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Objective To explore the effects of curcumin on pro-inflammatory factors in the lung microvascular endothelial cells (LMVEC) model stimulated by thrombus. Methods The LMVECs were divided into six groups according to the random number table method. No treatment was given to the blank control group ; the model group was cultured for 7 hours in normal medium; the curcumin group was treated with 40 μmol/L curcumin for 72 hours ; the shRNA group was infected with shRNA adenovirus for 72 hours; the irregular chemokines (CX3CL1) overexpression group was infected with CX3CL1 overexpressing adenovirus for 72 hours; the shRNA+curcumin group infected with shRNA adenovirus and treated with 40 μmol/L curcumin together for 72 hours; CX3CL1 overexpression +curcumin group infected with CX3CL1 overexpressing adenovirus and treated with 40 μmol/L curcumin together for 72 hours. After each group was given the corresponding pretreatment, the thrombus natural precipitation was added each group for 12 hours. The contents of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), the mRNA expression levels of CX3CL1, CX3CL1 receptor (CX3CR1), IL-6, TNF-α and the protein expression levels of CX3CL1/CX3CR1, CX3CR1/NF-κB in various groups were observed, repeat 3 times in each group. Results The contents and mRNA expression of IL-6, TNF-αand protein expression of CX3CR1, NF-κB in the LMVEC group were significantly higher than those in blank control group [IL-6 (ng/L): 207.90±16.69 vs. 85.93±20.32, TNF-α (ng/L): 239.60±15.27 vs. 101.23±11.92; IL-6 mRNA: 0.66±0.05 vs. 0.11±0.02, TNF-α mRNA: 1.06±0.04 vs. 0.02±0.01; CX3CR1 protein:3.94±0.58 vs. 1.00±0.31, NF-κB protein: 1.20±0.07 vs. 1.00±0.10; all P < 0.05]; the contents of IL-6 in shRNA group, CX3CL1 overexpression group, shRNA + curcumin group, CX3CL1 overexpression + curcumin group were all obviously lower than those in LMVEC group (ng/L: 183.60±11.52, 159.27±15.02, 117.03±7.91, 119.97±11.43 vs. 207.90±16.69, all P < 0.01); the content of TNF-α was markedly increased in shRNA group compared with that of LMVEC group (ng/L: 282.00±5.63 vs. 239.6±15.27), while the contents of TNF-α in CX3CL1 overexpression group, shRNA+ curcumin group, CX3CL1 overexpression + curcumin group were all lower than those in LMVEC group (ng/L: 216.97±9.20, 203.97±19.03, 191.97±17.50 vs. 239.6±15.27, all P < 0.05). The mRNA expression levels in CX3CL1 overexpression group and CX3CL1 overexpression + curcumin group were significantly higher than those in the blank control group and the LMVEC group (CX3CL1 mRNA: 55 210.3±1 209.2, 165 296.3±8 082.4 vs. 3.3±0.6, 2.0±0.0, all P < 0.01). The mRNA expression level of IL-6 in shRNA group was higher than that in LMVEC group (0.82±0.17 vs. 0.66±0.05), the mRNA expression level of IL-6 in CX3CL1 overexpression was lower than that in LMVEC group (0.29±0.03 vs. 0.66±0.05), the changes after pretreatment with curcumin were more significant (1.06±0.03 vs. 0.66±0.05 and 0.15±0.01 vs. 0.66±0.05); the mRNA expressions of TNF-α in shRNA group, CX3CL1 overexpression group, shRNA+ curcumin group were significantly lower than those in LMVEC group (0.41±0.04, 0.88±0.07, 1.01±0.02 vs. 1.06±0.04), the mRNA expression level of TNF-α in CX3CL1 overexpression + curcumin group was significantly higher than that in LMVEC group (1.36±0.01 vs. 1.06±0.04). The protein expression of CX3CL1, CX3CR1, NF-κB in shRNA group, CX3CL1 overexpression group, shRNA + curcumin group, CX3CL1 overexpressing + curcumin group were significantly higher than those in the LMVEC group (CX3CL1 protein: 0.41±0.07, 0.59±0.09, 0.69±0.61, 1.02±0.23 vs. 1.33±0.33, CX3CR1 protein: 0.85±0.18, 1.10±0.16, 1.32±0.18, 1.54±0.08 vs. 3.94±0.58, NF-κB protein: 0.33±0.07, 0.41±0.08, 0.41±0.07, 0.63±0.08 vs. 1.20±0.07). Conclusion Curcumin can inhibit the secretion of IL-6, TNF-α, CX3CR1 and NF-κB in thrombus-stimulated LMVEC model.
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Objective To discuss the influence of Tanshinone ⅡA on the tight junction protein of intestinal mucosal epithelial cells in rat severe septic models. Methods Seventy-five Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group and Tanshinone ⅡA injection high (20 mg/kg), medium (10 mg/kg) and low (5 mg/kg) dose groups, each group 15 rats. Sepsis rat models were established by cecal ligation and puncture (CLP) method, in sham operation group, only switched abdominal surgery was performed without CLP. In Tanshinone ⅡA injection groups, different doses of Tanshinone ⅡA were injected intraperitoneally after modeling for 10 minutes and 6 hours; in sham operation and model groups, equal volume of normal saline was injected intraperitoneally at the same times as above. After operation, 3 L/kg of normal saline was injected into the caudal vein in all rats for fluid resuscitation.Twelve hours after operation, the rats were killed, the abdominal lymph nodes, liver, spleen and kidney tissues were taken for bacterial culture and calculating the rate of bacterial translocation; under microscope, the histopathological changes of ileum mucosal tissues were examined and Chiu scoring was carried out; TdT-mediated dUTP nick end labeling (TUNEL) was applied to detect the ileum mucosal epithelial cell apoptosis and calculating the index (AI);fluorescence immunoassay and Western Blot methods were used to measure the contents and protein expression levels of tight junction protein, junctional adhesion molecule-1 (JAM), Claudin-1, Zonula occludens-1 (ZO-1), Occludin, c-Fos and Tryptase. Results ① In bacterial cultures of abdominal lymph node, liver, spleen and kidney, the positive rate of mesenteric lymph node was the highest, followed by liver and spleen, mainly Escherichia coli, Proteus mirabilis, etc. The highest positive rate of bacterial culture was in model group (38.8%), followed by low dose of Tanshinone ⅡA injection group (35.0%), and the lowest was 16.6% in high dose Tanshinone ⅡA injection group, the differences being statistically significant in comparisons between any pair of groups (all P < 0.05). ② Pathological examination showed that the pathological changes of ileum mucosa were obvious and the Chiu score (4.17±0.98 vs. 0) and AI (11.70±2.87 vs. 2.17±0.80) in model group were significantly higher than those in sham group (all P < 0.05); with the increase of dosage of Tanshinone ⅡA injection, the pathological changes of rat ileum mucosa were improved gradually, the Chiu score and AI were decreased gradually, and the degrees of decrease in high dose Tanshinone ⅡA group were more significant than those in model group (Chiu score: 1.12±0.79 vs. 4.17±0.98, AI: 3.65±1.98 vs. 11.70±2.87, both P < 0.05).③ Immunofluorescence staining showed that the positive staining of protein JAM, ZO-1 and c-Fos were all green in color, Claudin-1, Occludin and Tryptase were all red in color, the localizations of all of them were in the cytoplasm, the protein expression of JAM, Claudin-1, ZO-1, Occludin from strong to weak in turn were Sham group, high, medium, low dose Tanshinone ⅡA group and model group, the expression of c-Fos, Tryptase from strong to weak in turn were model group, low, medium, high dose Tanshinone ⅡA group and Sham group. ④ Western Blot showed that the expressions of ileum tissue JAM, Claudin-1, ZO-1 and Occludin in model group were all significantly lower than those of the sham group, while the expressions of c-Fos, Tryptase were obviously higher than those of the sham group, with the increase of dosage of Tanshinone ⅡA, the expressions of JAM, Claudin-1, ZO-1 and Occludin were increased gradually and the protein expressions of c-Fos and Tryptase were gradually decreased, and the changes in high dosage group of Tanshinone ⅡA were more significant than those in low and moderate groups [JAM (gray value): 25.39±1.82 vs. 12.41±1.34, 19.45±1.66, Claudin-1 (gray value): 28.44±1.56 vs.17.26±1.46, 21.23±1.34, ZO-1 (gray value): 28.84±1.59 vs. 16.45±1.21, 24.22±1.46, Occludin (gray value): 25.49±1.63 vs. 13.34±1.45, 19.45±1.37, c-Fos (gray value):15.76±1.36 vs. 27.84±1.36, 21.22±1.73, Tryptase (gray value): 14.44±1.41 vs. 28.14±1.38, 22.32±1.57], all the above comparisons of different dosage groups were statistically significant (all P < 0.05). Conclusion Tanshinone ⅡA injection may improve intestinal wall structure and reduce bacterial translocation by improving the intestinal mucosal tight junction protein in sepsis model rats, and this effect is positively correlated to Tanshinone ⅡA dosage.
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Objective To observe the effects of curcumin on B-type natriuretic peptide (BNP),cardiac troponin Ⅰ (cTnI) and D-dimer in rats with acute pulmonary embolism (APE).Methods The irregular cbemotactic factor (CX3CL1)-shRNA and overexpression vector were designed and synthesized,and adenovirus packages were respectively carried out.Sixty Spargue-Dawley (SD) rats were divided into normal control group,sham-operation group,model group,curcumin group,curcumin+CX3CL1-shRNA group and curcumin+CX3CL1 overexpression vector group by random number table (each group 10 rats).The APE rat models were duplicated by autologous blood embolus method.in the pharmacological intervention groups,1 day and again 40 minutes before operation,a dose of curcumin 100 mg/kg was administered by gavage respectively;in virus intervention groups,3 days before modeling,intravenous injection of 109 pfu into a tail vein was performed once;in sham-operation group and model group,an equivalent dose of normal saline 2 mL daily was administered by gavage;the normal group did not accept any intervention.Six hours after modeling,the change of lung tissue were observed under confocal laser scanning microscope,the lung tissue was taken for hematoxylin-eosin (HE) staining,histopathological changes of lung tissue were observed,and the levels of serum BNP,cTnI,D-dimer were detected by enzyme-linked immunosorbent assay (ELISA).Results Virus infected lung tissue was green,nucler was blue.The HE staining in lung tissue showed:histopathological changes in the model group were more obvious than those in normal group and sham-operation group,after curcumin intervention,some emboli in pulmonary arteries in all treatment groups were partially dissolved,and the inflammation was alleviated.The results of ELISA shows:the contents of cTnI,BNP,D-dimer were significantly higher in model group than those in shamoperation group [cTnI (ng/L):224.97 ± 37.96 vs.149.13 ± 36.65,BNP (ng/L):53.66 ± 21.72 vs.24.62 ± 10.44,D-dimer (ng/L):78.68 ± 13.10 vs.42.00 ± 9.30,all P ＜ 0.05];the contents of cTnI,BNP,D-dimer were significantly lower in curcumin group,curcumin+shRNA group and curcumin+CX3CL1 overexpression vector group than those in model group (cTnI was 162.98 ± 28.65,143.32 ± 34.55,159.50 ± 37.80 vs.204.97 ± 37.96,all P ＜ 0.05,BNP was 27.45 ± 11.00,26.95 ± 6.66,33.05 ± 7.05 vs.53.66 ± 21.72,all P ＜ 0.05,D-dimer was 54.34 ± 4.57,53.38 ± 11.01,56.33 ± 6.80 vs.78.68 ± 13.10,all P ＜ 0.05);there were no statistical significant differences in comparisons among all intervention groups (all P ＞ 0.05).Conclusion Curcumin is able to improve serum BNP,cTnI,D-dimer in APE rats,but its relation with CX3CL1 is not obvious.
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Klebsiella pneumoniae carbapenemases-producing Klebsiella pneumoniae (KPC-Kp) has caused a global public health crisis, and the severity of its infection is associated with high mortality in hospitalized patients. Therefore, the KPC-Kp prevention methods and the corresponding treatment strategy exploration are imminent. The risk factors and the treatment progress of KPC-Kp colonization or infection are reviewed in this paper to explore corresponding preventive measures and treatment strategies for clinical prevention and treatment.
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Objective To evaluate the therapeutic effect of acupoint catgut embedding on gastrointestinal dysfunction after surgery in patients with severe traumatic brain injury.Methods Sixty-eight consecutively hospitalized patients with gastrointestinal dysfunction after surgery in patients with severe traumatic brain injury admitted to the Department of Intensive Care Unit (ICU) in the First Affiliated Hospital of Zhejiang University of Chinese Medicine from January 2015 to December 2015 were enrolled.Six of those patients were withdrawn from the course of observation,and actually only 62 patients entered the study.They were divided into an observation group (32 cases) and a control group (30 cases) by random number table.All the patients of two groups were given conventional treatment of western medicine for consecutive 7 days.The observation group was additionally treated with acupoint catgut embedding.The control group took mosapride tablets via a nasogastric tube 5 mg,3 times a day.Intra-abdominal pressure (IAP),gastric residual volume (GRV),time for reaching standard enteral nutrition,serum diamine oxidase(DAO),D-Lactate activity,incidence of hospital-acquired pneumonia (HAP),hospitalization days in ICU,28-day mortality and adverse reactions of acupoint catgut embedding were observed in the two groups.Results After treatment,the levels of IAP,DAO,D-lactic acid were lower than those before treatment,GRV was less than that before treatment in the two groups,and after treatment for 7 days the changes were more significant in the observation group [IAP (cmH2O,1 cmH2O =0.098 kPa):12.42±3.11 vs.17.70 ± 2.98,GRV (mL):129.54 ± 57.54 vs.242.91 ± 99.29,DAO (U/L):9.06± 2.84 vs.13.60 ± 7.16,D-Lactate (mg/L):16.47 ± 5.39 vs.26.29 ± 9.62,all P ＜ 0.01].Days of mechanical ventilation (days:6.97 ± 4.13 vs.9.23 ± 4.61),time for reaching standard enteral nutrition (days:4.25 ± 3.15 vs.5.50 ± 3.12) and hospitalization days in ICU (days:9.50± 4.03 vs.12.20 ± 6.38) in observation group were significantly shorter than those in control group (all P ＜ 0.05).The incidence of HAP [15.6％ (5/32) vs.40.0％ (12/30)] and 28-day mortality [3.1％ (1/32) vs.6.7％ (2/30)] in observation group were obviously lower than those in control group.The patients in observation group had no significant adverse reactions of acupoint catgut embedding.Conclusions Acupoint catgut embedding therapy is not only easy to operate,but also can effectively improve gastrointestinal motility and intestinal barrier function in patients with gastrointestinal dysfunction after surgery of severe head injury,thus it is beneficial to reaching the enteral nutritional goal in less time,reducing the incidence of HAP,shortening the hospitalization time in ICU and promoting the recovery of the patients.
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Objective To investigate the risk factors of prognosis of gram-positive bacteria associated nosocomial bloodstream infections,and to investigate the drug resistance of the strains.Methods A total of 132 patients with gram-positive bacteria associated nosocomial bloodstream infections were collected from the First Affiliated Hospital of Zhejiang Chinese Medicine University during January 2010 and December 2012.Clinical data including demographic characteristics,underlying diseases,risk factors and use of antibacterial agents were retrospectively analyzed.According to 28-day prognosis,patients were divided into survival group (n =97) and death group (n =35).Binary logistic regression was used to identify the risk factors of 28-day fatality.Results Among 132 patients,49 (37.12％) were infected with coagulase-negative Staphylococcus,46 (34.85％) were infected with Staphylococcus aureus,37 (28.03％)were infected with Enterococcus.The rates of methicillin resistant coagulase negative Staphylococci (MRCNS) and methicillin-resistant Staphylococcus aureus (MRSA) were 77.55％ (38/49) and 54.35％ (25/46),respectively.The rate of linezolid resistant coagulase negative Staphylococci was 8.16％ (4/49) ; Four out of 37 strains (10.81％) of Enterococcus were both resistant to vancomycin and linezolid.Binary logistic regression showed that septic shock (OR =34.344,95％ CI:6.539-180.389,P =0.000),deep venous catheterization (OR =13.411,95％ CI:1.877-95.832,P =0.010),no catheter removal after infection (OR =8.759,95％ CI:2.197-34.911,P =0.002),parenteral nutrition (OR =3.684,95％ CI:1.072-12.663,P =0.038),inappropriate antibacterial therapy in early stage (OR =12.951,95％ CI:2.075-80.836,P =0.006) and Enterococcus associated bloodstream infections (OR =4.227,95％ CI:1.090-16.394,P =0.037) were independent risk factors of 28-day fatality in patients with gram-positive bacteria associated nosocomial bloodstream infections.Conclusions The predominant pathogens are coagulase-negative Staphylococcus,Staphylococcus aureus and Enterococcus in gram-positive bacteria associated nosocomial bloodstream infections.Patients with septic shock,deep venous catheterization,no catheter removal after infection,parenteral nutrition,inappropriate antibacterial therapy in early stage and Enterococcus associated bloodstream infections are likely to have high fatality rate.
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<p><b>OBJECTIVE</b>To develop an UPLC method for the determination of leonurine in traditional Chinese medicines.</p><p><b>METHOD</b>An Acquity UPLC BEH C18 column (2.1 mm x 100 mm) with 1.7 microm particle size was used. The mobile phase was composed of methanol and ammonium formate buffer (pH 4.0) in gradient mode. The flow rate was 0.3 mL x min(-1) and the chromatograpic run time was 18 min for one sample.</p><p><b>RESULT</b>The results showed that there was significant difference in the content of leonurine in the leonurus products from different pharmaceutical companies. The leonurine content in those products is in the range of 45.6-193 microg x g(-1).</p><p><b>CONCLUSION</b>The method is simple, reproducible and reliable. It can be used to control the quality of related drugs.</p>
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Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Gallic Acid , Leonurus , ChemistryABSTRACT
0.05).The successful rates of weaning from MV of embedment group(60%,75%) were significantly higher than those of control group(30%,35%) and traditional needle group(25%,30%) in the 2 nd and 4 th week after treatment(all P
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OBJECTIVE To investigate the clinical value of human leukocyte antigen DR(HLA-DR) expression on monocytes in prediction of prognosis in patients with hospital acquired pneumonia(HAP).METHODS A total of 68 patients suffered from HAP and transferred to intensive care unit(ICU) were enrolled in the study and divided into survivors group and nonsurvivors group according to prognosis.HLA-DR expression on monocytes,acute physiology,age and chronic health evaluation((APACHE)) Ⅱ score,serum cortisol level,T lymphocytes(subpopulation) and natural killer(NK) cell were observed.Relationship between HLA-DR expression on(monocytes) and APACHE Ⅱ score and 28-day mortality rates was analyzed through Logistic regression model.(RESULTS) HLA-DR expression on monocytes in nonsurvivors was significantly lower than that in survivors (P