1.Inhibitory effect of erythropoietin on high glucose-induced transition of human kidney proximal tubular epithelial cells and its possible mechanism
Yanxia CHEN ; Xiaohua QIN ; Xiangdong FANG ; Chong HUANG ; Weiping TU
Journal of Medical Postgraduates 2014;(10):1038-1041
Objective The core mechanism of renal insterstitial fibrosis (RIF) is epithelial-mesenchymal transition.This study aimed to investigate the effect of erythropoietin on high glucose-induced epithelial-mesenchymal transition ( EMT) of normal hu-man kidney proximal tubular epithelial cells (HK-2) and its possible mechanism. Mothods HK-2 cells cultured in vitro were ran-domly divided into a blank control group , a high glucose induction group , a mannitol induction group , an EPO induction group , an EPO (5, 10, and 20U/mL) inhibition group, and an Rho kinase inhibitor group.After 24 hours of intervention, the mRNA levels of RhoA and ROCK were determined by RT-PCR, those of E-cadherin and α-smooth muscle actin (α-SMA) proteins detected by immu-nofluorescence staining , and the expression of FN proteins in the supernatant measured by ELISA . Results Compared with the blank control group , the expressions of RhoA and ROCK 1 mRNA were significantly increased in the high glucose induction group (0.945 ±0.132 vs 1.400 ±0.022, 1.007 ±0.002 vs 1.913 ±0.011, P<0.05), but markedly decreased in the 5, 10, and 20U/mL EPO inhibition groups (1.400 ±0.022 vs 1.278 ±0.006, 1.400 ±0.022 vs 0.770 ±0.005, 1.400 ±0.022 vs 0.334 ±0.009, P<0.006) in comparison with the high glucose induction group , and the effects were related to the concentration of EPO .Compared with the blank control, the expression of E-cadherin protein was increased in the high glucose induction group (0.644 ±0.006 vs 0.107 ± 0.004, P<0.05), but remarkably decreased in the 5, 10, and 20 U/mL EPO inhibition groups (0.236 ±0.006, 0.433 ±0.010, 0.521 ±0.010) in comparison with the high glucose induction group (P<0.05), and the effects were also related to the concentration of EPO.Pearson correlation analysis showed a positive correlation between the mRNA expressions of RhoA and ROCK 1 in the high glu-cose induction and EPO inhibition groups . Conclusion EPO can inhibit high glucose-induced epithelial-mesenchymal transition of normal human kidney HK-2 cells and thus delay renal fibrosis , which mignt be related to the RhoA/ROCK signaling pathway .
2.The mechanism of RCE-4, an active ingredient of Reineckia carnea , in combination with celecoxib on the anti-proliferation of cervical cancer Ca Ski cells
Hong-heng ZHEN ; Fang-fang YOU ; Fan CHENG ; Kun ZOU ; Chong-xu CHEN ; Jian-feng CHEN
Acta Pharmaceutica Sinica 2021;56(7):1911-1920
This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-
3.Anti-cicatricial effect of tetrandrine drug delivery system in glaucoma filtration surgery in rabbit
Ying-ying, ZHENG ; Hong-bo, CHENG ; Fang-wei, YING ; Ming, LI ; Chong, WEN ; Qing, CHEN
Chinese Journal of Experimental Ophthalmology 2011;29(4):328-331
Background Scarring of the filtering bleb is a main cause of filtering surgical failure in glaucoma.It has been reposed that tetrandrine could suppress the proliferation of cultured human fibroblast of Tenons capsule in vitro and thus has the potential effect to prevent scarring after the filtering surgery. Objective Present study was to investigate the anti-cicatricial effect of tetrandrine drug delivery system(Tet DDS)during filtration surgery. Methods Filtration surgery was performed in bilateral eyes of 18 New Zealand white rabbits.The Tet DDS with 0.3 mg Tet,0.2 mg Tet or free-Tet were implanted subcunjunctially during the surgery.The filtering blebs were scored in 1 day,4,7,10,14 days after referring to the corneal thickness and bleb range under the slit-lamp biomicroscopy.The morphology of filtering bleb was assessed by in vivo confocal microscopy in 7 and 14 days after operation.The filtering bleb specimen was prepared in 7 and 14 days for the histopathological examination. Results The filtering bleb scores in Tet DDS implantation groups were significantly higher than those in free-Tet DDS group from 4 days through 14 days after trabeculectomy(P<0.01),and the scores showed a considerably increase in 0.3 mg Tet DDS group compared with 0.2 mg Tet DDS group from 7 days through 14 days after trabeculectomy(P<0.05).The filtering blebs of Tet DDS implantation groups were found with distinct subepithelial cystic spaces under the light microscopy and in vivo confocal microscopy on the 7th day and 14th day after surgery.Compared with free-Tet DDS group,the numbers of subepithelial mierocysts were much more(P<0.01)and the area of microcysts was larger(P<0.01)in Tet DDS group.The filtering tissue presented with more subepithelial microcysts and larger microcysts range in 0.3 mg Tet DDS group than 0.2 mg Tet DDS group in 7 and 14 days after operation(P<0.05).The inflammatory cell infiltration wag milder in 0.3 mg Tet DDS group in comparison with 0.2 mg Tet DDS group and free-Ted DDS group.Conclusion Tet DDS has strong inhibitory effects on inflammatory cells activity and fibroblagt activity the early stage after filtering surgery and therefore improve the surgery success rate.
4.Diagnostic significance of differential cell count in induced sputum to chronic cough
Wei LUO ; Ru-Chong CHEN ; Chun-Li LIU ; Ke-Fang LAI ; Nan-Shan ZHONG ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To explore the diagnostic significance of differential cell count in induced sputum to chronic cough and assessment of airway inflammation.Methods The sputum of 335 chronic cough patients were induced.Differential cell counts were measured in these samples.The side effects were observed during the induced procedure.The final diagnosis was made based on clinical manifestation and examination findings including pulmonary function tests,provocation test,induced sputum cell differentials, etc.Results The cause of chronic cough was defined in 322 patients.The six most important causes of cough were typical asthma(TA,n=84),eosinophilic bronchitis (EB,n=62),atopic cough (AC,n= 42),cough variant asthma (CVA,n=40),gastroesophageal reflux cough(GERC,n=37),rhinitis and/ or paranasal sinusitis (PNDs,n=32),and others and indefinite cause (n=25,13).Percentage of eosinophils were significantly increased in the induced sputum of AC,EB,CVA,and GERC patients (0.005,0.052,0.059,0.234) compared with those in other causes and the healthy controls (0) (P
5.Effects of erythropoietin in high glucose induced proliferation and apoptosis of human kidney proximal tubular epithelial cells and the possible mechanism
Yanxia CHEN ; Xianfeng WU ; Xiangdong FANG ; Xiaohua QIN ; Chong HUANG ; Weiping TU
Tianjin Medical Journal 2015;(1):25-29
Objective To study the effects of erythropoietin (rhEPO) in high glucose induced proliferation and apopto?sis of human kidney proximal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells cultured in vitro were divided into several groups randomly:blank control group, high glucose group, mannitol group, rhEPO control group, different concentrations of rhEPO treatment groups (5, 10, 20 U/mL) and Rho kinase group. The reverse tran?scription polymerase chain reaction (RT-PCR) was used to evaluate the mRNA levels of RhoA and ROCK after 24 hours. Tetrazolium salt method (MTT) was used to determine the cell proliferation. Cell apoptosis was detected by flow cytometry. Results Compared with blank control group the expression levels of RhoA and ROCK1 mRNA were significantly in?creased in high glucose group (P < 0.05). RhoA, ROCK1 mRNA expressions significantly decreased in rhEPO group than those of high glucose group (P<0.05). There was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in high glucose group and rhEPO group. MTT method showed that rhEPO significantly promoted the prolifer?ation of HK-2 cells (P<0.05). Flow cytometry analysis showed that high glucose induced apoptosis in HK-2 cells, which was significantly inhibited in rhEPO group and Rho kinase group as compared to that of high glucose group in a concentra?tion dependent manner (P<0.05). Conclusion rhEPO can promote HK-2 cell proliferation and inhibit apoptosis, which may be related to RhoA/ROCK signaling pathway.
6.Research progress and thinking on improving physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine based on crystal structure
Ying-chong CHEN ; Wei-cheng ZHOU ; Fang-fang LEI ; Li-zhen HUANG ; Qin ZHENG ; Ming YANG ; Peng-fei YUE
Acta Pharmaceutica Sinica 2021;56(8):2059-2069
The active ingredients in traditional Chinese medicine have been reported to possess significant pharmacological activity and played an important role in clinical treatments. However, lots of the active ingredients in traditional Chinese medicine suffer from disadvantages such as low solubility, high melting point and low stability that results in low bioavailability and limit its clinical application. Crystal structure plays an important role in improving physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine. This review concludes the research advances of several crystal forms used in the active ingredients in traditional Chinese medicine in terms of polymorph, cocrystal, amorphous/coamorphous and nanocrystal. And the effects of crystal forms on the physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine were reviewed. This research may be useful for the formulation preparation and development of the active ingredients in traditional Chinese medicine.
8.The effect of Th17 cells on A(H1N1) influenza virus clearance
Mutong FANG ; Yutian CHONG ; Guilin YANG ; Mingfeng LIAO ; Yingxia LIU ; Mingxia ZHANG ; Weilong LIU ; Xinchun CHEN ; Boping ZHOU
Chinese Journal of Infectious Diseases 2010;28(10):593-596
Objective To investigate the phenotype, frequency of Th17 cells and the association between Th17 cells and viral clearance in patients with H1N1 influenza A. Methods Three groups including 70 confirmed patients with H1N1 influenza A, 30 patients with seasonal influenza as well as 68 healthy subjects as controls were enrolled in this study. The percentages of Th1, Th2, Treg and Th17 lymphocytes in the peripheral blood were determined by intracellular staining and flow cytometry. The levels of interferon-γ (IFN-γ), transforming growth factor-beta (TGF-β),interleukin-6 (IL-6) in plasma and supernatant of the peripheral blood mononuclear cell (PBMC)culture were quantified by enzyme-linked immunosorbent assay (ELISA). Viral load in nasopharyngeal swabs was detected by real time quantitative reverse transcription-polymerase chain reaction (RTPCR). Data were analyzed by one way ANOVA and liner correlation analysis. Results The percentage of Th17 cells in H1N1 influenza A patients was (2. 740±0. 210)%, which the percentage of was significantly decreased compared to healthy subjects (3. 443 ±0. 154)% and seasonal influenza patients (3. 443±0. 277) % (F=4. 242, P<0. 05); while the percentage of Thl, Th2 and Treg cells were not significantly different among these groups. Moreover, the TGF-β level in plasma of H1N1 influenza A patients was (10±8) ng/mL, which was significantly lower than healthy subjects (43 ±32 ) ng/mL and seasonal influenza patient ( 18 ± 10) ng/mL ( F= 17.72, P<0.01 ). The TGF-β level in the supernatant of PBMC culture of H1N1 influenza A patients was (782 ± 736) pg/mL, which was significantly lower than healthy subjects (1462±315) pg/mL and seasonal influenza patients (1481 ±348) pg/mL (F=5. 730, P<0.01). Additionally, the viral clearance period was inversely correlated with the percentage of Th17 cells (r=-0.38, P=0.02). Conclusions The proportion of Th17 cells in patients with H1N1 influenza A is significantly decreased, which is closely correlated with the level of TGF-β. This decrease may results in the delayed viral clearance.
9.Detection of Viral Etiology of Children with Acute Respiratory Infection in Wenzhou Area from 2005 to 2006
shu-yan, CAO ; xiao-fang, CHEN ; xiao-hong, CAI ; meng-rong, LI ; jian, GONG ; xiu-di, WANG ; chang-chong, LI ; lin, DONG
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate the 7 kinds of respirovirus etiology of children with acute respiratory infection(ARI) in Wenzhou area from 2005 to 2006.Methods Three thousand nine hundred and seventy children with ARI visited the Yuying children's hospital were chosen,including 308 children with acute upper respiratory infection(URI) and 3 662 children with lower respiratory infection(LRI).Direct immunofluorescence(DIF) was used to detect the respiratory syncytial virus(RSV),adenovirus(ADV),influenza virus(IV) A and B,parainf-luenza virus(PIV) type 1,2,3 from nasopharyngeal secretions(NPS) collected from these patients.Results Among the 3 970 samples,1 773(44.7%) positive results were determined and the positive rate of RSV(36.2%) was the highest.The isolating rate of respirovirus were all conspicuous difference in sex(?2=9.2 P
10.The diagnostic value of comparison of ~(18)F-FDG PET/CT with contrast-enhanced CT in detecting primary hepatic carcinoma and postoperative recurrence
Xiao-yan, WANG ; Xiang-song, ZHANG ; Zhi-feng, CHEN ; Fang, LI ; Zi-ping, LI ; Guo-hui, RAO ; Xin-chong, SHI ; ping, HU
Chinese Journal of Nuclear Medicine 2010;30(1):15-18
Objective To compare diagnostic value of ~(18)F-fluoredeoxyglucose (FDG) PET/CT with contrast-enhanced CT in detecting primary hepatic carcinoma and postoperative recurrence.Methods Twenty-five cases of primary hepatic carcinoma or postoperative recurrent tumor underwent whole-body ~(18)F-FDG PET/CT and contrast-enhanced CT within one week's interval.They were retrospectively reviewed and the difierences between these two modalities were investigated.Results Of these 25 cases,there were 13 cases with primary hepatocellular carcinoma.1 case with intrahepatic cholangiocarcinoma and 11 cases with postoperative recurrence.The sensitivity of 18 F-FDG PET/CT and contrast-enhanced CT in diagnosing primary hepatic carcinoma was 78.6%(11/14) and 92.9%(13/14),and sensitivity in diagnosing postoperative recurrent was 100.0%(11/11) and 63.6%(7/11) respectively.Conclusion Contrast-enhanced CT may have a slight advantage over PET/CT in detecting primary hepatic carcinoma,but ~(18)F-FDG PET/CT combined with contrast-enhanced CT has even greater accuracy.Meanwhile,~(18)F-FDG PET/CT has better diagnostic accuracy in detection of postoperative recurrent tumor.