OBJECTIVE:To determine the plasma concentration of mycophenolic acid (MPA) in renal transplantation patients by HPLC-Fluoremetry. METHODS: The sample was subjected to precipitation of proteins using 5% Zinc Sulfate methanol saturated solution,and the supernatant (20 ?L) was taken for sample injection and determination on Zorbax Eclipse XDB C18 with mobile phase consisted of acetonitrile-methanol-0.2 mol?L-1 glycine buffer(18∶2∶80,pH=9.0) at a flow rate of 1.0 mL?min-1. The column temperature was of 25℃;the excitation wavelength(EX) was 342 nm and the emission wavelength(EM) was 425 nm. RESULTS: The linear range of MPA was 0.5～40 mg?L-1,with its lowest limit of quantitation at 0.5 mg?L-1. The methodology recovery was 98.23%～101.00%;the extraction recovery of MPA was 91.56%～94.46%;the intra-day RSD was 0.64%～3.22% and the inter-day RSD was 5.12%～6.10%. CONCLUSION: The method is sensitive,rapid,accurate,convenient,and applicable for the quantitative determination of plasma concentration of MPA in renal transplantation patients.

OBJECTIVE:To study the pharmacokinetics of triptolide in Tripterygium glycosides tablet in normal rats and adju-vant arthritis model rats in vivo,and provide reference for clinical rational drug use. METHODS:12 SD rats were randomly divid-ed into normal group and model group,6 in each group. Model group was subcutaneously injected complete Freund's adjuvant 0.1 mL to induce adjuvant arthritis model,normal group was subcutaneously injected the same volume of saline. After 14 d model-ing,2 groups were given Tripterygium glycosides tablet suspension 96 mg/kg intragastrically,the blood sample of eyes 0.4 mL were respectively taken before and 10,30,45,60,90,120,150,180,240,300,420 min after administration. The plasma con-centration of triptolide was determined by HPLC,the pharmacokinetic parameters were calculated by DAS 2.0 software,and the parameters were compared. RESULTS:The pharmacokinetic parameters of triptolide in normal group were cmax of(1.139±0.114)μg/mL,tmax of(2.167±0.606)h,t1/2α of(5.500±3.610)h,AUC0-7 h of(5.052±0.371)μg·h/mL,MRT0-7 h of(3.224±0.119)h, and CL of(11.616±2.986)mL/h;and those in model group were cmax of(0.916±0.103)μg/mL,tmax of(3.083±0.801)h,t1/2αof(5.593±1.795)h,AUC0-7 h of(4.707±0.347)μg·h/mL,MRT0-7 h of(3.429±0.139)h,and CL of(11.246±2.638) mL/h. Compared with normal group,cmax in model group was significantly decreased,tmax and MRT0-7 h were significantly prolonged(P<0.05). CONCLUSIONS:Adjuvant arthritis can affect the pharmacokinetics of triptolide in rats in vivo,and promote its absorption and removal.

Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.

Objective To clone and express HCMV UL123 gene exon 2,3(ie1-exon2,3) in bacterial two-hybrid bait plasmid,and identify its self-activation property.Methods HCMV ie1-exon2,3 carried on pTWIN1/ie1 recombinant was amplified by PCR and cloned into the pBT plasmid,which was transformed into E.coli XL1-Blue MRF' Kan host strain.The positive recombinant was identified by PCR,restriction enzyme digestions and sequencing analysis.The verified plasmid was transformed into bacterial two-hybrid system reporter strain.The soluble fusion protein was analyzed by SDS-PAGE and Western blot.The self activation effect of the recombinant was then tested.Results Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid was successfully constructed.The corresponding soluble fusion protein rIE1-N_(85)/?C1 was expressed in bacterial two-hybrid system reporter strain,and didn't show self-activation property.Conclusion Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid without self-activation property was successfully constructed,and it can be used to screen the library of fetal brains.

Objective To investigate the value of hepatic T2 value in evaluation of chronic HBV-related acute-on-chronic liver failure (HBV-ACLF).Methods The HBV-ACLF group,chronic hepatitis B group and control group who underwent liver MRI (M-GRASE sequence) were enrolled.The T2 map was produced from the post-processing software,and the mean T2 and R2 value of liver was calculated.The blood biochemical indexes from HBV-ACLF and chronic hepatitis B group were collected in 2 days pre-MR scaning.The differences of T2 and R2 values among 3 groups and the correlation between biochemical indexes and T2 value were analyzed.ROC curve was conducted to evaluate diagnostic efficiency of T2 value for HBV-ACLF.Results There were significant differences of T2value (x2 =19.074,P＜0.001) or R2 value (F=10.411,P＜0.001) among the 3 groups.The AUC of T2 value for diagnosing HBV-ACLF was 0.86 (P＜0.001),with the cut-off value 57.73 ms (R2=0.017).Moderate positive correlation was shown between T2 values and international normalized ratio (INR),prothrombin time (PT),haluronicacid (HA) values (rs =0.65,0.67,0.39,all P＜0.05),and moderate negative correlation was shown between T2 values and prothrombin activity (PTA),albumin (ALB),prealbumin (PA) values (rs =-0.67,-0.48,-0.37,all P＜0.05).Conclusion T2 or R2 value could reflect the liver function,and were correlated with some biochemical indexes,which illustrated a good diagnostic efficiency for diagnostic of HBV-ACLF.

Objective To examine the inter- rater reliablity of Myoton-3 Myometer in assessment of muscle tone in healthy subjects.Methods 20 healthy volunteers were assessed their muscle tone of biceps brachii and flexor carpi radialis muscles in relaxed state with Myoton-3 by 2 testers within 24 h. The frequency of damping oscillations (F value) measured by Myoton-3 was as the characteristic of the muscletone. The intraclass correlation coefficient (ICC) and Bland-Altman analysis were performed. Results The ICC was 0.72~0.88 and 0.79~0.89 in triple scan and ten-time scan pattern, respectively. The Bland-Altman analysis revealed no systematic errors between testers. ConclusionThe Myoton-3 Myometer is reliable between testers for measuring the muscle tone in healthy adults.

OBJECTIVETo evaluate the effect of electro-acupuncture (EA) on gastric mucosal oxygenation and systemic inflammatory response in patients undergoing endoscopic sinus surgery with controlled hypotension (CH), and to explore its protective effect on gastric mucosa.

METHODSFifty-four patients, 18-65 years old, grade I-II of American Society of Anesthesiology (ASA), who were scheduled for endoscopic sinus surgery were randomly assigned to two groups, group A (general anesthesia group) and group B (general anesthesia combined EA anesthesia group), 27 in each group. Conrolled hypotension was executed during operation, and mean arterial pressure (MAP) was maintained at 55-65 mmHg. After tracheal intubation gastric tesiometer catheter was indwelled through nasal cavity or oral cavity. After successful indwelling, it was connected with gastric mucosa monitoring mode of multifunctional parameters monitor. Patients' MAP and heart rate (HR), pHi, partial pressure of carbon dioxide (PgCO2), arterial partial pressure of carbon dioxide (Pg-aCO2) and endtidal pressure of carbon dioxide (Pg-etCO2) were measured and recorded at T, (immediately before induced hypotension), T, (20 min following induced hypotension to target MAP), T2 (40 min following induced hypotension to target MAP), T3 (20 min after ending induced hypotension), and T4(40 min after ending induced hypotension). Blood samples were intravenously collected, TNF-alpha, IL-1, and IL-6 were detected by ELISA 24 h before operation, during operation (T3), and 24 h after operation.

RESULTSAfter hypotension was induced, Pg-CO2, Pg-aCO2 and Pg-etCO2 increased significantly (P < 0.01, P < 0.05), while pHi decreased significantly (P < 0.01) in both groups at T1-T4 than those at T0. During T1-T4, PgCO2, Pg-aCO2, and Pg-etCO2 were higher (P < 0.01, P < 0.05), while pHi was lower in group A than in group B (P < 0.01). Furthermore, TNF-alpha, IL-1, and IL-6 increased significantly in both groups during operation and 24 h after operation, when compared with those 24 h before operation (P < 0.01, P < 0.05). TNF-alpha and IL-1 in group A were higher than those in group B (P < 0.05) during operation and 24 h after operation, but with no significant difference in the plasma concentration of IL-6 (P > 0.05).

CONCLUSIONEA exerted obvious protective effect of gastric mucosal injury in endoscopic sinus surgery with controlled hypotension, which might be achieved by increasing gastric mucosal blood flow, maintaining oxygen supply and demand, inhibiting inflammatory response, and alleviating injury of gastric mucosal barrier.

Acupuncture Analgesia ; methods ; Adolescent ; Adult ; Aged ; Anesthesia, General ; Arteries ; Blood Pressure ; Electroacupuncture ; methods ; Endoscopy ; Female ; Gastric Mucosa ; surgery ; Heart Rate ; Humans ; Hypotension, Controlled ; Interleukin-1 ; Interleukin-6 ; Male ; Middle Aged ; Tumor Necrosis Factor-alpha ; Young Adult

To observe the effect of Ligusticum wallichii-containing serum on the expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells. Clean-grade SD rats were randomly divided into 5 groups and orally given L. wallichii decoction, colchicine and normal saline for 7 d to prepare L. wallichii-containing serums. Except for the blank group, all of the remaining groups were stimulated with LPS 1 mg x L(-1) for 24 h. After being intervened, the L. wallichii-containing serums were cultured in 5% CO2 incubator at 37 degrees C for 24 hours. The expression of TLR4 and MyD88 were detected by RT-PCR and Western blot. After HSC was stimulated with LPS, TLR4 and MyD88 mRNA and protein expressions were significantly higher than the blank control group (P < 0.01). After being intervened with L. wallichii-containing serum, TLR4 and MyD88 mRNA and protein expressions were notably lower than the model group (P < 0.05 or P < 0.01). In conclusion, L. wallichii-containing serum could regulate the TLR4 signaling pathway and show the anti-fibrosis effect by inhibiting the expression of TLR4 and MyD88 in LPS-induced HSCs.
Animals ; Female ; Hepatic Stellate Cells ; drug effects ; metabolism ; Ligusticum ; Lipopolysaccharides ; pharmacology ; Liver Cirrhosis, Experimental ; drug therapy ; Myeloid Differentiation Factor 88 ; genetics ; physiology ; Phytotherapy ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; genetics ; physiology

Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as a novel human coronavirus and posed great threat to public health world wide,which calls for the development of effective and safe vaccine urgently. In the study, peptide epitopes tagrgeting spike antigen were predicted based on bioinformatics methods. Nine polypeptides with high scores were synthesized and linked to keyhole limpet hemocyanin (KLH). Female BALB/C mice were immunized with individual polypeptide-KLH, and the total IgG was detected by ELISA as well as the cellular mediated immunity (CMI) was analyzed using ELIs-pot assay. The results showed that an individual peptide of YVDVGPDSVKSACIEVDIQQTFFDKTWPRPIDVSKADGI could induce the highest level of total IgG as well as CMI (high frequency of IFN-γ secretion) against MERS-CoV antigen in mice. Our study identified a promising peptide vaccine candidate against MERS-CoV and provided an experimental support for bioinformatics-based design of peptide vaccine.
Animals ; Antibodies, Viral ; immunology ; Computational Biology ; Coronavirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Middle East Respiratory Syndrome Coronavirus ; genetics ; immunology ; Peptides ; administration & dosage ; genetics ; immunology ; Spike Glycoprotein, Coronavirus ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Objective To study the relationship between activity of thorax and each spinal intervertebral angle in patients with ankylosing spondylitis. Methods Each spinal intervertebral angle of 41 patients with ankylosing spondylitis were measured by Spinalmouse in different postures. And the activity of thorax was measured. Correlation between activity of thorax and shape of spinal were analyzed. Results The activity of thorax positively correlated with the entire lumbar spinal column in flexion (P<0.01), as well as the intervertebral angle of L1/2, L2/3 and L4/5 in flexion (P<0.05), but negatively correlated with the intervertebral angle of L1/2 and L2/3, curvature of the entire lumbar spinal column in upright and the intervertebral angle of L1/2, L3/4, curvature of the entire lumbar spinal column in extension. Conclusion There was a significant relation between activity of thorax and lumbar vertebra motor ability.