1.A study on the thethymidylate synthase gene polymorphisms and its relationship with clinical characteristics in breast cancer
Rencheng SUN ; Keli TIAN ; Zhongbing MA ; Qinye FU ; Qiang ZHANG ; Dezong GAO ; Yuyang LI ; Liang LI ; Zhigang YU
Chinese Journal of General Surgery 2010;25(12):973-976
ObjectiveTo investigate the distribution of thethymidylate synthase (TS) gene polymorphisms inChinese femaleHan-nationality breast cancer patients,including tandemrepeat polymorphisms in the enhancer region ( ER), 6 bp deletion/insertion (del6/ins6) polymorphism in the 3′untranslated region (3′-UTR) and G→C single-nucleotide polymorphism (SNP). The association of TS gene polymorphism with pathological results was retrospectively analyzed.MethodsThis study included 83 breast cancer patients who received no preoperative chemotherapy. The polymorphisms of TS, ER and TS 3′2UTR were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method.Results ( 1 ) Of the 83 patients, the frequencies of the TSER 3R/3 R, 2R/3 R and 2R/2R were 68.7% ,27. 7% and 3. 6%, respectively; (2) Due to existence of the G→C single nucleotide polymorphism,TSER genotypes were divided into 2R/2R, 2R/3C, 2R/3G, 3G/3G,3C/3C and 3G/3C,their frequencies were 3. 6%, 19. 3%, 8.4%, 19. 3%, 37. 3% and 12. 1%, respectively; ( 3 ) Of the 83 patients, the frequencies of the TS 3′-UTR ins6/ias6、ins6/del6 and del6/del6 were 8.4% 、50.6% and 41.0%, respectively; (4) TSER polymorphisms were correlated with lymph node metastasis ( P = 0.019 ),Ki67 ( P = 0.022 ) and histological grade ( P = 0.042 ). The TS 3′-UTR polymorphisms were correlated with age ( P = 0.002) and histological grade (P = 0.042). G→C SNP were correlated with lymph node metastasis ( P = 0.021 ) and histological grade ( P = 0.028 ).ConclusionsChinese female Han-nationality breast cancer patients have more genotypes frequencies of the TSER 3R/3R and 3′-UTR del6/del6; The TS polymorphisms are correlated with pathological factors of breast cancer.
2.Latest research progress in airway stenosis after lung transplantation
Yujie ZUO ; Menggen LIU ; Jiaxin WAN ; Yuxuan CHEN ; Wenlong HU ; Junjie ZHANG ; Yuyang MAO ; Jing CHEN ; Ailing ZHONG ; Lingzhi SHI ; Bo WU ; Chunrong JU ; Dong TIAN
Organ Transplantation 2024;15(3):474-478
With the optimization of surgical technologies and postoperative management regimens, the number of lung transplantation has been significantly increased, which has become an important treatment for patients with end-stage lung disease. However, due to the impact of comprehensive factors, such as bronchial ischemia and immunosuppression, the incidence of airway stenosis after lung transplantation is relatively high, which severely affects postoperative survival and quality of life of lung transplant recipients. In recent years, with the improvement of perioperative management, organ preservation and surgical technologies, the incidence of airway stenosis after lung transplantation has been declined, but it remains at a high level. Early diagnosis and timely intervention play a significant role in enhancing clinical prognosis of patients with airway stenosis. In this article, the general conditions, diagnosis, treatment and prevention of airway stenosis after lung transplantation were reviewed, aiming to provide reference for comprehensive management of airway stenosis after lung transplantation and improving clinical prognosis of lung transplant recipients.
3.Research progress on immunomodulatory properties of periodontal ligament stem cells
WEN Wen ; TIAN Yuyang ; XIE Xudong
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(1):76-80
Periodontal ligament stem cells (PDLSCs) have the potential for multidirectional differentiation and are the preferred seed cells for periodontal tissue regeneration. In recent years, a large number of studies have confirmed that PDLSCs also possess broad immunomodulatory properties. Therefore, in-depth exploration of their specific molecular mechanisms is of great significance for the treatment of periodontitis. The aim of this paper is to summarize the research progress on the regulation of PDLSCs on various immune cells and the effect of the inflammatory environment on the immune characteristics of PDLSCs to provide an important theoretical basis for the allotransplantation of PDLSCs and improve the therapeutic effect of periodontal tissue regeneration. Studies have shown that PDLSCs possess a certain degree of immunosuppressive effect on both innate and acquired immune cells, and inflammatory stimulation may lead to the impairment of the immunoregulatory properties of PDLSCs. However, current studies are mainly limited to in vitro cell tests and lack in-depth studies on the immunomodulatory effects of PDLSCs in vivo. In vivo studies based on cell lineage tracing and conditional gene knockout technology may become the main directions for future research.
4.Enzyme-linked Immunosorbent Assay in Quality Control of Chinese Medicines: A Review
Yuansheng XU ; Jiao LIU ; Tian ZHANG ; Yuyang ZHAO ; Hui TIAN ; Tiegui NAN ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):12-20
In the quality control of Chinese medicine, the detection of active components and toxic and harmful components are two important links. Although conventional methods such as high performance liquid chromatography and liquid chromatography-mass spectrometry can accurately quantify the above substances, they have shortcomings such as complicated operation, high costs, inability of detection at any time, difficult detection of insoluble and macromolecular substances. Enzyme-linked immunosorbent assay (ELISA) can adsorb antigens or antibodies on the surface of solid carriers and realize qualitative or quantitative analysis of targets by using the specific reactions of antigens and antibodies. This method is praised for the simple operation, high sensitivity, strong specificity, simple requirements for experimental equipment, a wide application range, and low costs. In recent years, ELISA has been widely used in the quality control of Chinese medicine, especially in the content determination of mycotoxins represented by aflatoxin and the qualitative and quantitative analysis of active components. ELISA plays an increasingly important role with its unique advantages, providing new methods and ideas for the rapid quality examination of large quantities of Chinese medicines. This paper reviews the research progress in ELISA for the quality control of Chinese medicine in recent years and prospects its technical development and application prospects, aiming to provide reference and research ideas for further using this method to ensure the quality, safety, and controllability of Chinese medicine.
5.Identification of Scolopendra Dispensing Granules by Allele-specific PCR
Yuansheng XU ; Li HU ; Chao JIANG ; Yuyang ZHAO ; Tianyun CHEN ; Hui ZHANG ; Hui TIAN ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):48-54
ObjectiveTo establish an allele-specific polymerase chain reaction (PCR) method for identifying Scolopendra dispensing granules, so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3(COX-3) gene sequence of Scolopendra, and the single nucleotide polymorphism (SNP) loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature, cycles, Taq enzymes, DNA template amount, PCR instruments, and primer concentrations, and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix, 0.4 μL forward primer (10 μmol·L-1), 0.4 μL reverse primer (10 μmol·L-1), 2.5 μL DNA template, and 9.2 μL sterile double distilled water. PCR parameters: Pre-denaturation at 94 ℃ for 3 min, 30 cycles (94 ℃ for 20 s, 62 ℃ for 20 s, 72 ℃ for 45 s), and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above, the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials, decoction pieces, and standard decoction freeze-dried powder of Scolopendra, as well as the intermediates and final products of Scolopendra dispensing granules, which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.
6.Non-targeted Metabolomics Analysis of Fuling Yunhua Granules in Treatment of Type 2 Diabetes Mellitus Rats
Mengyao TIAN ; Keke LUO ; Mengxiao WANG ; Tianbao HU ; Hongmei LI ; Zongyuan HE ; Lixin YANG ; Liyu HAO ; Nan SI ; Yuyang LIU ; Baolin BIAN ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):195-204
ObjectiveBased on non-targeted metabolomics, to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus(T2DM) rats by Fuling Yunhua granules, and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats, half male and half female, were randomly divided into the control group, model group, and high, medium, low dose groups of Fuling Yunhua granules(20.70, 10.35, 5.18 g·kg-1 in raw drug amount) and the positive drug group(pioglitazone hydrochloride tablets, 8.1 mg·kg-1). Except for the control group, other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ) to establish a T2DM rat model. After successful modeling, the treatment groups were administered the corresponding drugs by gavage, and the control group and model group were treated with an equal volume of saline by gavage, once/d, for 28 d. Fasting blood glucose(FBG) and glycosylated hemoglobin A1c(GHbA1c) levels were measured in all groups of rats during the administration period, and hematoxylin-eosin(HE) staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-LTQ-Orbitrap MS), and the data were processed using principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Differential metabolites were identified by the Human Metabolome Database(HMDB) and the Kyoto Encyclopedia of Genes and Genomes(KEGG), and screened for differential metabolites with variable importance in the projection(VIP) value>1, P<0.05, and fold change(FC)<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0, then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic(ROC) curves. ResultCompared with the control group, the FBG level of rats in the model group increased significantly(P<0.01), the GHbA1c content tended to increase, but the difference was not statistically significant, and the pancreatic tissue of rats was obviously damaged, the number of pancreatic islets decreased, and the pancreatic β-cells were obviously reduced, atrophied and enlarged. Compared with the model group, the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration(P<0.05, P<0.01), the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced(P<0.05), and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid, D-amino acid metabolism, steroid hormone biosynthesis, and glycerophospholipid metabolism in rats. Compared with the model group, the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted, and the pathway enrichment analysis found that D-amino acid metabolism, retinol metabolism, glycine, serine and threonine metabolism, tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol, D-piperidinic acid, D-serine, and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.