1.Study on UHPLC Fingerprints of Psoralea corylifolia L.
Xiaochuan CHEN ; Ziwei LI ; Huixian TANG ; Xiaomei ZHANG ; Na LI
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):865-868
This study was aimed to establish the ultra high performance liquid chromatography (UHPLC) fingerprints of Psoralea corylifolia L. The separation was achieved on a Shim-pack XR-ODS Ⅲ column (50 mm í 2.0 mm, 1.6μm) by gradient elution with acetonitrile-0.2% glacial acetic acid solution as the mobile phase. The flow rate was 0.1 mL·min-1 and the measurement wavelength was 246 nm. The temperature of the column was 45oC. The results showed that the UHPLC fingerprint of P. corylifolia L. was established and 10 characteristic common peaks were found, among which 6 peaks were recognized by comparing with reference substances. It was concluded that the method was rapid, reliable and reproducible. The established fingerprint can provide references for the study of sub-stance basis and quality control of P. c orylifolia L.
2.Plasma levels of leukocyte cell-derived chemotaxin 2 in patients with newly diagnosed type 2 diabetes complicated with non-alcoholic fatty liver disease and its association with insulin resistance
Huixian ZENG ; Zhen ZHANG ; Jianghong LIN ; Yinghui HU ; Hong CHEN
The Journal of Practical Medicine 2017;33(7):1090-1094
Objective To observe the changes in levels of plasma leukocyte cell-derived chemnotaxin 2 (LECT2) in patients with newly diagnosed type 2 diabetes (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD),and to investigate the clinical significance.Methods A total of 137 subjects were enrolled in the study,including 50 patients with newly diagnosed T2DM complicated with NAFLD,47 patients with newly diagnosed T2DM with non-NAFLD and 40 healthy subjects.The level of plasma LECT2 was determined by ELISA.Anthropometric data and other biochemical indicators were measured in three groups.The body mass index (BMI) and waist-to-hip ratio (WHR) were calculated.Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment (HOMA-IR,HOMA-%β).Results Plasma concentration of LECT2 in patients with newly diagnosed T2DM complicated with non-NAFLD was higher than that in type 2 diabetic patients with non-NAFLD [(32.95 ± 10.11 vs 29.08 ± 7.54) ng/mL,P < 0.01].Plasma LECT2 levels in both groups were significantly higher than that in normal control group [(22.38 ± 4.40) ng/mL,P < 0.01].Plasma LECT2 level was positively correlated with BMI,FPG,FINS,C peptide,HbA1c,GGT,TG and HOMA-IR,while negatively with HDL-C and HOMA-% β (all P < 0.01).Multivariate regression analysis showed that levels of BMI,FPG and HDL-C were important factors affecting plasma LECT2 level.Conclusions Plasma LECT2 concentration significantly elevates in patients with newly diagnosed T2DM complicated with non-NAFLD.Plasma level of LECT2 is closely correlated with insulin resistance and glycolipid metabolism.LECT2 may play an important role in the patho genesis of insulin resistance and T2DM.
3.Effects of TNF-?,sTNFRⅠon IL-8Production and Proliferation of Psoriatic Keratinocytes
Xianjin CHEN ; Kaiming ZHANG ; Guohua YIN ; Huixian LI
Chinese Journal of Dermatology 1995;0(04):-
Objective To study the effects of TNF -?and sTNFRⅠon IL -8production and proliferatio n of psoriatic keratinocytes.Methods In vitro cultured keratinocytes were treated with TNF -?,sTNFRⅠand an-ti -TNFRⅠ.IL -8levels in supernatants were me asured by enzyme-linked immunosobent assay.Expression of IL -8mRNA in keratinocytes was observed by semiquantitive RT -PCR and Southern blotting.Proliferation index wa s measured by MTT colorimetry.Results IL -8levels,mRNA expression and pro liferation index were significantl y higher in psoriatic patients than th ose in control group(P
4.Reform of endocrinology and metabolism course integration based on organ-system based learning
Huixian ZENG ; Li YANG ; Zhen ZHANG ; Rongping CHEN ; Rui YANG ; Yanzhen CHENG ; Hong CHEN
Chinese Journal of Medical Education Research 2017;16(1):75-79
Systems-based integrated course is the core and hot spot in current advanced medical education reform.An integrated organ-system oriented curriculum system of endocrinology and metabolism was applied in eight year clinical medicine and five year excellent doctor education.The teaching contents of endocrinology and metabolism from traditional Basic Medicine,Internal Medicine and Surgery were integrated and optimized to compile the integrated syllabus and teaching cases.Curriculum integration oriented PBL teaching and comprehensive morphology experimental teaching were implemented into the integrated endocrinology and metabolism system curriculum.This endocrinology and metabolism course integration based on organ-system based learning is conducive to establishing the organic connection between Basic Medicine and Clinical Medicine,and cultivating high-quality medical talents.
5.Determination of the Platelet Activating Factor in Silicotic Patients and its Effect on Fibroblasts
Qunwei ZHANG ; Yiqun MO ; Jinpin LOU ; Xinqiang ZHU ; Zhimin CHEN ; Linyun HE ; Huixian ZHONG
Environmental Health and Preventive Medicine 2000;5(4):134-137
Platelet-activation factor (PAF), one of the potent proinflammatory mediators, is produced from a large range of cells, including polymorphonuclear neutrophils, monocytes, and natural killer cells. To study the role of PAF in the pathogenesis of silicosis, we determined the PAF in silicotic patients and in healthy persons. The results showed that the concentration of PAF in the plasma of silicotic patients was significantly higher than that of healthy persons. Our in vitro experimental results showed that the total numbers of fibroblasts were markedly raised with added PAF from 0 to 1 μ g/ml. Adding 1 μ g/ml PAF significantly increased the total numbers of fibroblasts after culture for 48, 72, 96 hrs. Therefore, we suggest that PAF be possibly involved in the pathogenesis of silicosis. However, the mechanism remains to be further elucidated.
Platelet Activating Factor
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Effective
6.The application and effect of PBL teaching method in Hebei Medical University
Yuming WU ; Min ZHANG ; Xiaoxin HUANG ; Xiaojie CHEN ; Yuyan HU ; Huixian CUI
Chinese Journal of Medical Education Research 2015;(12):1271-1275
In order to promote the integration between disciplines, the convergence between basic course and clinical teaching, increasing students ability including the active learning and life-long learning ability, finding problem and solving problem ability, teamwork spirit and so on. After nearly 3 years preparation, Hebei Medical University successfully carried out the PBL teaching in Seven-year Clinical Medicine Science. Combining with the teaching activities, formative assessment was carried out, and PBL teaching website was established. The reform has already achieved initial results, got good responds from teachers and students. Through the study, it has been confirmed that the PBL teaching method in Hebei Medical University is effective and worthy to reference.
7.Performance of transfection with a complex plasmid encoding green fluorescent protein tagged cathepsin D in researches on chronic photodamaged fibroblasts
Yue ZHENG ; Haiyan CHEN ; Qingfang XU ; Congxiu YE ; Huixian LIU ; Jinling YI ; Wei LAI
Chinese Journal of Dermatology 2015;(12):876-879
Objective To evaluate the performance of transfection with a complex plasmid encoding green fluorescent protein tagged CatD (GFP-CatD)in researches on chronic photodamaged fibroblasts. Methods Human dermal fibroblasts (HSFs)were irradiated with ultraviolet A (UVA)at 25 J/cm2 once a day for 21 consecutive days to establish a chronic photodamaged cell model. A plasmid encoding GFP-CatD was constructed and transfected into some chronic photodamaged fibroblasts (experimental group). The photodamaged HSFs receiving no treatment served as the blank control group, and those transfected with the negative plasmid encoding GFP only as the negative control group. After additional culture, fluorescence microscopy and Western-blot analysis were performed to observe and measure the expression of GFP-CatD in HSFs respectively, flow cytometry and methyl thiazolyl tetrazolium (MTT)assay to evaluate the apoptosis and proliferation of chronic photodamaged fibroblasts respectively. Results Fluorescence microscopy showed the expression of GFP-CatD in cytoplasm of chronic photodamaged fibroblasts at 96 hours after transfection with the GFP-CatD-encoding plasmid. Western-blot analysis revealed that the expression of CatD in the experimental group was 1.28 times that in the blank control group. There were no significant differences in the apoptosis rate(4.29% ± 1.30%vs. 3.03% ± 1.70% , P > 0.05)or proliferative rate (45.20% ± 4.70% vs. 43.60 ± 3.90% , P > 0.05)between the experimental group and blank control group. Conclusion CatD could be traced in chronic photodamaged fibroblasts with no changes in biological activity or cell cycle after transfection with the GFP-CatD-encoding complex plasmid.
8."Reproduction of a model of ""two-hit"" sepsis model with complication of pneumonia in rat"
Yingying CHEN ; Huixian LI ; Shuai MA ; Bo DENG ; Jianxin LU ; Feng DING
Chinese Critical Care Medicine 2015;(10):805-810
ObjectiveTo reproduce a clinically relevant two-hit model of sepsis complicated by pneumonia and to explore the correlation between two-hit and immune state.Methods Eighty-one male Sprague-Dawley ( SD ) rats were divided into groups according to the random number table. Forty-five male rats were assigned respectively to sepsis-alone group, pneumonia 4 days and 7 days after sepsis groups, respectively. Survival rate of each group was observed. Another group of 36 male rats were divided into normal control group, sepsis-alone for 1, 4 and 7 days groups, and sepsis complicated by pneumonia for 4 days and 7 days after sepsis groups, each group consisted of 6 rats. Cecal ligation and puncture (CLP) was done in rats, andStreptococcus pneumoniae suspension (bacteria count 1×1010 cfu/mL) was injected via the nose on the 4th day or 7th day after CLP. Rats were sacrificed at corresponding time points, and 1 day after challenge ofStreptococcus pneumoniae on the 4 days or 7 days post CLP for the collection of blood and tissue samples to make bacterial count of the blood, splenocyte count, biochemical indices, cytokines concentration, pathological changes in spleen and apoptotic cells.Results① Compared with the rats of sepsis-alone group, the rats in pneumonia 4 days after CLP group had poor survival rate (4 vs. 11,χ2 = 6.533,P = 0.011), while no difference was found between pneumonia 7 days after CLP group and sepsis-alone group (9 vs. 11,χ2 = 0.600,P = 0.439).② The blood bacterial count and all the biochemical indexes were sharply increased on 1 day post-CLP in the rats of sepsis-alone group, and then they gradually lowered. Compared with the rats of 1 day post-CLP, the proportion of splenocytes were decreased on the 4th day post-CLP [dendritic cells (DC): (0.69±0.09)% vs. (0.87±0.31)%, CD4+T cells: (21.05±2.89)% vs. (24.84±4.59)%, CD8+ T cells: (10.62±1.79)% vs. (13.40±1.31)%, allP< 0.05], but T-regulatory cell (Treg) count was higher on the 4th day after CLP compared with sepsis-alone rats [(3.14±0.74 )% vs. (2.87±1.08)%,P< 0.05]. The biochemical indices, including alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and serum creatinine (SCr) were obviously lowered on 7 days post-CLP compared with 1 day after CLP [ALT (U/L): 35.33±11.52 vs. 81.00±38.40, AST (U/L): 70.33±42.16 vs. 156.00±28.11, BUN (mmol/L): 5.30±2.27 vs. 9.13±4.04, SCr (μmol/L): 55.33±10.67 vs. 96.67±45.79, allP< 0.05]. The serum levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-6, IL-1β) peaked on the 1st day after CLP [TNF-α:(18.03±2.88) ng/L, IL-6: (10.37±4.20) ng/L, IL-1β: (102.44±51.46) ng/L], and high mobility group box-1 (HMGB1) peaked on the 4th day after CLP [(1.76±0.71)μg/L]. The levels of anti-inflammatory cytokines transforming growth factor-β1 (TGF-β1 ) and soluble tumor necrosis factor receptor-Ⅰ (sTNFR-Ⅰ) maintained at high levels [7 days post-CLP: TGF-β1 was (0.90±0.56) ng/L, sTNFR-Ⅰ was (1.56±0.39) ng/L]. The spleen pathology became more marked with the time in the group of sepsis-alone, meanwhile the number of apoptotic spleencytes increased 4 days post-CLP as compared with that of the 1st day post-CLP (cells/HP: 52.99±20.79 vs. 16.05±3.28,P< 0.05).③ Compared with the same period of sepsis-alone group, the rats with pneumonia 4 days post-CLP group showed a higher blood bacterial count (log cfu/mL: 1.78±0.54 vs. 0.25±0.18,P< 0.05), while no difference was found between 7-day of post-CLP pneumonia group and sepsis-alone group (log cfu/mL: 0.57±0.46 vs. 0.13±0.12,P> 0.05). The same trend of changes, with slight reduction in splenocytes and biochemical indices were found between the groups of sepsis followed by pneumonia and sepsis-alone, but no significant difference was found. The level of HMGB1 in the 4-day group of sepsis with complication of pneumonia was further decreased compared with sepsis-alone group (μg/L:1.17±0.74 vs. 1.76±0.71,P< 0.05), and IL-1β in the 7-day group of sepsis complicated pneumonia was further higher than those of sepsis-alone group in the same period (ng/L: 105.73±25.06 vs. 61.04±31.29,P< 0.05), while there were no differences in levels of other cytokines between two-hit group and sepsis-alone group. Apoptosis of spleencytes in the 4-day group of sepsis complicated pneumonia was more marked than that of sepsis-alone group at the same period (cells/HP: 74.48±22.47 vs. 52.99±20.79,P< 0.05), while no difference was found between the 7-day groups of sepsis complicated pneumonia and the sepsis-alone group (cells/HP: 28.70±4.13 vs. 30.43±14.55, P> 0.05).Conclusions The mortality of this two-hit model with complication of pneumonia 4 days after CLP was significantly higher than that of single sepsis model. The ability of bacteria clearance was decreased, and immunocyte apoptosis was exacerbated. These findings may be with the result of the occurrence of immunoparalysis in the mid stage of sepsis. The two-hit model reproduced on 7 days after CLP might suggest reconstruction of host immune function, and maybe associated with the recovery of immune response.
9.A preliminary application of dynamic ~(13)N-NH_3 positron emission tomography imaging in hypopituitarism
Xiangsong ZHANG ; Zuoxiang HE ; Huazhang YANG ; Hongmei CHEN ; Anwu TANG ; Huixian QIAO
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Dynamic ~(13)N-NH_3 positron emission tomography (PET) imaging was performed in 8 normal subjects and 6 cases of hypopituitarism. The pituitary images were small and delayed, and blood perfusion and radioactive uptake were decreased in hypopituitarism. ~(13)N-NH_3 PET imaging shows diagnostic value in hypopituitarism.
10.Advanced glycation end products and its receptor induce apoptosis of L cells through NADPH oxidase mediated signaling pathway
Yinghui HU ; Zhen ZHANG ; Lei LEI ; Rui YANG ; Jianghong LIN ; Huixian ZENG ; Hong CHEN
The Journal of Practical Medicine 2017;33(3):358-362
Objective To investigate the effects of AGEs-RAGE on the apoptosis of GLUTag cells and explore the possiblc mechanism.Methods GLUTag cells treated with 0、100、200、300μg/ml of AGEs for 24h were examined for gene and protein expression of RAGE using RT-PCR and western blotting,respectively.GLUTag cells were randomly divided into four groups:control,200μg/ml AGEs,AGEs+siRNA-RAGE and AGEs+apocynin.The protein expression of p22phox、p47phox 、Bcl-2、Bax in the cells were detected with western blotting.The reactive oxygen species (ROS) levels were examined using 2'7'-dichlorodihydroflur-rescein diacetate (DCFH-DA) and the apoptosis of L cells were tested by AnnexinV-FITC/PI.Results AGEs increased thc cxpression of RAGE in a dose dependent manner.Treatment with AGEs induced a significant increase in the expression of p22phox,p47phoxand the activity of ROS,caused up-regulation of Bax and down-regulation of Bcl-2,which enhanced the apoptosis of GLUTag cells.Apocynin,the inhibitor of NADPH oxidase,prevented those responses and the effects caused by AGEs were abolished by inhibition of RAGE activity with siRNA.Conclusion AGEs positively regulate the exprcssion of NADPH oxidase-derived ROS and its down-steam signaling pathway p53/Bax by targeting RAGE,leading to the apoptosis of GLUTag cells.