This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
Aconitine ; analogs & derivatives ; chemistry ; pharmacokinetics ; Animals ; Biological Availability ; Dogs ; Drug Stability ; Female ; Male

Human hepatitis B virus surface antigen (HBsAg) binding protein(SBP) shows a specific binding ability to HBV surface antigen HBsAg. Previous work proved an ability of SBP to enhance the immune response of HBsAg vaccine. To investigate the function and mechanism of this protein, we constructed SBP-expression strains with Pichia pastoris expression system. We screened these strains and have got an expression strain with high protein expression quantity. Fermentation product was collected and purified to gain a large amount of purified protein. Identification of purified SBP with SDS-PAGE, High performance liquid chromatography, Western blotting and mass spectrometry suggested that the protein was highly purified and with a good integrity. ELISA test of purified SBP showed a significant binding ability to HBsAg, suggesting a good protein activity. This work offers a solid foundation to the research of SBP function and mechanism of immune enhancement.
Binding Sites ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; metabolism ; Humans ; Pichia ; genetics ; metabolism ; Receptors, Virus ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification

Objective To study the FMS-like tyrosine kinase-3 (FLT3) gene, NPM1 gene and c-kit gene mutations in acute myeloid leukemia (AML) by extracting DNA from the storage of bone marrow slides, and to investigate the relationship between the three gene mutations and clinical features in AML. Methods The bone marrow slides of 55 patients diagnosed with AML were enrolled in this study. The PCR, DNA sequencing and molecular cloning were used to detect and analyse the FLT3-ITD, NPM1 and c-kit gene mutations. Patients' remission, progression and survival time were also recorded. Results The DNA was successfully extracted from the bone marrow slides with -20 ℃ frozen storage without Wright stained, chemically fixed, and room temperature storage Wright stained discoloured by phenol ∶ chloroform ∶ isoamyl alcohol method, which can be used in PCR, direct sequencing and molecular cloning sequencing analysis. 10 of the 55 cases (18.2 %) were FLT3-ITD positive, including 9 cases with heterozygous mutations and 1 case with homozygous mutation. FLT3-ITD positive group had lower complete remission (CR) rate, shorter event-free survival (EFS) time and overall survival (OS) time than the negative group (P< 0.05). 9 of the 55 cases (16.4 %) had NPM1 heterozygous gene mutations, all belonging to type A. The EFS rate of the patients with NPM1 mutation was higher in 10 months and the OS rate was higher in 19 months (P< 0.05). 3 of 9 NPM1 mutations patients were FLT3-ITD positive. The CR rates of the four groups after initial remission induction therapy in order were NPM1+FLT3-ITD-, NPM1-FLT3-ITD-, NPM1-FLT3-ITD+, NPM1+FLT3-ITD+(P<0.05). Besides, NPM1-FLT3-ITD+was a risk factor affecting the OS (RR=1.250, P=0.005). 2 of the 55 cases (3.6 %) had c-kit gene mutations, namely mutant D816H and mutant D816V. The c-kit gene mutations were not found in patients with FLT3-ITD and NPM1 mutations. Conclusions The FLT3-ITD mutation is a poor prognosis molecular marker in AML, and NPM1 mutation is a good factor for the prognosis. NPM1-FLT3-ITD+is a risk factor affecting OS. The rate of c-kit gene mutation is low in AML, without the overlap of FLT3 and NPM1 mutations.

Objective:To compare the safety and effectiveness of two minimally invasive approaches for multi-vessel coronary revascularization.Methods:From August 2014 to February 2017,70 consecutive patients who underwent minimally invasive coronary artery bypass grafting in Peking University Third Hospital were randomly divided into two groups.In one group,40 patients underwent staged-hybrid coronary revascularization (staged-HCR) treatment;in the other group,30 patients underwent minimally invasive total arterial revascularization with bilateral internal thoracic artery (BITA).In staged-HCR group,the patients underwent minimally invasive direct coronary artery bypass grafting (MIDCAB) and percutaneous coronary intervention (PCI) procedure for treatment of multi-vessel disease.In BITA group,the patients underwent total arterial coronary artery bypass grafting with composite "Y" BITA graft.Preoperative and postoperative data of the two groups,including postoperative blood usage,mechanical ventilation time,domiciling duration in intensive care unit (ICU),major adverse cerebral and cardiovascular event (MACCE),and postoperative coronary angiography results were compared,in order to evaluate the safety and effectiveness of these surgical approaches.Results:The preoperative characteristics of 70 patients in the two groups showed no significant difference.All the patients underwent successfully,elective minimally invasive multi-vessel coronary artery bypass grafting as scheduled preoperatively.Postoperative result showed the patients in staged-HCR group took advantages in less postoperative mechanical ventilation time [Staged-HCR group (11.2 ± 8.7) h vs.BITA group (18.3 ± 9.1) h,P =0.013],shorter domiciling duration in ICU [Staged-HCR group (26.29 ± 4.05) h vs.BITA group (44.74 ± 28.75) h,P =0.022],and less total drainage [Staged-HCR group (695.57 ± 250.46) mL vs.BITA group (1 103.26±547.44) mL,P =0.03] than the patients in the group of minimally invasive total arterial revascularization with BITA.Postoperative in hospital coronary angiography showed satisfactory graft patency rates in both groups [97.5％ in Staged-HCR group vs.97.8％ in BITA group].No MACCE occurred in both groups during hospitalization.Conclusion:Staged-HCR is a feasible method for the treatment of multi-vessel revascularization involving right coronary artery.Minimally coronary revascularization with BITA is associated with superior long-term graft patency and it's recommended for patients who could not tolerate dual-antiplatelet therapy.This study shows that both minimally invasive surgical approaches are safe and effective for treatment of patients with multi-vessel coronary artery disease.

OBJECTIVE: To observe the demethylation effect of demethylation inhibitor 5-azacytidine (5-Zac) on programmed death receptor 1 (PD-1) in Molt-4 cells (T lymphocyte cell line) and to investigate the relationship between DNA demethylation and expression of PD-1. METHODS: Molt-4 cells were cultured in the medium containing different concentrations of 5-Zac(0, 5, 10 μmol/L) for 72 h. According to the concentrations of 5-Zac, the Molt-4 cells were divided into a 0 μmol/L 5-Zac group, a 5 μmol/L 5-Zac group, and a 10 μmol/L 5-Zac group. The expression of PD-1 in Molt-4 cells was detected by flow cytometry and the apoptosis rate was calculated. The mRNA transcription level of PD-1 was detected by real-time polymerase chain reaction; Molt-4 cell DNA in all groups were treated by sodium bisulfite. The PD-1 promoter fragment was amplified by PCR, the amplification fragments were transformed into E. coli., the positive clones were selected for equencing, and the methylation status of the fragments of PD-1 promoter was examined. RESULTS Seventy-two hours after the 5-Zac treatment, the expression rate of PD-1 in the Molt-4 cells in the 0 μmol/L 5-Zac group, the 5 μmol/L 5-Zac group, and the 10 μmol/L 5-Zac group was (1.13 ± 0.01)%, (18.96 ± 1.87)%, and (63.09 ± 6.25)% respectively, in a low concentration-dependent way. The PD-1 mRNA expression level was increased significantly with the 5-Zac treatment. Cells apoptosis showed that:compared with the 0 μmol/L 5-Zac group, the apoptosid rate in the 5 μmol/L 5-Zac group and 10 μmol/L 5-Zac group was signficantly increased, which was (1.9 ± 0.06)%, (8.89 ± 1.36)%, and (24.50 ± 3.68)% in the 0 μmol/L 5-Zac group, the 5 μmol/L 5-Zac group, and the 10 μmol/L 5-Zac mol/L group respectively. The bisulfite genomic sequencing showed that the demethylation probability of CpG points on -601 bp and -553 bp was significantly increased in the 5-Zac treated cells compared with those untreated. CONCLUSION 5-Zac can result in the increase of PD-1 expression in the human lymphoid cell series Molt-4 in vitro, and the apoptosis rate increases, which is related to PD-1 gene promoter demethylation.
Apoptosis ; genetics ; Azacitidine ; pharmacology ; Cell Line ; CpG Islands ; genetics ; DNA Methylation ; drug effects ; genetics ; Enzyme Inhibitors ; pharmacology ; Humans ; Programmed Cell Death 1 Receptor ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism ; T-Lymphocytes ; cytology ; metabolism

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Objective: To evaluate the effectiveness of therapeutic massage (tuina) for treating knee osteoarthritis (KOA). Methods: Six English and Chinese databases, including Chinese National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), China Biology Medicine Disc (CBM), Cochrane Library and PubMed databases, were independently searched to identify appropriate randomized controlled trials (RCTs) studying therapeutic massage for KOA compared to oral non-steroidal anti-inflammatory drugs (NSAIDs) alone. The main outcome measures were total effectiveness and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) score. Results: A total of 8 RCTs were included and they were of average quality. The results showed that therapeutic massage was more effective than NSAIDs comparing total effectiveness [risk ratio (RR)=1.14, 95％ confidence interval (CI) (1.07, 1.21), P<0.0001]; compared with NSAIDs, therapeutic massage produced more significant improvements in pain [mean difference (MD)=-2.06, 95％CI (-2.75, -1.36), P<0.00001], stiffness intensity [MD=-0.90, 95％CI (-1.05, -0.75), P<0.00001] and joint function [MD=-12.48, 95％CI (-13.91, -11.05), P<0.00001]. Conclusion: Therapeutic massage was more effective than oral NSAIDs in treating KOA. In relieving pain and stiffness and improving the function of knee joint, therapeutic massage was superior to NSAIDs.

Aged ; Female ; Humans ; Hypertriglyceridemia ; blood ; epidemiology ; Male ; Middle Aged ; Prevalence ; Uric Acid ; blood

OBJECTIVETo investigate the current demands for professional continuing medical education (CME) in ophthalmic nurses and their influential factors, and to provide evidence for properly setting the curriculum of professional CME for nurses.

METHODSQualitative and quantitative analysis was performed on 60 nurses in an ophthalmic hospital by questionnaire survey and group interview.

RESULTSOf the nurses surveyed, 49 (81.7%) thought the training time of professional CME needs to be adjusted, 40 (66.7%) referred to the training pattern, and 53 (88.3%) the training contents. The demanded courses of training mainly included professional nursing knowledge, professional nursing skills, and ophthalmic knowledge. Most nurses considered the influential factors for training were unreasonable training time and lack of applicability of some courses.

CONCLUSIONTo improve the quality of professional CME for ophthalmic nurses, the training curriculum should be designed in consideration of ophthalmic characteristics, different work experiences of nurses, and different nursing positions, the training time should be arranged properly, and various training patterns should be adopted flexibly.

Adult ; Cross-Sectional Studies ; Education, Nursing, Continuing ; Female ; Humans ; Needs Assessment ; Nurses ; Ophthalmology ; education ; Young Adult

Based on the literatures about the optimized effectiveness of the acupoint sticking therapy for diseases which are attacked in winter but are treated in summer, the main constituent elements of plaster, acupoints, the dose and the application time are reviewed and analyzed. It is suggested that the stepwise procedures are adoped to accumulate clinical evidence, standardize the technique instructions based on the clinical evidences, and perform high quality clinical trials.
Acupuncture Points ; Clinical Trials as Topic ; Drug Therapy ; methods ; standards ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Seasons ; Treatment Outcome