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OBJECTIVE：To stud y the mass spec trometry fragmentation regularity and establish rapid identification method of macrocyclic polyamine alkaloids from Tripterygium wilfordii. METHODS ：The determination was performed on Hypersil GOLDTM C18 column with 0.1％ formic acid solution （A）-methanol（B）as mobile phase （gradient elution ）at the flow rate of 0.25 mL/min. The column temperature was 40 ℃，and sample size was 2 μL. The mass spectrometric condition included ESI，positive ion mode ， breakdown voltage of 100 V，collision energy of 35 eV，and scanning range of m/z 100-2 000. UPLC-Q-TOF-MS/MS combined with PeakView 1.2 software were used to analyze mass spectrum of 4 macrocyclic polyamine alkaloids control as celafurine ， celabenzine，celacarfurin，celacinnine and total extract of T. wilfordii ，and summarized the mass fragmentation regularity of alkaloids. According to retention time ，accurate molecular weight and secondary mass fragment ion information ，referring to the above mass spectrometry fragmentation regularity ，macrocyclic polyamine alkaloids in the total extract of T. wilfordii were quickly identified，and whether it was a new compound was determined by consulting Scifinder database and literature comparison. RESULTS：There were three characteristic cleavage fragments of macrocyclic polyamine alkaloids ，i.e. m/z 160.11（base peak ），m/z 188.10 and m/z 100.07 in positive ion mode. According to the precise molecular weight and mass spectrometry fragmentation regularity，10 macrocyclic polyamine alkaloids were identified rapidly ，and compound 1-6，8 were 2-phenyl-1，5，9-triazacy- clotridecan-4-one， 9-acetyl-2-phenyl-1，5，9-triazacyclotridecan-4-one， 9-nicotinoyl-2-phenyl-1， 5， 9-triazacyclotridecan-4-one， celafurine，celabenzine，celacarfurine，celacinnine，respectively. Compounds 3，7，9 and 10 might be new compounds. CONCLUSIONS：The macrocyclic polyamine alkaloids canform three common characteristic fragment ions ，m/z 160.11（base peak ），m/z 188.10 and m/z 100.07，which can be used for the rapid identification of these active components.
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Objective To investigate the effect of abdominal vibration therapy on chronic obstructive pulmonary disease (COPD) patients with constipation. Methods In the respiratory department from August 2013 to July 2014, eighty-three COPD patients with constipation were divided into the experiment group (n=35) and the control group (n=48) using random digit table. The control group were treated with conventional western medicine and the experiment group received impingement abdomen vibration therapy on the basis of western medicine treatment. The two groups were compared in terms of symptoms of constipation and curative effect. Results The curative effect of the experiment groups was significantly better than that of the control group (P<0.05). The score on constipation in the experiment group was significantly lower than that of the control group (P<0.05). Conclusion The abdominal vibration therapy is effective in the treatment of COPD patients with constipation. It can improve their quality of life and be worthy of clinical application.
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Objective To investigate the afflication of non-humidified low flow nasal oxygen inhalation in the ICU patients. Methods Four hundred patients hospitalized in ICU and needing continuous low-flow oxygen inhalation for more than 24 h were randomly divided into experiment group and control group equally.The experiment group were managed with dried humidification bottle oxygen inhalation and the control group conventional humidified bottle oxygen inhalation.The two groups were compared in terms of 24 h respiratory reactions,bacterial contamination of the humidified bottle,complaints of patients about oxygen inhalation noises and time for nurses to change the humidified bottles.Results The two groups had no significant difference in adverse symptoms(all P>0.05).The rate of bacterial contamination in the experimental group was significantly lower than the control group(P<0.05),the noises were lower and the time for changing the bottle shorter(P<0.05).Conclusion Non-humidified low flow nasal oxygen inhalation in ICU patients is good for reducing the rate of bacterial contamination of humidified bottle and lowering workload and oxygen-inhaling noises.
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The PCR product of PvpA gene was cloned into prokaryotic expression vector pET41a(+) and the recombinant expression vector was then transformed into E.coli DH5a after identified by restriction enzyme digestion and PCR.The positive recombinant plasmid was transformed into E.coli BL21 (D3) and induced to express PvpA protein.The obtained protein was analyzed by SDS-PAGE and Western blotting,purified by Ni-NTA affinity chromatography.The results showed that the purified PvpA fusion protein was obtained successfully.The expressed protein reacted to the high anti-PvpA immune serum from rabbit specially by western blotting.This study would be helpful to established a new diagnostic method for the detection of M.gallisepticum.
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Objective To investigate the therapeutic effect of Shenmai injection(SMI)on neonatal hypoxic-ischemic encephalopathy(HIE)and it's possible mechanism.Methods 45 neonates with HIE were randomly divided into the SMI treatment group and routine treatment group.At the basic of routine treatment,the SMI treatment group was treated with 10 ml SMI added into 10% GS 30~50 ml via injecting into veins once a day for 7~10 d.The levels of serum S-100?,NSE were measured at 1 d,3 d,6 d after treatment.The development quotients(DQ)was evaluated at 3-month-old infants.Results Compared with routine treatment group,symptoms of nervous system in SMI treatment group were recovered obviously faster,the hospitalization time was shorter(all P
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Objective In order to research the best fermentation conditions of recombinant bacteria pThioHisA-B1/DH5? which contains gene of esterase B1.Methods Recombinant bacteria pThioHisA-B1/DH5? was induced in different culture media, induction temperatures, different inductor concentrations, different induction time in order to obtain the best expression effect.The expression products of different conditions were detected by SDS-PAGE and thin-layer gel scanning analysis.Results Based on the experiments, the optimized fermentation condition were that the culture medium was TB, inducing temperature was 28 ℃ and inducing time was 6 h,the concentration of IPTG was 0.4 mmol/L.The expression quantity of target protein attained 40.3% in total bacterial protein, and the soluble fusion protein accounted for 66.2% in total target proteins at the optimum conditions.Conclusion The optimized fermentation conditions of expressing fusion protein in recombinant bacteria have been found and in the conditions, soluble esterase B1 fusion protein of high yield can be obtained.