The highly virulent PRRSV isolate strain HN-1/06 was cultivated on Marc-145. To study the viral entry mechanisms, the GP5 gene of PRRSV isolate was amplified by RT-PCR and cloned into pcDNA3.0 to generate the expressing plasmid pcDNA-GP5. pcDNA-GP5 was transfected into 293T by the calcium phosphate precipitation method. Analysis of flow cytometry confirmed that the GP5 proteins were expressed in surface of the 293T cells. Then 293T cells were transfected with pcDNA-GP5, pHIT60 and pHIT111 plasmids to generate pseudotyping virus. The pseudotyping virus supernatant was harvested 48 hours post-transfection and was detected by Western blotting and infection assay. Western blotting indicated that the GP5 glycoproteins were incorporated into the retroviral pseudotyped virus. Infection assay showed that the pseudotyped virus infected 293T and Mark-145 cell. The pseudotyped virus could be used to further study infectious mechanism of PRRSV.
Animals ; Cell Line ; Cloning, Molecular ; Endothelial Cells ; cytology ; metabolism ; virology ; Leukemia Virus, Murine ; genetics ; metabolism ; Mice ; Porcine respiratory and reproductive syndrome virus ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transfection ; Viral Envelope Proteins ; biosynthesis ; genetics ; Virion ; genetics ; metabolism

OBJECTIVETo investigate the mechanisms of intestine absorption of astragaloside IV in rat.

METHODThe K(a) and P(app) of astragaloside IV was investigated using single-pass intestinal perfusion technique in rats. HPLC was used to determine the concentration of astragaloside IV. The effect of absorption site, drug concentration and the inhibitors of P-glycoproteon on the absorption had been studied.

RESULTBy the testing of the statistics, the K(a) and the P(app) values of the duodenum, jejunum, ileum, colonic had significant differences (P < 0.05). The concentration from 20-80 mg x L(-1) had no distinctive effect on the K(a) and P(app) of small intestine. The inhibitors of P-glycoproteon had no distinctive effect on the absorption of small intestine.

CONCLUSIONAstragaloside IV is absorbed by typical passive diffusion mechanism.

ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Chromatography, High Pressure Liquid ; Intestinal Absorption ; drug effects ; Intestines ; metabolism ; Kinetics ; Male ; Perfusion ; Rats ; Rats, Wistar ; Saponins ; metabolism ; pharmacokinetics ; Triterpenes ; metabolism ; pharmacokinetics ; Verapamil ; pharmacology

Duck IL-18 gene was amplified from plasmid pGEM-DuIL-18 by PCR. The PCR product digested with Pst I and Xho I was inserted into eukaryotic express vector pcDNA3.1(+) to generate an recombinant expression plasmid pcDNA3.1/DuIL-18 (pDuIL-18), and transformed into Escherichia coli JM109. The recombinant colonies were identified by restriction enzyme digestion, PCR and sequencing. DNA sequence confirmed the correct sequence of the recombinant eukaryotic expression plasmid pDuIL-18 in the reading frame and the ligation part. After the transfection of pDuIL-18 into Cos7 cells, duck IL-18 mRNA was expressed in Cos7 cell. The SDS-PAGE analysis showed that the expressed duck IL-18 protein had molecular weight of 23 000 D. The results of methyl thiazolyl tetrazolium (MTT) assay showed that duck IL-18 protein expressed in Cos7 cell could induce significantly transformation of duck T lymphocytes. Immunoenhancement effect of recombinant expression plasmid pDuIL18 on avian influenza vaccine was observed by proliferation response of the T lymphocytes from spleen. It can obviously enhance the cell-mediated immune response.
Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Ducks ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunity, Cellular ; Interleukin-18 ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology ; T-Lymphocytes ; immunology ; Transfection

OBJECTIVETo investigate the endovascular treatments for the ruptured aneurysms located at anterior communicating artery complex (ACoAC).

METHODSThe data of patients with ruptured ACoAC aneurysms treated in Department of Neurosurgery, First Affiliated Hospital to Fourth Military Medical University from May 2013 to December 2014 was retrospectively analyzed. Sixty-six cases were recruited including 50 male and 16 female patients. The patients aged from 31 to 69 years old, averaging (51±8) years. The Hunt-Hess grade at admission were 13 cases with grade Ⅰ, 36 cases with grade Ⅱ, 11 cases with grade Ⅲ, and 6 cases with grade Ⅳ. The most diameter of aneurysms sac: 14 cases less than or equal to 3 mm, 36 cases more than 3 mm but less than or equal to 7 mm, and 16 cases more than 7 mm. The height diameter/neck width ratio: 8 cases with absolute wide neck, 50 cases with relatively wide neck, and 8 cases with narrow neck. There were 28 cases underwent single micro-catheter embolization, 18 cases underwent double micro-catheters embolization, 14 cases underwent stent-assisted embolization and 6 cases underwent balloon-assisted embolization. The patients were followed up for 6 to 12 months and evaluated by modified Rankin score (mRS) and digital subtraction angiography (DSA). The ratio of total embolization, recurrence rate, and time from operation to reexamination of four groups managed by different endovascular treatment were compared by χ(2) test or F test.

RESULTSSixty cases were totally embolized, 3 cases subtotally embolized, 3 cases incompletely embolized. Mild hemiparalysis and aphasia occurred in 2 cases, and 1 case died of infarction induced by subarachnoid haemorrhage. The mRS at six months after operation were 0 in 31 cases, 1 in 22 cases, 2 in 8 cases, 3 in 2 cases, 4 in 2 cases, 6 in 1 case. All the included cases reexamined the DSA at averaging (7.5±1.0) month post-operatively and 4 cases recurred. There were not significant differences of the ratio of total embolization, recurrence rate, time from operation to reexamination among four groups (all P>0.05).

CONCLUSIONThe endovascular treatment maybe an ideal management for ruptured ACoAC aneurysms.

Adult ; Aged ; Aneurysm, Ruptured ; therapy ; Catheters ; Embolization, Therapeutic ; Female ; Humans ; Intracranial Aneurysm ; therapy ; Male ; Middle Aged ; Postoperative Period ; Recurrence ; Retrospective Studies ; Stents ; Treatment Outcome

Objective: Rheumatoid arthritis (RA) is one of the most common inflammatory arthropathy diseases, characterized by synovium hyperplasia and progressive destruction of articular cartilage, which is significantly associated with dysfunction and shortening of life span. Drug therapy is the main intervention for RA, but the glucocorticoid and immunosuppressant have many shortcomings in treatment, such as great side effect, slow onset and poor efficacy, while biological agents are too expensive. RA belongs to the category of "arthralgia syndrome" in traditional Chinese medicine (TCM). TCM treatment for RA has a well-established history and multiple advantages such as good curative effect and less side effects, but its mechanism needs to be further studied. Signaling pathway is involved in the pathogenesis and progression of RA, so it is one of the main targets in research on the pathogenesis of RA and related pharmacological research of therapeutic drugs. In recent years, a large number of studies have been carried out on the regulation effect of active components of TCM on RA signaling pathways. These signaling pathways include Wnt signaling pathway, Janus tyrosine kinase (JAK)-signal transducers and transcription activator (STAT) signaling pathway, mitogen-activated protein kinase (MAPKs) signaling pathway, phosphoinositide-3-kinase (PI3K)/serine/protein kinase B (Akt) signaling pathway, nuclear factor-kappa B (NF-κB) signaling pathway, Toll like receptor (TLRs) signaling pathway, receptor activator of the nuclear factor kappa B ligand (RANKL)/receptor activator of the nuclear factor kappa B (RANK)/Osteoprotegerin (OPG) signaling pathway and so on. By reviewing the research results in recent years, we hope to provide ideas and reference for the basic research, development of new drugs and clinical treatment of RA.

OBJECTIVE: To establish an interference mode of group counseling for diabetic patients with depression and to evaluate the effectiveness of this mode on depression, treatment compliance and blood sugar level in the patients.
 METHODS: One hundred diabetic patients with depression were randomly divided into a counseling group and a control group (n=50 per group). Self-Rating Depression Scale (SDS) was applied to all the patients. The interference mode of group counseling was established through literature review, expert consultation or interview. The counseling group received counseling for 8 times within 2 months.
 RESULTS: There was a significant difference in the SDS scores at 0, 3, 6 or 12 months after the intervention between the 2 groups (P<0.001). For the counseling group, there was a significant difference in the SDS scores between pre-intervention and 3, 6 or 12 months after intervention (P<0.001). However, there was no significant difference in the SDS scores between any two time points after the intervention (P>0.05). There was a significant difference in the compliance between any two time points after the intervention (P<0.05). Fasting blood glucose (FBG), 2 h postprandial blood glucose (2hPG) or glycosylated hemoglobin (HbA1c) was significantly different at any two time points after the intervention (P<0.05).
 CONCLUSION Group counseling can improve depression, compliance and blood sugar control in the diabetic patients.
Blood Glucose ; analysis ; Counseling ; Depression ; therapy ; Diabetes Mellitus, Type 2 ; psychology ; therapy ; Glycated Hemoglobin A ; analysis ; Humans ; Patient Compliance ; Psychotherapy, Group