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Objective:To evaluate the data quality of Shenzhen Type 1 Diabetes Alliance (SZT1D), and to provide a basis for evaluation and improvement for the continuous improvement of data quality.Methods:From December 2018 to July 2021, 697 first-visit type 1 diabetes (T1DM) patients (including 501 in Shenzhen and 196 out-of-Shenzhen) and 120 re-visited T1DM patients (including 113 in Shenzhen and 7 out-of-Shenzhen) who were registered by SZT1D in collaborative research platform network of China Type 1 Diabetes Alliance (hereinafter referred to as China T1D). The data quality was evaluated from three dimensions: data completion, accuracy and revisit. The data completion degree was evaluated by the overall data completion degree and the key indicator completion degree; the data accuracy was evaluated by the probability of abnormal blood glucose value; the patient′s return visit was evaluated by the return visit rate.Results:The main characteristics of T1DM in SZT1D were young and middle-aged adults [age: (34.4±17.1)years] with thin body [BMI: (19.80±3.52)kg/m 2)], half of male and female patients [proportion of male: 52.4%(365/697)]; the main types of diagnosis were classical T1DM [65.22%(150/230)] and latent autoimmune diabetes in adults(LADA) [26.08%(60/230)], and the fasting blood glucose (FPG) [(10.93±6.98)mmol/L] and glycosylated hemoglobin (HbA 1c) [(10.63±3.01)%] were high. The average completion rate of the overall data of the first diagnosed patients in SZT1D was only 60% [(62.9±31.5)%]: the number of patients with overall data completion ≥80% in SZT1D was only 50.2%(350/697); the number of patients with overall data completion ≥80% in Shenzhen was less than that outside Shenzhen [44.3%(222/501) vs 65.3%(128/196), P<0.001]. The key indicators with better completion rate of first-visit were disease course [76.2%(531/697)], age of onset [75.8%(528/697)], family history of diabetes [74.9%(522/697)], etc., but none of them had a completion rate of more than 80%, and the diabetes self-management behavior assessment questionnaire and scale score were completely missing; the frequency of daily blood glucose monitoring [46.1%(231/501) vs 64.3%(126/196), P<0.001], current insulin regimen [44.3%(222/501) vs 63.3%(124/196), P<0.001], number of diabetic ketoacidosis (DKA) since the onset of the disease [45.7%(229/501) vs 64.8%(127/196), P<0.001] and the number of symptomatic hypoglycemia in the past 1 month [39.3%(197/501) vs 63.8%(125/196), P<0.001] were higher in Shenzhen than those reported outside Shenzhen. In addition, the probability of abnormal FPG and postprandial glucose (PPG) [5.2%(24/466); 3.8%(19/236)] were low. The revisit rate was not high [17.2%(120/697)], and the revisit rate in Shenzhen was higher than that outside Shenzhen [22.6%(113/501) vs 3.6%(7/196), P<0.001]. The first revisit rate was 16.2%(113/697) and the second revisit rate was seriously insufficient [1.0%(7/697)]. Conclusions:The data quality of T1DM patients recorded by SZT1D needs to be further improved. Improving the information interconnection between China-T1D and SZT1D, employing quality control personnel and building a systematic data quality evaluation analysis and feedback mechanism are methods to promote the comprehensive, accurate and efficient input of T1DM data and continuously improve the evaluation methods to improve the overall data quality.
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Objective To investigate the effect of EP4 gene silencing on the growth and migration of K1 cells. Methods K1 cells with stable knockdown of EP4 were constructed with lentiviral vector. QRT-PCR and western blot analysis were used to detect the expression of EP4 mRNA and protein in K1 cells. CCK8 assay and flow cytometry were employed to measure cell viability and apoptosis. Transwell assay was applied to detect cell migration. Results Compared with the negative control group,the mRNA and protein expression of EP4 were sig-nificantly decreased in K1 cells with stable knockdown of EP4. Furthermore,shRNA-mediated silencing of EP4 gene remarkably suppressed cell viability and induced apoptosis of K1 cells.The migration of K1 cells with knock-down of EP4 was decreased compared with the negative control group. Conclusions EP4 gene silencing can in-hibit growth and induce apoptosis of K1 cells.Downregulation of EP4 can significantly reduce migration of K1 cells.
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Objective To explore the effect of ACE-inhibitor perindopril on the expression of scavenger receptor A (SR-A) gene in the kidney of diabetic rats.Methods Diabetes were induced in male Sprague-Dawley rats by peritoneal injection with streptozotocin (60mg/kg).The rats were then random di vided into normal control group, diabetes group and ACEI treatment group [4mg/(kg·d) for 24 weeks].Blood glucose concentration and 24h urinary albumin excretion were determined.The renal morphological change was observed.Immunohistochemistry was used to analyze CD68 positive macrophages,and the Mrna of SR-A in renal tissue was detected by quantitative real-time PCR.Results Compared with normal control group,blood glucose concentration,24h urinary albumin excretion and the number of CD68 positive macrophages were significantly increased [(5.3 ± 0.6) mmol/L vs (26.7 ± 3.3) mmol/L;(2.7 ± 1.3) mg/24h vs (26.7 ± 1.8)mg/24h;(0.77 ±0.24)/gcs vs (2.55 ±0.46)/gcs;(6.13 ±0.50)/HPF vs (11.9 ±2.12)/HPF;P ＜0.05],and the expression of SR-A Mrna were significantly up-regulated in diabetes group [ (5.6 ± 1.2 vs 1.5 ±0.2),P ＜0.05].After intervention with ACE-inhibitor,the up-regulations of the above mentioned parameters,except blood glucose concentration,were all significantly inhibited [ (3.6 ±1.4)mg/24h;(1.03±0.37)/gcs;(8.28±1.19)/HPF;3.4±0.7;P ＜0.05].Conclusion ACE-inhibitor might have renoprotective effects of diabetic nephropathy,it probably was associated with inhibiting the expression of SR-A gene.
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The rats were assigned to blank control group, classical induction group, and drynaria total flavonoid group. Whole bone marrow culture method was applied to isolate and purify rats bone mesenchymal stem cells (BMSCs). Akaline phosphatase activity, calcium nodes, TGF-β1 and BMP-2 secretion in the process of bone mesenchymal stem cells osteogenic differentiation were detected. The results showed that compared to the blank group and classical group, drynaria total flavonoid promoted osteogenic differentiation accompanied with increased TGF-β1 and BMP-2 secretion (all P＜0. 05). Drynaria total flavonoid may promote osteogenic differentiation of BMSCs via upregulating TGF-β1 and BMP-2 expressions, and play an active role in the treatment of osteoporosis.
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<p><b>OBJECTIVE</b>This study was to investigate the growth and proliferation characteristics of rat bone mesenchymal stem cells (BMSCs) isolated by the method of whole bone marrow culture and to explore the effect of cell inoculation density and incubation period on cell proliferation, with an aim to provide multipotential seed cells for preventing from degenerative disease.</p><p><b>METHODS</b>Bone mesenchymal stem cells were isolated by the method of whole bone marrow culture and then cultured in vitro. The cell morphologic features were observed by inverted microscope. The cell surface antigens were identified by flow cytometry. The effect of cell inoculation density and culture period on cell growth and proliferation was explored by analyzing the characteristics of a ten-day cell growth curve in 96-well plates.</p><p><b>RESULTS</b>Flow cytometry results showed the detection rates for CD29, CD34 and CD45 were 97.68% (7607/7788), 7.93% (661/8340) and 2.76% (215/7788) respectively, which was consistent with the expression characteristics of BMSCs surface antigens. BMSCs became uniform after three cell passages, existing in a typical shape of whirlpool or radial colony. The senescent cells started to appear at 7(th) passage, and more senescent cells were found at 10(th) passage. The growth curve for moderate inoculation density was typically S-shaped. Lag phase was found during the first two days, and logarithm growth phase was in the following three days. Plateau phase started from the 6(th) day and cell numbers decreased slightly from the 8(th) day.</p><p><b>CONCLUSION</b>The whole bone marrow culture is an effective way to obtain BMSCs. A moderate inoculation density was more advantageous to cell proliferation, by which more seed cells could be obtained.</p>
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Animals , Male , Rats , Bone Marrow Cells , Cell Biology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Flow Cytometry , Mesenchymal Stem Cells , Cell Biology , Rats, Sprague-DawleyABSTRACT
Objective To investigate the relationship between human leptin receptor(LEPR)gene G3057A polymorphism and type 2 diabetes complicated with non-alcoholic fatty liver disease(NAFLD).Methods 216 cases of newly diagnosed type 2 diabetes(104 cases complicated with NAFLD)and 108 cases of normal glucose tolerances(NGT)were recruited.Hemi-nested PCR-RFLP and PCR direct sequence analysis were conducted to detect the polymorphisms of LEPR G3057A polymorphism.The plasma leptin and insulin levels were measured by ELISA kit.Plasma lipid and glucose metabolic parameters were measured routinely.Liver ultrasound scanning Was carried out among all subjects.Results Type 2 diabetic patients complicated with NAFLD had higher plasma alanine aminotransferase(ALT),triglycerides(TG),low density lipaprotein cholesterol(LDL-C),leptin levels and lower plasma insulin levels than those cages without NAFLD and NGT.The variant frequency at nucleotide 3057 G to A transversion Was 76.0% in type 2 diabetic patients complicated with NAFLD,which Was also significant higher than those cases without NAFLD(62.1%)or NGT cases(53.2%)(x2=14.63,P＜0.01).Conclusions The polymorphism of LEPR gene 3057 probably contributes to the onset of NAFLD in type 2 diabetes by regulating lipid metabolism and affecting insulin sensitivity.
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Objective To investigate the effect of smad7 on AGEs-induced NRK52E cell tranedifferentiation. Methods NRK52E cells were transferred with Smad7 gene and the cell lines of doxcycllne (Dox) -regulated Smad7 expression were selected for the study. The NRK52E cells were random divided into two groups, Dox - group, the NRK52E cells were cultured with medium only containing AGEs, Dox +group, the NRK52E cells were cultured with medium containing AGEs and Dox. Nuclear translation of p-Smad2/3 were examined with immunocytochemistry. The expression of SmadT, α-SMA, E-cadherin protein were detected by western blot. Results Expression of Smad7 protein in the Dox + group was increased higher than that in Dos- group( t = 3.05, P<0.01). High expression of Smad7 caused a marked inhibition of p-Smad2/3 transnuclear location at 30min and 24h(t=4. 5,t= 3.2, P<0.01). Overexpression of Smad7 dramatically de- creased protein expression of α-SMA, while evidently increased protein of E-cadherin in the NRK52E cells stimulated by AGEs(t=3.47, t=3.25, P<0.01). Conclusion Overexpression of Smad7 can inhibit AGEs - induced transdifferentiation in NRK52E cells.
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Objective To investigate the molecular mechanisms that high glucose stimulate collagen Ⅰ synthesis in renal tubular epi-thelial cells. Methods Normal rat pwximal tubular epithelial (NRK52E) cells were cultured grown in RPMI-1640 medium and were divid-ed four groups: mannitol group, high glucose group, high glucose + neutralizing TGF-β1 antibody group, high glucose + IgG1 group. TGF-β1 in the supematant of cultured cells were measured by enzyme-linked immunosorbent assay (ELISA). Nuclear expression of p-Smad2/3 were examined by immunocytochemistry. Expression of collagen Ⅰ mRNA was detected by RT-PCR. Expression of collagen Ⅰ pro-tein was detected by Western blot. Results High glucose up-regulated the expression of collagen Ⅰ mRNA by time-dependent manor. Com-pared with mannitol group, high glucose markedly increased the level of TGF-β1 in supernatant of cultured cell after 24h and 48h and upreg-ulated p-Smad2/3 nuclear expression(t =4. 2, t = 3.25, P <0.01). Neutralizing TGF-β1 antibody inhibited high glucose- induced p-Smad2/3 nuclear expression, downregulated high glucose-induced collagen Ⅰ mRNA and protein expression(t = 3.12, t =3.02, P < 0.01). Conclusion High glucose stimulated collagen Ⅰ synthesis in renal tubular epithelial cells via activated TGF-β/Smad signaling path-way.
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13 2 mmol/L. Results:The incidence of diabetes in the treated group was 31 6% which was obviously less than 72 2% in control group ( P
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Objective To investigate the effects of smad7 on transdifferentiation and collagenⅠsynthesis in advanced glyeosylation end-products(AGE)-stimulated NRK52E cells.Methods NRK52E cells were transferred by pTet-on plasmid system and the cell lines of doxycycline(Dox)-regulated Smad7 expression were selected for the study.Transnuclear location of p-Smad2/3 was examined with immunocytochemistry.The mRNA and protein expressions of Smad7,?-SMA,E-cadherin,collagenⅠwere detected with RT-PCR and Western blot. Results AGE-induced expressions of Smad7 mRNA and protein were further increased in NRK52E cells by the addition of Dox in a dose-dependent manner.Overexpression of Smad7 caused a marked inhibition of p-Smad2/3 transnuclear location at 30 min(68.3% vs 31.2%,P
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AIM: To investigate the effects of advanced glycation end products on activation of Smad signaling pathway and collagenⅠ synthesis in proximal tubular epithelial cells.METHODS: Advanced glycation end products(AGE-BSA) were prepared by incubation of bovine serum albumin(BSA) with D-glucose.Normal rat proximal tubular epithelial(NRK52E) cells were cultured in RPMI-1640 medium with AGE-BSA.Phosphorylation and nuclear translocation of Smad2/3 were examined by immunocytochemistry.Levels of TGF-?_1 in supernatant of cell culture were measured by enzyme-linked immunosorbent assay(ELISA).Expression of TGF-?_1,Smad2,Smad3 and Smad7 mRNA were detected by RT-PCR.Expression of ?-SMA,E-cadherin and collagenⅠproteins were detected by Western blotting.RESULTS: AGE-BSA induced Smad2/3 phosphorylation and nuclear translocation,two peaks occured at 30 min(68% vs 16%,P
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AIM: To observe the effect of antibiotic treatment on the inflammatory mediator expression in peritoneum and the peritoneal transport function in rats with acute peritonitis, and explore its mechanisms. METHODS: Eighty-six SD rats were randomly divided into three groups. Control group (n=28) were treated with PBS (ip), peritonitis group (n=28) and treatment group (n=28) were challenged with the E.coli (ip), but at 3 h and 9 h gentamicin was given (ip) in treatment group. Seven rats of every group were randomly sacrificed at 24 h, 48 h, 72 h and 7 d. Peritoneal equilibration test (PET) was did before they were killed. Leukocyte count, pathological changes and the expression of CD45, NF-?B, IL-1?, TNF-? in peritoneum were examined. RESULTS: (1)The blood leukocytes in peritonitis group decreased strikingly, but did not change obviously in other two groups. The peritoneal fluid leukocytes in peritonitis group increased significantly from 24 h to 72 h, while in treatment group, it enhanced more strikingly than peritonitis group at 24 h, and recovered earlier. (2) Both in peritonitis group and treatment group, the expression of activated NF-?B, IL-1?, TNF-? and CD45 increased significantly, but the treatment group was lower than model group at 48 h and 72 h. The mRNA level of IL-1? and TNF-? had the same trend as their protein expression. (3) Compared with the control group, UF and D/D_0 Glu decreased significantly in model group and treatment group, and D/PTP increased dramatically. The D/P TP in treatment group lowered obviously compared with peritonitis group, while the net UF and D/D_0 Glu had not significant difference between treatment group and model group. CONCLUSION: Antibiotic treatment can partly decrease the expression of inflammatory mediators in peritoneum of rats with acute peritonitis and also can improve the protein transport ability to some extent, but can not improve the peritoneal ultrafiltration and the glucose transport function.
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AIM:To investigate the effect and the mechanism of Zhibai Dihuang Pill(Radix Rehmanniae praeparata,Fructus Corni,Rhizoma Dioscoreae,Rhizoma Anemarrhenae,Cortex phellodendri Chinensis,etc.) on patients with hyperthyroidism. METHODS: Eighty-five hyperthyroid patients were randomly divided into two groups.The control group(40 cases) were treated with propylthiouracil,while the treatment group(45 cases) were treated with propylthiouracil and Zhibai Dihuang Pill;in the 12-week long treatment period,the heart rate,body weight,thyroid free FT_3、FT_4 and TSH and the serum level of FAA,resistin,adiponectin,leptin of patients in both groups were measured. RESULTS: After treatment,the heart rate,the serum of FT_3,FT_4 decreased and the body weight,TSH increased in both groups(P0.05).(CONCLUSION): Zhibai Dihuang Pill can improve the abnormal metabolism of sugar and fat in patients with hyperthyroidism by its actions on the serum level of FFA,resistin,adiponectin,leptin.
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0.05). group 1 and group 4 could significantly reduce the level of lipid、FINS, increase the level of IAI (P0.05). After treatment, the clinical symptom score dropped in group 1、2、3 (P0.05); serum levels of resistin decreased and adiponectin increased in group 1、4 (P0.05). More significant difference was observed in group 1、4 between the four groups about the serum level of resistin、adiponectin、Leptin、TNF-?、CRP、IL-6 (P
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Objective:To investigate expression of cytokines and apoptosis factors in islets of female NOD mice at different weeks of age.Methods:54 female NOD mice were randomly divided into three groups(C1,C2,C3)and each group consisted of 18 mice.The mice were killed respectively at 4th,15th,30th week (of onset of diabetes),and their panreases were removed.The levels of expression of cytokines and apoptosis factors mRNA of the pancreas were detected by RT-PCR.The expressions of cytokines and apoptosis factors protein were measured by using immunohistochemical method.Insulitis scores were observed by HE staining.Results:Expressions of Fas and IFN-? mRNA in pancreas were stronger in C3 vs C1 or C2,in C2 vs C1(P0.05).Expressions of Fas and CPP32 were stronger in islet cells in C2 and C3 vs C1(P