1.Genotyping of Clinical Isolates of Malassezia from Patients with Tinea Versicolor by PCR-RFLP
Fan CUI ; Shiqin TAO ; Yongnian SHEN ; Guixia LU ; Wei CHEN ; Xiaofang LI ; Suquan HU ; Lijia YANG ; Weida LIU
Chinese Journal of Dermatology 2003;0(08):-
Objectives To develop a rapid genotyping method of clinical isolates of Malassezia from patients with tinea versicolor by PCR-RFLP,and to evaluate reliability of the approach as compared with biochemical classification.Methods Tween assimilation test and catalase reaction were carried out to identify 74 isolates of Malassezia species from patients with tinea versicolor and 7 Malassezia reference strains.The sequence of 28S rDNA of Malassezia species was amplified by PCR,and then the product was analyzed by RFLP with Eco88I,Bsp143Ⅱ and BshNⅠ,respectively.Results M.restricta,M.obtusa and M.pachydermatis were successfully identified by three restriction endonucleases.M.restricta was found to be more diverse from the other 6 species in genetic homology.By comparison with PCR-RFLP technique,a possible mistake was discovered with biochemical method.Conclusion PCR-RFLP is a promising molecular biological technique,which could rapidly and correctly classify Malassezia species.
2. Four-dimensional flow MRI quantification of pulmonary regurgitation in patients with repaired tetralogy of Fallot: a study on reproducibility and consistency
Shiqin YU ; Minjie LU ; Gang YIN ; Xinling YANG ; Chen CUI ; Xiuyu CHEN ; Shihua ZHAO
Chinese Journal of Radiology 2019;53(9):761-766
Objective:
To investigate the feasibility and consistency of four-dimensional flow (4D flow) quantification of pulmonary regurgitation in patients with repaired Tetralogy of Fallot (ToF) by comparing with conventional two-dimensional flow (2D flow) and echocardiography.
Methods:
Both the 4D flow and 2D flow imaging were acquired with repaired ToF (a total of 21 patients) consecutively on 3.0 T MR scanner from May 2018 to August 2018. Pulmonary flow and regurgitant fraction were measured by a commercial post processing software Circle CVI42. All patients underwent echocardiography within one week after or before MR examination. The inter/intra-observer variability by 2D/4D f1ow and agreement between the two methods were investigated by interclass correlation coefficients (ICC) and Bland-Altman analyses. The agreement between MR and echocardiography were analyzed by weighted Kappa coefficient. The correlation between pulmonary regurgitation and cardiac function was also investigated by Pearson analysis.
Results:
All patients were included and completed the examinations successfully. Both inter-observer and intra-observer agreement by 4D flow for total forward volume (ICC=0.993, 0.996, respectively,
3.Study on Protective Effect and Its Mechanism of Thymoquinone on Myocardial Fibrosis Induced by Lipopolysaccharide in Rats
Wei CAI ; Jiadan YANG ; Shan GAO ; Shiqin LI ; Shurong WANG
China Pharmacy 2019;30(4):464-469
OBJECTIVE: To investigate the protective effect and its mechanism of thymoquinone (TQ) on myocardial fibrosis (MF) induced by lipopolysaccharide in rats. METHODS: Totally 40 male Wistar rats were randomly divided into control group, model group, TQ low-dose and high-dose groups [5, 10 mg/(kg?d)], with 10 rats in each group. Except that control group was given constant volume of normal saline intraperitoneally, other groups were given LPS intraperitoneally to induce MF model, once a day, for consecutive 3 weeks. Since the first day of modeling, administration group was given relevant medicine intraperitoneally, once a day, for consecutive 3 weeks. After last medication, ELISA method was used to detect the contents of serum inflammation factors (IL-1β, IL-6, TNF-α) in rats. Cardiac mass parameters (HW/BM, LVW/BW) were weighed and calculated. HE staining was used to observe the histopathological changes of myocardial tissue. The contents or activities of oxidative stress indicators (MDA, SOD) and myocardial collagen indexes (HYP, Col-Ⅰ, Col-Ⅲ) were detected by chemical analysis, xanthine oxidase method or ELISA. mRNA expression of regulation genes (Col-Ⅰ, Col-Ⅲ, MMP-3, MMP-9, TGF-β1 and Smad3) related to myocardial fibrosis were determined by real-time fluorescence quantitative PCR. RESULTS: Compared with control group, there were swollen myocardial cells, disordered nuclei of different sizes and visible fiber hyperplasia in model group; the levels of serum inflammatory factors and LVW/BW, cardiac contents of MDA, HYP, Col-Ⅰand Col-Ⅲ, mRNA expression of Col-Ⅲ, TGF-β1 and Smad3 in model group were increased significantly (P<0.05), while SOD activity was decreased significantly (P<0.05). Compared with model group, the degree of myocardial fibrosis was improved in TQ groups to different extents; serum content of IL-1β and the contents of MDA, HYP and Col-Ⅰ in cardiac tissue in TQ low-dose group, serum contents of IL-1β, IL-6 and TNF-α, LVW/BW and the contents of MDA, HYP, Col-Ⅰ and Col-Ⅲ in cardiac tissue of TQ high-dose group as well as mRNA expression of Col-Ⅰ, Col -Ⅲ, TGF-β1 in TQ groups were decreased significantly (P<0.05). The activities of SOD in cardiac tissue were increased significantly in TQ groups (P<0.05). There was no statistical significance in other indexes among groups (P>0.05). CONCLUSIONS: TQ can protect against MF model rats to certain extent, the mechanism of which may be associated with the inhibition of inflammation reaction and oxidant stress reaction, and down-regulation of mRNA expression of Col-Ⅰ, Col-Ⅲ and TGF-β1.
4.Clinical and cardiac MR characteristics of heart involvement in patients with Fabry disease
Yangfei XU ; Kai YANG ; Xiaofeng LIU ; Xiuyu CHEN ; Yanyan SONG ; Yihui WANG ; Yucong ZHENG ; Shiqin YU ; Shujuan YANG ; Jiaxin WANG ; Zhixiang DONG ; Minjie LU ; Shihua ZHAO
Chinese Journal of Radiology 2022;56(2):168-174
Objective:To investigate the clinical and cardiac magnetic resonance (CMR) characteristics of heart involvement in patients with Fabry disease (AFD).Methods:From January 2018 to March 2021, eight AFD patients [3 males and 5 females, mean age (50±11) years old, range 26-60 years old] confirmed by genetic testing or pathology in Fuwai Hospital were retrospectively included in this study. At the same time, sixteen patients with hypertrophic cardiomyopathy (HCM) [6 males and 10 females, mean age (46±15) years old] and 16 healthy individuals [6 males and 10 females, mean age (51±11) years old] were included as controls. The clinical baseline data and CMR data of the patients were collected and analyzed. The CMR data were analyzed using the software CVI42, with the corresponding parameters automatically generated. One-way ANOVA or Kruskal-Wallis test was used to compare the differences in the parameters among the three groups. Independent-samples t test, Fisher precise test or Mann-Whitney U test were used for the comparison between each two groups. Results:Statistically significant difference was found in renal insufficiency between the HCM group and the AFD group; No other significant difference was found in other clinical factors and ECG results (all P>0.05). CMR results showed that in the AFD group, there were 5 cases with symmetric or roughly symmetric hypertrophy, and 3 with asymmetric hypertrophy. The late gadolinium enhancement (LGE) showed myocardial enhancement in 5 patients, mainly presenting as multiple intermural enhancement, and partially as local subendocardial enhancement. In the HCM group, fourteen cases suffered mainly asymmetric ventricular septal thickening, with or without thickening of other parts of left ventricular wall; and 2 cases had thickening of middle and distal part of the left ventricle. The LGE showed myocardial enhancement in 14 patients, which manifested as focal or patchy enhancement in hypertrophic myocardium, including focal enhancement in the right ventricular insertion of ventricular septum (more common) and subendocardial enhancement in the middle and far segments of left ventricle. Statistically significant difference was found in the differences between the left atrial anterior posterior diameter, the maximum wall thickness of left ventricular, the left ventricular myocardial mass index (LVMI) and the native T 1 value among the three groups (all P<0.001). However, there was no statistically significant difference in the left atrial anterior posterior diameter and the maximum wall thickness of left ventricular between AFD group and HCM group ( P>0.05). The LVMI in AFD group was higher than that in healthy group and HCM group (all P<0.05). Significant difference was found in the native T 1 value among the three groups, with the native T 1 value of the AFD group [(1 177.4±46.0) ms] was significantly lower than that of the healthy group [(1 244.5±34.3) ms] and the HCM group [(1 278.8±41.6) ms], with ( F=13.10, P<0.001). Conclusions:The clinical characteristics of AFD and HCM are quite similar. When AFD is suspected, CMR imaging should be the first choice for imaging examination. Especially, T 1 mapping imaging can provide important information for the diagnosis of AFD.
5. Research progress of lactate dehydrogenase A in digestive system tumors and related drugs
Siyu WANG ; Jiawei LI ; Chenghao LI ; Ling LI ; Qingyang GUO ; Lu QIU ; Shiqin ZHOU ; Yongqi LIU ; Jiawei LI ; Ling LI ; Yongqi LIU ; Jiawei LI
Chinese Journal of Clinical Pharmacology and Therapeutics 2023;28(4):445-454
Malignant tumors of digestive system are highly prevalent malignant tumors that seriously threaten human health around the world. At present, the curative efficacy and prognosis of traditional treatment methods cannot reach the expectation, so it is urgent to find new targets for cancer treatment and realize targeted therapy for tumors. Abnormal energy metabolism in tumor cells is regarded as a hallmark of cancer, and malignant tumor cells absorb glucose through aerobic glycolysis pathway, and obtain a small amount of energy and produce lactate under the catalysis of a series of enzymes. Lactate dehydrogenase A (Lactate dehydrogenase A, LDHA), as a key enzyme in the aerobic glycolysis pathway of tumor cells, plays an important role in the metabolic changes of tumor cells. Studies have demonstrated that LDHA has high expression characteristics in a variety of tumor cells,and its high expression in clinic is often related to the poor prognosis and high metastasis rate of tumors, which is expected to be a new target for cancer therapy. This article reviews the role of LDHA in the development of digestive system tumors and the research progress of related drugs.
6.Improvement effects of Runchang granules on the constipation in mice and its mechanism
Mengqin HUANG ; Xuesong WANG ; Yuhan GAN ; Shiqin LU ; Qiqi DENG ; Qing ZHU ; Jiao GUO
China Pharmacy 2024;35(2):160-165
OBJECTIVE To investigate the improvement effects of Runchang granules on the constipation in mice and its potential mechanism. METHODS The mice were randomly divided into normal control group, model group, Runchang granules low-dose and high-dose groups (5, 10 g/kg), mosapride group (0.003 g/kg, positive control), with 6 mice in each group. The latter 4 groups were given loperamide intragastrically (0.004 g/kg), twice a day, for 3 consecutive days. Normal control group and model group were given purified water intragastrically, and administration groups were given relevant medicine intragastrically for 7 consecutive days. After the last medication, fecal moisture content and intestinal motility of mice were determined, while the structures of colon and ileum, and the secretion of colonic mucus were observed. Protein expressions of tyrosine kinase receptor (c-kit), mucin 2 (MUC2) and stem cell factor (SCF) were determined in colon; meanwhile, the mRNA expression levels of inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, inducible nitric oxide synthase (iNOS)] as well as factors related to promoting intestinal motility [neuronal nitric oxide synthase (nNOS), smooth muscle myosin light chain kinase (smMLCK), 5-hydroxytryptamine 4 receptor (5-HT4R), MUC2, SCF, c-kit] were determined. RESULTS Compared with model group, the fecal water content, intestinal propulsion rate, protein expression of c-kit in colon, relative expressions of MUC2 and SCF protein, and mRNA expressions of factors related to promoting intestinal motility (except for nNOS and SCF in Runchang granules low-dose group) were all increased significantly in Runchang granules low-dose and high-dose groups, and mosapride group (P<0.05 or P<0.01). mRNA expression levels of inflammatory factors were decreased significantly(P<0.05 or P<0.01). Both colon and ileum injuries improved, and the secretion of colon mucus was increased significantly in Runchang granules high-dose group (P<0.01). CONCLUSIONS Runchang granules have laxative effect and can improve constipation in mice, and its mechanism may be related to the promotion of the secretion of colon mucus and MUC2 expression, and the activation of SCF/c-kit signaling pathway.