Objective To build an HPLC-ELSD method for simultaneous determination of 8-O-acetylharpagide and harpagide inAjuga decumbens Thunb.;To determineAjuga decumbens Thunb. samples from different area and of different batches.Methods Chromatographic separation was achieved on a Dikma C18 column (4.6 mm×150 mm, 5μm). The mobile phase consisted of water (A) and acetonitrile (B), using a gradient elution of 5% B at 0-6 min, 5%-15% B at 6-11 min, 15% B at 11-17 min, and the flow rate was 1.0 mL/min. An evaporative light scattering-detector (ELSD) was used with the temperature of drift tube at 103℃ and the gas flow rate of air was at 1.6 L/min.Results Good linearity was shown at the concentration range of 0.017-17.32μg for 8-O-acetylharpagide, and 0.003-1.62μg for harpagide. The validated method was of great precision and accuracy. The contents of 8-O-acetylharpagide and harpagide inAjuga decumbens Thunb. from Fujian province were higher than the contents in other localities.Conclusion The content of harpagide can be used as index component to monitor quality control ofAjuga decumbens Thunb..

Objective:To investigate the role of TSLP and NF-κB in the pathogenesis of adenomyosis.Methods:Immunohisto-chemistry was employed to detect the expression of TSLP and NF-κB p65 in the endometrial glandular cell and stromal cell of 35 adeno-myosis patients and 20 hysteromyoma patients as control , and analyze the correlation of the two proteins.The concentration of TSLP was measured by specific ELISA in the serum of healthy subjects besides of the patients with adenomyosis and hysteromyoma .Results:TSLP and NF-κB p65 were both positively expressd in the glandular cells and stromal cells of ectopic and eutopic endometrium of patients with adenomyosis , their expression was significantly higher than that of the control group ( P<0.01 ) , and the expression of TSLP and NF-κB p65 in ectopic endometrium was significantly higher than that in eutopic endometrium ( P<0.01 ).There was positive correlation between TSLP and NF-κB p65 in ectopic endometrium.The concentration of TSLP was higher in the serum of patients with adenomyosis.Conclusion:The TSLP is highly expressed in endometrial glandular and stromal cells of patients with adenomyosis , and there is higher level of TSLP in serum , and there is correlation between the expression of TSLP and NF-κB in ectopic endometrium.

Objective To investigate the evolution of surgical treatment and it's influence on prognosis in reptured liver cystic echinococcosis.Method Clinical data of 110 surgically treated cases of reptured liver cystic echinococcosis from January 1990 to December 2008 were retrospectively analysed,and these cases were divided into three groups by different surgical approach;Group A(22 cases):removal of internal capsule,with formaldehyde or hydrogen peroxide regional anthelminthic,with residual cavity side edge of lock stitching or greater omentum filling or drainage after residual cavity closure;Group B (65 cases):Removal of intemal capsule,with hypertonic saline regional anthelminthic,with external capsule extrahepatic partial resection,with residual cavity bile leakage sutured,and with residual cavity open drainage,Group C(23 cases):the total or subtotal external capsule stripping operation,with hypertonic saline regional anthelminthic and drainage.Result The rate of overall postoperative complications of residual cavity was 40.9%、16.9%、0.0%(P<0.05)respectively and the recurrence rate was 18.2%、4.6%、0.0%(P<0.05)respectively;drainage time needed was the longest in group A,and shortest in group C,whereas the operation time was the longest in group C and it costed higher blood loss.Intraabdominal implantation of the hydatid significantly decreased with hypertonic saline treatment.Conclusion Total or near total removal of the external capsule of the hydatic cyst and hypertonic saline treatment with postoperative external drainage is the therapy of choice for reptured liver cystic echinococcosis.

Objective To establish a rabbit VX2 liver tumor model by transplanting tumor tissue mass into the rabbit's liver,to analyze and observe the growing features of the liver tumor.Methods The tumor tissue mass(about 106-108 VX2 liver tumor cells)was inoculated into the left hepatic labe in 20 rabbits to establish rabbit VX-2 hepatic carcinoma model.The observation included the following two respects.(1)The tumor's volume at 7,10,14,17 and 21 days after the procedure was measured by ultrasonography and the growth rate of tumor was calculated.(2)The morphological feature of the tumor was inspected both macroscopically and microscopically.Results The growing pattern of the tumor was compatible with the exponential curve.Seventeen days after transplantation the increase rate of the tumor volume was much higher than that of the tumor diameter. Histopathologjcally,the growing pattern of the tumor took the form of infiltrative way,with its appearance being quite similar to the VX2 squamous cell carcinoma.Conclusion Transplantation of tumor tissue mass is the technique of first choice to establish the VX2 liver carcinoma model in rabbits.This experimental model is a very ideal animal form for both clinical and fundamental studies of liver carcinoma.

BACKGROUND: Many operations for isolating, purifying and identifying bone marrow mesenchymal stem calls (BMSCs) are complicated and cost much. Also they have great effect on cell activity. Whether whole bone marrow adherent culture can avoid above-mentioned disadvantages remains unclear. At present, many studies huve been done to confirm an effective and low cost method for isolating, purifying and identifying such cells.OBJECTIVE: This study is to in vitro induce and differentiate rat BMSCs by whole bone marrow adherent culture,and to identify the cells.DESIGN: A controlled observational experiment.SETTING: Qingdao University Medical College.MATERIALS: This study was carried out in the Laboratory of Oral Cavity and Laboratory of Molecular Biology (provincial level) Qingdao University Medical College between November 2005 and March 2007. Twenty Wistar rats of either gender, aged 3 to 4 weeks, of SPF grade, weighing 120-150 g, were provided by the Qingdao Laboratory Center. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Fetal bovine serum (FBS, Hangzhou Sijiqing Bioengineering Material Research Institute), alkaline phosphatase (ALP) kit (Nanjing Jiancheng Bioengineering Research Institute), reverse transcription kit (American Promega Corporation) and primer (Shanghai Bioengineering Co.,Ltd.) were used in this study.METHODS: Adult rat BMSCs were isolated and cultured by whole bone marrow adherent culture. They were digested with 2.5 g/L trypse and inoculated at a density of 5 ×107 L-1 in 6-well culture plate. Then, the cells were divided into experimental group and control group. Inducing culture medium was added to experimental group, and the same amount of basic culture medium was added to control group. ① Cell differentiation and calcium tuberculation were observed under the inverted microscope. ② Biological characteristics of induced cells were detected by calcium tubercle Von Kossa and alizarin Bordeaux. ③ALP activity was detected by diazo salt staining. ④Human core binding factor alpha subunit-1 (Cbf α-1), osteocalcin (OCN) and osteoblast-specific Osterix (OSX) mRNA expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: ① Induction and differentiation results of cells. ② Biological characteristics of cells induced by rat BMSCs. ③ ALP activity. ④ Cbf α-1, OCN and OSX expressions.RESULTS: ①Inducing culture medium was added in the serial subcultivation. About 9 days later, cell clones were connected to each other. On about 21 to 28 days, some pykno-round mineralized tubercles appeared. Meanwhile,control cells were connected to each other, but they did not form the tubercle. ② In the experimental group, when MSCs were induced for 21 to 28 days, obvious round or oval calcified tubercles were seen by naked eyes. The results of Von Kossa staining exhibited black sediments, and those of alizarin Bordeaux staining exhibited salmon tubercles. Calcium tubercles were not found in the control group. ③The ALP activity after 2 weeks of induction was obviously increased in the experimental group, but was relatively weak in the control group. ④In the experimental group,Cbf α-1, OCN and OSX expressions were significantly increased after induction.CONCLUSION: After being in vitro induced and differentiated by whole bone marrow adherent culture, rat BMSCs exhibited morphological and biological characteristics similar to typical osteoblasts.

Objective To establish a suitable and stable metastatic rabbit VX2 liver tumor model for the use in experimental study,to discuss the successful rate of different tumor transplanting methods,and to analyze the digital subtract angiography(DSA)imagining features of the transplanted liver tumor.Methods Sixty male New Zealand white rabbits were randomly and equally divided into 3 groups with 20 rabbits in each group.For the rabbits of two groups,receiving injection methods and used as control groups,VX2 carcinoma particle(containing about 5×107 carcinoma cells) was inoculated into the left hepatic lobe through injection via hepatic artery or through direct injection with a syringe needle.For the rabbits in the retrofit group tumor tissue particle(containing about 106-108 carcinoma cells)was directly transplanted into liver through puncturing of the Glisson's capsule.The observation included the following two respects. (1)The tumor's survival rate of the three groups was evaluated.(2)The DSA imaging feature of the transplanted tumor was observed.Results The survival rate of the transplanted tumor in three groups was 7/20,10/20 and 19/20 respectively,with the survival rate of the retrofit group being the highest in 3 groups(P＜0.05).On DSA the transplanted tumors were rich in blood supplying.Conclusion For the establishment of rabbit VX2 liver carcinoma model,the direct transplantation of the tumor tissue particle is obviously superior to the injection method(direct injection or through hepatic arterial injection)in obtaining higher successful rate.This technique provides clinical and fundamental liver cancer studies with a reliable,stable and mature tumor animal model.

8-O-acetylharpagide and harpagide are two kinds of effective component of Ajuga decumbens extract. A sensitive LC-MS/MS method has been established for pharmacokinetics of 8-O-acetylharpagide and harpagide in beagle dog after oral administration of from A. decumbens extract. Female beagle dogs received orally 12.9, 25.7 mg x kg(-1) p. o. Concentrations of 8-O-acetylharpagide and harpagide in plasma were determined by LC-MS/MS method at different time points and all pharmacokinetic parameters were estimated by non-compartment analysis. The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B), which was run at a flow rate of 0.3 mL x min(-1). Chromatographic separation was achieved on an Agilent ZORBAX XDB-C18 column (2.1 mm x 50 mm, 3.5 microm) using a gradient elution of 5% B at 0-2 min, 95% B at 2. 1-5 min and 5% B at 5. 1-10 min. All analytes, including the IS, were monitored under positive ionization conditions and quantified in MRM mode with transitions of m/z 429.2-369.2 for 8-O-acetylharpagide, m/z 387.2-207.2 for harpagide, and m/z 149.2-103.1 for IS. High purity nitrogen was employed as both the nebulizing and drying gas. Other parameters of the mass spectrometer were optimized as follows: drying gas flow 10.0 L x min(-1); drying gas temperature 300 degrees C; capillary voltage 4 000 V. Results showed that 8-O-acetylharpagide and harpagide showed a dose-dependence profile. T(max) of 8-O-acetylharpagide is 1.7 h, and T(max) of harpagide is 1.57 h, which was higher than T(max) of 8-O-acetylharpagide and harpagide after oral administration of from A. decumbens extract in rats. The different pharmacokinetic parameters may be due to the species differences of rat and beagle dog.
Administration, Oral ; Ajuga ; Animals ; Dogs ; Female ; Iridoid Glycosides ; pharmacokinetics ; Plant Extracts ; metabolism ; Pyrans ; pharmacokinetics ; Rats ; Species Specificity

Objective To evaluate the safety and effectiveness of endovascular balloon dilatation in removing incarcerated tunnel cuffed catheter (TCC) . Methods The clinical data and the imaging materials of 4 hemodialysis patients with incarcerated TCC, who received endovascular balloon dilatation at Taizhou Hospital of Zhejiang Province, China, during the period from January 2017 to March 2018, were retrospectively analyzed. After the treatment, the patient's vital signs were monitored, and the procedure-related complications such as hemopneumothorax, subcutaneous hematoma, arrhythmia, etc. were documented. Results The mean age of the 4 patients was (73.3±6.4) years. The average indwelling time of TCC was 5.5 years (4-8 years) . Successful treatment of incarcerated TCC was achieved in all 4 patients. In one patient, both 5-mm and 6-mm balloons were used to simultaneously dilate the dual cavities of TCC.Temporary arrhythmia occurred in one patient during operation. No serious postoperative complications occurred. Conclusion For the remove of long-term indwelling incarcerated TCC, endovascular balloon dilatation is minimally-invasive, safe and effective, although more researches are needed to further confirm its safety and reliability.

Objective To compare the long term effectiveness of tonsillotomy(TT)vs. tonsillectomy (TE)by hypercator in treatment of obstructive sleep apnea hypopnea syndrome(OSAHS)in children. Methods 119 children with tonsil hypertrophy were diagnosed with OSAHS and included in the study. Among them,57 children underwent TT by hypercator and 62 TE only after preoperative laboratory polysomnography(PSG). All participants were interviewed 5 years after operation to assess the satisfaction of their caregivers,investigate the frequency of respiratory infections,the rate of tonsil regrowth and evaluate their quality of life by Obstructive Sleep Apnea-18(OSA-18). Results Some of them received postoperative PSG. All parents reported alleviation of breathing obstruction,but the postoperative satisfaction in TT group was higher than TE group mainly because of the shorter time return to normal diet. The rate of the respiratory tract infections was not increased in all cases after surgery. Four cases of children were observed with tonsillar regrowth,but no surgery was needed for further treatment. The rate of the dry throat symptoms in TT group was lower than TE group. Conclusions Tonsillotomy by hypercator can effectively eliminate the obstruction symptoms in children with OSAHS,superior to TE for its shorter recovery period and smaller oral mucosal injury and lower rate of tonsils regrowth and it is worth to clinical promotion.

Objective To investigate the dynamic changes of peripheral blood lymphocyte subsets and their correlation with renal function in recipients with stable graft status after renal transplantation. Methods Forty-five recipients who underwent renal transplantation for the first time and had stable graft function within postoperative 6 months were selected. The proportion and absolute value of lymphocyte subsets were detected by flow cytometry (FCM) in 180 peripheral blood samples from recipients at 15 d, 1, 3 and 6 months after renal transplantation. The dynamic changes of lymphocyte subsets with the extension of postoperative time and their correlation with serum creatinine (Scr) and blood urea nitrogen (BUN) were analyzed. Results The Scr levels did not significantly differ at 4 time points after renal transplantation (all P > 0.05). The BUN levels significantly differed between 15 d and 1 month after renal transplantation, and between 1 and 3 months after renal transplantation (P=0.002, P=0.001). The proportion of CD3⁺CD8⁺T cells, CD3⁺CD4⁺T cells, natural killer (NK) cells and CD4/CD8 ratio at postoperative 15 d significantly differed from those at 1 month after operation (P=0.009, P=0.004, P < 0.001, P=0.004). The proportion of B cells significantly differed between 15 d and 1 month, and between 1 and 3 months after renal transplantation (both P < 0.001). The absolute values of CD3⁺T cells, CD3⁺CD8⁺T cells, CD3⁺CD4⁺T cells and NK cells at postoperative 15 d significantly differed from those at 1 month after renal transplantation (P=0.001, P=0.002, P=0.003, P < 0.001). The absolute values of CD3⁺CD8⁺T cells significantly differed between 3 and 6 months after operation (P=0.015). The absolute value of B cells at 1 month after renal transplantation significantly differed from that at 3 months after renal transplantation (P=0.001). The proportion and absolute value of lymphocyte subsets were not significantly correlated with the Scr level (both P > 0.05). The proportion and absolute value of CD3⁺CD8⁺T cells and NK cells were negatively correlated with BUN (P < 0.001-0.05), whereas the proportion of CD3⁺CD4⁺T cells and B cells was positively correlated with the BUN level (P < 0.001-0.05). The absolute value of CD3⁺T cells was negatively associated with the BUN level (P < 0.05). Conclusions T cells and NK cells in the lymphocyte subsets of stable recipients raise to the stable state within 1 month after renal transplantation, whereas B cells decrease to stable state within 3 months renal transplantation. The dynamic changes of lymphocyte subsets are correlated with the BUN level.