To provide insights into the mechanism for the attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue, a contrasted study was carried out on the pharmacokinetics of brucine and strychnine in mice plasm, which are active and toxicant ingredient in the Tibetan medicine Renqing Mangjue. LC-MS/MS was used to detect simultaneously the concentrations of brucine and strychnine in mice plasm at-different time intervals after administration parallelly and randomly, and the pharmacokinetic software Kinetica 5. 0 was selected to non-compartmental analysis (NCA) for data, and statistical analysis software SPSS 19. 0 was used for significance test on the pharmacokinetic parameters. A reliable LC-MS/MS method was established for the determination of brucine and strychnine in blood plasma, which are consistent with the requirements of the preclinical pharmacokinetic study confirmed by the methodology. The linear concentration ranges of brucine and strychnine were 0.301-104.4 µg · L(-1) (r = 0.999 5) and 0.305-106 µg · L(-1) (r = 0.999 7), respectively; The intra-day and inter-day variable coefficients were both less than 10.0% with good precision; The average extraction recoveries of brucine and strychnine were 116.23% and 112.82%, and RSD were 3.2% and 2.3% separately;The average matrix effects of brucine and strychnine were 122.48% and 116.36%, and RSD were 7.7% and 4.4%, respectively. The pharmacokinetic results showed that AUCtot of brucine and strychnine in Zuota group were both increased remarkably (P < 0.05), and the Cmax of brucine in Zuota group was about 5.25-fold higher than that of brucine in non-Zuota group (P < 0.05). The Tmax of brucine and strychnine reduced to one-eighth and one-quarter respectively compared with those in Non-Zuota group. In addition, the eliminations of brucine and strychnine in vivo were accelerated after the compatibility of Zuota. A significant difference (P < 0.05) occurred at the MRT0-t, of brucine, while the MRT0-∞ and Lz of strychnine were statistically significant upon the inspection level α = 0.1. It was found that the absorption degree of brucine and strychnine in Zuota group increased in the range of the safe dose (or concentration), while their elimination rates were accelerated, which may be one of the mechanisms for attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue.
Animals ; Female ; Male ; Medicine, Tibetan Traditional ; Mice ; Strychnine ; analogs & derivatives ; pharmacokinetics

According to foreign and domestic standards of clinical quality and the author team’ experience summary of long-term clinical study, it has been found that quality control in clinical acupuncture operators is closely related to operator’s ability, good communication between doctors and patients, signature for informed consent, implementation of inclusion and exclusion criteria, practice of blind method and random concealment, patient’s compliance, standardized writing and keeping of CRF, management of instruments and consumables, and coordination and association between researchers. The implementation method of quality control in clinical acupuncture operators is explored by the clinical practice of the author team in “973” project, and analysis and discussion of the above key points.

Objective To develop self-microemulsifying drug delivery system(SMEDDS) of Acanthopanax senticosus total saponins(ASTS) and the determination method.Methods The equilibrium solubility of ASTS in different compositions of oils,emulsifier and assistant emulsifier was investigated.The self-microemulsion formula was optimized by constructing the pseudo-ternary phase diagrams of blank SMEDDS and the studying the self-microemulsifying efficiency and the stability of drug-loaded SMEDDS.Taking isofraxidin,an effective component of A.senticosus,as the content determination index,the content was determined by HPLC.Results The optimal self-microemulsion formula was composed of Caf,propanediol,and ethyl linolenate.The ratio of them was 16∶4∶5.The average particle size was 40 nm.The average content of isofraxidin was 92.6 ?g/mL.Conclusion The acquired microemulsion with small particle size is stable.The content determination method of taking isofraxidin as the quality control index is accurate and reliable.

BACKGROUND:It is discovered recently that miRNA is a new regulator that is able to have an impact on gene expression and miRNA contributes to proliferation, differentiation and self-renewal of pluripotent stem cels.
OBJECTIVE:To investigate the mechanism by which miR-155 regulates chondrogenic differentiation of bone marrow mesenchymal stem cels.
METHODS: Sixty healthy Sprague-Dawley aged 12 weeks were randomized into study group and control group, with 30 in each group. Under anesthesia, rats were sacrificed to harvest bone marrow of the lower limbs. Then bone marrow mesenchymal stem cels were isolated, cultured, and transfected with miR-155 mimics in the study group and a negative control sequence in the control group. After chondrogenic induction, RT-PCR was used to detect the expressions of Sox9, Colagen II, Aggrecan and Colagen X gene, and western blot assay to detect the expression of Sox9 and Runx2 proteins.
RESULTS AND CONCLUSION:Compared with the control group, the mRNA expressions of Sox9, Colagen II and Aggrecan were higher, but the mRNA expression of Colagen X was lower in the study group (P < 0.05); the protein expression of Sox9 was higher, but the protein expression of Runx2 was lower in the study group (P < 0.05). These findings indicate that miR-155 promotes the chondrogenic differentiation of bone marrow mesenchymal stem cels and moreover, it can suppress the hypertrophy trend of bone marrow mesenchymal stem cels differentiating into chondrocytes.

Objective To investigate the effect of testosterone on mitotic orientation in rat prostate epithelial cells and the relative differential gene expression.Methods Twenty SPF male SD rats were divided into 2 groups at random and then subjected to castration.One group of rats was administrated with testosterone 3.7 mg daily for 30 days and the control group was only injected with olive oil.Microscopic analysis was performed using immunohistochemistry.Differential gene expression analysis was conducted by gene microarray and RT-PCR techniques.Results In the testosterone-adminis-trated group, there was a significant mitosis orientation parallel to the basement membrane.But in the control group, mito-sis orientation was oriented perpendicular to the basement membrane.Using the gene microarray and RT-PCR techniques, the cell proliferation genes such as Ran, Tgm4 and Wnt2 in Wnt signal pathway were up-regulated in the testosterone group.Conversely, suppressor cell proliferation genes such as Dkk3 and Fas were down-regulated.Conclusions Mitotic orientation of prostate epithelia cells is changed after testosterone administration.Wnt signal pathway and AR singling path-way also have an influence on the mitosis orientation and cell proliferation.

Objective To study the characteristics of hepatic artery anatomy and master the method of arterial repair,especially the method and technology of repair of anomalous donor arteries.Methods Sixty-four(human) liver arteries were dissected and repaired,and 31 of these were used in liver transplantation.Results Twelve of 64 cases had anatomical anomaly of hepatic artery(12/64,18.75%).Five of 24 fetal(liver) donors(20.83%) had anatomical anomaly of hepatic artery,one arose from the superior mesentery(artery) as a replaced right hepatic artery,3 from accessory right hepatic artery,and in 1 case the hepatic(artery) arose from superior mesentery artery directly.Seven adults had hepatic artery anomalies:2 cases of right hepatic artery being replaced by artery derived from SMA,2 cases of left hepatic artery deriving from left gastric artery and 3 cases of accessory right hepatic artery derived from SMA.In 31 hepatic(transplantations),4 grafts with anatomical anomaly of right hepatic artery were used.Conclusions (Reconstruction) of hepatic artery plays an important role in hepatic transplantation.Avoidance of injury to(aberrant) hepatic avteries during removal of donor liver is a key to ensure successful arterial repair.During(repair) of very small accessory hepatic arteries,ligation or repair of the artery may be determined after(observation) of backflow from collateral vessels.

Objective To compare the health related quality of life (HRQoL) of ileal conduit versus orthotopic ileal neobladder using the FACT-BL scale.Methods One hundred and thirty patients underwent radical cystectomy and urinary diversion for invasive bladder cancer from Jan 2006 to Dec 2010 at our hospital.According to different urinary diversions,patients were divided into ileal conduit (IC) and orthotopic ileal neobladder (NB).HRQoL was assessed using FACT-BL.The following scores calculated from the FACT-BL questionnaire and list:physical well-being ( PWB ),social/family well-being ( SWB ),emotional well-being(EWB),functional well-being( FWB),overall bladder-special subscale and total FACT-BL were included.A high FACT score indicated a high level of HRQoL.The medical records of each patient were then reviewed.Clinical parameters including age,sex,type of diversion and pathological status were recorded and analyzed.Results A total of 94 questionnaires were collected,of these patients 50 had an IC and 44 had an NB.Among the respondents,the age at surgery was significantly younger in NB group than IC group (P =0.014 ).While the percent of males,follow-up years and pathological stage T3 or greater revealed no significant difference.The NB group got significantly higher scores in PWB,SWB,EWB and FACT-BL than the IC group.Which indicated that the HRQoL was higher in patients following orthotopic ileal neobladder.Conclusions The HRQoL was better in patients who followed orthotopic ileal neobladder than those who followed ileal conduit.This may indicate that orthotopic ileal neobladder should be considered first when making a decision about which urinary diversion should be used.

BACKGROUND: Exhaled nitric oxide (eNO) is one of the airway condensate derived markers, reflecting mainly airway inflammation in asthma and other lung diseases. The changes of eNO levels as pathophysiology of neonatal hypoxemic respiratory failure (HRF) in early postnatal life have not been thoroughly studied. The present study was to establish a method for measuring eNO concentrations in neonates with or without HRF. METHODS: Twenty-two newborn infants with HRF and 26 non-NRF controls were included within the first 24 hours of postnatal life. Their eNO levels were detected with a rapid-response chemiluminescence analyzer daily during the first week of their postnatal life, and lung mechanics and gas exchange efficiency were monitored at the same time, such as pulse oxygen saturation (SpO2), inspired fraction of oxygen (FiO2) and other parameters. RESULTS: During the first two days of postnatal life, eNO values of HRF neonates were significantly higher than those of the control neonates (day 1, 7.9±3.2 vs. 5.8±1.8 parts per billion [ppb], P<0.05; day 2, 8.8±3.2 vs. 6.0±2.4 ppb, P<0.05), but there were no significant differences in the following days. With SpO2/FiO2 increasing, difference of eNO values between the HRF and non-HRF neonates became narrowed, but there was still a two-fold difference of eNO/[SpO2/(FiO2×100)] on days 5-7. CONCLUSION: We established a method for measuring eNO and found difference in neonates with or without HRF, which diminished with prolonged postnatal days, reflecting pathophysiological characteristics of HRF.

Objective To explore the curative effect of deep-brain magnetic stimulation (DMS) on learned helplessness behavior in the chronic restrained stress (CRS) rat model. Methods Twenty-nine Sprague-Dawley rats were randomly divided into control group (n=8) and CRS group (n=21). CRS group was exerted chronic restrained stress, while the control group did not receive any stress, for three weeks. Then learned helplessness behavior was tested using Forced Swimming Test (FST) and the hopeless rats of the CRS group were divided ran-domly into sham group (n=6), DMS group (n=8) and citalopram group (n=7), that received corresponding treatment respectively. They were evaluated with FST again after one-week treatment. Results The immobile time in FST was longer in CRS group than in the control group after three-week stress (F=11.260, P=0.002). After one-week treatment, no significant improvement was found in the citalopram group (F=1.565, P=0.235), however, the immobile time in DMS group decreased (F=6.277, P=0.025), and was shorter than that in the sham group (F=5.560, P=0.036). Conclusion CRS could result in learned helplessness behavior, which could be alleviated with one-week DMS.