1.The Effect of Rennin-angiotensin System on Endothelin-1 and Nitric Oxide in Rat Glomerulosclerosis
Zhequan JI ; Cuiwen HUANG ; Jiecheng LIANG
Journal of Chinese Physician 2001;0(09):-
Objective To investigate the correlation among glomerulosclerosis and endothelin-1(ET-1) and nitric oxide(NO), and the effect of blocking renin-angiotensin system(RAS) on rat glomerulosclerosis. Methods 30 SD rats underwent unilateral nephrectomy plus adriamycin (6mg/kg) injection through caudal vein to establish rat glomerulosclerosis models. These rats were divided randomly into glomerulosclerosis group (group D), Benazepril treatment group (group DB) and Losartan treatment group (group DL). Another 10 SD rats served as sham-operation group (group C). 6 weeks after treamtent, the mRNA and protein expressions of ET-1 and iNOS in renal cortex were detected using RT-PCR and Western blotting analysis respectively, and the content of fibronectin(Fn) was measured using immunohistochemical method. Results Group D occurred massive proteinuria, hypoalbuminemia and hypercholesterolemia, which had significant differences compared with group C (P
2.Tanshinone IIA protects against triptolide-induced liver injury via Nrf2/ARE activation.
Cuiwen GUAN ; Jing JIN ; Jia LI ; Zhongxiang ZHAO ; Zhiying HUANG
Acta Pharmaceutica Sinica 2013;48(9):1397-402
The aim of this study is to investigate the protection effect of tanshinone IIA (Tan) against triptolide (TP)-induced liver injury and the mechanisms involved. Acute liver injury was induced by intraperitoneal injection of TP (1 mg x kg(-1)) in mice. The activities of AST, ALT and LDH in serum and the levels of GSH, GST, GSH-PX, SOD, CAT and MDA in liver tissue were detected. The histopathological changes of liver tissues were observed after HE staining. Nrf2 translocation in liver tissue was detected by Western blotting, and real-time PCR was used to measure the expression levels of GCLC, NQO1 and HO-1 mRNA. The results showed that pretreatment with Tan significantly prevented the TP induced liver injury as indicated by reducing the activities of AST, ALT and LDH (P < 0.01). Tan pretreatment also prevented TP-induced oxidative stress in the mice liver by inhibiting MDA and restoring the levels of GSH, GST, SOD and CAT (P < 0.05). Parallel to these changes, pretreatment with Tan could attenuate histopathologic changes induced by TP. Furthermore, the results indicated that Tan pretreatment caused nuclear accumulation of Nrf2 as well as induction of mRNA expression of antioxidant response element (ARE)-driven genes such as GCLC, NQO1 and HO-1. These results indicated that Tan could protect against TP-induced acute liver injury via the activation of Nrf2/ARE pathway.
3.Schisandrin B protects against nephrotoxicity induced by cisplatin in HK-2 cells via Nrf2-ARE activation.
Mei LI ; Jing JIN ; Jia LI ; Cuiwen GUAN ; Wenwen WANG ; Yuwen QIU ; Zhiying HUANG
Acta Pharmaceutica Sinica 2012;47(11):1434-9
This study is to investigate the protection effect of schisandrin B (Sch B) against oxidation stress of HK-2 cells induced by cisplatin and the mechanisms involved. HK-2 cells were cultured and divided into different groups: solvent control group, cisplatin exposure group, positive group, Sch B treatment group. Cell viability and toxicity were evaluated by MTT and LDH assay. GSH level and SOD enzymes activities were also measured. DCFH-DA as fluorescence probe was used to detect ROS level by fluorescence microplate reader. Nrf2 translocation was detected by Western blotting. Real time Q-PCR was used to detect expressions of NQO1, HO-1 and GCLC mRNA level. The results showed that Sch B could significantly inhibit the decline of cell viability induced by cisplatin treatment (P < 0.05) and the protective effect was in a dose dependent manner. Furthermore, Sch B treatment significantly inhibited the increase of ROS level induced by cisplatin and reversed the decrease of GSH level (P < 0.05). When Sch B concentration was up to 5 micromol x L(-1), SOD enzyme activities were also enhanced significantly compared with that of the cisplatin group (P < 0.05). It was shown that Sch B could cause nuclear accumulation of Nrf2 in association with downstream activation of Nrf2 mediated oxidative response genes such as GCLC, NQO1 and HO-1. These results suggested Sch B could protect against the oxidative damage of HK-2 cells induced by cisplatin via the activation of Nrf2/ARE signal pathway.
4.Effect of human recombinant erythropoietin on inflammation of hyperoxic lung injury in neonatal rats
Qili HUANG ; Jing REN ; Yanmei XIE ; Zequan JI ; Haiyan LIU ; Cuiwen HUANG
Chinese Journal of Applied Clinical Pediatrics 2020;35(5):383-388
Objective:To explore the effects of human recombinant erythropoietin (rhEPO) on inflammation of hyperoxic lung injury in neonatal rats.Methods:Seventy-two neonatal rats were randomly divided into 4 groups: control group, BPD group, hyperoxia + low-dose recombinant erythropoietin [EPO(L)]group, and hyperoxia + high-dose recombinant erythropoietin [EPO(H)]group.The neonatal rats in BPD group, hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were exposed to 850 mL/L oxygen.Then the neonatal rats in hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were given 800 IU/kg and 2 000 IU/kg rhEPO by subcutaneous injection respectively at 1 d, 3 d and 7 d, while the control group and BPD group were given the same amount of 9 g/L saline water.Initially, the body weight of each group was recorded at 3 d, 7 d and 14 d. The morphological structure changes of lung tissues were observed by HE staining under light microscope, and the radial alveolar count(RAC) in lung tissues were detected.The expression of nuclear factor kappa B(NF-κB) was detected by immunofluorescence staining; Western blot was applied to determine the protein expression of phosphorylated NF-κB(pNF-κB), inhibitor protein(IκB) and Caspase-3 in lung tissues; and the expression of interleukin-1β(IL-1β) in bronchoalveolar lavage fluid was determined using enzyme linked immunosorbent assay (ELISA).Results:(1) On the 14 th day, the body weight of neonatal rats in the BPD group was lower than that in the control group [(18.97±3.21) g vs.(27.97±2.30) g], and the difference was statistically significant( P<0.01); however, the body weights of neonatal rats in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group[(24.16±2.15) g, (26.04±1.97) g] was much heavier than that in the BPD group, and the differences was statistically significant(all P<0.05). (2) The morphological structure of lung tissues which was observed by HE staining showed that in the BPD group, there were a few inflammatory cells infiltration in alveolar septum on the 3 rd day, the inflammatory response was more evident on the 7 th day, when exudation could be seen in the alveolar cavity; and on the 14 th day, the number of pulmonary alveoli decreased, pulmonary bulla formed, and septa were thickened.Besides, it was also observed that compared with control group, RAC was significantly decreased in BPD group on the 14 th day(5.50±1.29 vs.14.33±2.80), and the difference was statistically significant( P<0.01). Pathological changes were ameliorated and the infiltration of inflammatory reaction cells was reduced in the hyperoxia+ EPO(L)group and hyperoxia + EPO(H)group.RAC was remarkably higher in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group than that in the BPD group on the 14 th day, and the differences were statistically significant (all P<0.05). (3)Immunofluorescence results showed that: the number of NF-κB p65 positive cells increased significantly and fluorescence intensity increased in the BPD group, while EPO could greatly reduce the number of NF-κB p65 positive cells and lower the fluorescence intensity.(4)Western blot results indicated that compared with the control group, the expressions of pNF-κB p65 and Cleaved Caspase-3 was significantly increased, and the differences were statistically significant (all P<0.05); and IκB was significantly lower, and the difference was statistically significant ( P<0.05). The expression of NF-κB p65 and Cleaved Caspase-3 was significantly lower, but IκB was significantly higher in the hyperoxia+ EPO(L)group and the hyperoxia+ EPO(H)group than those in the BPD group, and the differences were statistically significant (all P<0.05). (5) ELISA results revealed that the expression of IL-1β in the BPD group was significantly higher than that in the control group, and the difference was statistically significant ( P<0.05); Compared with BPD group, the expression of IL-1β was significantly lower in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group, and the differences were statistically significant (all P<0.05). Conclusions:EPO can reduce hyperoxia-induced lung tissue apoptosis and protect newborn rats against hyperoxic lung injury by decreasing the inflammatory response of cells through the NF-κB pathway on BPD.
5.Evaluation on the performance of clinical laboratory automation system
Minghong TONG ; Xuemei ZUO ; Hui DING ; Zhonghua HUANG ; Xin WEI ; Tinying ZHANG ; Chenxia DENG ; Xiangming CUI ; Cuiwen CHEN
Chinese Journal of Laboratory Medicine 2017;40(10):810-815
Objective By analysis of the key performance indexes of the clinical laboratory automation system, to clarify the advantage and optimize the comprehensive performance of the laboratory automation system.Methods Key performance indexes were Collected from January 2017 to April 2017 in biochemistry and immunoassay group of Clinical Laboratory of Shanghai Tong Ren Hospital.(1)The data were collected and compared by the before-and-after method,the starting time of the automation system and initial sample test were analyzed.(2)Key performance indexes were analyzed for the time of specimen registration,inspection,and reporting.(3)The specimen turnaround time(TAT)was analyzed based on two months operation of the laboratory automation system.In view of disadvantage of infectious assays, setting up priority sample absorption, then TAT performance was re-evaluated.(4)By the assessment of total serum dosage required in the automation system, the number of blood vacuum tubes were reduced reasonably.The pros and cons of laboratory automation system were analyzed and the potential improvement were proposed.Results (1)According to the sample peak shift forward,the system start time could move forward 30 minutes earlier.(2)With the adopting of railway logistics,the specimens were sent to the lab and the registration time was at 7:25 am,and the time required for specimen delivery was greatly reduced which made specimen test,report and audit time all moved forward accordingly.(3)Data has shown that specimen TAT declined dramatically based on the performance of the first two month operation of the automation system,biochemical items were shortened 2 h,and the immunoassay shortened 4 h,respectively.Moreover the trend keeps better gradually.With setting up priority absorption infectious tests,the TAT was improved greatly,TAT reduced the average by 40 min.(4)500 μl(including the sample in dead space of vacuum tube)were needed for all the 65 biochemical items included in the system, and 1 495 μl serum were used for the 28 immunoassay.As a result, a total of 2 000 μl serum will be enough for sample analysis by the system, which provided the feasibility to reduce 3 vacuum tubes averagely.Considering the current automation system does not include all the analysis items in our lab directory, a few tests remain to be performed on offline instruments respectively.The methodology for some infectious agents are different from previous method, therefore some test results may need a period of time for comprehensive clinical appreciation.Furthermore,due to the parallel connection of multiple instruments included in the system, more rigorous and frequent quality control becomes a necessity,which may rely on more strict quality control procedure to guarantee the quality.Conclusions The application of the automation system significantly enhanced the efficiency of clinical laboratory all round.In addition, by the quantitative indicators, it is possible to monitor the system operation performance real time, which may feedback and facilitate the improvement constantly,and result in auto confirmation the majority results,eventually.