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ObjectiveTo establish a rapid method for evaluating the heterozygosity of Murraya paniculata germplasm materials and provide as a foundation for developing germplasm breeding and innovation measures for M. paniculata. MethodSingle nucleotide polymorphisms （SNPs） were screened from the genome resequencing data of 65 plants of M. paniculata. A self-written script was used to transform 20 SNPs into restriction fragment length polymorphism （RFLP） markers. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was employed to detect the 20 RFLP markers in 12 M. paniculata germplasm accessions， and the heterozygosity of M. paniculata germplasm accessions was calculated based on the number of enzyme-cutting bands at the 20 RFLP marker sites. Plink was used to calculate the whole genome heterozygosity of 12 M. paniculata germplasm accessions， and the results obtained with different methods were compared. ResultThere was no significant difference in the heterozygosity calculated by the PCR-RFLP method and the genome resequencing method. The PCR-RFLP and genome resequencing methods identified 8 and 9 germplasm accessions， respectively， with a heterozygosity level less than 30%. Seven germplasm accessions with heterozygosity less than 30.00% were calculated by both methods. ConclusionThe PCR-RFLP method established in this study for evaluating the heterozygosity of M. paniculata germplasm demonstrates the precision of 87.5% and the accuracy of 77.8%. This method serves as a reference for developing heterozygosity evaluation methods in other medicinal plant germplasm resources.
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Objective:To study and analyze the uncertainty of active breathing coordinator (ABC) technology for liver and lung cancer therapy using proton and heavy ion.Methods:Before each treatment, each patient received a verification radiograph through the supporting imaging frame in treatment room. 200 verification radiographs were taken for 20 lung cancer patients and 200 for 20 liver cancer patients. Ipiodol markers, which were fixed relative to the location of the tumor, were injected into the liver cancer patients. The position changes of ipiodol markers could reflect the position changes of liver tumors. Verification radiographs were registered with the vertebral body as the main target, and the change value of tumor location was recorded.Results:For liver cancer cases, the values of position change in the left and right, head and foot, and dorsal abdomendirection were (-0.05± 0.28) cm, (0.15±0.33) cm, (-0.12±0.27) cm, and (-0.03±0.13) cm, (-0.05±0.14) cm and (0.02±0.16) cmfor lung cancer cases, respectively ( P=0.280, <0.001, <0.001). For liver cancer cases, the dispersionin the left and right, head and foot, and dorsal abdomendirectionwas (0.20±0.09) cm, (0.25±0.06) cm, (0.19±0.09) cm, and (0.09±0.03) cm, (0.10±0.03) cm and (0.13±0.03) cm for lung cancer cases, respectively ( P<0.001, <0.001, 0.008). The proportion of tumor location changes of≤5 mm in three directions in liver and lung cancer patientswas (92%, 83%, 93%) vs. (99%, 99%, 100%)( P=0.030, 0.002, 0.007). Conclusion:The application of ABC technology in the proton heavy ion therapy of lung and liver cancer has good reproducibility, and the stability of ABC technology in the treatment of lung cancer is better than that of liver cancer.
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ObjectiveTo establish a polymerase chain reaction（PCR） method to accurately discriminate the crude materials of Murrayae Folium et Cacumen， Murraya exotica and M. paniculata. MethodBased on the difference in chloroplast genome sequences of M. exotica and M. paniculata， species-specific identification primers P03 and P04 of M. exotica and M. paniculata were designed according to single nucleotide polymorphism （SNP） on the chloroplast genome. A multiplex allele-specific PCR identification method was established for the identification of M. exotica and M. paniculata following the optimization of annealing temperature， number of cycles， and primer concentration ratio. The established PCR method for identification was explored and verified in terms of tolerance and feasibility by investigating the type of Taq polymerases and PCR system model. ResultIn this multiplex allele-specific PCR identification method， about 330 and 230 bp of specific fragments were amplified from DNA templates of M. exotica and M. paniculata， respectively， under the following conditions：cycle number of 31， annealing temperature of 60 ℃， and primer concentration ratio of P03 and P04 of 1∶2. Consistent results were obtained for samples from different sources. ConclusionThe multiplex allele-specific PCR identification method established in this study can accurately identify the origin of Murrayae Folium et Cacumen， which can be used for the simultaneous identification of M. exotica and M. paniculata by the length of fragments in a single identification assay.
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AIM To compare liver-protective and anti-inflammatory effects of extracts from root,stem,leaf,and flower of Gentiana rigescens.METHODS The mouse model for the immunological liver injury induced by Concanavalin A,and mouse ear swelling model for inflammation caused by dimethylbenzene were used for the comparison of the liver-protective or anti-inflammatory effects of four parts individually.RESULTS Four aqueous extracts of Gentiana rigescens showed the dose-dependent decrease in the activity of ALT and AST in serum and liver index,and alleviation of hepatic tissue injury induced by Concanavalin A in mice.The effects of the extracts from the leaf and root were better than those from the stem and flower.These extracts presented dose-dependent inhibition against the ear swelling caused by dimethylbenzene in mice.The effects of the extracts from the leaf and stem were better than those from the flower and root.CONCLUSION Extracts from the root and leaf of G.rigescens have liver-protective effect,and parts from the stem and leaf have anti-inflammatory effect.