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

BACKGROUND:Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE:To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS:Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION:Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.

BACKGROUND:Osteogenic differentiation is a complex process involving transcriptional and post-transcriptional regulation by multiple signaling pathways, and the specific mechanisms remain unclear. It is of great significance to study the role of critical miRNAs in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in the treatment of osteoporosis and bone defects. OBJECTIVE: To explore the regulatory ability of miR-21/Sprouty1 function axis in the osteogenic differentiation of BMSCs in patients with postmenopausal osteoporosis. METHODS:BMSCs were isolated from healthy people (H-hBMSCs) and patients with postmenopausal osteoporosis (PMOP-hBMSCs), and their osteogenic ability was compared. Expression of miR-21 and Spry1 at gene and protein levels was detected by real-time RT-PCR and western blot assay, respectively. miR-21 expression was upregulated via transfection in PMOP-hBMSCs, and the osteogenic ability and Spry1 expression of the cells were detected, while real-time RT-PCR and western blot were used to detect the expression of osteogenic marker genes, Runx2 and Osterix. RESULTS AND CONCLUSION:Compared with H-hBMSCs, PMOP-hBMSCs osteogenic ability was weakened significantly, miR-21 expression decreased, and Spry1 expression increased, indicating an inhibition to the miR-21-Spry1 function axis. Through the transfection of miR-21 and down-regulation of Spry1, the expression levels of Runx2 and Osterix were increased, and PMOP-hBMSCs osteogenic ability was partially restored.

Objective Acute respiratory dysfunction (ARD) can occur after aortic surgery with the use of cardiopulmonary bypass and deep hypothermic circulation arrest, but relatively little is known about acute respiratory dysfunction in the patients with type A aortic dissection. This study aims to analyze the independent risk factors of acute respiratory dysfunction after A type aortic dissection surgery and to assess possible prevention and treatment option in the future. Methods Clinical data of the 252 patients including 193 male patients and 59 female patients who underwent type A aortic dissection surgery from February 2009 to October 2010 were collected. The mean age was 47 years. Postoperative acute respiratory dysfunction was defined as oxygenation impairment (PaO2/FiO2 ＜ 150) that occurred within 72 h of surgery except pleural effusion, cardiogenic pulmonary edema, pneumonia, pulmonary embolism and haemato-/ pneumothorax. There were 187 acute A type aortic dissection patients and 65 chronic type A aortic dissection patients. Clinical characteristics including age, gender, weight, height, history of hypertension, history of smoking, preoperative complications such as preoperative shock and acute renal failure, pericardial effusion, previous cardiac surgery, time from event to surgery, malperfusion syndrome, cardiopulmonary time, cross-clamp time,deep hypothermia circulation arrest time, surgical procedure, duration of intensive care unit stay and postoperative complications including tracheotomy, dialysis dependent renal failure and hospital mortality were gathered. Arterial blood analysis, chest X ray, ventilator parameters, number of blood transfusion and flood balance were assayed after operation. All the factors were evaluated by means of univariate and multivariate logistic regression analysis to identify relative risk factors of ARD. Results Acute respiratory dysfunction occurred in 32 (12.7% ) patients. The in-hospital mortality was significant difference between acute respiratory dysfunction group and non- acute respiratory dysfunction group (P ＜ 0.05). The value of BMI, incidence of acute aortic dissection, preoperative SBP level, cardio-pulmonary bypass time, aortic clamp time and total arch replacement in acute respiratory dysfunction group were significantly higher than the values in non- acute respiratory dysfunction group. Multivariate Logistic regression analysis showed blood transfusion more than 10 units and cardio-pulmonary bypass time more than 160 minutes were independent risk factors of early stage acute respiratory dysfunction after type A aortic dissection surgery.Conclusion Acute respiratory dysfunction after type A aortic dissection was a severe early stage postoperative complication and was associated with in-hospital mortality. The patients in acute aortic dissection were prone to have acute respiratory dysfunction. The independent risk factors of acute respiratory dysfunction included blood transfusion more than 10 units and cardio-pulmonary bypass time more than 160 minutes.

AIM:To explore the diagnosis and treatment methods of radioaction-induced optic neuritis ( RION) through the clinical dates of 17 patients.
METHODS: It was a retrospective case series study. From August 2008 to October 2013, 17 cases (24 eyes) of Rion clinical dates from Chinese PLA General Hospital were studied. The diagnosis methods including visual acuity, pupil, fundus, visual field, fundus fluorescein angiography (FFA), visual electrophysiological testing, and head MRI. To analysis the clinical date of patients with diagnosis of RION by statistical description.
RESULTS: The deterioration degree of vision: 13 eyes were classified as Ⅳ, 9 eyes as Ⅲ, 2 eyes as II. Ten eyes RAPD ( + ) , visual electrophysiology is extinguished. The retina of 5 eyes showed flame hemorrhages and cotton wool spots exudation. Optic nerve head edema in one eye. T1 - weighted MRI enhanced in 19 eyes which showed optic nerve of the intracranial and intratubal segments abnormal changed, optic chiasm and pituitary stalk signal abnormalities and enhancement of the optic nerve. Tortuous optic nerves and rough edges were observed in 5 eyes. Treatment effect: 4 eyes of visual acuity improved, 1 eye from blindness to light perception,1 eye from 0. 08 to 0. 2, 1 eye from 0. 4 to 0. 6,1 eye from 0. 04 to 0. 15, the rest of the cases did not see any improvement.
CONCLUSION: The unique clinical manifestation of RION can provide objective basis for clinical diagnosis in time, but there have not been proven any effective treatments.

Poly(β-amino esters) (PBAE) are used for drug carrier and have many advantages, such as pH-sensitivity, low toxicity, structural diversity and the synthetic method of PBAE is easy. Therefore they are possessed broad application prospect in tumor-targeted drugs delivery systems. In this paper, the structural features and target drugs delivery property of PBAE are reviewed. The application forms of PBAE and different anti-cancer drugs loaded in the copolymer for tumor-targeted drugs delivery systems are introduced particularly.
Antineoplastic Agents ; chemistry ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Esters ; chemistry ; Hydrogen-Ion Concentration ; Neoplasms ; drug therapy ; Polymers

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

Coprecipitation method was used to prepare triiron tetroxide magnetic nanoparticles enclosed in L-DOPA, and then EDC was used to activate the carboxyl group of L-DOPA after the nanoparticles were synthesized. The carboxyl group of L-DOPA formed amide bond with specific amino on the aptamer by dehydration condensation reaction. The surfaces of magnetic nanoparticles were modified with aptamer and L-DOPA. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), nanoparticle size analysis (SEM), magnetic measurement (VSM) and other testing methods were used to detect the magnetic nanoparticles in different stages. The endothelial progeni-tor cells (EPCs) were cocultured with the surface modified magnetic nanoparticles to evaluate cell compatibility and the combination effect of nanoparticles on EPCs in a short period of time. Directional guide of the surface-modified magnetic nanoparticles to endothelial progenitor cells (EPCs) was evaluated under an applied magnetic field and simulated dynamic blood flow condition. The results showed that the prepared magnetic nanoparticles had good magnetic response, good cell compatibility within a certain range of the nanoparticle concentrations. The surface modified nanoparticles could combine with EPCs effectively in a short time, and those nanoparticles combined EPCs can be directionally guided on to a stent surface under the magnetic field in the dynamic flow environment.
Endothelial Progenitor Cells ; cytology ; drug effects ; Ferrosoferric Oxide ; chemistry ; Humans ; Levodopa ; chemistry ; Magnetite Nanoparticles ; chemistry ; Spectroscopy, Fourier Transform Infrared ; X-Ray Diffraction

In order to study the accumulation of Fritillaria thunbergii cultivar, peimine content in Xiaye, Kuanye, Duozi and Xiaosanzi bulbs of different sizes and parts was determined by HPLC-ELSE. The results indicated that the peimine content varied significantly with the cultivar type, the size and part of bulb. The distribution laws of peimine were as follow: Xiaosanzi > Duozi > Xiaye > Kuanye, small-size bulb > big-size bulb, core bud > scale. The peimine yield per plant in Duozi was the highest.
Cevanes ; analysis ; Chromatography, High Pressure Liquid ; Fritillaria ; chemistry ; growth & development

Objective To establish an appropriate preterm hypoxic-ischemic brain injury animal model. Methods A total of 32 pregnant New Zealand white rabbits at gestational day 25 were selected. The uterine blood supply in pregnant rabbits was blocked for 30, 35, 37, 40 minutes respectively, while in the control group it was not blocked. Then the pregnant rabbits were subjected to cesarean section 24 hours (at embryonic day 26, A group) or 5 days (at embryonic day 30, B group) after the experimental procedure. The general conditions of the newborn rabbits were recorded. The degree of neurobehavioral impairment in newborn rabbits was evaluated. The histological changes of brain tissue were observed. Results In A group, all newborn rabbits survived with ischemia for 30 minutes, while the stillbirth rates increased from 31.0% to 100% with ischemia from 35 to 40 minutes. In survived nowborn rabbits, the brain water content and the number of apoptotic brain cells were increased with prolonged ischemia. All these differences were statistically significant (all P<0.05). In B group, the stillbirth rates increased to 50.0% and 65.7% respectively with ischemia for 35 or 37 minutes. The birth weight of survived newborn rabbits were significantly lower than that in the control group. The neurobehavioral test scores were significantly lower in ischemic groups than that in the control group. All these differences were statistically significant (all P<0.05). The pathological examination of brain tissue found that the white matter damage in B group was more obvious than that in A group. Conclusions Continuous blockage of uterine blood supply in pregnant rabbits at gestational day 25 causes stillbirth, neurobehavioral damages and white matter injury as well as fetal rabbit intrauterine growth restriction, which can be used for the preparation of preterm hypoxic-ischemic brain injury animal model.