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Partial least square (PLS) combining with Raman spectroscopy was applied to develop predictive models for plasma paclitaxel concentration detection. In this experiment, 312 samples were scanned by Raman spectroscopy. High performance liquid chromatography (HPLC) was applied to determine the paclitaxel concentration in 312 rat plasma samples. Monte Carlo partial least square (MCPLS) method was successfully performed to identify the outliers and the numbers of calibration set. Based on the values of degree of approach ( ), moving window partial least square (MWPLS) was used to choose the suitable preprocessing method, optimum wavelength variables and the number of latent variables. The correlation coefficients between reference values and predictive values in both calibration set ( ) and validation set ( ) of optimum PLS model were 0.933 1 and 0.926 4, respectively. Furthermore, an independent verification test was performed on the prediction model. The results showed that the correlation error of the 20 validation samples was 9.36%±2.03%, which confirmed the well predictive ability of established PLS quantitative analysis model.
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In order to make the experimental teaching adapt to the development of modern teaching idea and to meet the needs of the pharmaceutical industry for high quality talents, pharma-cology experiment teaching method has been reformed. Single teaching method has turned to diversi-fied teaching method according to the experimental contents, difficulty and characteristics of teaching situation, such as leading method of using theory after experiment in validation experiments and single blind method in multidrug efficacy experiment in the early stage, case-based teaching in comprehensive experiments in the middle stage , and student teaching method in designing experiments in the later stage. The study results show that students' interest in learning has been inspired,their experimental enthusiasm has been mobilized,and their operation, analysis and problem-solving ability has been improved, which is advantageous to the comprehensive quality education.
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Objective To establish a model of full-thickness avascular meniscal defect to assess outcome of bone-marrow mesenchymal stem cells (BMSCs) modified with human insulin-like growth factor Ⅰ (hIGF-Ⅰ) gene and compounded with injectable calcium alginate gel in repair of meniscal defect.Methods Models of full-thickness defect were created in the anterior comer of meniscus in goats,an area lacking of blood supply.The trial categorized the models to four groups:gene-ehanced tissue engineering (GETE) group (hIGF-Ⅰ transfected BMSCs were mixed with calcium alginate gel),BMSCs group (BMSCs were mixed with calcium alginate gel),empty group (calcium alginate gel was used alone) and control group (the defect was excluded from repair).Macroscopy was done at 4,8,and 16 weeks after operation.Variation of repair tissue was observed by light and scanning electric microscopy and aggrecan in repair tissue was determined as well.Results Meniscal defect was on the mend at 4-16 weeks after operation in GETE group,with the defect area being thoroughly filled with the white,elastic and tight repair tissue similar to normal meniscal tissue.Macroscopic examination showed a better result in GETE group than that in other groups.Light microscopy showed that repair tissue which was mainly fibrochondrocytes was arranged in line with calcium alginate fibers and that space between the fibers was mostly crammed with the matrix secreted by those cells.At the same time,those cells were tightly arranged and the matrix secreted by those cells was equally distributed according to light microscopy.Electroscopy demonstrated neat and tight arrangement of fibers and tight extracellular matrix in fiber space in GETE group.Aggrecan concentration in GETE group was relatively higher than in other groups,but still had difference from the normal meniscus.Conclusion hIGF-Ⅰ gene-transfected BMSCs combined with injectable calcium alginate gel can improve the effect in repair of full-thickness meniscal defect.
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A approach to segment the color image based on Hue-Saturation-Value (HSV) space was proposed, and it was used to segment the color image digitized from the bone sections stained with Masson-Goldner's Trichrome by CCD camera. According to HSV approach, color image was transformed from RGB space to HSV space at first, and then the image was segmented by using the threshold value of hue (90 < H < 150) and saturation (S > 0.25) to find the image of the green color stained trabecular. And then used the threshold value of saturation (S < 0.2) and value (V > V(background) x 0.95) to find the unstained high brightness field. At last, unstained high brightness fields which were concluded in stained trabecular image were filled with the color of trabecular, so the completely trabecular image could be drawn. The results showed that HSV approach was fast and simple, and it could be an efficient automatic algorithm.
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Humans , Algorithms , Bone Density , Bone and Bones , Pathology , Color , Image Processing, Computer-Assisted , Osteoporosis , Pathology , Staining and LabelingABSTRACT
Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.
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BACKGROUND: The existed repair method for cartilage defects has shortcomings of insufficient repairing tissue numbers, poor biomechanical properties, as well as donor site complication. Thus it is deficient to repair large-sized osteochondral defects using one method.OBJECTIVE: To investigate the therapeutic effect of tissue engineering modified Mosaicplasty on repairing large-sized osteochondral defects.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Center Laboratory of Qingdao University Medical College from January to September 2009.MATERIALS: The hircine bone mesenchymal stem cells (BMSCs) were in vitro cultured, and resuspended with algin solution to obtain BMSCs-calcium alginate gel.METHODS: Totally 12 goats were prepared for osteochondral defects models and were divided into 3 groups.BMSCs-Mosaicplasty group, BMSCs compound with injectable alginate calcium gel was then applied to fill the "dead space" after Mosaicplasty. In the Mosaicplasty group, the defects were repaired by Mosaicplasty. There was no treatment in the control group.From 4 to 16 weeks postoperatively, the animals were sacrificed and the in gross and under electromicroscopy.MAIN OUTCOME MEASURES: ①Gross observation: the joint was exposed to observe the repair effect at weeks 4, 8,16 after operation. ②Histological examination: specimens were harvested at 16 weeks after operation and observed by haematoxylin-eosin staining, toluidine blue staining under light microscopy. ③Transmission electron microscope was used at 16 weeks after operation.RESULTS: The transplanted subchondral bone and superficial cartilage was integrated hardly with each other or with recipient sites in tissue engineering modified Mosaicplasty groups at 16 weeks after operation. The quality and appearance of the transplanted and regenerated cartilage was similar to normal hyaline cartilage. Under microscopy, the regenerated cartilage was integrated with neighbor tightly in regular arrange. ECM distributed evenly and deeply stained by alcian blue. There was no obviously repaired in the control group.CONCLUSION: Tissue engineering can ameliorate the outcome of Mosaicplasty to repair the osteochondral defects.
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AIM: To improve on the acute cumulative death rates method in animal by intravenous (iv) administration. METHODS: A mathematic model was established to simulate the pharmacodynamic and pharmacokinetic process by the program written by Visual Basic.net. This program evaluated the acute cumulative death rates (ACD method) for use of pharmacokinetics of traditional Chinese medicine (TCM). RESULTS: The ACD method related on its first dosage, and an unsuitable dosage could lead to a wrong results. It also had been proved that, by adjusting the 2nd dosage it was a more suitable to wider range of dosage. CONCLUSION: By suitable 2nd dosage and enough animal number, ACD method can be used in the pharmacokinetics of TCM.
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Objective:To compare the antihypertensive effect of L-amloipine with that of a mlodipine.Methods:Blood pressure and heart rate of SHRs were measured with blood press ure meter of rat (SBP-1) by administrating L-amlodipine and amlopdipine through mo uth.Hemodynamic parameter of anaesthetic dogs was determined by administrating drugs through duodenum with Polygraph System (RM6000).Results:L-amlodipine could slowly reduce the blood pressure and heart rates of clearing SHRs,it could also reduce the blood pressure of normal anaesthetic dogs and the action of L-amlodipine was similar to that of amlodipine.Conclusion:L-amlodipine as a new antihypertensive drug can significantly re duce blood pressure.
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AIM: To evaluate the effects of compound salivia miltorrhiza injection on an experimental model of kidney thrombus induced by lipopolysaccharide(LPS).METHODS: The model of microvascular thrombosis in the rabbits' kidney was performed by the method of Hermida,which was induced by infusing LPS.Treatments were begun simultaneously with LPS infusion,through the contralateral marginal ear vein.Six different groups were established: NS 10(ml?h~(-1)) was infused as the negative control group,compound salivia miltorrhiza injection was infused with the dosage of(0.1)(Low-dose),(0.2)(medium-dose),and 0.4(high-dose)(ml?kg~(-1)?h~(-1)),heparin 600,000(IU?kg~(-1)?h~(-1)) as positive control group.The further rabbits, which were given neither LPS nor compound salivia miltorrhiza injection,were infused with saline solution through both marginal ear veins.The measurement of fibrinogen concentrations and platelet counts were used to assess the degradation of microvascular thrombosis.Kinney sections were examined for the presence of fibrin microthrombi.RESULTS: Compound salivia miltorrhiza injection was infused with the dosage of(0.1)(Low-dose),(0.2)(medium-dose),and(0.4)(high-dose)(ml?kg~(-1)?h~(-1)),and the fibrinogen concentrations and blood platelet counts were improved,and the fibrin deposition was degraded.CONCLUSION: Compound salivia miltorrhiza injection can inhibit effectively LPS-induced renal microvascular thrombosis.
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AIM: To improve the acute cumulative death rates method (ACD method) in animal by oral administration. METHODS: A mathematic model was established to evaluate the dose-toxic effect relationship by twice oral administration and the experimental steps were improved too. The methodological quality was explored using the simulation data from computer program and the real experimental date from the reference paper. RESULTS: The results showed that the experimental data could be fitted to its theoretical data from LD_5/2 to LD_ 95/2. Concentration-time curve after po ordinary powder of Semen Strychni in mice were fitted to a one-compartment with T_ 1/2(ka)= 1.136 h,T_ 1/2(ke)= 7.100 h,and T_ max= 3.576 h. CONCLUSION: The improved ACD method can be used in the pharmacokinetics of TCM by oral administration.
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AIM To observe the synergical effects of ginsenoside of stem and leaf(GSL)in combination with choline on learning and memory of Alzheimers disease(AD). MEHODS AD animal models were made by damaging nucleus basalis of Meynert with quinolinic acid. One time training passive aviodance step-down and water-maze spatial localization task were used to observe the ability of learning and memory.RESULTS Treatment with combination of GSL(400 mg?kg -1 ?d -1 ,ig) and choline(200 mg?kg -1 ?d -1 ,ig) on AD rats decreased significantly the number of errors on step down (ig for 13 days) and the training times to reach the criterion on water maze (ig for 16 days). The effect of GSL in combination with choline was more remarkable than that of GSL or choline and proved no obvious difference compaired with that of 1,2,3,4-tetrahydroacridine (10 mg?kg -1 ?d -1 ,ig). Value Q was more than one after administrition of both GSL and choline. CONCLUSION GSL in combination with choline may synergically improve the impairement of learning and memory of AD.