Objective: To establish a method for the content determination of ginsenoside Rg1, Re, and Rb1 in Piwang Granules. Methods: HPLC-ELSD was used. The analysis was carried on Shim-pack VP-ODS (150 mm × 4.6 mm, 5 μm) column with mobile phase consisted of acetoneitrile-water, and the gradient elution (0-20 min, acetoneitrile 20%, 20-35 min, acetoneitrile 20%→30%, 35-55 min, acetoneitrile 30%, 55-56 min, acetoneitrile 30%→45%) was used. The flow rate was 0.8 mL/min. The column temperature was 25℃. The drift tube temperature was 100℃. The flow rate of gas was 3 L/min. Results: The responses of ginsenoside Rg1, Re, and Rb1 were liner in the ranges of 125.44-470.40 (r = 0.999 6), 88.96-333.60 (r = 0.999 2), and 157.12-589.20 μg/mL (r =0.999 5), respectively. The average recoveries were 100.27% (RSD = 1.713%, n = 9), 101.29% (RSD = 1.220%, n = 9), and 101.00% (RSD = 1.668%, n = 9). Conclusion: The method is simple, rapid, and with good repeatability. It can be used for the quality control of Piwang Granules.

Venezuelan equine encephalitis (VEE) is a zoonotic disease caused by the Venezuelan equine encephalitis virus (VEEV) complex. This disease has not yet been reported in China, and it is therefore essential to establish a rapid and accurate method for detection of the virus in order to prevent and control this disease. In this study, a one-step real-time quantitative RT-PCR method was developed for the detection of the VEEV complex. A pair of specific primers and a Taqman probe were designed corresponding to a conserved region of the VEEV gene nspl, allowing the detection of all known strains of different sub- types of the virus. Using RNA synthesized by in vitro transcription as template, the sensitivity of this method was measured at 3.27 x 10(2) copies/microL. No signal was generated in response to RNA from Chikungunya virus (CHIKV), nor to RNA encoding the nsp1 fragment of Eastern equine encephalitis virus (EE-EV) or Western equine encephalitis virus (WEEV), all of which belong to the same genus as VEEV. This indicates that the method has excellent specificity. These results show that this one-step real-time quantitative RT-PCR method may provide an effective tool for the detection of VEEV in China.
China ; DNA Primers ; genetics ; Encephalitis Virus, Venezuelan Equine ; classification ; genetics ; isolation & purification ; Encephalomyelitis, Venezuelan Equine ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

We analyzed the surveillance data of measles from January 1999 to October 2000 in Jiangxi Province for the aim of measles control. The results showed that 3*!184 measles cases were reported from the province and 1 reported case died. Most of the reported measles cases were under 15 years old. The incidence of measles in age group between 7-10 years old was higher than that in other age groups and most of the reported cases having vaccination histroy were 4-10 year old children. Both the sporadic and epidemic measles in some districts existed at the same time and the reported measles cases extensively distributed in all districts of Jiangxi Province. The outbreaks of measles in some districts had affected the morbidity of measles of the whole province To prevent and control measles outbreak are the main effective measures to control this disease.

The main producing areas of Cistanche spp. in Neimongol, Ningxia, Gansu and Xingjiang were surveyed. Plant specimens and samples of 4 species and a new variety, named C. salsa var.albiflora P.F.Tu et Z.C.Lou, were collected and identified. Their distribution and abundance of resources are reported,and measures of exploiting and protecting comdined with sand-control are suggested.

Various column chromatography, such as silica gel, Sephadex LH-20, ODS, and semi-preparative HPLC was used to isolate and purify the chemical constituents from Cinnamomum cassia. The structures were determined on the basis of NMR and MS spectral data analysis, together with the comparison with literature data. Fifteen compounds were isolated from the 85% aqueous ethanol extract of C. cassia, and their structures were identified as (2R, 3R)-5,7,3',4'-tetramethoxyflavan-3-ol( 1), (2R, 3R)-5,7-dimethoxy-3',4'-methylenedioxyflavan-3-ol (2), coumarin (3), cinnamic acid (4), (E)-2-hydroxy-phenylpropionic acid cinnamoyl ester (5), 3, 3', 4, 4'-tetrahydroxy biphenyl (6), methylstictic acid (7), epi-boscialin (8), (1R,2S,3S,4S)-2,3-epoxy-1, 4-dihydroxy-5-methyl-5-cyelohexene (9), 4,5-dihydroxy-3-methyl cyclohex-2-enone (10), cis-4-hydroxymellein (11), and 2-hydroxy-4-methoxyl-cinnamaldehyde (12). Compounds 5-11 were obtained from this genus plants for the first time.
Cinnamomum aromaticum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure

Objective To discuss Extracorporeal Membrane Oxygenation(ECMO) management method and effect during inter-hospital transport of potential cardiac death donors after cardiac death (DCD).Methods 8 potential donors after cardiac death with brain injury were supported by ECMO for inter-hospital transport.All donors were inserted Medtronic overall cannula into one side femoral artery and venous.The position of catheters were guided by ultrasound.The front-end of venous catheter located in the junction of atrium and inferior vena cava,meanwhile the front-end of artery catheter was below renal artery.100 IU/kg heparin was injected before inserting cannulas.Flow of ECMO maintained at 2.0～3.0 L/min,and oxygen flow was 2～3 L/min during ECMO supporting.When hemodynamics of potential donors were stable,patients were moved into ambulance with ECMO for inter-hospital transport.Results A total of 8 ECMO transports were performed for central circulatory collapse caused brain injury.Patients were previously cannulated and on ECMO prior to transport and transported a distance of more than 100 kilometer from our institution by ambulance.ECMO running times were 120 min,and operation process circulatory stable.Conclusion ECMO can ensure inter-hospital transport of potential donors after cardiac death safety.

Objective To investigate the effect of operation for hyperthyroidism assoiciated with hypokalemic periodic paralysis(HPP).Methods We retrospectively analysed the clinical data of 121 cases of hyperthyoidism associated with HPP.Among them 81 patients received subtotal thyroidectomy after taking Lugol solution for 2 weeks;40 patients received non-operative therapy.Results The plasma potassium,T3,T4,TSH and BMR levels of patients who received subtotal thyroidectomy were all normal 1 week post-operatively,Only 2 patients suffered symptoms of relapse at follow up of 0.5-10 years,with cure rate of 97.5%;8 of the patients who received non-operative therapy recovered,with cure rate of 20.0%.Conclusions Operation for hyperthyroidism associated with HPP could cure HPP and hyperthyroidism simultaneously.The therapeutic efficacy of operation is rapid and stable,and is markedly better than that of non-operative therapy.

To explore the relationship between the perioperative change in erythrocytic pyruvate kinase (PK) activity and surgical hyperglycemia in the patients undergoing upper abdominal surgery under intravenous procaine balanced anesthesia. Seventeen patients with ASA grade Ⅰ～Ⅱ, being scheduled for selective cholecystectomy or subtotal gastrectomy, were randomly enrolled in our study. PK activity of erythrocytes and its pertinent modulators, such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), inorganic phosphate (Pi), magnesium, plasma glucose and serum insulin were dynamically assayed in the surgical patients receiving intravenous procaine balanced anesthesia. The results showed that PK activity was decreased significantly at 10 min and 24 hours after operation as compared with that of preoperative period. Changes in PK activity were positively correlated with ATP/ADP ratio( r =0 680, P 0 05). It is our supposition that the decreased PK activity and the disturbance of glycolytic pathway might be directly or indirectly induced by anesthesia and surgical trauma, leading to hindrance of utilization of glucose and Pi, as well as synthesis of ATP. Therefore, an inhibition of glycolytic reaction is one of the important mechanisms of "surgical hyperglycemia".

Objective To express the recombinant human canstatin protein, and to examine its biological activity. Methods Canstatin cDNA was cut off from the plasmid pUCm-T/canstatin with restriction enzymes BamHⅠ and Hind Ⅲ. The cDNA fragment was then ligated into the correspondence sites of plasmid pET-22b(+) by T4 DNA ligation enzyme and transformed into E.coli BL21 which was induced to express proteins with isopropyl-1-thio-b-dgalactopyranoside (IPTG). The expressed proteins were analyzed by SDS-PAGE and purified through Ni-NTA column affinity chromatography. Chick chorioallantoic membrane (CAM) assay was performed to determine the activity of the recombinant protein. Results Canstatin cDNA from pUCm-T showed one clear objective DNA band with electrophoresis. Seven of positive colonies were selected and identified by restriction enzyme analysis with BamH Ⅰ and Hind Ⅲ. Electrophoresis revealed that all selected colonies had two specific bands,one near the location of primary plamid,the other near that of objective gene fragment. After IPTG induction, there was a new protein band about 24 000 on SDS-PAGE.The induced product over total bacterial proteins in 1,2, 3. and 4 hours after induction was 18. 2%, 18. 8%, 23.0% and 23.4%, respectively, by densitometry examination. CAM assay demonstrated that the recombinant canstatin protein significantly inhibited the embryonic neovascularization in a dose-dependent manner. Conclusion The prokaryotic expression vector of human canstatin gene has been successfully constructed, laying the foundation for further clinical study.

Objective To clone human canstatin gene, construct its prokaryotic expression vector, express and purify its recombinant protein. Methods The total RNA was extracted from human placenta tissues. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid pET-22b(+) and then transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG). The expression of protein was analyzed through SDS-PAGE. Cells after induced 3 hours by IPTG were harvested, sonicated briefly and the proteins were purified through affinity chromatography. Results (1)The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis and the concentration was 1.8 g/L. (2)The target sequences were specifically amplified through RT-PCR. (3)The purified RT-PCR product was cloned into pUCm-T vector and then sequenced, demonstrating to be the same as that of canstatin gene in GenBank. (4) the expression vector pET-22b(+) was constructed and verified by the method of BamH Ⅰ and Hind Ⅲ digestion. (5) After IPTG induction, there was a new protein band about Mr 24kD on SDS-PAGE. The percent expressed product over total bacterial proteins after 1, 2, 3, and 4 hours of induction was 18.2%, 18.8%, 23.0% and 23.4%, respectively, estimated by densitometry. (6)After affinity chromatography, SDS-PAGE showed only one clear band existed in 125 mmol/L or 250 mmol/L imidizone elution. Conclusion Human canstatin gene has been successfully cloned and its prokaryotic expression vector has also been successfully constructed. Further more, purified recombinant proteins are obtained through affinity chromatography, laying foundation for further study of its clinical application.