Objective To evaluate the status of P-selectin and platelet activating factor(PAF) in vivo in patients with deep venous thrombosis(DVT) and to observe their changes under interneving of medicines.Methods P-selectin and PAF of fourty patients and twenty normal subjects were fluorescence labled with corresponding monoclonal antibodies by flow cytometry( FCM ) and immunologic method respectively. Results P-selectin and PAF in patients with DVT were higher than that in normal subjects in early period of the disease, and they significantly decreased in different time after patients were treated. P-selectin was significantly different between patients who received sodium ozagrel treatment and those who not (P ＜0.05 ), but PAF was similar( P ＞ 0. 05 )after fourteen days. One month later, P-selectin and D-dimer in DVT patients were lower than before. However, the positive rate of P-selectin of DVT was still higher than normal subjects. Conclusions The platelet is activated in vivo in patients with DVT, so does fibrinolysis. Sodium ozagrel can decrease activity of platelet. P-selectin and PAF may be used diagnostic markers. Post-discharge patients are still at high-risk and must be regularly followed-up.

Rupture of anterior cruciate ligament (ACL) is a common sports injury.It will cause knee osteoarthritis because of joint instability and acceleration of degenerative changes after injury.Arthroscopic reconstruction with a tendon graft is a common procedure to achieve function recovery.A series of histological changes and structural modification happened between the tendon and bone tunnel after ACL reconstruction and complete tendon-bone healing was achieved finally.A number of factors affects the healing process which determined the long-term result of the treatment.Published literatures reported that about 11％-32％ patients underwent ACL reconstruction were not satisfied with the results and 10％ of them required reoperation.Studies on the tendon-bone healing have long been a research hotspot in sports medicine.Controversy still remains not only on the fundamental healing process but also on the stimulating factors despite of a large amount of researches on its physiological basis,influencing factors,et al.This article provides a review of the basis and influence factors of the healing process,and summarizes the methods to accelerate the process of tendon-bone healing.

Effects of six kinds of Chinese herb extracts, including Folium Crataegi extract, Herba Epimedii extract, Folium Acanthopanacis Senticosi extract, Trifolium pratense L. extract, Folium Ginkgo extract and Radix Puerariae extract, on the activities of CYP450 isozymes (CYP1A2, CYP2C, CYP2E1, CYP2D, CYP3A) in rat hepatic microsomals were studied by using a UPLC-MS/MS (MRM) and cocktail probe substrates method. The results showed that effects of six kinds of Chinese herb extracts on each CYP450 isozyme activity were inhibitory. The IC50 of Folium Crataegi extract for the inhibition of rat microsomal CYP2D activity was only for 4.04 microg x mL(-1), which showed the highest inhibition; Trifolium pratense L. extract had strong inhibitory action to CYP2D, the IC50 value was 5.73 microg x mL(-1); Folium Crataegi extract also had strong inhibitory action on CYP2E1, the IC50 value was 10.91 microg x mL(-1). Furthermore, the IC50 of Folium Ginkgo extract for the inhibition of rat microsomal CYP3A, 2D, 2E1 activities were 45.12, 35.45 and 22.41 microg x mL(-1), respectively, and the IC50 of Folium Acanthopanacis Senticosi extract on the inhibition of rat microsomal CYP2E1 activity was 32.89 microg x mL(-1). In addition, mechanism of inhibition experimental results showed that the inhibiting abilities of Folium Crataegi extract and Radix Puerariae extract on each CYP450 isozyme increased with the increasing of the preincubation time, therefore, the inhibitory effects were a mechanism-based inhibition.

Japanese encephalitis virus (JEV) is a single-stranded and positive-sense RNA, which has a single ORF (open reading frame), encoding a polyprotein precursor. Non-structural protein 3 (NS3) plays an important role in processing the polyprotein precursor and has become an important drug target of flavivirus. In this study, NS2BH-NS3 gene was amplified by PCR and subcloned to the prokaryotic expression plasmid, resulting pET30a-NS2BH-NS3. The fusion protein was expressed in Escherichia coli BL21 (DE3) in soluble form after induction by Isopropyl beta-D-1-Thiogalactopyranoside (IPTG). The recombinant protein was purified by Ni-NTA affinity column. Then a fluorescence resonance energy transfer (FRET) method was used to determine enzymatic activity and the assay conditions were optimized. After screening 113 compounds, we found two compounds inhibiting the activity of NS2BH-NS3. This study provides a convenient and cost-effective method for screening of JEV NS3 protease inhibitor.
Encephalitis Virus, Japanese ; enzymology ; Escherichia coli ; metabolism ; High-Throughput Screening Assays ; Protease Inhibitors ; chemistry ; RNA Helicases ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Serine Endopeptidases ; metabolism ; Viral Nonstructural Proteins ; metabolism

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.

Background:Capsule endoscopy( CE)has been widely used for the diagnosis of small intestinal diseases. How to enhance the complete examination rate( CER)has attracted more and more attention in recent years. Aims:To investigate the effect of different intervention on gastric transit time( GTT)and small bowel transit time( SBTT)in CE. Methods:Ninety patients undergoing CE from January 2012 to May 2013 at Changhai Hospital were enrolled and randomly divided into control group,right lateral position group and motion group. The patients in control group were allowed to keep quiet, patients in right lateral position group were asked to lie on their right side,and patients in motion group were asked to take rapid pace walking after swallowing the capsule until the capsule passing pylorus. GTT,SBTT,CER and disease diagnosis rate among the three groups were compared. Results:Compared with control group,GTT was significantly decreased in right lateral position group(P﹤0. 05),however,no significant difference in SBTT was found between right lateral position group and control group(P ﹥0. 05). GTT and SBTT were both significantly decreased in motion group than in control group(P﹤0. 05). No significant differences in CER and disease diagnosis rate were found among the three groups(P﹥0. 05). Conclusions:Right lateral position can shorten GTT,rapid pace walking can shorten GTT and SBTT,and both these two intervention have substantial clinical values in CE.

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.

Objective:To investigate the pathogenic bacteria distribution and risk factors of pulmonary infection after tracheotomy in patients with stroke coma,and to put forward preventive measures.Methods:96 patients with stroke coma from January 2016 to February 2017 in our hospital were retrospectively analyzed.The incidence of pulmonary infection and distribution of pathogenic bacteria of patients with stroke coma were analyzed.At the same time,the risk factors of pulmonary infection were analyzed by single factor and multiple factors logistic regression analysis,and corresponding preventive measures were put forward.Results:The incidence of pulmonary infection after tracheotomy in 96 patients with stroke coma was 48.96％ (47/96).A total of 104 pathogens were isolated and cultured,including gram negative bacteria 69 strains (66.35％),gram positive bacteria 20 strains (19.23％) and fungus 15 strains (14.42％).Single factor regression analysis results showed that pulmonary infection after tracheotomy in patients with stroke coma was closely related with age,basic diseases,time of tracheotomy,and bed time,use of broad-spectrum antibiotics,smoking history,artificial airway,times of sputum suction and inhalation(P＜0.05),and it was not related to the patient's gender,weight,stroke type (P＞0.05).Multivariate logistic regression analysis showed that age 45 years old,complicated with basic disease,time oftracheotomy 5 d,use of broad-spectrum antibiotics,smoking history and the establishment of artificial airway were risk factors of pulmonary infection after tmcheotomy in patients with stroke coma (P＜0.05).ROC analysis results showed that the critical point (threshold C) oftmcheotomy time was 4.3 days,and the sensitivity and specificity were 0.851 and 0.918 respectively.Conclusion:The main pathogenic bacteria of pulmonary infection after tracheotomy in patients with stroke coma is gram-negative bacteria,age 45 years old,complicated with basic disease,time of tmcheotomy 5d,use of broad-spectrum antibiotics,smoking history and the establishment of artificial airway can lead to pulmonary infection after tracheotomy in patients with stroke coma,and the risk of pulmonary infection in patients with stroke coma will increase considerably after the time of tracheotomy for more than 4.3 days.Targeted measures should be taken to reduce the risk of pulmonary infection according to pathogenic features and risk factors.

To observe the effect of pIgR on Salmonella Enteritidis induced inflammation in jejunum and ileum in Chicken,7 day Hyline chickens were taken orally with Salmonella Enteritidis (SE) and killed after 1 d,3 d,7 d and 14 d.The mRNA expression of Toll-like receptor 4(TLR4),MyD88,TRAF6,NF-κB and pIgR was detected by real-time RT-PCR and pIgR protein level was detected by Western blot.The results showed that TLR4 signaling pathway was activated and mR-NA level of the pIgR in jejunum and ileum was enhanced (P＜0.01) and protein level of the pIgR in jejunum and ileum was up-regulated by SE.The study proved that TLR4 signal pathway on mucosal cell surface of jejunum and ileum was activated and expression of pIgR was up-regulated and gut mucosal immunity of chicken was strenghtened.

0.05), while the differences between hepatocarcinoma and cirrhosis as well as hepatocarcinoma and hepatitis B were significant (P