2.Expression of Toll-Like Receptor in Peripheral Blood Mononuclear Cells of Rats with Nephrotic Syndrome Induced by Respiratory Syncytial Virus
jin, WU ; zheng, WANG ; yan-nan, GUO ; hong-yu, DUAN
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To explore the expression and the role of Toll-like receptor(TLR3 and TLR4) in rats with nephrotic syndrome induced by respiratory syncytial virus(RSV).Methods SD rats were inoculated intranasally and intraperitoneally with 6?106 plaque for-ming unit(PFU) RSV to construct RSV-induced nephropathy in rat model.Rats were anesthetized and blood was withdrawn from cardiac on day 4,14,30,60 after inoculation.The normal ones without intervention were set as control group.The renal histology was observed by light microscope and electron microscope.The urinary protein collected in 24 hours were measured.Meanwhile,the expressions of TLR3 and TLR4 were detected by indirect immunofluorescence staining and flow cytometry in peripheral blood mononuclear cells of rats.The results were analyzed by SPSS 13.0 softwore.Results After inoculation,the proteinuria increased and under the electron microscope the foot processes of glomerular epithelial cells were fused which resembled human minimal change nephrotic syndrome.Proteinuria reached the peak and the fusion of foot processes were most extensive in rats of RSV at 60 d.The expressions of TLR3 and TLR4 in each group of RSV-induced nephropathy in rat models were significantly higher than those in normal control group(Pa0.05).Conclusions TLR3 and TLR4 in peripheral blood mononuclear cells of RSV-induced nephropathy rat mo-dels had being significantly activated until 60 d after RSV inoculation.TLR signaling pathway may play an important role in nephrotic syndrome of rats induced by RSV.
3.Effect of propofol on the apoptosis of PC12 cell induced by glutamic acid
Zheng LI ; Jin-Chun SI ; Xiang-Yu GAO ; Zhe LIU ; Nan LIANG ; Zheng NAN
The Chinese Journal of Clinical Pharmacology 2016;32(12):1115-1117
Objective To explore the effect of propofol on the apoptosis of PC12 cell induced by glutamic acid.Methods PC12 cells were in-duced 10 mmol · L-1 glutamate for 48 h, and were divided into model group , propofol low , medium and high dose groups.The normal cells were used as control group.Control group and model group were received culture medium without any drugs for 48 h.Propofol low , medium and high dose groups were given 12.5 , 25.0 , 50.0 μmol · L-1 propofol for 48 h.The viability of PC12 cell was measured by MTT assay.PC12 cell apoptosis was measured by flow cytometry.The activity of Caspase -3 was determined by spectrophotometric method.The expression of FBJ osteosarcoma oncogene ( c -fos ) and early growth response protein 1 ( Egr -1 ) were detected by Realtime -PCR and western blot.Results Compared with the normal group , the cell apoptosis rate , the activity of Caspase -3 , the expression of c -fos mRNA and protein increased , and the cell viability , the expression of Egr -1 mRNA and protein decreased in the model group ( P<0.01 ).Compared with the model group, the acitivity of Caspase -3, the cell apoptosis rate , the activity of Caspase -3, the expression of c-fos mRNA and protein decreased , and the cell viability , the expression of Egr-1 mRNA and protein increased in propofol low -dose , medium -dose and high -dose group ( P <0.05 ).Conclusion Propofol suppressed the apoptosis of PC 12 cell induced by glutamic acid , which was related with the expression of c-fos and Egr-1.
5.The kinetics of in vitro lysozyme deposition on rigid gas-permeable contact lens for a short period
Jin ZHANG ; Ziyi WAN ; Kaihui NAN ; Qi ZHENG ; Jia QU ; Tingfei XI
Chinese Journal of Tissue Engineering Research 2010;14(3):473-476
BACKGROUND: The contact lenses were easily contaminated by adsorbing components from the tear film, particularly protein after wearing for a period of time. Lysozyme adsorption dynamics of fluorosilicone acrylate contact lenses has been studied in order to further improve data of protein adsorption, reduce adsorbing amount of surface protein, and prevent surface contamination of contact lenses.OBJECTIVE: To investigate the adsorption dynamics of fluorosilicone acrylate contact lenses to lysozyme in vitro. METHODS: A stock solution of lysozyme was prepared in Hanks balanced salt solution (2.0 g/L, solution Ⅰ) and different trifluoroacetic acid (TFA) concentrations were prepared. Recovery experiment, the contact lenses were placed in shaking incubator at 37 ℃ for varying time intervals. After incubation there was a single rinsing in Hanks balanced salt solution. Contact lenses in control group were placed in diluted water, and contact lenses in the other group were placed in different concentrations of TFA. For deposition, FSA contact lenses in experimental group were placed in shaking incubator at 37 ℃ for varying time intervals. After incubation there was a single rinsing in Hanks balanced salt solution. Then FSA contact lenses were immersed in 0.2% TFA solution. The amount of lysozyme was assayed with BCA method.RESULTS AND CONCLUSION: Lysozyme which attached to fluorosilicone acrylate contact lenses could be resolved by TFA, and the recovery was influenced by the immersed time and the concentration of TFA. The optimal time was 1 hour, and the optimum concentration was 0.2%. The adsorption dynamics of lysozyme on FSA contact lenses was a second-phased process, i.e., lysozyme adsorption increased rapidly during 10 minutes-1 hour, reached a plateau at 1 hour, stably adsorbed during 1-24 hours, and reached a saturation of 0.349 mg/cm~2. The recovery of lysozyme was lower at 10 and 30 minutes, but reached 90%-100% while the time of incubation was between 40 minutes and 24 hours.
6.TNF-αpromotes apoptosis of rat islet βcell line
nan Nan LIANG ; Qian ZHANG ; zheng Xiang WU ; hui Jia ZHAO ; Jin WANG ; ying Xiang JIAO
Basic & Clinical Medicine 2017;37(12):1668-1673
Objective To observe the effect of tumor necrosis factor-α( TNF-α) on islet cell apoptosis and TXNIP expression.Methods INS-1 cells were cultured in vitro, treating with TNF-α(0, 1, 5, 10 and 20 mg/L).We tested the effect of TNF-αon cell viability by CCK-8.INS-1 cells were treated with TNF-α( 5 mg/L, 24 h) for the proper concentration and incubation time; mRNA expression of TXNIP and ChREBP were measured by real-time PCR;in addition, protein levels of TXNIP , ChREBP and FOXO1 were analyzed by Western blot .Results TNF-αdecreased the survival rate of INS-1 cells in a dose-dependent manner ( P<0.05 ) , and induced apoptosis;protein and mRNA expression of TXNIP and ChREBP were significantly higher than that in control group ( P<0.05 );while the expression of protein level of FOXO 1 was down-regulated .Conclusions TNF-αinduces apoptosis in INS-1 cells and aggravates the cells damage .
8.Changes of left ventricular remodeling in hypertension patients with carotid atherosclerosis of phlegm-dampness syndrome.
Nan LI ; Hui-Ying YE ; Guan-Yi ZHENG ; Xiu-Ying CHEN ; Hua-Pin HUANG ; Jin-Guo LI ; Xu-Dong SUN
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(4):402-405
OBJECTIVETo study changes of left ventricular remodeling (LVR) in hypertension patients with carotid atherosclerosis (CAS) of phlegm-dampness syndrome (PDS).
METHODSDoppler ultrasonography data of CAS were observed in 223 hypertension patients with CAS (as the hypertension group, including 119 patients of the PDS group and 104 of the non-PDS group), 81 CAS patients with non-hypertension, and 19 non-hypertension non-CAS patients (as the control group). The difference in the degree of LVR was compared among the above groups.
RESULTSThe left ventricular posterior wall thickness (LVPWT), inter ventricular septum thickness (IVS), E/A were higher in the hypertension group than in the non-hypertension group (P < 0.05). The left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic diameter (LVESD), stroke volume (SV) were higher in the soft plaque hypertension group and the soft plaque non-hypertension group than in the hard plaque group, the thickening intimal group, and the normal intimal group (P < 0.01 , P < 0.05). The LVEDD, LVESD, and SV were higher, and the ejection fraction (EF) was lower in the PDS hypertension group than in the non-PDS hypertension group (all P < 0.05). Of them, LVEDD, LVESD, and SV were higher in the soft plaque group than in the hard plaque group (P < 0.01), the thickening intimal group (P < 0.01) and the normal intimal group (P < 0.05). There was no statistical difference in PDS hypertension between the soft plaque group and the hard plaque group (P > 0.05).
CONCLUSIONThe hypertension patients with CAS of PDS might be correlated to LVR, and LVR was more obviously in the soft plaque patients.
Aged ; Aged, 80 and over ; Carotid Artery Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Case-Control Studies ; Female ; Humans ; Hypertension ; diagnosis ; diagnostic imaging ; physiopathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ultrasonography ; Ventricular Remodeling
9.Selecting methods of controls concentration for internal quality control and continuity of control chart between different reagent lots for HBsAg qualitative detection.
Jin-ming LI ; Huai-jing ZHENG ; Lu-nan WANG ; Wei DENG
Chinese Journal of Hepatology 2003;11(4):228-231
OBJECTIVETo establish a model for one choosing controls with a suitable concentration for internal quality control (IQC) with qualitative ELISA detection, and a consecutive plotting method on Levey-Jennings control chart when reagent kit lot is changed.
METHODSFirst, a series of control serum with 0.2, 0.5, 1.0, 2.0 and 5.0ng/ml HBsAg respectively were assessed for within-run and between-run precision according to NCCLs EP5 document. Then, a linear regression equation (y=bx + a) with best correlation coefficient (r > 0.99) was established based on S/CO values of the series of control serum. Finally, one could choose controls with S/CO value calculated from the equation (y = bx + a) minus the product of the S/CO value multiplying three-fold between-run CV to be still more than 1.0 for IQC use. For consecutive plotting on Levey-Jennings control chart when ELISA kit lot was changed, the new lot kits were used to detect the same series of HBsAg control serum as above. Then, a new linear regression equation (y2 = b2x2 + a2) with best correlation coefficient was obtained. The old one (y1 =b1x1 + a1) could be obtained based on the mean values from above precision assessment. The S/CO value of a control serum detected by new lot kit could be changed to that detected by old kit lot based on the factor of y2/y1. Therefore, the plotting on primary Levey-Jennings control chart could be continued.
RESULTSThe within-run coefficient of variation CV of the ELISA method for control serum with 0.2, 0.5, 1.0, 2.0 and 5.0ng/ml HBsAg were 11.08%, 9.49%, 9.83%, 9.18% and 7.25%, respectively, and between-run CV were 13.25%, 14.03%, 15.11%, 13.29% and 9.92%. The linear regression equation with best correlation coefficient from a test at random was y = 3.509x + 0.180. The suitable concentration of control serum for IQC could be 0.5ng/ml or 1.0ng/ml. The linear regression equation from the old lot and other two new lots of the ELISA kits were y1 = 3.550(x1) + 0.226, y2 = 3.238(x2) +0.388, and y3 =3.428(x3) + 0.148, respectively. Then, the transferring factors of 0.960 (y2/y1) and 0.908 (y3/y1) were obtained.
CONCLUSIONThe results shows that the model established for IQC control serum concentration selecting and for consecutive plotting on control chart when the reagent lot is changed is effective and practical.
Enzyme-Linked Immunosorbent Assay ; methods ; standards ; Evaluation Studies as Topic ; Hepatitis B Surface Antigens ; blood ; Humans ; Quality Control ; Reagent Kits, Diagnostic ; standards ; Reproducibility of Results
10.The efficacy and safety of antiviral treatment in inactive hepatitis B surface antigen carriers
Zhenhuan CAO ; Yali LIU ; Lina MA ; Junfeng LU ; Yi JIN ; Zhimin HE ; Nan GENG ; Yanhong ZHENG ; Xinyue CHEN
Chinese Journal of Infectious Diseases 2017;35(7):387-392
Objective To evaluate the feasibility and safety profile of pegylated-interferonα-2a (Peg IFNα-2a) combined with adefovir dipivoxil (ADV) in inactive hepatitis B surface antigen (HBsAg) carriers (IHC).Methods This was a single center, prospective and open-label study.IHC were divided into therapeutic group (T, 112 subjects) and control group (C, 72 subjects) according to personal willingness.Patients with hepatitis B virus (HBV) DNA<20 IU/mL were treated with Peg IFNα-2a monotherapy, and those with HBV DNA ≥20-<2 000 IU/mL were treated with Peg IFNα-2a combined with ADV.Total therapy duration was 96 weeks.For patients who achieved HBsAg seroconversion and continued consolidation treatment for 24 weeks, the treatment duration could be less than 96 weeks.t test was used for continuous variable comparison between the two groups, while chi-square test or Fisher′s exact probability method was used for counting data analysis.The related factors affecting HBsAg clearance was analyzed by univariate or multivariate logistic regression analysis.Results A total of 194 patients were enrolled with 112 in therapeutic group and 72 in control group.The HBsAg clearance rate and seroconversion rate at week 48 in therapeutic group were 30.8% (32/104) and 26.0% (27/104), respectively.The rates at week 96 increased to 45.2% (47/104) and 38.5% (40/104), respectively.The HBsAg clearance rates at weeks 48 and 96 in control group were both 1.5% (1/68).HBsAg seroconversion was not achieved in control group.The HBsAg clearance rate in treatment group was significantly higher than that in control group (χ2=39.066, P<0.01).The quantitative HBsAg levels at baseline (OR=2.313, 95%CI: 1.258-4.251, P=0.007), week 12 (OR=3.159, 95%CI: 1.826-5.466, P<0.01) and week 24 (OR=3.347, 95%CI: 2.050-5.465, P<0.01), the decline of HBsAg at week 12 (OR=5.343, 95%CI: 2.085-13.689, P<0.01), and week 24 (OR=4.855, 95%CI: 2.380-9.902, P<0.01), and alanine transaminase (ALT) elevation at week 12 (OR=3.520, 95%CI: 1.369-9.052, P=0.009) were independent predictors for HBsAg clearance.Conclusions Peg IFNα-2a-based treatment for IHC could achieve higher HBsAg clearance rate and seroconversion rate, and has a safety profile.Decline of HBsAg at week 12 and week 24 with ALT elevation at week 12 could predict a higher HBsAg clearance rate.