1.Influence of experimental conditions on the test results of cryoglobulin
Hongliang RUI ; Hong CHENG ; Yipu CHEN
Chinese Journal of Laboratory Medicine 2016;39(12):901-905
Objective This study was to observe the effects of different test conditions on the qualitative and quantitative detection of cryoglobulin .Methods We prepared 5 blood samples of different types of cryoglobulinemia . We detect the cryoglobulin qualitatively and quantitatively at different temperatures (37 ℃and room temperature of 20-25 ℃), and with different observation time (3 days and 7 days) and with different amount of blood (5 ml and 20 ml) .Further we will categorize the type of cryoglobulin and detect the components of cryoglobulin by immunofixation electrophoresis ( IFE) and other laboratory tests.Results (1) Blood samples from two groups were clotting and the serums were separated at 37 ℃ and room temperature respectively , and cryoglobulins of two groups were all qualitatively positive . Quantitative detection of cryoglobulins showed that the concentrations of cryoglobulins of room temperature group are lower than that of 37℃group;(2) Compared with 7 days, observing for only 3 days may lead to false-negative results in qualitative detection of cryoglobulin , and concentrations of cryoglobulin are also decreased;(3) Compared with 20 ml blood sample,5 ml blood sample is not enough for qualitative and quantitative detection of cryoglobulins .It may lead to false-negative results;(4) After purification, IFE and other laboratory tests can be used to categorize the types and find the components of cryoglobulins .Such examinations are helpful for finding the potential causes of cryoglobulinemia .Conclusions The positive of serum cryoglobulin is a key indicator of cryoglobulinemia .Detection of cryoglobulin can be affected by temperature, observed time and the blood volume for measurement .In addition, IFE and other laboratory tests are helpful for finding the type and the components of cryoglobulin .
2.Adiponectin antagonizes angiotensin Ⅱ -induced extracelluar matrix production of human renal mesangial cells
Min TAN ; Yipu CHEN ; Gongyao TANG ; Hongliang RUI
Chinese Journal of Nephrology 2011;27(8):591-596
Objective To investigate the effects of adiponectin on angiotensin Ⅱ-induced extracellular matrix production of mesangial cells(MCs) and its possible signaling pathway.Methods RT-PCR and indirect immunofluorescence examination were performed to detect the adiponectin receptors in MCs.Quantitative real-time PCR and enzyme-linked immunosorbent assay (ELISA) were used to observe the effects of adiponectin on angiotensin Ⅱ-induced transforming growth factor β1(TGF-β1) and fibronectin production of MCs.Western blotting was used to measure the ratio of p-AMPK to total AMPK.Results(1)Adiponectin receptors 1 and 2 were found in MCs. (2)The up-regulated mRNA and protein expression of TGF-β1 and fibronectin in MCs induced with 10-7 mol/L angiotensin Ⅱ (Ang Ⅱ) was significantly inhibited by 10 mg/L adiponectin (P<0.05).(3)The p-AMPK/AMPK ratio was significantly increased after incubation with adiponectin for 15 min and 30 min(vs 0 min, P<0.05), which suggested that adiponectin could activate the AMPK signaling pathway in MCs.The activation of AMPK signaling pathway was blocked by 40 μmol/L compound C, a specific inhibitor of AMPK. (4)The inhibitory effects of adiponectinonangiotensin Ⅱ-upregulatedTGF-β1andfibronectinexpressioninMCswere significantly relieved by 40 μmol/L compound C (P<0.05).Conclusions There are adiponectin receptors 1 and 2 in MCs.Adiponectin has inhibitory effects on the angiotensin Ⅱ-upregulated TGF-β1and fibronectin expression in MCs.AMPK signaling pathway may play an important role in the effects of adiponectin above-mentioned.
3.Pathogenesis of anemia in chronic aristolochic acid nephropathy rats
Yingjin QIAO ; Yipu CHEN ; Hongliang RUI ; Hongrui DONG ; Zhangsuo LIU
Chinese Journal of Nephrology 2009;25(12):925-929
Objective To study the pathogenesis of anemia in chronic aristolochic acid nephmpathy(CAAN) rats. Methods The hemoglobin(Hb)values of sixty-two male SD rats were assayed to determine its normal range.Among them,24 rats with normal Hb value were randomly divided into 2 groups:model group (MG)in which rats received the extract of Aristololochia manshuriensis Kom (AmK) by gavage,and control group (CG) received tap water only by gavage.Body weisht(BW),Hb,24 h urinary protein excretion(UP)and creatinine clearance (Ccr)of 6 rats in each group were measured before administration and at the end of the 8th week, respeetively.then these rats were sacrificed.The relative area of renal interstitial fibrosis was measured by microscopy.The mRNA expression of erythropoietin (EPO)in kidney tissue Was determined by real-time RT-PCR;protein expression of type I collagen(Coll),aminopeptidase P (APP),hypoxia indHeible factor let and 2α(HIF-1α and HIF-2α)in kidney tissue Was examined by immunohistochemistry staining. Results Hb values of normal rats presented normal distribution. The normal Hb was (155.9±16.5) g/L. Rat anemia was diagnosed when Hb was below 123.6 g/L. There was no difference in all the examination results between CG and MG before administration (P>0.05). Compared with CG, the Hb and Cer in MG were significantly decreased [(121.66±15.68) g/L vs (169.00±12.89) g/L, (0.63±0.13) ml/min vs (1.27±0.18) ml/min, P< 0.01], and the UP in MG was significantly increased at the end of the 8th week [(27.04±9.40) mg/d vs (6.11±0.84) mg/d, P<0.01]; the relative areas of fibrosis and Col l in renal interstitium of MG were significantly enlarged [(12.89±2.33)% vs (0.55±0.10)%, (13.92±2.92)% vs (1.32±0.84)%, P<0.01]; the protein expression of APP and the mRNA expression of EPO in the kidney tissue of MG were significantly down-regulated [(0.55±0.23)% vs (3.77±1.06)%, 0.005±0.001 vs 0.032±0.013, P<0.01]; the protein expression of HIF-lα and HIF-2α in the kidney tissue of MG was significantly up-regulated (2.55±0.16 vs 1.12±0.46, 2.33±0.33 vs 1.15±0.27, P<0.01), at the end of the 8th week. Conclusions The pathogenesis of anemia in CAAN may be due to the decreased production of EPO caused by the destruction of peritubular capillary. The compensatory up-regulation of HIF-lα and HIF-2α expression can not prevent the anemia development.
4.Study of glomerular podocyte injury induced by aristolochic acid
Hong CHENG ; Yipu CHEN ; Hongrui DONG ; Yanyan WANG ; Hongliang RUI
Chinese Journal of Nephrology 2012;28(3):222-225
Objective To explore whether the glomerular podocytes can be damaged by aristolochic acid. Methods Thirty-two male SD rats were equally divided into the following 2 groups:model group in which the rats received the extract of Aristolochia manshuriensis Kom (AmK) by gavage; control group only received tap water by gavage.24 h urinary protein excretion was measured at the end of the 1st and 4th week,and SDS-PAGE gel electrophoresis was performed to detect the protein in urine.At the end of the 4th week,all the rats were sacrificed and the glomeruli were isolated by laser capture microdissection technique.The mRNA expression of nephrin,podocin,CDA2P,podocalyxin and podoplanin in isolated glomeruli was determined by RT-PCR,and the average width of glomerular foot process was measured by electron microscopy and image analysis. Results At the end of the 4th week,24 h urinary protein excretion in the model group was significantly higher than that in the control group (P<0.01) and the urinary albumin content in model group was also obviously increased.The average width of glomerular foot process in the model group was significantly larger than that in control group (P<0.01).The mRNA expressions of nephrin,podocin,CDA2P,podocalyxin and podoplanin in glomeruli were significantly down-regulated in the model group compared with the control group,which decreased by 34%,62%,56%,50%(P<0.01) and 27% (P<0.05),respectively. Conclusions Aristolochic acid can damage the glomerular podocytes,resulting in the down-regulation of nephrin,podocin,CD2AP,podoplanin and podocalyxin mRNA expression, the segmental widening of foot process, and increased urinary protein excretion.
5.Hirsutella sinensis antagonizes the effect of aristolochic acid on human proximal tubular epithelial cells
Mei ZHANG ; Gongyao TANG ; Hongliang RUI ; Yipu CHEN
Chinese Journal of Nephrology 2005;0(08):-
Objective To study whether Hirsutella sinensis (HS) can antagonize aristolochic acid (AA) induced fibrogenesis on human proximal tubular epithelial cells (HKC). Methods HKC were incubated with medium alone, medium with HS 10 mg/L, medium with AA-Na 40 mg/L or medium with AA-Na 40 mg/L and HS 10 mg/L, respectively. After 12 h (for mRNA) or 36 h (for protein) ,cells were lysed,and the mRNA and protein expression level of transforming growth factor-?1 (TGF-?1), connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase-1 (TIMP-1)and plasminogen activator inhibitor-1 (PAI-1) of HKC were measured by RT-PCR, ELISA (for TGF-?1, TIMP-1 and PAI-1) and Western blotting (for CTGF), respectively. Results The mRNA and protein expression of TGF-?1, PAI-1, CTGF and TIMP-1 were significandy up-regulated by AA-Na 40 mg/L. Compared with the control group, the mRNA expression of TGF-?1,.CTGF, TIMP-1 and PAI-1 was up-regulated to 1.24,1.31,1.27 and 1.36 times,respectively (P
6.Adipogenic and tenogenic differentiation of tendon-derived stem cells isolated from an animal model of chronic tendinopathy in vitro
Hui CHEN ; Yucheng LIN ; Hongliang XU ; Chen WANG ; Yunfeng RUI
Chinese Journal of Tissue Engineering Research 2014;(45):7320-7326
BACKGROUND:Chronic tendinopathy is a tendon disorder extremely common in athletes and in the general population with repetitive strain injuries of tendons. The pathogenesis of tendinopathy remains unclear and hence treatment of tendinopathy is usual y pal iative.
OBJECTIVE:To investigate the of adipogenic and tenogenic ability of patel ar tendon-derived stem cel s isolated
from chronic tendinopathy and healthy rats in vitro.
METHODS:Tendon-derived stem cel s were isolated from patel ar tendons of chronic tendinopathy and healthy rats respectively. The tendon-derived stem cel s were cultured to the 3rd passage in complete culture medium, and cel morphology was observed. The cel s were divided into adipogenic induction group and control group. Cel s in the adipogenic induction group were cultured in adipogenic induction medium, while those in the control group cultured in complete culture medium. The ability of adipogenic differentiation between tendon-derived stem cel s isolated from the tendon of chronic tendinopathy and healthy rats in vitro was examined by oil red O staining and quantification assay. The mRNA expressions of C/EBPαand PPARγ2 were detected by real-time quantitative PCR. When 70%-80%cel s were confluent, the mRNA expressions of Col1a1, Scx, Tnmd and Dcn were also detected by real-time quantitative PCR.
RESULTS AND CONCLUSION:At the third passage, slender spindle-shaped cel s were seen in both two groups, but there was a little change in the cel morphology in the chronic tendinopathy group. Lipid droplets were formed after the cel s were cultured in adipogenic induction medium for 21 days. This was not observed in the control group. We observed more oil red O-positive oil droplets in tendon-derived stem cel s from the tendons of chronic tendinopathy rats than healthy rats. The difference between them was statistical y significant (P=0.004). The results of real-time quantitative PCR showed that the mRNA expressions of C/EBPαand PPARγ2 in the tendon-derived stem cel s from the tendons of chronic tendinopathy rats were significantly higher than those in tendon-derived stem cel s from the tendons of healthy rats (P=0.004);the mRNA expressions of Col1a1, Scx, Tnmd and Dcn in the tendon-derived stem cel s from the tendons of chronic tendinopathy rats were significantly lower than those in tendon-derived stem cel s from the tendons of healthy rats (P=0.009). In conclusion, tendon-derived stem cel s from chronic tendinopathy rats showed a higher ability of adipogenic differentiation, but a lower capacity of tenogenic differentiation compared to tendon-derived stem cel s from healthy rats, which might contribute to better understand the pathogenesis of tendinopathy.
7.Effect of Medical Ethics on the Quality of Life and Treatment Compliance for Breast Cancer Patients
Nanlin LI ; Hui WANG ; Hongliang WEI ; Rui LING
Chinese Medical Ethics 2016;29(2):254-256
Objective:To analyze and discuss the influence of medical ethics on the quality of life and treatment compliance for breast cancer patients in clinical application, and to expound the positivehidden value of medical ethics to dealing with the doctor-patient relationship. Methods:176 cases of breast cancer patients in a hospital from December, 2012 to December, 2014 were selected to participate in this study. They were divided into experi-mental group (n=88) and control group (n=88) using the sequence number. The subjects in control group were treated according to the NCCN guidelines for breast cancer, while the experimental group was treated with medical ethics on the basis of the routine methods. The effect and safety were explored. Result:In evaluation of the quality of life, it was found that the excellent rate of patients in the experimental group was significantly higher than those in the control group (89. 8% vs. 68. 2%, P<0. 05). The incidence of complications in the study group was sig-nificantly lower than the control group ( P<0 . 05 ) . The complication incidence in the experimental group was sig-nificantly lower than that of the control group (73. 9% vs. 96. 6%,P<0. 05). Conclusion:Medical ethics has a great influence on the quality of life and treatment compliance for breast cancer patients, and its application has a positive indirect and clinical utility.
8.Data collection of signals in the multi-channel sEMG system of masticatory muscles and development and preliminary clinical application of an analytic system.
Hongliang DU ; Xin LI ; Shan LI ; Rui ZHANG ; Rong SONG ; Lan LI ; Wei WANG ; Hong KANG
Journal of Biomedical Engineering 2014;31(1):23-28
The aim of this study was to design a simple, economic, with high Common Mode Rejection Ratio (CMRR), preamplifier and multi-channel masticatory muscle surface electromyography (sEMG) signal acquisition system assisting to diagnose temporomandibular disorders (TMD). We used the USB interface technology in the EMG data with the aid of the windows to operate system and graphical interface. Eight patients with TMD and eight controls were analyzed separately using this system. In this system, we analyzed sEMG by an optional combination of time domain, frequency domain, time-frequency, several spectral analysis, wavelets and other special algorithms under multi-parameter. Multi-channel sEMG System of Masticatory Muscles is a simple, economic system. It has high sensitivity and specificity. The sEMG signals were changed in patients with TMD. The system would pave the way for diagnosis TMD and help us to assess the treatment effect. A novel and objective method is provided for diagnosis and treatment of oral-maxillofacial disease and functional reconstruction.
Algorithms
;
Computer Graphics
;
Data Collection
;
Electromyography
;
Humans
;
Masticatory Muscles
;
physiology
;
physiopathology
;
Sensitivity and Specificity
;
Signal Processing, Computer-Assisted
;
Temporomandibular Joint Disorders
;
diagnosis
;
User-Computer Interface
9.Wnt-7a inhibites epithelial to mesenchymal transition in mice of unilateral ureteral obstruction model
Peiling BAO ; Guoqin WANG ; Hongliang RUI ; Guolan XING ; Hong CHENG ; Yipu CHEN
Chinese Journal of Nephrology 2012;28(9):720-724
Objective To investigate the protective effects of Wnt-7a protein on renal interstitial fibrosis in mice of unilateral ureteral obstruction (UUO)model.Methods Eighteen male C57BL/6 mice were randomly divided into 3 groups:sham-operation group,the UUO model group and Wnt-7a treatment group.The body weight of mice was measured everyday.All the mice were sacrificed at thc seventh day after the operation.The left kidney was taken for histology evaluation and molecular biology assay.Masson's stain was performed as a main indicator of interstitial fibrosis.The expression of vimentin,α-smooth muscle actin,and E-adherin in renal tissue was detected by immunohistochemistry staining and the expression of α-smooth muscle actin and E-cadhe(nn) in renal tissue was detected by Western blotting.Results Compared with sham-operation group,body weight of the (,)odel group was significantly lower (P<0.05),and the relative area of interstitial fibrosis was significantly larger (P<0.05).Furthermore,the expression of vimentin and α-SMA was significantly up-regulated (P<0.05),and the expression of E-cadherin was significantly down-regulated (P<0.05).Compared with model group,all the above-mentioned abnormalities were restored to some extent and showed significant differences (P<0.05) in Wnt-7a treatment group.Conclusion Wnt-7a protein can decrease the interstitial fibrosis by inhibiting epithelial to mesenchymal transition in UUO mice.
10.Valproic acid suppresses SIRT1 inducing hepatotoxicity
Xiangyu HOU ; Jing JIN ; Hongliang LI ; Rui LIU ; Xiaomei FAN ; Min HUANG
Chinese Pharmacological Bulletin 2015;(1):31-34
Aim To detect the role of sirtuin1 ( SIRT1 ) in hepatotoxity caused by valproic acid ( VPA) . Methods The changes of SIRT1 expression of HepG2 cells were detected by Western blot. And then SIRT1 plasmid or siRNA was transfected to con-struct SIRT1 overexpressed or knocked-down HepG2 cells. Furthermore, SRB assays were taken to observe the changes of viability of these cells exposed to VPA. Results VPA suppressed SIRT1 expression in a time and dose-dependent manner. SIRT1 overexpression showed a protective effect to the cytotoxicity caused by VPA, and the IC50 before and after transfection was (4. 025 ± 0. 47) and (10. 87 ± 1. 50) mmol·L-1 re-spectively. Moreover, transfection of SIRT1 siRNA sensitized HepG2 cells to VPA, and the IC50 before and after transfection was (1. 938 ± 0. 16) and (0. 663 ± 0. 05) mmol·L-1 respectively. Conclusion VPA suppressed SIRT1 expression in HepG2 cells and over-expression of SIRT1 could reduce cytotoxicity induced by VPA.