Object To study the chemical constituents of Hibiscus mutabilis L.Methods Isolation and purification were carried out on silica gel, or polyamide column chromatography etc. Constituents were identified and structurally elucidated by physicochemical properties and spectral analysis.Results Ten compounds were obtained, nine of them were determined as tetracosanoic acid (Ⅰ), ?-sitosterol (Ⅱ), daucosterol (Ⅲ), salicylic acid (Ⅳ), emodin (Ⅴ), rutin (Ⅵ), kaempferol-3-O-?-rutinoside (Ⅶ), kaempferol-3-O-?-robinobinoside (Ⅷ) and kaempferol-3-O-?-D-(6-E-p-hydroxycinnamoyl)-glucopyranoside (Ⅸ).Conclusion All compounds are isolated from the plant for the frist time except Ⅱ and Ⅵ.

Objective To optimize the technology conditions of ultrasonic-enzyme-assisted extraction for sanguinarine from Zanthoxylum nitidum.Methods Extraction rate of sanguinarine determined by HPLC was served as an index.The applicability of the extraction solvent added with acid and enzymatic hydrolysis pretreatment to the ultrasonic-enzyme-assisted extraction of Zanthoxylum nitidum was investigated.Ultrasonic power,extraction frequency and solvent volume were optimized by orthogonal experiment.Finally,ultrasonic extraction time was optimized in dynamic process.Results The optimal process was as follows:Zanthoxylum nitidum powder was extracted 3 times by ultrasonic-wave (250 W) with 40％ ethanol (0.2％hydrochloride) as solvent (extracted for 15 rmin with 6-fold solvent at the first time,then extracted for 12 min with 3-fold solvent at the second and the third time,respectively).The extraction rate of sanguinarine was 88.6％.Conclusion The process is economic,efficient,energy-and time-saving,and provides experimental basis for industrial production of sanguinarine.

Objective To establish normal reference interval for four items of blood coagulation on ACL‐TOP Automatic coagu‐lation analyzer .Methods The fasting anti‐coagulation blood samples were collected from 1 268 inpatients and people conducted physical examination ,all subjects without liver disease ,history of blood disease and coagulation disfunction .The prothrombin time (PT) ,activated partical prothrombin time(APTT) ,thrombin time(TT) and serum levels of fibrinogen(FIB) were determined by u‐sing ACL‐TOP automatic coagulation analyzer which was producted by America IL company .And data of determination results were used to establish the normal reference intervals of indexes in this laboratory .Results The precision and accuracy of this analy‐zer was good .There were differences of normal reference intervals between which established in this laboratory and which provided by the manufacturer .Conclusion Each laboratory should establish its own normal reference interval ,not blindly refer to reference interval provided by regents manual .

Nanobodies are derived from the variable domain of the heavy-chain antibodies (HCAbs) that occur naturally in the serum of Camelidae. They are the smallest antibody fragments capable to bind antigens. With the characteristics of their increased solubility, increased domain stabilities, nanomolar affinities, easy crossing the blood-brain barrier, easy generation, engineering, optimization and tailoring, easy humanization, nanobodies have extensive application prospects in diagnosis and detection. Although nanobody has demonstrated tremendous success, a number of practical challenges limit its broader applications in disease diagnosis and detection, including construction of a phage library and selection of nanobody fragments with high affinity and immunogold labeling technique. Here, we review several recent findings on the use of nanobodies in molecular diagnostics and suggest some practical strategies in resolving the current challenges in this attractive research area, particularly to optimize the affinity, solubility, humanization of nanobodies.
Humans ; Immunoglobulin Heavy Chains ; chemistry ; Single-Domain Antibodies ; chemistry ; drug effects

Objective To investigate the role of dimethylarginine dimethylaminohydrolase-2 (DDAH-2)/asymmetric dimethylarginine(ADMA)in pathophiology of preeclampsia by detecting expression of DDAH-2 in placenta and serum plasma ADMA.Methods From Jan.2004 to Jan.2005,30 preeclampsia patients(PE group)were chosen in the Third Affiliated Hospital.Guangzhou Medical College matched with 10 normal third trimester women as control(control group).The placental DDAH-2 mRNA expression was detected by fluorescence quantitative polymerase chain reaction(FQ-PCR)and the plasma concentration of ADMA WSB determined by high performance liquid chromatography(HPLC).Results(1)The level of ADMA in PE group was significantly higher that than of control group[(18.0±7.2)mg/L vs.(10.3±1.7)mg/L,P<0.01].The expression level of ADMA in preeclampsia occurring before 34 gestatinal weeks WaS significantly higher than that of preeclampsia occurring after 34 gestational weeks[(22.0±7.0)ms/L vs.(12.7±2.8)mg/L,P<0.01].(2)The Placental DDAH-2 mRNA expression in preeclampsia patients was remarkably lower than that of control group[1×10(5.23±0.45)copy/μlvs.1×10(5.65±0.08)copy/μl,P<0.01].The Placental DDAH-2 mRNA in preeclampsia occurring before 34 gestatinal weeks was significantly lower than that of preeclampsia occurring after 34 gestational weeks [1×10(5.02±0.46)copy/μl vs.1×10(5.61±0.19)copy/μl,P<0.01].Conclusion Our results suggested that low expression of DDAH-2 in placenta and increased serum ADMA level might confer the susceptibility to preeclampsia.

Objective To investigate the effect of persistent hypoglycemia on the cerebral oxidative stress and matrix metalloproteinase-2 (MMP-2) activity of newborn rats and to explore the potential mechanism of hypoglycemia-induced brain injury of newborn.MethodsSeven-day-old rats were injected insulin subcutaneously to induce persistent hypoglycemia.Cerebral cortex,hippocampus and thalamus were collected after 36 hours' observation and stored in -80 ℃.Glutathione (GSH)and glutathione disulfide (GSSG) were detected by commercial kit and GSSG/GSH was calculated.MMP-2 activity was detected by gelatin zymography. The data were analyzed by t test,one or two-wayanalysis of variance and Pearson correlation analysis.Six rats were set as the normal glucose group. ResultsThirty-six hours after induction of hypoglycemia, the cerebral GSSG and GSSG/GSH of hypoglycemic pups elevated about 1.5 times higher than those of control group [GSSG:(15.89 ± 5.46) vs (6.15 ± 3.42) mg/g protein,t =3.704,P =0.004; GSSG/GSH:(5.58± 1.79) ％ vs (2.79±1.76) ％,t =2.712,P=0.022].The GSSG and GSSG/GSH in thalamus of hypoglycemia were significantly higher than those of control [GSSG:(15.93 ± 5.75) mg/g protein vs (5.03±5.14)mg/g protein,P＜0.05; GSSG/GSH:(6.50±3.25) ％ vs (2.41±3.12) ％,P＜0.05],whereas there were no significant differences in the cortex ard hippocampus.The total MMP-2 activity of hypoglycemic animals (2.22±0.59) was significantly higher than that of control (1.21± 0.17)(t=4.064,P=0.002),and significant differences were found between the two groups in cortex (2.14 ± 0.5 vs 1.17± 0.27),hippocampus (2.31± 0.72 vs 1.22 ± 0.37) and thalamus (2.22±0.68 vs 1.24±0.18) with all P＜0.01].The activity of MMP-2 was positively related to GSSG (r=0.575,P=0.0002) and GSSG/GSH (r=0.484,P=0.0003).ConclusionsOxidative stress might play an important role in the persistent hypoglycemia induced brain injury of 7-day-old rat pups and thalamus might be most vulnerable to hypoglycemia.Hypoglycemia might also elevate MMP-2 activity which is positively related to GSSG level and GSSG/GSH of the brain.

Objective To analyze the characteristic of auditory brainstem responses (ABR) evoked by chirp in normal hearing subjects .Methods Fifteen cases (30 ears) with normal hearing young people were recorded ABR by stimulation with two kinds of sounds :chirp and click .The data were compared .Results The response threshold of chirp-ABR were lower than that of click -ABR .The average difference was 8 .59 dB .At 90 dB nHL ,the wave V amplitude yield no significant difference between chirp -ABR and click-ABR .The wave V amplitude had a signifi-cant difference between chirp -ABR and click-ABR at 60 dB nHL .At 90 and 60 dB nHL ,the wave V amplitude of chirp-ABR had not a significant difference .The occurence rate was 40% for the wave I of chirp -ABR ,obvi-ously less than that of click -ABR .At 90 dB nHL ,the wave V latency of chirp -ABR was shorter than that of click-ABR .Conclusion The wave V response threshold of chirp -ABR is less than that of the click -ABR .The chirp-ABR is more advantageous than the click -ABR for assessing hearing threshold .

The hemolysin production of 19 strains of Streptococcus spp. isolated from pigs in the Jiangsu and Shanghai regions in recent years were examined. Eight isolates of Streptococcus suis type 2 from Jiangsu showed weak hemolysis on blood agar, but a stronger reaction in Todd-Hewitt broth(THB). The hemolysin belonged to the group of thiol-activated hemolysins. Nine strains of Streptoccus equi subsp. zooepidemicus showed strong hemolysis on blood agar and in THB containing 5% new-born calf serum, but no hemolysis in THB alone. This hemolysin was similar to streptolysin S(SLS). Another two isolates were atypical members of Lancefield group C Streptococcus and showed strong hemolysis on blood agar and in THB with 5% new-born calf serum, but the hemolysin was unlike either streptolysin O or SLS.

Aim To investigate the role of calcitonin gene related peptide(CGRP),ATP sensitive potassium channel(KATP)and spinal nerve in the protective effect of intrathecal morphine preconditioning against myocardial postischemia injury in rats.Methods Rats were established intrathecal catheter placement and myocardial ischemia reperfusion models were randomly assigned to 10 groups:control group(CON);intravenous dimethylsulfoxide group(DMSO);intravenous CGRP8-37 group(CGRP8-37,a selective CGRP receptor antagonist,3 nmol?kg~-1);intravenous glibenclamide group(GLI,a non-selective KATP channel inhibitor,0.3 mg?kg~-1);intrathecal 10 ?l of 1% lidocaine group(LID);intrathecal 3?1 ?g?kg~-1 morphine preconditioning group(MPC);intravenous 3 nmol?kg~-1 CGRP8-37+MPC group(CGRP8-37+MPC);intravenous 0.3 mg?kg~-1 GLI+MPC group(GLI+MPC);intravenous 0.3 mg?kg~-1 GLI+LID group(GLI+LID).Indicators to be observed were MAP,HR and RPP(MAP?HR),the volume of area at risk(AAR)and infarct size(IS),and the area of myocardial infarction,which was demonstrated by IS/AAR.Results Compared to CON group,the volume of IS and IS/AAR was reduced in MPC,LID,LID+MPC and GLI+LID group(P