OBJECTIVETo establish a system of cell suspension culture for Saussurea involucrate.

METHODThe effects of different factors on cell growth and flavonoids production of S. involucrate were systematically studied including the media, initial pH values of the medium, carbon sources, inoculum quantity, and plant growth substance.

RESULTThe optimum medium was N6, initial pH values of the medium was 5.8, sucrose concentration was 50 g x L(-1), inoculum quantity was 60-80 g x L(-1) FW. Medium supplemented with BA (0.5 mg x L(-1)) and NAA (3 mg x L(-1)) was suitable for cell growth, but medium containing BA (0.2 mg x L(-1)) and NAA (2 mg x L(-1)) was suitable for flavonoids production.

CONCLUSIONCell growth and flavonoids production in the suspention of S. involucrate culture cell should be optimized.

Carbon ; Cell Proliferation ; drug effects ; Culture Media ; Flavonoids ; biosynthesis ; Hydrogen-Ion Concentration ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; cytology ; growth & development ; metabolism ; Saussurea ; cytology ; growth & development ; metabolism ; Seeds ; cytology ; growth & development ; metabolism ; Sucrose ; Tissue Culture Techniques ; methods

OBJECTIVETo establish a protocol of rapid clonal propagation of Saussurea involucrate by using embryo as explants.

METHODMS medium was used as basal medium and BA and NAA were supplemented to find out the optimal hormone combinations for adventitious buds initiation, adventitious bud multiplication and rooting.

RESULTAll embryo explants started to grow adventitious buds within 45 days when they were cultured on the media supplemented with 0.02-0.1 mg x L(-1) NAA and 2 mg x L(-1) BA; The adventitious buds multiplicated within 30 days when they were transferred to the media containing 0.1-0.2 mg x L(-1) NAA and 2-3 mg x L(-1) BA; 92% of the adventitious buds rooted well after they were planted on the MS medium containing macroelements at half strength and 0.4 mg x L(-1) NAA for 25 days. The regenerated plantlets grew well after they were transplanted and the survival rate was up to 70%.

CONCLUSIONPlantlets of S. invducrate regenerated high frequence through adventitious bud.

Culture Media ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; chemistry ; Regeneration ; Saussurea ; growth & development ; Tissue Culture Techniques ; methods

Objective To investigate the effect of dexmedetomidine on general anesthesia maintenance and recovery on children receiving cardi-ac catheterization.Methods Eighty-four children undergoing cardiac catheterization ( 1 -11 years old ) were randomly divided into two groups:41 cases in dexmedetomidine combined with sevoflurane group (DS group) cases and 43 cases in sevoflurane group (S group).All of the children were injected with katemine 1 mg· kg-1.Then dexmedeto-midine was injected with an initial loading dose ( 1 μg · kg -1 ) for 10 minutes followed by a continuous infusion ( 2 μg · kg-1 · h-1 ) in DS group.Normal saline was injected in S group.The depth of anesthesia in all patients were monitored by bispectral index ( BIS) , which were main-tained between 55 and 65 through adjusting sevoflurane inhalation.During operation, the data of heart rate (HR), mean arterial pressure ( MAP) , respiration rate ( RR ) , BIS, pressure of end -tidal carbon dioxide ( Pet CO2 ) and concentration of end-tidal sevoflurane ( Cet Sev ) of children at the time to enter the operating room ( T0 ) , the time point of laryngeal mask ( LMA ) insertion ( T1 ) , 20 min after LMA ( T2 ) , the time point of LMA removed ( T3 ) were recorded, as well as the time of operation and recovery, complication including supplemental sedatives, and the rate of respiratory depression, nausea and vomit.Results The data of HR in DS group were significantly lower at T1, T2and T3 compared with S group (P<0.01).The exhaled sevoflurane concentra-tion of DS group was significantly lower at T2 and T3 than S group ( P<0.01 ).The recovery time of DS group was significantly longer than those of S group ( P<0.01 ).The cases requiring supplemental sedatives in DS group were less than that in S group(12 vs 37, P<0.01).Conclusion Dexmedetomidine is advantageous on general anesthesia maintenance and recovery in children receiving cardiac catheterization.

Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) is a novel phlebovirus, causing a life-threatening illness associated with the symptoms of severe fever and thrombocytopenia syndrome. The sequence and structure of the genome have already been illustrated in previous study. However, the characteristics and function of the structure and non-structure proteins is still unclear. In this study, we identified the density of the purified SFTSV virions as 1.135 g/mL in sucrose solution. Using RT-PCR method, we amplified the full coding sequence of RNA dependent RNA polymerase(RdRp), glycoprotein precursor (M), glycoprotein n (Gn), glycoprotein c (Gc), nuclear protein (NP) and non structural protein (NSs) of SFTSV (strain HB29). Respectively inserted the target genes into eukaryotic expression vector pcDNA5/FRT or VR1012, the target protein in 293T cell were successfully expressed. By analyzing the SFTSV virions in SDS-PAGE and using recombinant viral proteins with SFTS patients sera in Western blotting and Immunofluorescent assay, the molecule weight of structure and non-structure proteins of SFTSV were defined. The study provides the first step to understand the molecular characteristics of SFTSV.
Bunyaviridae Infections ; virology ; Cell Line, Transformed ; Fever ; virology ; HEK293 Cells ; Humans ; Orthobunyavirus ; genetics ; metabolism ; Thrombocytopenia ; virology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; Viral Structural Proteins ; biosynthesis ; genetics ; Virion ; genetics ; metabolism

Aim To investigate the effects of DAP on human rheumatoid arthritis synovial fibroblasts(RA-FLS)and its relationship with endoplasmic reticulum stress(ERS)PERK/ATF4/CHOP signaling pathway.Methods RA-FLS cells were cultured and identified by immunofluorescence assay for Vimentin and CD68.CCK-8 detected(0,5,10,20,30,40,50,60,70)mg·L-1 DAP on the proliferation activity of RA-FLS cells.According to the proliferation activity,the experiment was divided into blank group(Blank),low dose group(L-DAP),medium dose group(M-DAP),high dose group(H-DAP)and high dose+specific inhibitor 4-PBA group(H-DAP+4-PBA).The levels of TNF-α and IL-6 were detected by ELISA.Cell apoptosis was detected by flow cytometry.The invasion and migration of cells were detected by Transwell and scratches assays,and the expressions of endoplasmic reticulum stress-related proteins GRP78,PERK,P-PERK,ATF4,CHOP,Caspase-12,C-Caspase-12 and Bcl-2 were assessed by Western blot.Results CCK-8 results showed that compared with 0 mg·L-1 DAP group,the proliferation activity of each group in 20-70 mg·L-1 DAP group was significantly different(P<0.05),and the proliferation rate corresponding to 0,20,40 and 60 mg·L-1 DAP group was significantly different(P<0.05).This concentration was used as the basis for blank,low-dose,medium-dose,high-dose groups and high-dose+4-PBA experimental grouping.DAP could inhibit the release of inflammatory cytokines IL-6 and TNF-α from RA-FLS cells in concentration,reduce the invasion ability of cells,promote apoptosis,upregulate the expression of PERK,P-PERK,ATF4,GRP78,CHOP,Caspase-12, C-caspase-12 proteins and decrease the expression level of anti-apoptotic protein Bcl-2.Another set of experiments demonstrated that high dose DAP+4-PBA could up-regulate the expression of IL-6 and TNF-α,as well as the invasion of cells,and inhibit the expression of apoptosis and ERs-related proteins.Conclusions Daphresin regulates the secretion of inflammatory factors by activating the PERK/ATF4/CHOP signaling pathway,inhibits the proliferation and invasion of RA-FLS cells,and induces apoptosis,which is expected to be a potential therapeutic pathway for RA.