1.Angiotensin 1-7 attenuates left ventricular remodeling in deoxycorticosterone acetate salt hypertensive rats
Chinese Pharmacological Bulletin 2014;(7):994-999
Aim To investigate the effect of angioten-sin 1-7 (Ang 1-7 )on the cardiac hypertrophy and my-ocardial fibrosis in deoxycorticosterone acetate (DOCA)-salt hypertensive rats and its possible mecha-nism.Methods Male Sprague-Dawley rats were used to establish DOCA-salt hypertensive model,which un-derwent uninephrectomy surgery and were subcutane-ously injected with a DOCA,and replaced drinking water with 1% saline solution for 4 weeks.DOCA-salt animals were implanted with osmotic minipumps, which delivered Ang 1-7 chronically for 4 weeks (200 ng ·kg-1 ·min-1 ).Arterial blood pressure,left ven-tricular function in rats,the area of myocardial cells in HE stained specimens,and the area of myocardial fi-brosis Sirius red staining specimens were measured. Real time PCR was used to detect the expression of at-rial natriuretic factor (ANF ) and β-myosin heavy chain (β-MHC)mRNA in heart,and TGF-β1 protein expression was observed by Western blot in myocardial tissue.Results In the first week,DOCA salt rat had a significant increase in arterial blood pressure,and reached a peak in the fourth week;while the left ven-tricular end-systolic pressure (LVESP),left ventricu-lar end-diastolic pressure (LVEDP ) and ventricular contraction the maximum rate of pressure rise (+dp/dt)had also undergone significant changes in DOCA salt rats. After chronic infusion of Ang 1-7 for 4 weeks,the arterial pressure,LVESP and LVEDP were significantly reduced and +dp/dt were increased sig-nificantly in DOCA salt rats (P<0.05 ,n=7 ).Ang 1-7 significantly reduced the cardiac index and myocar-dial cell area,as well as the up-regulated expression of ANF and β-MHC mRNA in DOCA salt rats (P <0.05 ,n =7 ).Meanwhile,Ang 1-7 also significantly decreased the perivascular fibrosis and interstitial fibro-sis area, and significantly inhibited the increase of TGF-β1 expression in DOCA salt rats (P<0.05 ,n=7 ).Conclusion These results indicate that Ang 1-7 has a cardioprotective effects through reducing arterial pressure and improving cardiac fibrosis and hypertro-phy in the DOCA-salt model of hypertension.
2.Effect and safety of different concentrations of disintegrin Echistatin to early posterior capsule opacification in diabetic rabbits
Guangxia, QIAN ; Shaojian, TAN ; Hao, LIANG ; Yingying, CHEN
Chinese Journal of Experimental Ophthalmology 2013;32(11):975-979
Background Posterior capsular opacification (PCO) is a primary cause of blurred vision after extra-capsular cataract extraction (ECCE),and there is a higher incidence of PCO in the patients with diabetes mellitus.Echistatin (Ecs) can suppress the proliferation of lens epithelial cells (LECs) and thereby inhibit the formation of PCO.However,its mechanism and safe dose deserve to study.Objective The aim of this study was to observe the inhibitory effect of different concentrations of Ecs on LECs proliferation in the early stage of PCO in diabetic rabbits and explore the safe dose of Ecs.Methods Diabetic mellitus was induced by injection of 90 mg/kg alloxan via ear vein in 15 New Zealand white rabbis.ECCE was performed in the right eyes of the rabbits.The rabbits were randomized to the control group and 5.0,7.5,10.0 and 15.0 μg/ml Ecs group according to randomized number table method.Ecs of 0.2 ml in above doses was injected into the anterior chamber after ECCE in different concentrations of Ecs groups,and 0.2 ml distilled water was used in the same way in the only diabetic rabbits as the control group.Postopeartive response of ocular anterior segment was observed and PCO was graded under the slit lamp microscope.The corneal and retinal specimens were prepared 10 days after operation for the assay of preliferative cell nuclear antigen (PCNA) expression in LECs by immunochemistry to evaluate the effective dose of Ecs.Regular histopathological examination was performed,and apoptosis of corneal endothelial cells and retinal cells was detected by TUNEL method to assess the safe concentration of Ecs.Results Different degrees of corneal edema and exudation in anterior chamber were seen in the eyes of different groups.The inflammatory response disappeared 3-5 days in the control group and 5.0 μg/ml Ecs group and 7 days in the ≥7.5 μg/ml Ecs groups.PCO was 1-2 grade in the control group and 5.0 μg/ml Ecs group and 0 grade in the ≥ 7.5 μg/ml Ecs groups.The difference in the positive expression level (absorbance,A) for PCNA in LECs was significantly different among the control group and various Ecs groups (F=18.006,P=0.001),and the positive expression level of PCNA in the ≥ 7.5 μg/ml Ecs groups was markedly reduced in comparison with that in the control group (P =0.010,0.001).Hematoxylin and eosin staining showed an normal morphology and order arrangement in corneal endothelial cells and intact structure in retinal internal limiting membrane in the groups.TUNEL assay revealed that the apoptosis values (mean A value) of corneal endothelial ceils and retinal cells in the ≤ 10.0 μg/ml Ecs groups were not significantly changed in comparison with the control group (all at P>0.05),but the apoptosis values in the 15.0 μg/ml Ecs group were markedly higher than those in the control group (P=0.004,0.018).Conclusions Ecs can inhibit the early PCO in diabetic rabbits and show the optimal effect at the concentrations of 7.5 and 10.0 μg/ml without visible eytotoxicity to eye other tissues.Therefore,these two doses of Ees might be used for the study of long-term therapeutic effectiveness.
3.Effect of acupuncture using Xingnao Kaiqiao needling method on patients with post-stroke depression
Xijun HE ; Jilin TAN ; Benguo WANG ; Yingying LAI
Chinese Journal of Rehabilitation Theory and Practice 2005;11(6):467-468
ObjectiveTo investigate the effect of acupuncture using Xingnao Kaiqiao needling method on patients with post-stroke depression (PSD).Methods172 PSD patients were randomly divided into the acupuncture group and control group with 86 cases in each group. The acupuncture group was treated with acupuncture using Xingnao Kaiqiao needling method, and different symptoms were also punctured various acupoints; the control group was given fluoxetine treatment, 20 mg/d. Two groups were given the same routine treatment and early rehabilitation, 8 weeks being a course.ResultsThe total effective rates of the acupuncture group and control group were individually 89.54% and 73.26%, and there was a significant difference between two groups (P< 0.05).ConclusionAcupuncture treatment using Xingnao Kaiqiao needling method can effectively treat post-stroke depression without obvious side effect.
4.Upregulation of brain APJ receptor expression induced by high salt in pressure overload rats
Qi ZHANG ; Yingying TAN ; Dongmin LIU ; Xiaoming ZHAO
Chinese Pharmacological Bulletin 2015;(5):630-635
Aim To observe the effect of high salt stress on sympathetic nerve activity and brain Apelin and APJ system in pressure overload rats.Methods The suprarenal abdominal aorta was banded in male SD rats to create the pressure overload model.Four weeks later,the rats were fed a high-salt (8%NaCl)diet or a regular-salt (0.3% NaCl)diet for 1 week,and the hemodynamic changes of the rats were recorded.Sym-pathetic activity was evaluated by measuring 24 h uri-nary norepinephrine (NE)by ELISA.Real-time RCR and Western blot method were used to analyze the Ape-lin and APJ expression in the paraventricular nucleus of rats.In another protocol,ML22 1 or benzamil was intracerebroventricularly infused in the aorta banding rats fed with high-salt,and levels of 24 h urinary NE and cardiac index were recorded.In addition,normal SD rats were intracerebroventricularly infused with Na+-rich aCSF for 1 week,and the changes of blood pressure,24 h urine-NE and APJ expression levels were detected.Results In the pressure overload rats fed with high salt diet for 1 week,the mean arterial blood pressure (MAP),heart rate (HR),left ventric-ular end systolic pressure (LVESP),24 h urinary NE and APJ mRNA protein expression in paraventricular nuclear were significantly increased compared with those in control groups (P<0.05 ).However,the he-modynamics,24 h urinary NE and APJ expression showed no significant change in the sham rats fed a high salt diet.In addition,ICV infusion of APJ recep-tor antagonist ML221 or the ENaC antagonist benzamil significantly inhibited the 24 h urinary NE level and HW/BW ratio in the pressure overload rats fed high salt diet (P <0.01 ).Intracerebroventricular infusion of high Na+aCSF for 1 week in normotensive rats also significantly increased the MAP,HR,urinary-NE lev-els and brain APJ expression (P <0.0 1 ).Conclu-sions High salt increases the sympathetic nerve activ-ity in rats with pressure overload model, which is closely related to the brain APJ receptor expression and Na+sensitivity.Brain APJ receptor may be an impor-tant target in the development of salt sensitivity.
5.Diagnostic value of MSCT for appendix mucinous cystadenoma
Jing TAN ; Jun FENG ; Yingying DING ; Dafu ZHANG
Journal of Practical Radiology 2014;(12):2007-2009
Objective To discuss the MSCT performances of appendix mucinous cystadenoma in order to improve the preoperative diagnosis.Methods MSCT plain and enhanced findings of mucinous appendix mucinous cystadenoma proved by pathology in 6 pa-tients were analyzed retrospectively.Results CT showed cystic dilatation of the appendix in 2 patients with heterogeneous density, and cystic mass in the right iliac fossa in 4.As for the cystic wall,uniform thin wall was seen in 4,curvilinear mural calcification in 3 and septation in 2.Dynamic enhanced CT showed the ring mural enhancement in 4.In addition,the blurred surrounding fatty tis-sues were seen in 2.Conclusion MSCT findings of mucinous appendiceal cystadenoma appeared as cystic dilatation or cystic mass in the right iliac fossa,curvilinear mural calcification and enhanced wall.
6.CT Diagnosis of Osteolytic Metastases of the Vertebral Column
Yingying DING ; Depei GAO ; Jing TAN ; Kun LI
Journal of Practical Radiology 2001;0(08):-
Objective To study CT value of diagnosis and identified diagnosis in osteolytic metastases of the vertebral column through describing their CT manifestation.Methods In 46 patients, 72 vertebrae osteolytic metastases were analyzed and compared with X-ray findings of 28 cases.Results In 72 vertebrae osteolytic metastases, there were destruction of 74 corpus vertebrae in 45 patients, of pediculus arcus vertebraes in 22 cases, of processus transverses in 15 cases, of processus spinosus in 11 cases and lamina vertebrae in 11 cases. Micrometastases were concentrated in corpus vertebrae(45/49). CT found the rate of bone-destruction, affection of vertebrae canal and soft tissue around vertebrae were 100%, 67.8% and 71.4%, but the corresponding rates were only 53.6%, 14.3% and 32.1% by X-ray. Conclusion Vertebral column metastases destructed corpus vertebrae at first, vertebral metastases are the origin or base of destruction of the pediculus arcus vertebrae, lamina vertebrae, processus transverses and processus spinous. CT scanning is more sensitive in finding and evaluating the lesions than X-ray plain film.
7.Protective effects of nicotinamide mononucleotide on ethanol-induced DNA damage in L02 cells
DI Chunhong ; YIN Jie ; ZHONG Wenying ; ZHANG Yingying ; CAO Yuejia ; TAN Xiaohua
Journal of Preventive Medicine 2024;36(6):548-552
Objective:
To investigate protective effects of nicotinamide mononucleotide (NMN) on ethanol-induced DNA damage in L02 cells, so as to provide the evidence for adjuvant therapy of NMN on alcoholic liver diseases.
Methods:
L02 cells were pretreated with different concentrations of NMN (0, 1, 2, 4 and 8 mmol/L) for 6 h, and then were exposed to 0.4% ethanol for 12 h. The treated cells were divided into the control group, 0.4% ethanol group and different concentrations of NMN groups. Cell viability was analyzed using trypan blue staining for determining the concentration of NMN as a protective agent. The effects of NMN on ethanol-induced DNA damage in L02 cells were evaluated using immunofluorescence detection and reactive oxygen species (ROS) assay. L02 cells were exposed to 0.4% ethanol for 12 h, cultured in a medium containing a protective concentration of NMN, and divided into PBS group and NMN group. Cell viability was detected at 0, 2, 4, 8, 16 and 32 h, and the effects of NMN on repairing ethanol-induced DNA damage were evaluated by alkaline comet assay.
Results:
The cell viability was lower in 0.4% ethanol group than than in the control group, and was higher in different concentrations of NMN groups than in 0.4% ethanol group (all P<0.05), with no significant difference in the cells viability between 4 mmol/L and higher concentrations of NMN groups and the control group (all P>0.05). Therefore, 4 mmol/L NMN was selected as a protective agent. The cell tail moments, relative immunofluorescence intensities of γH2AX and relative levels of ROS were higher in 0.4% ethanol group than in the control group, and lower in 4 mmol/L and higher concentrations of NMN groups than in 0.4% ethanol group (all P<0.05). The cell viability was increased and the cell tail moment was shortened with the increase of 4 mmol/L NMN intervention time; and the cell viability in 4 h and more of NMN groups were higher, and the cell tail moment were lower than that in PBS group (all P<0.05).
Conclusions
NMN attenuates DNA damage in a dose-dependent manner and promotes the repair of DNA damage in a time-dependent manner. NMN has a protective effect on ethanol-induced DNA damage in hepatocytes.
8.Knockout and identification of the surface antigen 43 gene in escherichia Coli JM109
Yonghao HUANG ; Huange ZHAO ; Songlin ZHOU ; Yingying LIN ; Guanghong TAN ; Fengying HUANG
Chongqing Medicine 2015;(31):4330-4332
Objective To knockout and identify the Antigen 43 (Ag43) in the Escherichia Coli JM109 .Methods Mutation group Ⅱ introns RNA protein complexes (RNP) gene sequence was obtained by Sigma Company′s TargeTron Gene Knockout Sys‐tem and Ag43 gene specific designed PCR primers amplification ,then ,to acquired Ag43 specific recombinant RNP plasmid pACD4K‐Ag4 ,this gene sequence was inserted into the plasmid pACD4K‐C of RNA′s expression .Finally ,pEGFP‐Ag43 was trans‐formed into JM109 and inserted the group Ⅱ intron into the Ag43′s locus by IPTG inducing expression .Results The best insertion locus was between 1 812 and 1 913 .Through the agarose electrophoresis gel ,the RNP gene sequence was consistent with the expec‐ted value (350 bp) .The pEGFP‐Ag43 vector was correctly constructed which was proofed by endonuclease Nhe Ⅰ and Hind ⅡI di‐gestion as predicted products (3 646 and 4 029 bp;7 000 and 550 bp ,respectively ) .The PCR and gene sequence results indicated that the group Ⅱ intron was inserted into the locus between 1 812 and 1 913 in the Ag43 gene .Conclusion Successful knockout of the Ag43 in Escherichia Coli JM109 found basis to further study the Ag43′s function and regard the coli as host bacteria of Ag43 chimeric protein recombinant .
9.Acupuncture on the Basic Fibroblast Growth Factor and Type Ⅰ Collagen in Colons of Rats with Crohn's Disease
Chen ZHAO ; Juying DING ; Jindan MA ; Linying TAN ; Huangan WU ; Yingying ZHANG ; Linshan ZHANG ; Zhen WANG
Journal of Acupuncture and Tuina Science 2011;09(1):1-6
Objective:To observe the impacts of herb-partitioned moxibustion,warm moxibustion and electroacupuncture on the basic fibroblast growth factor(bFGF)and collagen type Ⅰ(Col Ⅰ)in colons of rats with Crohn' disease(CD),and discuss the mechanism of acupuncture therapy on the intestinal fibrosis in CD.Methods:The model rats were developed by TNBS as multiple proinflammatory method.The rats were randomly divided into 5 groups:a normal group,a model group,a warm moxibustion group,an electroacupuncture group and a herb-partitioned moxibustion group.The treatments were carried out at Tianshu(ST 25)(bilateral)and Qihai(CV 6)in different treatments.The immunohistochemistry was used to detect the expression position of Col Ⅰ and bFGF.Results:The expressions of Col Ⅰ and bFGF in colons of rots in the model group significantly increased(compared with the normal group,P<0.01).After the herb-partitioned moxibustion,warm moxibustion and electroacupuncture,the expressions of Col Ⅰ and bFGF reduced markedly in the rats with CD(P<0.01).The expression of bFGF and Col Ⅰ in the colons had an obvious correlation in the Spearman rank correlation analysis.Conclusion:Acupuncture treatment reduced the abnormally high levels of expressions for Col Ⅰ and bFGF in colons.Col Ⅰ and bFGF participated in the fibrosis.Acupuncture treatment may reduce the bFGF expression in colons to regulate the excessive deposition,treating the intestinal fibrosis in CD.
10.Hydrogen sulfide reduce renal ischemia/reperfusion injury by NOD-like receptor pathway
Zhicheng TAN ; Yuanyuan SHI ; Yan YAN ; Yingying WU ; Wenli LIU ; Guangyuan LI ; Rongshan LI
Chinese Journal of Nephrology 2014;30(8):604-608
Objective To investigate whether the nod-like receptor (NLR) pathway is involved in protection of hydrogen sulfide (H2S) preconditioning during renal ischemia reperfusion.Methods Male Wistar rats were randomly divided into 3 groups:sham operation (Sham) group,renal ischemia/ reperfusion (I/R) group subjected to occlusion of left renal pedicle for 45 min then reperfusion for 24 hours,and sodium hydrosulfide (NaHS) preconditioning group with continuous infusion of NaHS (300 nmol/min) by left renal artery for 15 min before I/R treatment.Renal injuries were evaluated by HE staining.The protein levels of NOD1,NOD2,nuclear NF-κB P65 and caspase-1 were analyzed by Western blot assay.The protein level of MCP-1 and IL-1β expressions was determined by immunohistochemical staining assay.Cell apoptosis were evaluated by Tunel staining assay.Results In I/R group,the renal NOD1 and NOD2 protein expressions were upregulated.Moreover,the nuclear NF-κB P65 expression was also elevated with an increase in its target genes-MCP-1 and IL-1β (All P < 0.01).HE staining revealed the existence of acute tubular necrosis in I/R kidney.TUNEL staining revealed more apoptotic cells in risk zone with the activation of caspase-1 of I/R-treated kidney(P <0.01).NaHS preconditioning reversed I/R-induced increase in the expression of NOD1 and NOD2(P <0.05).NaHS preconditioning also reduced I/R-induced activation of NF-κB P65 (P < 0.05) and upregulation of MCP-1 and IL-1β (P < 0.01).Moreover,NaHS preconditioning attenuated inflammation,repressed caspase-1 activation and reduced apoptotic cells after I/R.Conclusion Hydrogen sulfide preconditioning can alleviate renal ischemia/reperfusion injury by Nod-like receptor dependent on inflammatory pathway.