colon cancer 77.3% and rectal cancer 83.2%. Patients undergone radical excision in early gastric cancer accounted for 69.1%, advanced gastric cancer 79.9%, left colon cancer 91.9%, right colon cancer 83.9% and in rectal cancer for 88.3%. CONCLUSIONS: People aged 50 to 70 tend to get GI cancer in Heilongjiang province. Gastric cancer is the most common GI cancer. Radical excision is the main choice of therapy.

[Comparison of soil C and N in rubber plantation and seasonal rain forest].

With the rubber plantation and seasonal rain forest in Xishuangbanna of Southwestern China as test objectives, a comparative study was conducted on their litter input, soil total C and N contents, and seasonal changes of soil active C and N from 2006 to 2007. Comparing with seasonal rain forest, rubber plantation had lower amount of aboveground litterfall and higher amount of floor mass, reflecting the lower decomposition rate (turnover coefficient, K) of litters, and had higher C/N ratio of litters and soil, indicating that the organic matters in rubber plantation were more resistant to degradation. The surface soil total organic C, labile organic C, and microbial biomass C concentrations in rubber plantation accounted for 60%-70% of those in seasonal rain forest, and the soil NO3(-)-N concentration and pH value in rubber plantation were lower than those in seasonal rain forest, indicating that the conversion from seasonal rain forest to rubber plantation decreased the C and N inputs from aboveground litterfall and the availability of soil C and N, and caused soil acidification. Moderate land management strategies for rubber plantations were needed to prevent the degradation of soil quality and to maintain the productive sustainability.

Validation of VKORC1 and CYP2C9 genotypes on interindividual warfarin maintenance dose: a prospective study in Chinese patients.

OBJECTIVES: To develop a warfarin-dosing algorithm that could be combined with pharmacogenomic and demographic factors, and to evaluate its effectiveness in a randomized prospective controlled clinical trial. METHODS: A pharmacogenetics-based dosing model was derived using retrospective data from 266 Chinese patients and multiple linear regression analysis. To prospectively validate this model, 156 patients with an operation of heart valve replacement were enrolled and randomly assigned to the group of pharmacogenetics-guided or traditional dosing for warfarin therapy. All patients were followed up for 50 days after initiation of warfarin therapy. The log-rank test was compared with the time-to-event (Kaplan-Meier) curves. Cox proportional hazards-regression model was used to assess the hazard ratio of the time to reach stable dose. RESULTS: The linear regression model derived from the pharmacogenomic model correlated with 54.1% of warfarin dosing variance. The final multiple linear regression model included age, body surface area, VKORC1, and CYP2C9 genotype. The study showed that the hazard ratio for the time to reach stable dose was 1.932 for the traditional dosing group versus the model-based group and a close and highly significant relationship was observed to exist between the predicted and the actual warfarin dose (R=0.454). CONCLUSION: A pharmacogenetics-based dosing algorithm has been developed for improvement in the time to reach the stable dosing of warfarin. This model may be useful in helping the clinicians to prescribe warfarin with greater safety and efficiency.

Gastric cancer cell lines induced by trichostatin A.

AIM: To explore the effect of trichostatin A (TSA) on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS: The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS: TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups (37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line) and control group (0.85+/-0.14 vs 1.14+/-0.07, P=0.02; 0.94+/-0.07 vs 1.15+/-0.06, P=0.02). Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of G1-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group. CONCLUSION: TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.

Vegetation cover of forest, shrub and pasture strongly influences soil bacterial community structure as revealed by 16S rRNA gene T-RFLP analysis.

Bacterial community structure is influenced by vegetation, climate and soil chemical properties. To evaluate these influences, terminal restriction fragment length polymorphism (T-RFLP) and cloning of the 16S rRNA gene were used to analyze the soil bacterial communities in different ecosystems in southwestern China. We compared (1) broad-leaved forest, shrub and pastures in a high-plateau region, (2) three broad-leaved forests representing a climate gradient from high-plateau temperate to subtropical and tropical regions and (3) the humus and mineral soil layers of forests, shrub lands and pastures with open and restricted grazing activities, having varied soil carbon and nutrient contents. Principal component analysis of the T-RFLP patterns revealed that soil bacterial communities of the three vegetation types were distinct. The broad-leaved forests in different climates clustered together, and relatively minor differences were observed between the soil layers or the grazing regimes. Acidobacteria dominated the broad-leaved forests (comprising 62% of the total clone sequences), but exhibited lower relative abundances in the soils of shrub (31%) and pasture (23%). Betaproteobacteria was another dominant taxa of shrub land (31%), whereas Alpha- (19%) and Gammaproteobacteria (13%) and Bacteriodetes (16%) were major components of pasture. Vegetation exerted more pronounced influences than climate and soil chemical properties.

[L-arginine pretreatment lowers cerebral metabolism for cerebral protection during deep hypothermic circulatory arrest].

OBJECTIVE: To evaluate the effect of L-arginine pretreatment on cerebral metabolism for cerebral protection during deep hypothermic circulatory arrest (DHCA). METHODS: Fifteen healthy adult canines of either sex weighing 14.7-/+2.4 kg were randomly divided into 3 groups (n=5), namely the normal saline group, L-arginine pretreatment group (pretreated with 100 mg/kg L-arginine 60 min before DHCA), and L-arginine combined with 7- nitroindazole treatment group (pretreated with 100 mg/kg L-arginine and 25 mg/kg7-Ni 60 min before DHCA). For all the canines, extracorporeal circulation was established routinely to allow nasopharyngeal temperature reduction to 18 degrees celsius;, at which point DHCA commenced followed 90 min later by reperfusion. At 30 min before DHCA and 0, 45 and 90 min after DHCA as well as at 60 min after reperfusion initiation, blood samples were collected from the jugular vein and arterial to measure the plasma lactic acid, and the cerebral cortex of the parietal lobe was sampled determine the activity of Na(+)-K(+)ATPase. The cerebral water content was also determined after execution of the canines. RESULTS: In the two pretreatment groups, the level of lactic acid production (shown by the difference in lactic acid levels between the jugular venous and arterial blood) and the cerebral ATP consumption were similar (P>0.05), but both were significantly lower than those of the control group (P<0.05). The cerebral water content was the lowest in the combined treatment group, followed by exclusive L-arginine group, and the highest in the control group (P<0.05), with significant difference between the 3 groups (P<0.05). CONCLUSION: L-arginine pretreatment can lower cerebral metabolism during DHCA to offer protective effect on the brain.

[Impact of trichostatin A on gastric carcinoma cell line SGC-7901].

OBJECTIVE: To investigate the effect of trichostatin A(TSA) on SGC- 7901 cells. METHODS: Cytotoxicity and cell viability of gastric cancer cell line SGC- 7901 were assayed by MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope. Cell cycle and apoptosis rate were analyzed by flow cytometry. Histone H3 acetylation was detected by Western blot. RESULTS: TSA showed apparently cytotoxicity in SGC- 7901 cells. The growth curve showed the growth ratio decreased with the increase of TSA concentration. Apoptosis rate were significantly different between TSA treated group(75 ng/ml for 72 h)and control group (P < 0.05). Morphologic changes of apoptosis including nuclear chromatin condensation and fluorescence strength were observed with fluorescence microscope.TSA treatment (75 ng/ml for 72 h) sensitively induced apoptosis in the cell,which was demonstrated by the migration of many cells to the sub- G1 phase,the reduction of G1- phase cells and the increment of apoptosis rate (29.54%) in flow cytometric analysis. The expression of acetylated histone H3 was increased in TSA group(75 ng/ml) for 48 h compared with control group by Western blot. CONCLUSIONS: TSA can induce SGC- 7901 cell apoptosis. The expression of acetylated histone H3 may contribute to the apoptosis.

[Efficacy and safety of adjuvant post-surgical therapy with imatinib in gastrointestinal stromal tumor patients with high risk of recurrence: interim analysis from a multicenter prospective clinical trial].

OBJECTIVE: To evaluate the efficacy and safety of postoperative adjuvant chemotherapy with imatinib in gastrointestinal stromal tumor(GIST) patients who had high risk of recurrence. METHODS: A prospective, open-label, multi-center trial conducted in sixteen teaching hospitals in China was carried out. The criteria of the enrolled patients included age more than 18 years old, CD117 positive GIST, tumor size more than 5 cm, pathological mitosis counts more than 5/50 HPF, and treatment beginning within 4 weeks after complete resection and with imatinib (400 mg, once a day) for at least 12 months. The 1, 3 year recurrence rates, disease free survival, overall survival rate and quality of life were evaluated. RESULTS: From Aug. 16th 2004 to Sep. 13th 2005, there were totally 74 patients screened and 57 patients (34 men, 23 women) enrolled in the imatinib treatment group. The primary tumors were located in the stomach in 50.9%, the small intestine in 38.6% and the colorectum in 10.5% of the cases. All the patients received radical resection. Until the cut-off date of interim analysis, there was no evidence of tumor relapse or metastasis in all patients and no death was reported either. Among the 57 enrolled patients with intention to treat(ITT), twelve patients finished the protocol (per protocol, PP). The disease free survival was (268.3 +/-120.2) d in ITT analysis, and (396.7+/-38.2) d in the PP analysis. The incidence of adverse effect was 44.4% . The score in quality of life showed no statistically significant difference between the baseline visit and the follow-up visits. CONCLUSION: Imatinib is a promising postoperative adjuvant chemotherapy in GISTs patients with high risk of recurrence, and the adverse effects are receivable.

[Experimental study of cerebral protection by retrograde vs selective antegrade cerebral perfusion during deep hypothermic circulatory arrest].

OBJECTIVE: To compare the effect of cerebral protection between retrograde cerebral perfusion (RCP) and selective antegrade cerebral perfusion (SACP) during deep hypothermic circulatory arrest (DHCA) in canine models. METHODS: Fifteen healthy adult dogs were randomly divided into 3 groups (n=5), namely the simple DHCA group (group I), DHCA+RCP group (group II) and DHCA+SACP group (group III). Extrocorporeal circulatory was established routinely in the dogs, and DHCA commenced when the nasopharyngeal temperature was reduced to 18 degrees C. During DHCA, RCP and SACP were applied in groups II and III, respectively. All the models were rewarmed after 90 min of DHCA and the cerebral reperfusion continued for 90 min. Cerebral oxygenous metabolic function, cerebral temperature and ultrastructural changes of the neurons were observed in the 3 groups at different time points during the operation. RESULTS: The jugular venous oxygen saturation (SjvO(2)) increased with the temperature reduction, and then decreased after DHCA commencement, showing significant changes at different time points in groups I and II. SjvO(2) in group III were significantly higher than that in the other two groups after 90 min of DHCA (P=0.000). Brain temperature significantly increased in group I during DHCA as compared with that in groups II and III (P=0.000), but showed no significant difference between the latter two groups (P=0.195). The ultrastructure of the neurons underwent obvious changes after reperfusion for 30 min in group I. In group II the neuronal ultrastructure was basically normal at 60 min during DHCA and changed slightly at 90 min, but in group III no obvious changes were seen at 90 min during DHCA and only slight changes occurred at 30 min of reperfusion. CONCLUSIONS: RCP can not supply enough oxygen but can maintain low cerebral temperature, and provide short-term brain protection. DHCA+SACP provides better brain protection than simple DHCA and DHCA+RCP, and has a promising prospect in cardiac surgery.

[Study on the extraction of the active component in Atractylode macrocephala with ultrasonic wave technology optimized by the response surface method].

OBJECTIVE: To create Atractylode macrocephala inspissation decoction pieces. The effect of ultrasonic wave on extraction of the active components in A. macrocephala was studied in a water solution. METHOD: The factors including the ratio of material to liquid, ultrasonic power, ultrasonic time, soaking time, particle size etc, were studied. The best extraction method was found through the response surface method. RESULT: The best extraction method was found as follows: the granularity of material 0.1 mm, the repetition times of ultrasonic process 3 times, the soaking time before the ultrasonic process 30 min, the ratio of liquid to material 10:1, the soaking time after the ultrasonic process 2.6 h, the time of the ultrasonic wave 15.5 min, the power of the ultrasonic wave 531 W, the rate of reservation of active components 88.5%, the rate of inspissation 1.6. CONCLUSION: The ultrasonic wave can used in the extraction of the active components in A. macrocephala and a model equation that can be used to predict the experiment was get through the response surface method.

[Anatomic study of small intestinal vessel in piglet small intestine transplantation].

OBJECTIVE: To study the anatomy of the small intestine,and investigate the optimal selection of donors,recipients,and their small intestine vessels in piglet small intestine transplantation. METHODS: The weight and length of 30 piglets were measured. Angiography and pigments perfusion were used to observe the main vessels of the small intestine,and the length of the small intestine,and the external diameter of the main vessels of the small intestine were measured in vivo and ex vivo. RESULTS: The length of the small intestine was 11.5 times as long as the body length, and its weight accounted for 2.3% of the body weight. The outer diameters of abdominal aorta (AT), mesenteric anterior artery (MAA) and its 5(th)-6(th) branches in vivo and ex vitro were 4.3/4.6mm, 2.5/2.7mm and 1.9/2.2mm respectively. The total number of MAA's branches was 6-8 in general and its 5(th)-6(th) branches were the longest [(20.0 +/- 7.0) mm, (22.0 +/- 8.2) mm]. The outer diameter of mesenterial anterior vein (MAV) was 1-2 mm wider than that of MAA. CONCLUSIONS: AT, MAA and its 5(th)-6(th) branches are the preferable vessels for small intestine transplantation. In segmental small intestine transplantation, the length of the small intestine and body weight can be used to primarily select the suitable animals.

[Accelerated fatigue test in vitro of expanded polytetrafluoroethylene artificial heart valve].

OBJECTIVE: To study the durability of expanded polytetrafluoroethylene artificial heart valve (ePTFE AHV). METHODS: Six ePTFE AHVs were tested for 400 million times against accelerated fatigue using TH-2200 artificial heart valve exosomatic accelerated fatigue instrument. Hydromechanical parameters of fore-and-aft accelerated fatigue test of the 6 AHVs were obtained by TH-1200 artificial heart valve exosomatic pulsatile stream instrument. RESULTS AND CONCLUSION: The mean gradient pressure spanning the valve and the effective orific area of ePTFE AHVs did not undergo significant changes after fore-and-aft the fatigue test, but the regurgitation volume and regurgitation rate of ePTFE AHVs were reduced after the accelerated fatigue test, suggesting good durability of ePTFE AHV.

[Development study on model WY multi-functional thoracic cavity closed drainage system].

Based on the improved design of the existing thoracic cavity closed drainage system, a new multi-functional device is developed and is described here in detail. The device is more convenient and more efficient than the existing system. Besides, it has a function of autotransfusion. Animal experimental results show that it has attained the goal of the improved design.

[Ultrafiltration with different membranes and respiratory function after cardiopulmonary bypass].

OBJECTIVE: To investigate the protective effect of ultrafiltration on pulmonary function after cardiopulmonary bypass (CPB) by comparing two different membranes used in the ultrafiltration. METHODS: Thirty patients undergoing cardiac surgery with CPB were randomly divided into adsorption group (n=15) and control group (n=15), and in the former group, AN69 membrane was used for ultra-infiltration, with polysulfone (PS) membrane adopted in the control group during CPB. Plateau airway pressure (P(Plateau)), peak airway pressure (P(Peak)), static pulmonary compliance (Cst), dynamic pulmonary compliance (Cdyn) and respiratory index (RI) were measured or calculated before and 5, 60, 120, and 240 min after CPB in each group respectively. RESULTS: During the period of 5 to 240 min after CPB, the increase in P(Plateau), P(Peak), RI and decrease in Cst and Cdyn were much more obvious and lasted for longer time in the adsorption group than in the control group (P<0.05). No operative death or hemoglobinuria occurred in these cases. CONCLUSION: Ultrafiltration with AN69 membrane more effectively reduces CPB-induced lung injury and improves the postoperative respiratory function than with PS membrane.

[Surgical treatment of ascending aortic aneurysm by Bentall's procedure: report of 14 cases].

OBJECTIVE: To review our experience in surgical treatment of ascending aortic aneurysm (AAA) with aortic insufficiency in 14 cases of by Bentall's procedure. METHODS AND RESULTS: All the patients underwent replacement of the ascending aorta and aortic valve with composite valved vascular prosthesis and received direct implantation of the aortic graft of the coronary orifices (Bentall's procedure) with satisfactory results. Five patients had ascending aortic dissection and 4 had typical Marfan's syndrome. All the patients suffered severe aortic insufficiency while 2 had mitral insufficiency. Coronary orifices were implanted in situ in 13 cases while in 1 case, Cabrol's method was adopted. No death occurred during the surgery and in the subsequent follow-up, and all the patients had good recovery. CONCLUSION: Bentall's procedure can be effective in the treatment of AAA with aortic insufficiency.