A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

Objective To explore the expression change of death associated protein kinase(DAPK)and therapeutic time window for traumatic brain injury in rats.Method The traumatic brain injury models of rats were achieved by free drop impact model.The adult rats were randomly divided into normal control,sham-operation and TBI groups.Samples of"TBI group were acquired at the time point of2h,8h,24h,48h and 72h after brain injury.Pathological changes of injured neurons were demonstrated by H-E staining,the expression of DAPK mRNA W8.8 detected by RT-PCR,relative quantitation of neuron apoptosis Wa$detected by TUNEL.Results At the Sh after brain injury,the expression of DAPK increased evidently and neuron apeptosis Wag detected.The expression of DAPK and quantitation of neuron apeptosis reached peal[at the 24h after injury.Conclusion These obaervatiorm showed that the expression of DAPK in the cerebral tissues reached peak at the 24h after injury.The optimum therapeutic time window wag 24h after injury.

American Medical Association ; Congresses as Topic ; Hematology ; United States

To explore the effective components represented by fingerprint contributed to allelopathic effect of different Salvia miltiorrhiza aqueous concentration on seeds and seedlings of radish, grey relational analysis was used to establish the chromatography-efficacy relation. The results show that 15 peaks devote high allelopathic contribution to radish seeds and seedlings. The study will provide a new concept for allelochemicals screening and study.
Allelopathy ; Chromatography, High Pressure Liquid ; Pheromones ; analysis ; metabolism ; pharmacology ; Plant Extracts ; analysis ; metabolism ; pharmacology ; Raphanus ; drug effects ; growth & development ; Salvia miltiorrhiza ; chemistry ; metabolism

Described in the paper are contents and characteristics of building smart healthcare system at a research hospital,and practices of the hospital in exploring smart healthcare.Specific measures include staged building of an e-hospital emphasizing information security; closed-loop transformation with mobile and barcode technology for higher efficiency and patient safety;optimizing the service flow with smart interconnection to create an orderly outpatient environment; implementing healthcare regulations with information systems to ensure a homogeneous medical environment;and integrating the entire medical system to promote coordinated development of regional healthcare service. These measures help upgrade the information service effectiveness and quality of care of the hospital.

Objective To explore the importance of gene diagnosis and prenatal diagnosis of spinal muscular atrophy(SMA),and improve the clinical diagnosis of SMA by analyzing the electrophysiological and gene characteristics of SMA.Methods Fifteen cases with SMA including 9 male and 6 female were enrolled in this study.The age was 5 months to 12 years.The 15 cases were subdivided into 3 clinical types,5 cases of type Ⅰ including 3 male and 2 female aging 5-18 months;7 cases of type Ⅱ including 4 males and 3 females aging 5 months-3 years;3 cases of type Ⅲ including 2 male and 1 female aging 3-12 years.They were all characterized by symmetric muscle weakness(more proximal than distal)associated with atrophy,absence or marked decrease of deep tendon reflexes,loss of voluntary movement and inability to sit or stand.The clinical characteristics and changes of electromyography(EMG)and nerve conduction velocity were assessed in all cases by using Danish Medoc Keypoint myoelectricity and evoked potentials inducer.The survival of motor neuron(SMN)gene was detected by PCR and restriction endonuclease spectrum analysis in 10 cases.Results EMG analysis found 94% patients had spontaneous potential,90% patients had increased duration of motor unit,and amplitude increased in 89% patients.Motor nerve conduction velocity was determined in 78 nerves.Motor nerve compound action potential wave amplitude decreased in 52 nerves,among them,distal latent period prolonged and motor conduction velocity reduced slightly in 36 nerves.Sensory nerve conduction velocity was determined in 45 nerves and remained normal.The SMN gene detection revealed deletion of exon 7 and 8 in 9 cases,deletion exon 7 in 1 case.The SMN gene detection in 10 patients and their parents didn't find any deletion of exon 7 and 8.Conclusions The definite diagnosis of SMA will rely on the typical clinical characteristics,changes of EMG and gene deletion analysis.Gene diagnosis of SMA lays a basis for prenatal diagnosis.

Objective:To investigate the changes and clinical significances of serum levels of granulocyte macrophage colony-stimulating factor (GM-CSF),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels in patients with rheumatoid arthritis (RA).Methods:126 cases of patients with RA in our hospital from June 2013 to December 2016 were selected as RA group and divided into two subgroups as active RA group (69 cases) and remission RA group (57 cases) according to the severity of the disease.55 healthy cases who receive physical check in our hospital at same time were selected as the control group.The serum GM-CSF,IL-6 TNF-α levels were detected by enzyme linked immunosorbent assay (ELISA) and compared between different groups,the correlation of serum GM-CSF,IL-6 TNF-α levels with DAS28 were analyzed.Results:The serum GM-CSF,IL-6 TNF-α levels of RA group were significantly higher than those of the remission RA group and control group (P＜0.01),and the serum GM-CSF,IL-6 TNF-α levels of remission RA group were also significantly higher than those of the control group (P＜0.01).Positive correlation between serum GM-CSF,IL-6 and TNF-α levels were found(P＜.0.01).The level of DAS28 was significantly correlated with the levels of serum GM-CSF,IL-6 and TNF-α (r=0.473,0.584,0.675,all P＜0.01).Conclusion:The levels of serum GM-CSF,IL-6 and TNF-α were up-regulated in patients with RA and positively correlated with the activity of RA,which might be of important clinical significance in the early diagnosis,disease evaluation and prognosis of RA.

Objective To investigate the inhibitory effect and the effect on cell proliferation and apoptosis of artemether(ART) on human gastric adenocarcinoma cell lines and pancreatic cancer cell lines in vitro. Methods The inhibitory effect of ART on human gastric cancer cell lines(SGC-7901 and MKN-45) and human pancreatic cancer cell lines(SW-1990 and BXPC-3) were detected by MTT assay,and the effect on cell cycle and apoptosis were evaluated by flow cytometry. Results It was indicated by MTT assay that the killing effect of ART on the cancer cell lines were positively correlated to time and dosage(P

OBJECTIVETo understand the status of standard medication of antiretroviral therapy and identify its correlationship with preventing mother-to-child transmission of HIV (PMTCT).

METHODSQualitative and quantitative methods for measuring of standard medication towards antiretroviral therapy of PMTCT were used in a cross-sectional study of 167 pregnant women who had accessed to PMTCT services and 57 physicians providing service from January 2005 to June 2007 in 5 HIV epidemic counties in Yunnan province.

RESULTSA total of 167 mothers were recruited, of which 65.87% (110/167) demonstrated for standard medication. The percentage of rational selection of antivirus-therapy regimen was 88.02% (147/167). HIV positive mothers with rational using medicine during labor covered 81.37% (131/161). 87.42% (146/167) of women demonstrated good compliance. The main correlations with the standard medication level of HIV positive pregnant women and their infants were as follows: diagnose time (OR = 2.617; 95% CI: 1.184 - 5.783), place of delivery (OR = 0.064; 95% CI: 0.007 - 0.607), minorities (OR = 0.344; 95% CI: 0.162 - 0.730), understanding of HIV women for antiretroviral therapy (OR = 6.843; 95% CI: 1.449 - 32.312), and doctors' cognition about the regimen for antiretroviral therapy was not enough; 5 key knowledge points (purpose of PMTCT, the consequence of not requiring standard medication, the effect of knowing regimens, the relationship of CD4 levels and selected regimens, the side effect of the relevant medicine) score rate was 47.72% (136/285).

CONCLUSIONThe proportions of standard medication in PMTCT antiretroviral therapy were not high. Standard medication might be associated with multiple factors from doctors, patients and society. It is necessary to make some steps to improve standard medication of antiretroviral therapy in PMTCT.

Anti-HIV Agents ; therapeutic use ; China ; Cross-Sectional Studies ; Female ; HIV Infections ; drug therapy ; prevention & control ; transmission ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control ; Pregnancy ; Pregnancy Complications, Infectious ; drug therapy