Objective: To explore the active components of antiplatelet aggregation of Erigeron breviscapus, chemical fingerprints and bioactivity detection were used to carry out spectrum-effect correlation analysis. Methods: A fingerprinting method was established by ultra-high performance liquid chromatography with ultraviolet detection (UPLC-UV) and then used for fingerprinting of different batches of E. breviscapus. The antiplatelet aggregation biopotency of different batches of E. breviscapus was tested, and the possible active substances were deduced based on spectrum-effect correlation analysis. Furthermore, all five compounds were verified by antiplatelet aggregation in vitro, and the contribution value of relative activity of the five compounds was calculated according to the difference of the contents of five kinds of monomer compounds in E. breviscapus. Results: Through the spectrum-effect correlation analysis of chemical fingerprints and antiplatelet aggregation biopotency of E. breviscapus, five chromatographic peaks with higher bioactivity correlation coefficient were screened and identified, including chlorogenic acid, caffeic acid, scutellarin, isochromic acid A, and ischlorogenic acid C. Further in vitro experiments showed that the five compounds had different levels of antiplatelet aggregation at same concentration (inhibition rate: 16.5%-85.5%). The sequence of the relative activity contribution is scutellarin > ischlorogenic acid C > caffeic acid > ischlorogenic acid A > chlorogenic acid. In terms of activity contribution of five compounds, chlorogenic acid C and scutellarin was larger than other compounds. Conclusion: A method for the determination of antiplatelet aggregation biopotency of E. breviscapus in vitro was established. Moreover, scutellarin and ischlorogenic acid C are the main active substances of E. breviscapus in the aspect of antiplatelet aggregation in vitro.

Objective To explore the molecular regulation mechanism of asporin in the matrix synthesis and secretion of the intervertebral disc,and to clarify its role in degenerative lesions of intervertebral disc.Methods There were 8 cases of intervertebral disc tissue in patients with severe intervertebral disc herniation (including typical clinical symptoms,signs and Pfirrmann's grade Ⅲ).There were 6 male and 2 female with an average age of (20.25 ± 3.37) years old (ranged from 11 to 28 years).After primary culture and redifferentiation in alginate beads,cells were reseeded and treated with different concentrations of TGF-β1 for 6,12,18 and 24 h.Total RNA extracted from the cells and was subjected to real-time PCR analysis to examine the expression of asporin.After silencing the expression of endogenous asporin by siRNA,the cells stimulated 24 h with TGF-β1.Total RNA extracted from the cells and was subjected to real-time PCR analysis to examine the expression of asporin,collagen Ⅱ and proteoglycans.After treatment of specific p38 inhibitor or ERK inhibitor for 12 h,cells were stimulated with TGF-β1 for 24h.Protein extracted from the cells by protein extraction kit to examine the level of asporin.Results In the primary intervertebral disc cell experiment,TGF-β1 stimulation induced asporin transcription significantly in a dose and time dependent manner.After 24 h stimulation,a significant difference between different concentration groups (5,10 and 15 ng/ml) was observed,2.754±0.24,3.651 ±0.319 and 4.583±0.38,respectively (F=24.782,P=0.001).Knockdown of endogenous asporin led to the upregulated expression of aggrecan and collagen]Ⅱ (aggrecan:t=7.387,P=0.002,collagen Ⅱ:t=4.443,P=0.0113).Specific p38 inhibitor was used to block p38 phosphorylation,and TGF-β1 on asporin induction was significantly inhibited.Conclusion Our results have verified a functional feedback loop between TGF-β1 and asporin in human intervertebral annulus cells indicating that TGF-β1 can increase asporin expression,whereas asporin inhibits TGF-β 1 signaling pathway by negative feedback,thereby inhibiting TGF-β1 mediated synthesis of extracellular matrix,and TGF-31 can increase asporin expression by p38 in human intervertebral disc cells.

Objective To construct four types of glucagon-like peptide-1 (GLP-1) and human serum albumin (HSA) fusion proteins that can be realeased at different rate in vivo by introducing protease cleavage sites between these two moieties.The therapeutic effect and release rate are studied to achieve balanced pharmacokinetics ( PK) and pharmacody-namics ( PD) of GLP-1 and HSA fusion proteins.Methods The gene with different polypeptide joint of GLP-1 and HSA fusion proteins were synthesized by overlap extension PCR amplification, cloned into expression vector pPIC9 and transformed into Pichia pastoris GS115.Then, fusion proteins were obtained by protein purification after being induced by methanol.The preliminary PK and PD of the fusion proteins were studied after purification.Results The fusion protein Gly2-GLP-1-GGGGG-HSA showed no release while Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA showed a slow, medium and fast release rate, respectively, after incubation with furin.In vitro biological activity test results dispalyed that each type of fusion protein promoted insulin secretion of MIN6 cells.In vivo PK test indicated the half-life size of fusion proteins was the largest in Gly2-GLP-1-GGGGG-HSA, followed by Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA.In vivo PD test exhibited hypoglycemic activity that was the highest in Gly2-GLP-1-VTR-HSA, followed by Gly2-GLP-1-SARSVRA-HSA, Gly2-GLP-1-GRSRVTRSV-HSA, and Gly2-GLP-1-GGGGG-HSA.Conclusion GLP-1 can be released from fusion proteins with full activity after the introduction of protease cleavage sites.Releasable fusion proteins at an appropriate release rate have the most balanced PK and PD.

Objective To investigate the value of ultrasound in tethered cord syndrome (TCS) in infants. Methods From December 2005 to July 2013, 25 cases TCS were confirmed by surgery in Shenzhen children's hospital. The ultrasonogram and MRI of 25 infants were analysed retrospectively. The diagnostic accuracy of ultra-sound and its clinical significance were evaluated. Results In 25 cases of TCS confirmed by surgery, the coincidence rate of conus level detected by ultrasound [96%(23/24)] was lower than that by MRI (100%(25/25)). Ultrasound showed reduced spinal cord vibration in 4 cases, disappeared spinal cord vibration in 21 cases. Two cases of reduced spinal cord vibration detected by ultrasound were confirmed as disappeared spinal cord vibration by surgery. The coincidence rate of disappeared spinal cord vibration detected by ultrasound was 91%(21/23). The sacral spinal seg-ments form showed by ultrasound were entirely consistent with those of MRI, including 4 cases of enlarged spinal cord, 13 cases of spinal cord without enlargement and 8 cases of spinal cord ended with rat caudate. Twenty-five cases of TCS had malformations:7 cases meningocele (3 cases combined lipoma), 14 cases myelomeningocele (5 cases combined lipoma, 1 case combined hydromyelia), 3 cases spinal canal-epidermis fistula (all combined lipoma) and 1 case solitary lipoma. Compared with the operation findings, ultrasound misdiagnosed 2 cases of myelomeningo-cele as meningocele, missed 1 of case lipoma which combined with meningocele. MRI missed two cases of spinal canal-epidermis fistula. Conclusions Infantile spinal ultrasound examination can accurately locate the position of conus, accurately display the spinal cord. Compared with MRI, ultrasound examination can real-time visually display spinal cord vibration and help to diagnose tethered cord. Ultrasound examination are convenient, repeatable opera-tion with low cost, therefore it can be used as the preferred screening method to diagnose of tethered spinal cord.

OBJECTIVEEvaluating the accuracy and safety as well as the equivalence compared with the control kit of RIDA QUICK Norovirus detection kit(R-Biopharm, Germany).

METHODSBased on the results of commercially available IDEA Norovirus detection kit (ELISA), the sensitivity and specificity and accuracy of RIDA QUICK Norovirus detection kit (immunochromatographic assay) were evaluated.

RESULTSThe sensitivity and specificity of RIDA QUICK Norovirus detection kit were 98.4% and 92.4%, and the accuracy was 97.6% compared with the control kit.

CONCLUSIONRIDA QUICK Norovirus detection kit has good sensitivity and specificity for the detection of norovirus antigens.

Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunochromatography ; methods ; Norovirus ; isolation & purification ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

Objective To investigate the clinical value of coronary artery Z-scores on echocardiography in diagnosing coronary artery abnormalities. Methods The echocardiography results of 612 patients with Kawasaki disease (KD) at the acute and recovery phase were retrospectively studied. Coronary artery luminal diameters were converted to body-surface-area-adjusted Z-scores. According to coronary Z-scores classiifcation, all the subjects were divided to four groups:415 cases with no dilation (ND), 133 cases with small coronary artery abnormalities (SCAAs), 47 cases with large coronary artery abnormalities (LCAAs), and 17 cases with giant coronary artery abnormalities (GCAAs). Clinical features (gender, age, typical clinical manifestations, fever duration) and laboratory results (CRP, ESR, WBC, PLT) were compared among all the four groups. Coronary artery diameters and the Z-scores were compared between acute and convalescence phase. Results Along with the increase of coronary Z-score, fever duration was prolonged [ND group:(7.75±3.12) d, SCAAs group (8.50±4.12) d, LCAAs group: (8.57±3.58) d, GCAAs group: (11.88±4.33) d, F=22.375, P<0.05]. With coronary Z-score increasing, PLT also increased (F=22.029, P=0.000), and the highest PLT was observed in GCAAs group. There were no significant differences in the CRP, ESR and WBC among all the four groups (F=0.236, 1.116, 0.121, all P>0.05). No significant different coronary diameters were found in ND cases between recovery and acute phase [(2.24±0.34) mm vs (2.33±0.36) mm, t=1.926, P > 0.05]. But there were significant difference in the coronary Z-scores of ND patients between recovery and acute phase (0.41±0.82 vs 1.17±0.75, t=8.332, P < 0.05). The coronary Z-scores in SCAAs group (1.32±0.89 vs 3.40±0.62, t=11.073, P < 0.05), LCAAs group (3.12±2.27 vs 6.20±1.28, t=4.579, P<0.05) and GCAAs group (11.88±6.77 vs 20.4±9.70, t=3.480, P<0.05) at recovery phase were smaller than values at acute phase. Conclusions The KD coronary Z-scores are the body-surface-area-adjusted standard value, and not subject to the influence of children growth and development. Therefore, it may accurately evaluate the severity of coronary artery abnormalities and its recovery process. Accurate quantitative of the coronary artery luminal dimensions is important in KD clinical management and prognosis prediction.

OBJECTIVEAccording to test results of the Hospital of AIDS screening laboratory in 2008, after counting analysis to assess the prevalence of AIDS, we can early detect positive cases in the future and effectively control the spread of AIDS.

METHODSAll serum samples were screened by ELISA method and we reexaminated the samples by PA. As long as one result is positive by the two methods, then we sent the positive samples to Beijing Center for Disease Control and Prevention by Western Blot method to confirm the result.

RESULTSA total of 21 467 samples were detected and 29 (13.5% 0) were positive screening results. We confirm there were 7 (24.1%) positive samples and 12 (41.4%) suspected samples. We researched the epidemiology of the specimens by its source and age and sex.

CONCLUSIONApplication of ELISA method for HIV screening test has a practical significance, it is accurate and fit to record the results of the screening test for AIDS.

Acquired Immunodeficiency Syndrome ; blood ; diagnosis ; epidemiology ; Adolescent ; Adult ; Age Distribution ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Antibodies ; blood ; Hospitals ; statistics & numerical data ; Humans ; Infant ; Male ; Mass Screening ; Middle Aged ; Young Adult

OBJECTIVETo optimize the techniques for extracting hypotensive active peptides from Agrocybe aegerita.

METHODSThe effects of the liquid/solid ratio, extraction time and temperature, pH value of the initial liquid on the extraction percentage (EP) of the hypotensive active peptides were investigated, and inhibition percentage (IP) of the extracts on angiotensin I-converting enzyme was determined.

RESULTSOptimal extraction of the hypotensive active peptides from Agrocybe aegerita was achieved with the liquid/solid ratio of 40:1, extraction time of 3 h, extraction temperature at 30 degrees Celsius;, and pH=8 of the initial liquid. The EP of the hypotensive active peptides from Agrocybe aegerita could reach 87.7% with IP of the extracts on angiotensin I-converting enzyme of 54.0%.

CONCLUSIONThis method is simple and efficient for extracting hypotensive active peptides from Agrocybe aegerita.

Agrocybe ; chemistry ; Antihypertensive Agents ; isolation & purification ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Dietary Proteins ; isolation & purification ; pharmacology ; Peptides ; isolation & purification ; pharmacology

OBJECTIVETo provide DNA molecular marker for identification of Nelumbo nucifera by exploring the differences of nrDNA-ITS sequence of N. nucifera originated from different habitats.

METHODTo compare nrDNA-ITS base sequence using specific PCR-ITS.

RESULTThe completed sequence of ITS and 5.8 S rDNA, and the partial sequences of 18S rDNA and 26S rDNA, totally 750 bp, from N. nucifera were obtained. The differences among N. nucifera from different habitats and from different cultivars were found.

CONCLUSIONThe method can be used to identify N. nucifera among different species and to distinguish their fakes. It provided the basis for identifying N. nucifera from different geographical regions by comparison of their ITS sequences.

Base Sequence ; China ; DNA, Plant ; chemistry ; genetics ; metabolism ; DNA, Ribosomal Spacer ; classification ; genetics ; Deoxyribonuclease EcoRI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Drug Contamination ; prevention & control ; Geography ; Nelumbo ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo study the in vitro antiviral effect of ribavirin combined with an oral preparation of traditional Chinese medicine "Hu Fei" (protecting the lung) on respiratory syncytial virus (RSV).

METHODSCytopathic effects (CEP) of RSV on Hep2 cells were observed after adding different concentrations of ribavirin, Hu Fei and combination of both into the culture medium.

RESULTSThe minimum concentration of ribavirin and Hu Fei for complete inhibition of CPE caused by RSV was 7.80 microg/ml and 5.00 mg/ml, respectively. When the combination of ribavirin and Hu Fei was applied, their minimum concentrations needed for complete inhibition were decreased to 0.98 ?g/ml and 0.63 mg/ml.

CONCLUSIONSBoth ribavirin and Hu Fei showed in vitro anti-RSV effect, but the inhibitory effect of combined ribavirin and Hu Fei was more potent than either of the preparation alone.

Antiviral Agents ; pharmacology ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Microbial Sensitivity Tests ; Respiratory Syncytial Virus, Human ; drug effects ; Ribavirin ; pharmacology