OBJECTIVE:To establish a TLCS method for the determination of Sarsa sa pogenin in Kangbingdu oral liquid.METHODS:TLCS was used with benzol-acetone(9 ∶1)as developing agent Sawtooth scanning was at ?S=443nm,?R=600nm RESULTS :The linear range was 1 028?g～3 624?g(r=0 9 995) The average recovery was 100.89%,and the RSD was 3 36%(n=5) The stability and precision were go od CONCLUSION:TLCS method can be used for the determination of Sarsasapogenin in Kangbingdu oral liquid

BACKGROUND: The investigation of culturing, passaging and establishing human limbal stem cells can strengthen the recognition of the stem cells and provide the enough cellular reserve for the basic and clinical research of limbal stem cell transplantation.OBJECTIVE: To explore a method of pessaging and establishing cell line of human limbal stem cells cultured in vitro.DESIGN: Randomized controlled observation.SETTING: Gannan Medical College.MATERIALS: The experiment was performed at Scientific Center of Gannan Medical College and the National Key Laboratory of Ophthalmology Hospital Affiliated to Sun Yat-Sen University from June 2003 to April 2004. Fresh human limbus corneae were isolated from two healthy donors. Procedures were performed according to the informed consent of the donors. Main reagents contained RPMI-1640 (Sigma R8755, containing L-glutamine) and 200 g/L fetal calf serum (FCS) (Gibco 16140-071). DMEM medium, chondroitin sulfatase and human epidermal growth factor (hEGF) were purchased from Sigma Co. USA; HEPES and DMSO were bought from Gibco, USA; 100% glycerinum was purchased from Yunjia Huangpu Pharmaceutical Product Limited Company, PRC; glutaraldehyde was bought from E.Merk, Germany; Alcohol, chlorhydric acid, acetone and methyl aldehyde were purchased from Beijing Chemical Agent Company, PRC; 0.25% parenzyme was bought from Shanghai Xinhua Pharmaceutical Factory, PRC.Above-mentioned reagents were analytical pure grade.METHODS: After digestion, human limbal tissues in limbal basilar part with an abundant pigment were cultured in the culture flask containing RPMI-1640 and 200 g/L FCS and in culture dish containing amniotic extracellular matrix (AECM) as the cultural supporter. Primary and passage cells were observed under light microscope and scanning electron microscope (SEM). The revival ratio of stem cell refrigeration of every generation was calculated by the trypanblau exclusion experiment.MAIN OUTCOME MEASURES: ① Observational results of limbal stem cells during the primary culture and serial subcultivation in vitro, and ② revival ratio of stem cell refrigeration.RESULTS: ①Findings of primary culture: Most limbal stem cells in the culture flask had the adherence and were arrayed uniformly sparsely to form monolayer and adhered to the bottom of culture flask under the inverted phase contrast microscope after 1-day culture. ② Findings of serial subcultivation: After human epidermal growth factor (hEGF) was added into the second passage, cells were scattered into the monolayer and adhered to grow quickly.Morphological variability of all the cells increased obviously when passage the 30th generation. The cellular volume was obviously increasing, and the round or irregular round cells gathered together. The 33rd generation human limbal stem cells still could vigorously differentiate, proliferate and grow in ACEM. ③ The revival ratio of stem cell refrigeration was 82.2%.CONCLUSION: The human limbal stem cell lines were preliminarily established by culturing and freezing the cells of 33 generations in vitro. The human limbal stem cell lines preferred to grow in the culture dish containing AECM as the cultural supporter.

OBJECTIVE: The purpose of this study was to prospectively investigate the value of the myocardial extracellular volume fraction (ECV) in predicting myocardial functional outcome after revascularization of coronary chronic total occlusion (CTO). MATERIALS AND METHODS: Thirty patients with CTO underwent cardiovascular magnetic resonance (CMR) before and 6 months after revascularization. Three baseline markers of functional outcome were evaluated in the dysfunctional segments assigned to the CTO vessels: ECV, transmural extent of infarction (TEI), and unenhanced rim thickness (RIM). At the global level, the ECV values of the whole myocardium with and without a hyperenhanced region (global and remote ECV) were respectively measured. RESULTS: In per-segment analysis, ECV was superior to TEI and RIM in predicting functional recovery (area under receiver operating characteristic curve [AUC]: 0.86 vs. 0.75 and 0.73, all p values < 0.010), and it emerged as the only independent predictor of regional functional outcome (odds ratio [OR] = 0.83, 95% confidence interval [CI]: 0.77–0.89; p < 0.001) independent of collateral circulation. In per-patient analysis, global baseline ECV was indicative of ejection fraction (EF) at the follow-up examination (β = −0.61, p < 0.001) and changes in EF (β = −0.57, p = 0.001) in multivariate regression analysis. A patient with global baseline ECV less than 30.0% (AUC, 0.93; sensitivity 94%, specificity 80%) was more likely to demonstrate significant EF improvement (OR: 0.38; 95% CI: 0.17–0.85; p = 0.019). CONCLUSION: Extracellular volume fraction obtained by CMR may provide incremental value for the prediction of functional recovery both at the segmental and global levels in CTO patients, and may facilitate the identification of patients who can benefit from revascularization.
Collateral Circulation ; Coronary Vessels* ; Follow-Up Studies ; Humans ; Infarction ; Magnetic Resonance Imaging* ; Myocardial Infarction ; Myocardial Ischemia ; Myocardium ; Prospective Studies ; ROC Curve ; Sensitivity and Specificity

Objective To Pulse oximetry saturation has been wildly used clinically.It has been reported that pulse oximetry plethysmographic waveform (POP) reflected the peripheral tissue perfusion.In this study,we parameterized POP,observed the value of POP parameters in normal adults,and established the normal reference value range.Methods A multi-center prospective descriptive study.Total of 1 019 adult volunteers with normovolemia from 7 cities were enrolled in this study.Sex,age,height,weight and pulse oximetry data in awake and spontaneous breathing under in quiet conditions in the room temperature were collected.POP parameters and perfusion index were analyzed using MATLAB 2012a software.The normal reference value ranges of POP parameters,including the amplitude of POP (Amp) and the area under the curve of POP (AUC),were formulated.Results Statistical differences of POP parameters were detected between men and women in the normal adult.The 95％ confidence reference value of POP parameters in normal population was as follows:Amp (104.8-2298.7) PVA and AUC (3265.8-6028.5) PVPGin total,Amp (129.4-2433.6) PVA and AUC (3319.0-5862.2) PVPG in male;Amp (89.5-2138.2) PVA and AUC (3163.9-5929.9) PVPG in female.Conclusions POP,including the amplitude of POP (Amp) and the area under the curve of POP (AUC),had normal reference value ranges in normal adults.

Objective Myocardial functional outcome after revascularization of coronary chronic total occlusion (CTO) was prospectively predicted using extracellular volume fraction (ECV) based on cardiovascular magnetic resonance (CMR). Methods Thirty patients with CTO underwent CMR before and 6 months after percutaneous coronary intervention (PCI) were enrolled. The CMR scan protocol included cine, pre?contrast and post?contrast T1 mapping and late gadolinium enhancement (LGE). Ejection fraction (EF) and segmental wall thickening (SWT) were calculated using CVI 42 software. SWT less than 45% indicated myocardial segment dysfunction. According to the American Heart Association (AHA) scientific statement, the dysfunctional segments assigned to CTO vessel were selected, and three baseline imaging markers, ECV, transmural extent of infarction (TEI) and unenhanced rim thickness (RIM) were respectively evaluated. The myocardial segments were divided into two subgroups, group with well?developed collaterals and group with poorly?developed collaterals, based on the collateral circulation using Rentrop classification. Baseline and follow?up values of SWT and EF were evaluated using paired Student′s t?test. Using an increase in SWT>10% as standard reference, ROC analysis was conducted to describe the predictive performance of baseline markers. A mixed linear model was used to probe the relationship between collateral circulation and SWT. Stepwise logistic regression analysis was used to determine the independent predictors of regional functional recovery. The differences of EF between poorly?developed and well?developed collaterals were compared by Student t test. Results The baseline mean segmental wall thickening (SWT) of the dysfunctional segments increased from 21.6% (9.7%, 33.3%) to 38.4% (19.0%, 51.2%) after PCI (Z=-6.869, P<0.001), and EF was also significantly higher compared with baseline (54.5%±8.5 % vs. 50.7%± 6.6%, t=-5.706, P<0.001). ECV showed good performance in predicting functional recovery with cutoff value 34.7%, area under ROC curve (AUC) 0.86, sensitivity 91%, and specificity 66%. The AUC of ECV was superior to TEI and RIM (AUC: 0.75 and 0.73, all P value<0.01). The segments with well?developed collaterals were associated with a higher SWT at follow?up [46.6% (36.6%, 64.2%) vs. 33.5% (12.8%, 47.8%),F=5.791, P=0.02]. Logistic regression analysis demonstrated that mean segmental ECV was the only independent predictors of regional functional outcome after PCI (OR=0.83, 95% confidence interval: 0.77—0.89; P<0.001). Conclusions ECV by CMR may provide incremental value for the prediction of regional functional recovery in CTO patients, and baseline collateral circulation correlates with the regional systolic function after revascularization.