Establishment of a rapid method for quality assessment of cardiotonic effect of Shengfupian / 药学学报

Based on the similar structure of adrenaline shared by higenamine (HI), salsolinol (SA) and coryneine (CO), a photochemical colorimetric sensor based on the displacement reaction of o-diphenol hydroxyl group and alizarin red S-phenylboric acid system was constructed to quickly distinguish and identify the cardiac strength of Shengfupian. The results show that the optimal condition of the sensor is: the molar ratio of alizarin red S (ARS) to phenylboric acid (PA) is 1∶3, reaction temperature is 0 ℃; The preparation method of the sample solution is optimized as follows: 2.5 g of Shengfupian powder was taken, 10 times the amount of methanol was added, and 300 W, 40 kHz ultrasound was carried out for 15 min; methodological studies showed that the method had good precision, repeatability and stability. The |△G| value (G is green, |△G| = |G after - G before|) of each sample was obtained by response values determination of 14 batches of Shengfupian. LC-MS/MS was used to determine the contents of three cardiac components in Shengfupian. It was found that the order of the total contents of cardiotonic components was basically consistent with |△G|. Then the correlation was analyzed, and the correlation coefficient R2 was as high as 0.87, which proved the scientificity and accuracy of this method. This study fills the methodological gap of rapid evaluation of the quality of Shengfupian, and provides the key technical support for the high quality and good price of Shengfupian in the market circulation and clinical application.

Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells

BACKGROUND AND OBJECTIVES: The imperative role of dental pulp stem cells (DPSCs) in regenerative therapy demands an in-vitro expansion which must deal with the safety and ethical problems associated with fetal bovine serum (FBS). The primary aim of this study was to compare the effects of human platelet rich fibrin (hPRF) exudate Vs FBS on proliferation and osteodifferentiation of human dental pulp stem cells (hDPSCs). The secondary one was to determine the optimum concentration of hPRF exudate inducing hDPSCs proliferation and osteodifferentiation. METHODS: The direct method was used to prepare hPRF exudate. hDPSCs were isolated from impacted mandibular third molars of twelve donors by the outgrowth method. For cell viability and proliferation rate testing, 96 well plates were used and the assay was done in duplicate and the trial repeated four times under the same conditions. Six wells were used to contain 10% FBS, serum free media, 1%, 5%, 10% and 20% concentrations of hPRF exudates, respectively. The proliferation assay was carried out by MTS tetrazolium cell proliferation assay kit and Elisa reader. The study design for osteodifferentiation protocol was exactly as the proliferation one and instead the assay was carried out by alizarin red with Elisa reader. RESULTS: Compared to 10% FBS, 10% hPRF exudate was the optimum concentration for hDPSCs proliferation, while 1% hPRF exudate was the optimum concentration for osteodifferentiation of hDPSCs. CONCLUSIONS: Avoiding the risk of zoonosis which may be occurred with FBS, it is recommended to use 10% hPRF exudate for proliferation and 1% for osteodifferentiation.

Corneal Endothelial Damage after Deep Excimer Laser Ablation

To investigate the effect of deep excimer laser ablation on the corneal endothelium, excimer laser photo refractive keratectomy(PRK) with different ablation depths were performed to obtain various residual corneal thickness(range: 90-250 micrometer) in white rabbit corneas(N=50). Corneal endothelium was stained with Alizarin red S(pH 4.2) for 2 minutes three days after excimer laser ablation and was analyzed morphometrically using digitizer the morphometric parameters according to residual corneal thickness. In the corneas of residual corneal thickness(RT) over 200 micrometer and untreated controls(N=6), endothelial damages were rarely seen. With the decrease of residual corneal thickness, hexagonality and shape factor decreased, and coefficient of variation of the cell area(CV) increased(p<0.01). In corneas of RT less than 200 micrometer, endothelial damages were found and become more severe in corneas of RT between 90~149 micrometer. Hexagonal shaped cells were rarely observed, and the shapes of most cells were changed. Deep excimer laser PRK might affect corneal endothelial cells if the RT is less than 200 micrometer, and these findings suggest that care is recommended when doing deep excimer laser corneal ablation, especially excimer laser assisted in situ keratomileusis.

Subepidermal Calcified Nodule

Subepidermal calcified nodule(SCN) is a form of calcinosis cutis which is usually present as a single small, raised, hard nodule with verrucous surface. A 13-year-old girl presented with a 10-month history of a ricegrain-sized, hard, yellowish white papule on both upper eyelids. The lesions were asymptomatic and had increased slowly in size. An excisional biopsy specimen of the left upper eyelid showed acanthosis and narrow pointed rete ridges of the epidermis, and closely aggregated deposition of basophilic material in the uppermost dermis. The material in the dermis did not stain with von Kossa. It was confirmed as calcium deposition by staining with alizarin red S which is far more specific for calcium than the von Kossa stain.

Morphometric Analysis of the Rabbit Corneal Endothelium Specular Microscopy Versus Alizarin Red S Staining

Analysis of endothelial morphology by computer assisted digitizer and image analysis program provides very useful indices of cell shape and size which appear to correlate to the monolayer's functional status. To study the influences of the Alizarin red S staining, which is commonly used in animal experiments of corneal endothelium and vital staining, on the morphologic characteristics of corneal endothelium, morphometric data(density, area, coefficient of variation, perimeter, shape factor, hexagonality, lengths) obtained by specular microscopy of the endothelium are compared to data obtained by Alizarin red S staining of the endothelium of the excised cornea. Mean endothelial cell area was measured as 389.58 +/- 37.14 micrometer2, density was 2588 +/- 251 cells/mm2. The corresponding values measured after Alizarin red S staining, cell area was 407.42 +/- 45.3 micrometer2 and density was 2484 +/- 294 cells/mm2. But no significant differences were noted in comparing all morphometric data obtained by staining to that obtained from specular microscopy. Therefore, Alizarin red S staining combined with cell morphometric analysis could provide valuable data in a cornea which lacks clarity limits or precludes specular microscopy.

COMPOUND STAINING METHOD SHOWING OSSIFIED SKELETAL STRUCTURES AND CARTILAGE IN FETAL, NEONATAL AND ADULT RAT / 解剖学报

Objective To compare compound staining of Alizarin red S and Alcian blue showing ossified skeletal structures and cartilage in fetal,neonatal and adult rat. Methods All specimens were skinned and fat tissue removed completely and eviscerated,then fixed in 95% ethanol;Acetone was used remove fats; 0.1% Alizarin red S and 0.15%-0.3% Alcian blue were used in ossified bone and cartilage;Differentiation and clearing in 0.5%-2% aqueous KOH. Results Ossified bones showed rose red;cartilage showed blue;the others were transparent.Conclusion The best results of Alizarin red S and Alician blue compound staining could be obtained in all rat skinned,eviscerated,and removed the adipose tissues.