ABSTRACT
Objective:To determine lysophosphatidic acid receptor 6 (LPAR6) expression in patients with mycosis fungoides (MF) , a variant of cutaneous T-cell lymphoma (CTCL) , and to investigate its role and mechanism of action in the development and prognosis of CTCL.Methods:A total of 110 patients with confirmed MF were collected from Department of Dermatology, Peking University First Hospital from 2011 to 2020, including 24 with large-cell transformation (LCT) and 25 with non-large cell transformation (NLCT) in the discovery cohort, and 24 with LCT and 37 with NLCT in the validation cohort. RNA sequencing and RT-PCR were conducted to determine the LPAR6 expression in patients in the discovery cohort and validation cohort respectively. LPAR6 expression was compared between patients with LCT and those with NLCT, and its effect on the prognosis of patients was evaluated. Two LPAR6-overexpressing CTCL cell lines MyLa and Sz4 were constructed to evaluate the effect of LPAR6 overexpression on proliferative activity of MyLa and Sz4 cells, with the cells normally expressing LPAR6 as the control group; after the treatment with LPAR6-related ligand lysophosphatidic acid (LPA) , 2S-OMPT, adenosine triphosphate (ATP) or adenosine (ADO) , the effects of LPAR6 activation on the proliferative activity and apoptosis of LPAR6-overexpressing MyLa and Sz4 cells were evaluated by the MTS method and flow cytometry respectively. Log-rank test was used for prognostic analysis, and t test or Mann-Whitney U test was used for comparisons between two groups. Results:As RNA sequencing showed, LPAR6 was one of the significantly underexpressed genes in the LCT group in the discovery cohort; in the validation cohort, LPAR6 expression (median[ Q1, Q3]) was significantly lower in the LCT group (204.90[81.90, 512.70]) than in the NLCT group (809.40[417.50, 1 829.20], U= 242.00, P= 0.002) ; in the two cohorts, the underexpression of LPAR6 was significantly associated with increased risk of poor prognosis (both P < 0.01) . Cell proliferation assay showed no significant difference in the proliferative activity of MyLa or Sz4 cells between the LPAR6 overexpression group and control group at 0, 24, 48 and 72 hours during the experiment (all P > 0.05) ; 48 hours after activation of LPAR6 by LPA, 2S-OMPT, ATP and ADO in MyLa cells, the LPAR6 overexpression group showed significantly decreased cellular proliferative activity (1.38 ± 0.01, 1.04 ± 0.01, 1.09 ± 0.03, 1.23 ± 0.01, respectively) compared the control group (1.73 ± 0.04, 1.23 ± 0.01, 1.24 ± 0.01, 1.42 ± 0.03, t= 30.33, 18.38, 4.78, 5.75, respectively, all P < 0.05) , but significantly increased cell apoptosis rate (17.93% ± 0.88%, 17.75% ± 0.35%, 23.97% ± 0.57%, 31.44% ± 0.34%, respectively) compared the control group (3.98% ± 0.03%, 7.81% ± 0.59%, 11.95% ± 0.85%, 12.02% ± 0.48%, t= 15.93, 14.49, 11.74, 33.01, respectively, all P < 0.05) ; 48 hours after activation of LPAR6 by 2S-OMPT and ADO in Sz4 cells, compared with the control group, the LPAR6 overexpression group also showed significantly decreased cellular proliferative activity (2S-OMPT: 1.29 ± 0.04 vs. 1.48 ± 0.01; ADO: 1.27 ± 0.01 vs. 1.51 ± 0.02; both P < 0.05) , but significantly increased cell apoptosis rate (2S-OMPT: 41.70% ± 0.70% vs. 29.35% ± 0.55%; ADO: 37.05% ± 0.15% vs. 24.60% ± 1.00%; both P < 0.05) . Conclusions:LPAR6 was underexpressed in the patients with LCT, and its underexpression was significantly associated with increased risk of poor prognosis. In vitro activation of LPAR6 could inhibit the proliferation of CTCL cells and promote their apoptosis, suggesting that the decrease of LPAR6 expression may be one of the important mechanisms underlying disease progression in patients with LCT.
ABSTRACT
Objective:To investigate molecules involved in the occurrence of pruritus in patients with mycosis fungoides (MF) .Methods:Totally, 522 patients with MF were enrolled from Peking University First Hospital from October 2009 to August 2021, and the incidence of pruritus was calculated. The patients were grouped according to whether they suffered from pruritus or not. RNA sequencing data on biopsied skin lesions of 49 patients were analyzed to identify differentially expressed genes between patients with pruritus and those without; enzyme-linked immunosorbent assay and immunohistochemical techniques were performed to determine the protein expression of CC chemokine ligand 17 (CCL17) in serum samples from 88 MF patients, and in tissue samples from 81 MF patients, respectively; flow cytometry was conducted to detect markers for T lymphocyte activation and differentiation in peripheral blood samples from 46 MF patients to identify peripheral blood lymphocyte subsets associated with pruritus. Statistical analysis was carried out by using chi-square test, Mann-Whitney U test, and Spearman correlation analysis. Results:Among the 522 patients with MF, 305 were males and 217 were females; 347 were diagnosed with early-stage MF, and 175 with advanced MF. The incidence of pruritus was 67.2% (351/522) in the patients with MF, and significantly higher in the patients with advanced MF (81.7%, 143/175) than in those with early-stage MF (59.9%, 208/347; χ2 = 25.03, P < 0.001) . RNA sequencing showed that CCL17 mRNA expression was significantly higher in the MF patients with pruritus than in those without (fold change = 10.09, P < 0.001) . The serum CCL17 concentration was significantly elevated in the patients with pruritus (1 017.05[377.12, 4 831.80] pg/ml) compared with those without (361.66 [180.47, 500.08] pg/ml; Z = -4.57, P < 0.001) , and correlated with pruritus scores ( r = 0.57, P = 0.010) . In both early and advanced stages of MF, the serum CCL17 concentration was significantly higher in the patients with pruritus than in those without ( Z = -3.68, P < 0.001; Z = -2.54, P = 0.011, respectively) . Immunohistochemical staining revealed that there was no significant difference in the relative quantification value of CCL17 between the patients with pruritus and those without ( Z = -1.84, P = 0.066) . The percentage of CD3 +CD4 +CD26 -CCR4 + malignant T cells significantly increased in the MF patients with pruritus than in those without ( Z = -2.03, P = 0.043) , and was positively correlated with serum CCL17 concentrations ( r = 0.49, P < 0.001) . Conclusions:Both CCL17 mRNA expression in lesional tissues and serum CCL17 concentrations increased in MF patients with pruritus, and CCL17 was associated with the occurrence of pruritus. CCL17 may be involved in the occurrence of pruritus through the recruitment of CD3 +CD4 +CD26 -CCR4 + malignant T cells.
ABSTRACT
Objective:To observe the changes in the biomechanical properties of the cornea of diabetic retinopathy (DR), and analyze its relationship with the degree of DR.Methods:A retrospective study. From September 2020 to February 2021, 83 patients with type 2 diabetes (T2DM) combined with DR treated in the Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, 83 eyes (DR group), 30 patients with T2DM without DR recruited from the outpatient clinic 30 eyes (NDR group) and 30 eyes of non-diabetes patients (NDM group) were included in the study. All left eyes were chose as the study eye. Among the 83 eyes in the DR group, 39 eyes were non-proliferative DR (NPDR) and 44 eyes were proliferative DR (PDR). Based on this, they were divided into NPDR group and PDR group. There was no statistically significant difference in age ( t=1.10) and sex ratio ( χ2=0.46) among patients in the DR group, NDR group, and NDM group ( P>0.05); body mass index ( t=3.74), glycosylated hemoglobin ( t=35.02) and the length of the eye axis ( t=5.51), the difference was statistically significant ( P<0.05). The eye response analyzer (ORA) was used to measure the corneal hysteresis (CH), corneal resistance factor (CRF), Goldman related intraocular pressure (IOPg), and corneal compensatory intraocular pressure (IOPcc). The corneal topography was used to measure the central corneal thickness (CCT) of the examined eye. The differences of CCT, IOPcc, IOPg, CH, CRF among multiple groups were compared by one-way analysis of variance. Multiple linear regression was used to analyze the relationship between CH, CRF and related influencing factors in DR patients. Results:There were statistically significant differences in CCT, IOPcc, IOPg, CH, and CRF among the eyes of the DR group, NDR group, and NDM group ( F=3.71, 5.60, 9.72, 9.02, 21.97; P<0.05). Pairwise comparisons were between groups, CH, CRF: the difference between the DR group and the NDM group and the NDR group was statistically significant ( P<0.05); CCT: the difference between the DR group and the NDM group was statistically significant ( P<0.05), and The difference in the NDR group was not statistically significant ( P>0.05). CCT, CH, CRF: the difference between the NDR group and the NDM group was not statistically significant ( P>0.05). The results of multiple linear regression analysis showed that CCT and IOPcc in DR patients were independent influencing factors of CH [CCT: β=0.01, 95% confidence interval ( CI) 0.01-0.03, P=0.013; IOPcc: β=-0.15, 95% CI -0.25--0.05, P=0.005]; Age, CCT, IOPcc were independent influencing factors of CRF [Age: β=-0.06, 95% CI -0.09--0.03, P<0.001; CCT: β=0.01, 95% CI 0.00-0.02, P=0.049; IOPcc: β=0.16, 95% CI 0.07-0.25, P=0.001]. The comparison of CCT, CH, CRF, adjusted CH, and adjusted CRF of the eyes in the NDR group, NPDR group, and PDR group were statistically significant ( F=3.76, 5.36, 12.61, 6.59, 10.41; P<0.05). Pairwise comparison between groups, CH, CRF, adjusted CH, adjusted CRF: the difference between the NPDR group, the PDR group and the NDR group was statistically significant ( P<0.05), and the difference between the PDR group and the NPDR group was not statistically significant ( P>0.05); CCT: The difference between NPDR group and NDR group, PDR group and NPDR group was not statistically significant ( P>0.05), and the difference between PDR group and NDR group was statistically significant ( P<0.05). Conclusion:The CH and CRF of eyes with T2DM and DR are elevated; CCT and IOPcc are independent influencing factors of CH, and age, CCT and IOPcc are independent influencing factors of CRF.
ABSTRACT
Objective To investigate the effects of Epigallocatechin gallate (EGCG) on cardiac protection in diabetic rats and the expression of TGF-1/Smad3 signaling pathway. Methods The influence of clean level 40 SD rats, weight 200-220 g, divided into four random groups:control (Sham) group, diabetic cardiomyopathy model (DC) group, EGCG group, and metformin positive control group (Met).Post 8 weeks of high-fat-diet administration, the rats were injected intraperitoneally with STZ to establish the diabetes cardiomyopathy model. Upon successful model establishment, the EGCG group was intraperitoneally injected with EGCG and the cardiac function of the rats was measured after 28 days of drug administration. Then, the pathological results of the myocardial tissue were analyzed. Triglyceride (TG), total cholesterol (TC), glycosylated hemoglobin (HbA1 c), and blood glucose (FBG) concentrations were also measured. Further, the concentrations of superoxide dismutases (SOD), malondialdehyde (MDA), catalase (CAT), and glutathione peroxidase (GPX) in serum were measured by ELISA. The expression of TGF-β1 and Smad3 in kidney tissues of the rats was measured by Western blotting analysis. Results EGCG could reduce the glucose, lipid, and MDA levels in the blood of the diabetic rats, enhance cardiac systolic and diastolic functions, inhibit TGF-β1 and Smad3 protein expression, enhance the activity of SOD, CAT and GPX, and reduce myocardial tissue fibrosis. Conclusion EGCG can protect diabetic rat hearts by improving metabolic disorder, and its mechanism may be related to the oxidative-stress mediated by the TGF-β1/Smad3 signaling pathway.
ABSTRACT
A new method for the rapid determination of total phthalates (PAEs) in edible oils was developed. The PAEs in edible oils all were hydrolyzed to phthalic acid with tetrabutylammonium chloride (TBAC) as catalyst. Then phthalic acid was extracted by the supramolecular solvent ( SUPRAS) made up of octanol, tetrahydrofuran and aqueous solution, and detected by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS). As a result, hydrolysis time was 10 min. The linear range of phthalic acid was 0. 05- 2. 0 mg / L with a good correlation coefficients ( r > 0. 999). The limits of detection ( LOD) and quantification (LOQ) were 5. 41 and 18. 05 μg / kg, respectively. The recoveries of target analyte at three spiked levels were in the range of 84. 6% - 104. 5% . The repeatability, expressed as relative standard deviation (RSD), was 2. 6% for intra-day and 3. 7% for inter-day. The total PAEs content of 12 edible oils was found in the range of 0. 30-1. 09 mg / kg.
ABSTRACT
Objective To evaluate the effect of the clinical nursing path integrated with the holistic nursing on advanced schistosomiasis patients with ascites. Methods A total of 226 advanced schistosomiasis patients with ascites were randomly di?vided into a control group and an experimental group (113 cases each group). The subjects in the experimental group were nursed by the clinical nursing path integrated with the holistic nursing,while those in the control group were nursed only by the holistic nursing. Then the clinical relevant indexes of the two groups were observed,and the quality of life of the patients before and after hospital discharge was assessed. Results The improvement rate,satisfaction degree,and awareness rate of health knowledge of the patients in the experiment group were 93.8%,100%and 97.4%,respectively,which were significantly higher than those of the control group(all P<0.05). The mortality rate and the complication rate of the patients in the experimental group were 0 and 2.7%,respectively,which were significantly lower than those of the control group(both P<0.05). In addi?tion,the average hospitalization days and the hospitalization cost of the experiment group were(12.2 ± 0.7)d and(4 725.0 ± 310.1)Yuan respectively,which were less than those of the control group(both P<0.01). When 6 months after the discharge from hospital,the quality of life of the patients in the experimental group in various fields was significantly better than that of the control group(all P<0.05). Conclusions The clinical nursing pathway integrated with holistic nursing can effectively improve the improvement rate and decrease the mortality of the advanced schistosomiasis patients with ascites;meanwhile,it can short?en the hospitalization time and save the hospitalization cost. Therefore,this nursing model is suitable for popularization and ap?plication in the treatment and nursing work of the advanced schistosomiasis assistance.
ABSTRACT
In this paper is reported a new approach for the treatment of sarcoma--electroporation therapy. Electroporation can accelerate pharmacal molecules into cytoplasm by transient electromagnetic pulses. We have utilized the phenomenon of electroporation treating the S-180 sarcomas in the hind legs of the Kunming mice by intratumoral injection of anti-tumor agent at low dose. From the experiment, we learned that this approach can bring about remarkable effect. The technical procedure is easy to do and easy to control. Especially, it is useful in curing the flat tumor and has little untoward side effect. It deserves to be recommended as a new approach to treating the tumor in clinics.
Subject(s)
Animals , Mice , Antineoplastic Agents , Therapeutic Uses , Apoptosis , Cell Line, Tumor , Cyclophosphamide , Therapeutic Uses , Electrochemotherapy , Methods , Neoplasm Transplantation , Sarcoma 180 , Drug Therapy , PathologyABSTRACT
We chose Hela cells as research object and studied the cytotoxicity generated by cyclophosphamide, an antitumor drug, after cell electroporation by the use of electromagnetic pulses. Comparison between the electroporation group and the contrast group revealed the greatly enhanced cytotoxicity of the electroporation group, indicating that under some conditions electromagnetic pulses can enhance the cytotoxicity of antitumor drugs. The results of this study provide reliable evidences and a feasible approach for clinical treatment of tumor.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Cell Survival , Cyclophosphamide , Pharmacology , Drug Screening Assays, Antitumor , Electromagnetic Phenomena , Electroporation , Methods , HeLa CellsABSTRACT
This article reports the experiment studies on treating the S-180 sarcomas of KM mice with high-intensity electric pulse and antitumor drug (cyclophosphamide). The results showed that the experimental group of electric field combined with drug has the best effect on tumor, compared with the control group. In addition, electric field can inhibit the formation of vas Capillaries and decrease the supply of nutrition for tumor cell. In conclusion, electric field has inhibited the growth of tumor.