ABSTRACT
OBJECTIVE: To extract microamounts of animal-derived DNA from the products of Colla Corii Asini boiled at high temperature, establish and optimize a rapid identification method of donkey-derived components in Colla Corii Asini by polymerase chain reaction(PCR), and establish a new molecular biological method for assessing the quality of Colla Corii Asini. METHODS: The donkey-derived genomic DNA was extracted by DNA purification column instead of phenol, chloroform and other toxic organic solvents in SDS-PK method, and the SDS-PK method was optimized with the donkey-derived genomic DNA. RESULTS: The optimum sample size of Colla Corii Asini was 0.20 g. High quality genomic DNA of Colla Corii Asini could be obtained quickly after digestion in water bath for 1 h and then purified by DNA purification column. The purity ranged from 1.70 to 1.80, and the concentration of Colla Corii Asini could reach (187.8±0.56)ng•μL-1. PCR amplification, cloning, and sequencing were performed using specific primers, and the similarity to GenBank's registered Donkey species (MG931481.1) was 100%. CONCLUSION: This study provides animal-derived genomic DNA fragments from deep-processed Colla Corii Asini and Colla Corii Asini products within 90 min. The purity and concentration of extracted DNA can meet the requirements of molecular biological identification of Colla Corii Asini. The established PCR method can quickly identify the scorpion-derived components in Colla Corii Asini. The cloned donkey specific gene fragment can be used as a standard positive control to identify the authenticity of Colla Corii Asini. It is expected that it will be widely used in the quality supervision of Colla Corii Asini and related products.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Astragalus membranaceus (APS) on the proliferation, osteogenic capacity and structure of periodontal ligament cells (PDLCs) in vitro.</p><p><b>METHODS</b>PDLCs were cultured in vitro with APS of 0.08, 0.1, 0.2, 0.4 mg x mL(-1). Methyl thiazolyl tetrazolium (MTr), alkaline phosphatase (ALP) and cell structure were detected to determine the proliferation and differentiation of PDLCs proliferation and differentiation.</p><p><b>RESULTS</b>When the APS was 0.2 mg x mL(-1), the absorbance of MTT and ALP exhibit significantly increased as compared to the control (P < 0.05). The cells cultured in vitro with APS of 0.2 mg x mL(-1) had the normal structure.</p><p><b>CONCLUSION</b>APS with proper concentration in short-term culture may promote the proliferation and differentiation of PDLCs.</p>
Subject(s)
Alkaline Phosphatase , Astragalus propinquus , Cell Differentiation , Cell Proliferation , In Vitro Techniques , Periodontal LigamentABSTRACT
<p><b>OBJECTIVE</b>Carboplatin (CBP)-resistant cell line (Tca8113/CBP) and pingyangmycin (PYM)-resistant cell line (Tca8113/PYM) were established in vitro. Ginkgolic acids' influence over multidrug resistance (MDR) of drug-resistant cells was discussed by ginkgolic acids coupled with chemotherapy drugs.</p><p><b>METHODS</b>The expression of P-glycoprotein (P-gp) was detected by immunohistochemistry. MTT assay was applied to ascertain the resistance index of drug-resistant cells. The effect of different concentrations of ginkgolic acids on the proliferation of drug-resistant cells and parental cell was measured by MTT assay. Making sure the non-toxic concentration of ginkgolic acids and observing the reversal effect of ginkgolic acids on drug-resistant cells. Resistance index was redetermined by MTT assay after ginkgolic acids coupled with chemotherapy drugs induced the cell lines for some time.</p><p><b>RESULTS</b>Immunohistochemistry showed that P-gp positive expression rate of drug-resistant cells was significantly higher than parental cells. The non-toxic concentration of ginkgolic acids which was determined by MTT assay was 10 microg x mL(-1). The reversal folds of Tca8113/CBP cell line to CBP and Tca8113/PYM cell line to PYM were 2.94 and 2.43 respectively. Before coupled with ginkgolic acids, the resistance indices of Tca8113/CBP and Tca8113/PYM cell lines were 3.24 and 11.9 respectively. When ginkgolic acids was added with chemotherapy drugs for some time, the resistance indices of Tca8113/CBP and Tca8113/PYM cell lines were 2.18 and 4.43 respectively.</p><p><b>CONCLUSION</b>This experiment successfully induced the drug-resistant cell lines of Tca8113/CBP and Tca8113/PYM. The method of chemotherapy drugs coupled with ginkgolic acids further confirmed the effect on proliferation of Tca8113/CBP and Tca8113/PYM cell lines was reducing. Non-toxic concentration of ginkgolic acids can partially reverse the drug resistance of Tca8113/ CBP and Tca8113/PYM cell lines. Furthermore, MDR level of drug-resistant cells decreased somewhat when they were induced by ginkgolic acids coupled with chemotherapy drugs for some time.</p>
Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Antineoplastic Agents , Bleomycin , Carcinoma, Squamous Cell , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Mouth Neoplasms , SalicylatesABSTRACT
<p><b>OBJECTIVE</b>To study the expression of vascular endothelial growth factor-C (VEGF-C) in oral squamous cell carcinoma (OSCC) and its relation to angiogenesis, lymphangiogenesis, as well as lymph node metastasis.</p><p><b>METHODS</b>Sixty-seven archival specimens from patients with oral squamous cell carcinoma were investigated, whose clinicopathologic data were completely conserved. Immunohistochemical staining was performed to detect the expression of VEGF-C, microvessel density (MVD), lymphatic vessel density (LVD). The correlations between VEGF-C expression and MVD, LVD, as well as other clinicopathological features were measured.</p><p><b>RESULTS</b>Although no correlation between VEGF-C expression and tumor location, histological grade, or gender of the patients was observed (P > 0.05), OSCC patients with more advanced clinical stages and lymph node metastasis were prone to have high expression of VEGF-C (P = 0.015 and P < 0.001, respectively). Cases with high-expression of VEGF-C also showed significantly more often higher LVD (P = 0.001) but not MVD (P = 0.125). In addition, cases with lymph node involvement presented higher LVD than other cases (P = 0.026).</p><p><b>CONCLUSION</b>VEGF-C may promote lymph node metastasis by inducing lymphangiogenesis in OSCC.</p>