This study aimed to investigate the drug susceptibility of wild-type and mutant avian influenza A (H7N9) virus neuraminidase (NA) to oseltamivir and zanamivir. Codon optimized DNA of H7N9 (A/ Hangzhou/1/2013) NA was synthesized and constructed into the pcDNA3.1/His vector (NA(H7N9-WT)). Mutant NA(H7N9-H274Y) and NA(H7N9-R292K) plasmids were constructed by directed mutagenesis PCR using NA(H7N9-WT) plasmid as the template followed by sequencing. NA plasmids were transfected into 293T cells and cell lysates containing NAs were collected 48 h post-transfection. Wild-type and mutant NAs were analyzed by Western blotting and their activities were tested by the 4-MUNANA-based assay. All three NAs were expressed and enzymatic activities were confirmed. The effects of oseltamivir and zanamivir on all three NAs were then tested. It showed that the half maximal inhibitory concentrations (IC50s) of oseltamivir carboxylate on NA(H7N9-WT), NA(H7N9-H274Y) and NA(H7N9-R292K) were 1.6 nM, 15.1 nM, and > 1 000 nM with fold changes of 9 and > 625, respectively. The IC50 values of zanamivir on NA(H7N9-WT), NA(H7N9-H274Y), and NA(H7N9-R292K) were 1.1 nM, 1.4 nM, and 38.0 nM with fold changes of 1.3 and 34, respectively. These results indicated that oseltamivir and zanamivir could significantly inhibit NA(H7N9-WT). NA(H7N9-R292K) showed high-level resistance to both drugs (34-fold and 625-fold) and NA(H7N9-H274Y) was sensitive to both (1.3-fold and 9-fold). These results indicated that both oseltamivir and zanamivir could be used for patients infected with the H7N9 virus. However, when patients carried the H7N9 virus with a NA R292K mutation, other medications would be preferred over oseltamivir or zanamivir.
Antiviral Agents ; pharmacology ; Humans ; Influenza A Virus, H7N9 Subtype ; drug effects ; enzymology ; genetics ; Influenza, Human ; virology ; Microbial Sensitivity Tests ; Mutation ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Oseltamivir ; pharmacology ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Zanamivir ; pharmacology

Objective To summarize the clinical features of neonatal lower respiratory tract infection of respiratory syncytial virus (RSV). Methods Data of newborns admitted into Deparrment of Neonatology of Yuying Children Hospital of Wenzhou Medical College from January to December 2010 with RSV lower respiratory tract infection were retrospectively analyzed. The characteristics of clinical manifestation,examination,treatment and prognosis were studied.Factors affecting the duration of hospitalization were analyzed with x2 test. Results Of 354 cases of neonatal lower respiratory tract infection,75 (21.2％) were RSV positive,whose attack time was from 5 to 25 days with an average of 13.2 days; and there were two preterm infants (2.7％).The chief clinical manifestations were cough (100.0％,75/75),followed by shortness of breath (56.0％,42/75),spittle (52.0％,39/75) and fever with temperature ≥38.0 ℃ (25.3％,19/75).The white blood cell count of one newborn was higher than 20.0× 109/L; 52 cases (69.3％) had higher blood lymphocyte percentage (＞ 0.48) ; eight cases (10.7％) had higher level of C-reactive protein (≥ 8 mg/L).Among the 58 patients who underwent blood gas analysis,28 (48.3％) had carbon dioxide retention (partial pressure of carbon dioxide≥ 45 mm Hg). Chest X-ray results within 48 hours after hospitalization suggested that 52.0％ patients (n =39) had pneumonia and 20.0％ (n=15) had bronchitis.Among 69 patients who had sputum cultures,20 cases were positive.And for the 11 cases who had blood cultures,three were positive for Staphylococcus epidermidis.Finally,73 cases were cured,two were improved and no death was reported.The duration of hospitalization was from 3 to 17 days with an average of 8.4 days.Neonates with RSV lower respiratory tract infection who had higher C-reactive protein (≥8 mg/L),carbon dioxide retention,nasal oxygen catheter or assisted ventilation had higher percentage of longer hospital stay (≥7 days). Conclusions Compared with infants,the clinical characteristics of RSV lower respiratory tract infections in neonates are atypical,which are mainly menifested by cough,shortness of breath,spittle,but rare wheezing.There are no effective interventions against RSV infection except for supportive therapy.

Objective Previous studies indicated that activation of Toll-like receptor4 (TLR4) was involved in the progression and instability of atherosclerotic plaque.Anti-inflammatory effects were shown in statins. However,the mechanisms underlying these effects have not been well explored.We test the hypothesis that a por- tion of these anti-inflammatory effects are mediated by regulation of TLR4 expression.Methods One hundred twenty-one subjects (22 normal persons,17 patients with stable angina and 82 patients with ACS) were recruited. 41 patients with ACS were randomized to atorvastatin 10 mg/d or atorvastatin 40 mg/d on top of routine anti-anginal treatment.Serum level of hsCRP,blood lipids,TLR4 expression on CD_(14)~+ monocytes were measuered before and after one month treatment.TLR4 expression on CD_(14)~+ monocytes were quantified via flow-cytometry.Results hsCRP and TLR4 expression on CD_(14)~+ monocytes in patients with ACS were higher than patients with stable angina and normal persons(hsCRP,ACS:11.1?14.3 vs stable angina:2.5?2.7 mg/L vs normal:2.3?4.2 mg/L,P

Female ; Humans ; Infant, Newborn ; Mastocytosis, Systemic ; diagnosis ; pathology

OBJECTIVETo study the relationship between pharmacological effects and properties of traditional Chinese medicine by the decision tree algorithm.

METHODBased on of pharmacological effects of traditional Chinese medicine, the decision tree algorithm was applied in the study on the relationship between pharmacological effects and properties of traditional Chinese medicines. A model was established with the decision tree algorithm for the purpose of predicting the properties of traditional Chinese medicine components.

RESULTThe established model was reliable and stable, and could be used to predict the properties of traditional Chinese medicine components.

CONCLUSIONThe prediction for the properties of traditional Chinese medicine components with a decision tree model could reflect the theoretical connotation of the properties of traditional Chinese medicine components to some extent and provide a new method for studying the properties of traditional Chinese medicine components.

Objective To investigate the effect of lamivudine (LAM) on serum interferon (IFN)-γ and interleukin (IL)-4 levels in patients with chronic hepatitis B(CHB). Methods Serum IFN-γ and IL-4 levels were measured with enzyme-linked immunosorbent assay (ELISA) in 66 CHB patients at baseline and 3, 6, 9 and 12 months after LAM treatment respectively. And 20 healthy volunteers served as healthy control. The comparision of pretreatment and post-treatment was done using t test and numberation data were analyzed by non parametric rank sum test. Results In HBeAg positive patients, the serum level of IFN-γ was (21.03±4.44) ng/L in complete response group, which was higher than partial response group [(13.85±3.92) ng/L] and non-response group [(10.63± 3.11) ng/L] (t=7.56,t=11.87, both P<0.01). Take that IFN-γ is 15.66 ng/L as boundry patients with high baseline IFN-γ level showed much higher complete response rate (31.0% vs 8.7%, x2 =8.391, P<0.01) and lower non-response rate (13.8% vs 52.2%,x2=4. 256, P<0.01) than those with low baseline IFN-γ levels. After LAM treatment, the IFN-γ/IL-4 ratios in complete response patients were approximate or even higher than those in healthy group, whereas the IFN-γ/ IL-4 ratios of patients with partial response and non response were lower than those in healthy group. In HBeAg negative patients, the IFN-γ/IL-4 ratios increased slowly but didn't reach the same levels of healthy group. Conclusions It is suggested that LAM treatment can increase IFN-γ release and reduce IL-4 release in CHB patients. The response type to LAM therapy is associated with the recovery of T helper cell (Th) 1/Th2 balance post-treatment and pretreatment IFN-γ levels.

OBJECTIVE:To explore the protective effect and its mechanisms of Sinomenium acutum polysaccharide on mice with acute alcoholic liver injury. METHODS:60 mice were randomly divided into blank control group (normal saline),model group (normal saline),bifendate group (positive control,150 mg/kg) and S. acutum polysaccharide low-dose,medium-dose, high-dose groups(100,200,400 mg/kg),10 in each group,intragastrically administrated,once a day,for continual 7 d. 1 h af-ter last administration,mice received 50% ethanol (0.1 mL/10 g) intragastrically to induce acute alcoholic liver injury model ex-cept for those in blank control group. After 12 h,alanine aminotransferase(ALT),aspartate aminotransferase(AST)levels in se-rum,malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),glutathione peroxidase(GSH-Px)levels in liv-er tissue of mice were determined;hematoxylin-eosin staining was conducted to observe the pathological changes in liver tissue;flow cytometry was used to detect the cell apoptosis rate. RESULTS:Compared with blank control group,mice in model group showed pathological changes in edema,disordered cell arrangement and local necrosis;ALT and AST levels in serum,MDA level in liver tissue and hepatocyte apoptotic rate were significantly increased,while the SOD,GSH and GSH-Px levels in liver tissue were significantly decreased,with statistical significances (P<0.01). Compared with model group,cell degeneration and necrosis degree of mice were improved in S. acutum polysaccharide medium-dose and high-dose groups;except for cell apoptosis rate of liver in S. acutum polysaccharide low-dose group was not decreased significantly,the above-mentioned indicators in other treatment groups were significantly improved(P<0.05 or P<0.01). CONCLUSIONS:S. acutum polysaccharide shows obvious protective effect on mice with acute alcoholic liver injury,its mechanism might re-late to anti-oxidation stress and inhibiting hepatocyte apoptosis.

Actins ; metabolism ; Child, Preschool ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Gastrointestinal Stromal Tumors ; pathology ; Humans ; Immunohistochemistry ; Myofibroma ; metabolism ; pathology ; surgery ; Stomach ; chemistry ; pathology ; surgery ; Stomach Neoplasms ; metabolism ; pathology ; surgery

According to the principle of evidence-based medicine (EBM), Chinese medical literatures based descriptive statistical analysis of common Chinese medical syndrome types were performed. By data extraction, standardization, and frequency calculation of disease names and syndrome types from 286 literatures in line with the inclusion criteria, the frequencies of diseases and syndromes were obtained to analyze common syndrome types in clinical practice, to analyze the distribution features of disease related syndromes and syndrome related diseases, to analyze the distribution of basic Chinese medical syndrome types in clinical common diseases as a whole, thus providing reference for clinical and basic researches.
Evidence-Based Medicine ; Humans ; Medicine, Chinese Traditional

BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.