1.Effect of endogenous nitric oxide on superoxide dismutase-1 activity and apoptosis of vascular endothelial cells
Zaifeng ZHANG ; Xiuli WANG ; Shangyue ZHANG ; Xiaoyu TIAN ; Lulu ZHANG ; Junbao DU ; Hongfang JIN ; Yaqian HUANG
Chinese Journal of Applied Clinical Pediatrics 2021;36(15):1176-1180
		                        		
		                        			
		                        			Objective:To investigate the regulatory effects of endogenous nitric oxide (NO) on the activity of superoxide dismutase-1 (SOD1) and apoptosis of human umbilical vein endothelial cells (HUVECs).Methods:HUVECs were taken as the research object.The endothelial NO synthase (eNOS) short hairpin RNA(shRNA) lentivirus was employed to transfect HUVECs to knock down eNOS.HUVECs were divided in 4 groups: the scramble group, the eNOS shRNA group, the eNOS shRNA + sodium nitroprusside(SNP) group and the eNOS shRNA+ SNP+ tris (2-carboxyethyl) phosphine hydrochloride (TCEP) group.The protein expressions of eNOS and SOD1 dimer/monomer in cells were detected by western blot.The activity of SOD was detected by the enzyme-linked immunosorbent assay.The NO content in cells was detected with NO fluorescence probe.The level of superoxide anion in HUVECs was detected with dihydropyridine (DHE). The terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay was adopted to detect the apoptosis of HUVECs in situ.Results:Compared with the scramble group, the endogenous NO content (2.690±0.420 vs.15.029±2.193, P<0.01), eNOS protein expression (1.000±0.778 vs.3.141±0.199, P<0.01), SOD1 dimer/monomer ratio (4.6±1.0 vs.7.6±2.0, P<0.05) and SOD activity [(0.432±0.254) Carmen′s unit/10 4 cell vs.(1.000±0.116) Carmen′s unit/10 4 cell, P<0.01] were significantly decreased, while the level of intracellular superoxide anion (11.180±1.560 vs.6.146±1.007, P<0.01) and HUVECs apoptosis [75.0 (55.0, 100.0)% vs.0 (0, 0)%, P<0.01] were significantly increased in the eNOS shRNA group.Compared with the eNOS shRNA group, the content of endogenous NO (16.705±0.116 vs.2.690±0.420, P<0.01), the ratio of SOD1 dimer/monomer (7.3±2.0 vs.4.6±1.0, P<0.05) and the activity of SOD [(0.737±0.060) Carmen′s unit/10 4 cell vs.(0.432±0.254) Carmen′s unit/10 4 cell, P<0.05] were significantly increased, while the level of superoxide anion (6.897±1.648 vs.11.180±1.560, P<0.01) and the HUVECs apoptosis [0 (0, 0)% vs.75.0 (55.0, 100.0)%, P<0.01] were significantly decreased in the eNOS shRNA+ SNP group.Compared with the eNOS shRNA + SNP group, the ratio of SOD1 dimer/monomer (4.4±0.9 vs.7.3±2.0, P<0.05) and the activity of SOD [(0.214±0.084) Carmen′s unit/10 4 cell vs.(0.737±0.060) Carmen′s unit/10 4 cell, P<0.01] were significantly decreased, while the level of superoxide anion (10.917±1.552 vs.6.897±1.640, P<0.01) and the apoptosis level of HUVECs[63.6 (55.0, 90.0)% vs.0 (0, 0)%, P<0.01] were significantly increased in the eNOS shRNA+ SNP+ TCEP group.However, there was no significant difference in the NO content (16.112±0.926 vs.16.705±0.116, P>0.05). Conclusions:Endogenous NO could effectively antagonize the apoptosis of endothelial cells by increasing the cysteine-dependent SOD1 dimer/monomer ratio, enhancing SOD activity and inhibiting the accumulation of reactive oxygen species.
		                        		
		                        		
		                        		
		                        	
2.Optimization of Anti-infective Program after Surgery for a Case of Infant Patient with Intussusception and Enterobrosis by Clinical Pharmacists
Changhai SU ; Junbao LIU ; Peitao XIE ; Duo HAO ; Xiaojuan ZHANG
China Pharmacy 2020;31(21):2672-2676
		                        		
		                        			
		                        			OBJECTIVE:To provide th e ideas a nd for individualized anti-infective treatment of infection after surgery for infants and young children with intussusception and enterobrosis ,and to provide reference for clinical pharmacists participating in the clinical treatment. METHODS :Clinical pharmacists optimized the anti-infection program for an 11-month-old infant patient infected after surgery with intussusception and enterobrosis in Ordos Central Hospital ;they put forward medication suggestions in respects of the selection of initial anti-infection treatment program ,drug replacement ,the selection of anti-infection treatment program after blood culture showed Enterococcus coli and Enterococcus faecium ,and dosage adjustment. RESULTS :According to the judgment of the common pathogens and the hospital or community infections in the infant patient with intussusception and enterobrosis,cefoperazone sulbactam 1.0 g,q12 h was adjusted to cefoperazone sulbactam 0.5 g,q8 h combined with Metronidazole chloride sodium injection 20 mL,q8 h;when the blood culture showed E. coli (ESBL-)and E. faecium ,it was recommended to add vancomycin 0.15 g,q12 h. After poor treatment ,it was recommended to adjust the vancomycin dose to 0.2 g,q8 h. All the above suggestions were adopted by doctors. And the child ’s body temperature dropped after treatment ,the blood culture turned negative and laboratory indicators returned to normal. The child was discharged smoothly. CONCLUSIONS :Infants and young children are special groups. Therefore ,before using antibiotics ,clinical pharmacists should evaluate the age ,body weight ,liver and kidney functions of infants and young children. They should also help doctors select and adjust drugs ,frequency and dosage on the basis of pharmacokinetic characteristics and safety ,so as to avoid adverse drug reactions while ensure curative effect.
		                        		
		                        		
		                        		
		                        	
3.Special issues in diagnosis of Kawasaki disease
Chinese Journal of Applied Clinical Pediatrics 2020;35(13):961-964
		                        		
		                        			
		                        			Kawasaki disease (KD) is an acute, self-limiting, and medium-sized vasculitis, which has been the commonest cause of acquired heart disease in children in developed countries.Without timely diagnosis and treatment, up-to 25% of the affected children may develop coronary artery abnormalities (CAA). Due to the lack of the specific diagnostic method, KD is mainly diagnosed according to the clinical criteria.As a result, typical KD is recognized easily, but it is a big challenge to diagnose KD patients with incomplete or atypical symptoms.The pandemic of novel coronavirus disease 2019 (COVID-19) around the world makes the diagnosis of KD even more complex.In this review, hot issues in diagnosing KD were discussed according to the 2017 guidelines for diagnosis and treatment of KD recently published by the American Heart Association (AHA), expecting to provide help for diagnosis of KD children.
		                        		
		                        		
		                        		
		                        	
4.Changes of facial volume maintenance rate after single autologous fat grafted for repaired progressive facial hemiatrophy
Junbao CHEN ; Binghang LI ; Jianjian LU ; Jiajie XU ; Chao ZHANG ; Fang XIE ; Liya YANG ; Lu YANG ; Shuyuan LI ; Li TENG
Chinese Journal of Medical Aesthetics and Cosmetology 2019;25(4):302-305
		                        		
		                        			
		                        			Objective To measure the change of facial volume maintenance rate after autologous fat grafted for repaired progressive facial hemiatrophy by using three-dimensional digital technology.Methods 3D scanner was used to acquire facial data in 10 patients with progressive facial hemiatrophy before operation;Mimics 17.0 software was used to reconstruce patients' facial 3D model and to calculate the volume of facial tissue defect;autologous fat was grafted to repair facial deformity.The facial volume maintenance rate was calculated in all the patients 3 months and 6 months after operation.Results We had performed facial 3D data acquisition and facial repaired with autologus fat grafted in 10 patients;patients' facial morphology was improved.The mean facial volume maintenance rate was (35.80±3.44)% in 3 months and (27.82±3.80)% 6 months after surgery.Conclusions The mean facial volume maintenance rate in postoperative 3 months is inferior to that in 6 months in single autologous fat grafted for repairing progressive facial hemiatrophy.
		                        		
		                        		
		                        		
		                        	
5. Effects of CD34+ selected stem cells for the treatment of poor graft function after allogeneic stem cell transplantation
Xinhong FEI ; Junbao HE ; Haoyu CHENG ; Yuming YIN ; Weijie ZHANG ; Shuqin ZHANG ; Xiaocan WANG ; Jingbo WANG
Chinese Journal of Hematology 2018;39(10):828-832
		                        		
		                        			 Objective:
		                        			To evaluate the efficacy and safety of purified CD34+ stem cell boost in the treatment of poor graft function (PGF) after allogeneic hematopoietic stem cell transplantation (HSCT) .
		                        		
		                        			Methods:
		                        			12 patients with poor graft function, reported in our hospital during January 2014 to March 2018, were retrospectively analyzed; The donors of 12 patients were HLA mismatched family members, and all treated with donor purified CD34+ stem cell after G-CSF mobilization, calculating and statistical analyzing the purity of separation and the recovery rate of CD34+ stem cells. The related complications and the recovery of blood cells after infusion were observed.
		                        		
		                        			Results:
		                        			The purity of CD34+ cells in the separation products was 92.0% (44.0%-97.0%) , and the recovery rate was 55.0% (45.0%-96.7%) . The median number of CD34+ cells was 1.9 (0.9-4.4) ×106/kg with CD3+ cells as 0.6 (0.3-2.0) ×104/kg. The median durations of white blood cells, platelet and red blood cells recoveries were 18 (14-39) , 29 (16-153) and 60 (9-124) days, respectively. All 12 patients didn’t experience serious adverse reactions in the process of infusion, 10 patients achieved hematopoietic recovery, 1 case partial remission, 1 case no recovery, without occurrence of aggravated infection, graft versus host disease and other complications.
		                        		
		                        			Conclusion
		                        			The infusion of donor purified CD34+ stem cell was a safe and effective method for PGF after allogeneic HSCT. 
		                        		
		                        		
		                        		
		                        	
6.Establishing cases library of renal diseases and the feedback after the application of the cases library
Lian HE ; Xinkui TIAN ; Wen TANG ; Song WANG ; Junbao SHI ; Zhen ZHUANG ; Aihua ZHANG
Chinese Journal of Medical Education Research 2018;17(2):210-213
		                        		
		                        			
		                        			Clinical teaching is the critical stage for medical students turning to qualified doctor.In order to overcome the objective problems of insufficient clinical case resources,using the electronic medical record system to collect cases,the real case library of kidney disease was initially constructed,which was presented in the form of network resources.This case database was applied to assist teaching in the probation of eight year medical students at the undergraduate stage,and the application of the case database was evaluated in the form of questionnaire.It is found that case database is helpful to students' clinical learning,and has the necessity of further improvement and good prospects for popularization.It provides a new idea for clinical teaching.
		                        		
		                        		
		                        		
		                        	
7.Changes of autophagy-related genes in the brains of the rats in the developmental stage with epilepsy
Qin-Rui LI ; Ying HAN ; Junbao DU ; Hongfang JIN ; Jing ZHANG ; Manman NIU ; Jiong QIN
Chinese Journal of Applied Clinical Pediatrics 2018;33(12):909-912
		                        		
		                        			
		                        			Objective To explore the changes of Beclin-1,P62/SQSTM1,microtubule-associated protein 1 light chain 3 (LC3)and unc-51 like autophagy activating kinase 1 (ULK-1)in the brains of the rats in the deve-lopmental stage with epilepsy. Methods Seventy-two male Sprague Dawley (SD)rats aged 21 days were randomly divided into the control group and the epilepsy group. The rats in 2 groups were randomly subdivided into 4 groups according to the time intervals (3 h,6 h,12 h and 48 h),respectively,with 9 rats in each group. The rats in the epilep-sy group were injected with kainic acid (12 mg/kg)to induce epilepsy,and the rats in the control group were injected with equal volume of saline. The rats in 2 groups were anaesthetized and sacrificed. Then,the brain tissues of the rats were quickly removed according to the time intervals. The brain damages were determined by adopting Nissl staining method. The apoptotic cells were detected by Terminal - deoxynucleoitidyl transferase mediated nick end labeling (TUNEL)assays. The expressions of Beclin-1,P62/SQSTM1,LC3 and ULK-1 mRNA levels in cortex were mea-sured by using real-time quantitative polymerase chain reaction (qPCR)analysis. Results Nissl staining indicated that many neurons were damaged performing vague outline,irregularly aligned,pyknotic nuclei and shrunken somata in the epilepsy 48 h group. In addition,there was a huge loss of neurons in cortex in the epilepsy 48 h group [(82 ± 8)num-bers],compared with the control group [(122 ± 8)numbers],and the difference was statistically significant (F=3. 768, P=0. 01). The apoptotic cells tremendously increased in the epilepsy 48 h group [(13 ± 7)numbers],compared with the control group [(2 ± 1)numbers]by TUNEL analysis,and the diffe-rence was statistically significant (t= -3. 821, P=0. 003). qPCR showed the mRNA levels of Beclin-1,P62/SQSTM1,LC3 and ULK-1 were upregulated in the epi-lepsy 12 h group (1. 70 ± 0. 75,1. 75 ± 0. 77,1. 52 ± 0. 43,7. 48 ± 6. 12)and the epilepsy 48 h group (1. 63 ± 0. 43, 1. 48 ± 0. 74,1. 74 ± 0. 55,7. 69 ± 5. 65),compared with the control group (1. 00,1. 00,1. 00,1. 00),and the differences were statistically significant (F=2. 820,3. 452,5. 811,5. 002,all P<0. 05). Conclusion The autophagy activates be-fore apoptosis occurs,and autophagy-related genes probably are involved in epilepsy-induced brain damage.
		                        		
		                        		
		                        		
		                        	
8.Effect of endogenous sulfur dioxide on the apoptosis induced by cobalt chloride in the human pulmonary arterial endothelial cells
Xin LIU ; Da ZHANG ; Kun LI ; Xiaoqi YU ; Chaoshu TANG ; Junbao DU ; Hongfang JIN ; Yaqian HUANG
Chinese Journal of Applied Clinical Pediatrics 2018;33(13):999-1003
		                        		
		                        			
		                        			Objective To explore the effect of endogenous sulfur dioxide (SO2)on the apoptosis induced by cobalt chloride (CoCl2)in the human pulmonary arterial endothelial cells (HPAECs).Methods CoCl2was used in the primary HPAECs to mimize hypoxia-induced cell apoptosis.The aspartate aminotransferase 1(AAT1),and the key enzyme generating endogenous SO2 were over -expressed by transfecting HPAECs with lentivirus containing AAT1 cDNA.HPAECs were divided into 4 groups:vehicle group,vehicle + CoCl2 group,AAT1 group and AAT1 + CoCl2 group.The expressions of AAT1,B-cell lymphoma-2 (bcl-2),bcl-associated X protein (bax),Caspase-3 and activated Caspase-3 (cleaved Caspase-3)in the HPAECs were measured by Western blot.The AAT activity was assessed with colorimetry method.The SO2 content in the HPAECs was in situ observed by SO2-specific fluorescent probe.The HPAECs apoptosis was investigated by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL)assay.Results There were significant differences in the endogenous SO2 content,the expre-ssions of AAT1 and bcl-2,and the ratio of cleaved Caspase-3/Caspase-3 among 4 groups of HPAECs were (F=147.364,23.738,6.521,64.884,all P<0.05).However,there was no difference in the expression of bax among 4 groups of HPAECs (F=1.620,P>0.05).Compared with vehicle group,AAT activity [(0.96 ± 0.24)Carmen's unit/μg vs.(2.21 ± 0. 60)Carmen's unit/μg],endogenous SO2 content (40.71 ± 7.72 vs.105.60 ± 16.20)and bcl-2 expression (0.59 ± 0.19 vs.1.02 ± 0.20)in the HPAECs of vehicle +CoCl2 group were significantly de-creased,while the cell apoptosis assessed by TUNEL and the ratio of cleaved Caspase-3/Caspase-3 (1.56 ± 0.25 vs.0.95 ± 0.13)were significantly increased (all P<0.05).However,there were no differences in the expression of AAT1 (0. 50 ± 0.12 vs.0.53 ± 0.11)in the HPAECs between vehicle group and vehicle+CoCl2 group (P>0.05). The SO2 content (351.50 ± 42.43 vs.105.60 ± 16.20)and AAT1 expression (1.22 ± 0.33 vs.0.53 ± 0.11)in the HPAECs of AAT1 group were higher than those of vehicle group (all P <0. 05 ). Compared with AAT1 group, endogenous SO2content (333.50 ± 46.22 vs.351.50 ± 42.43)and the expression of AAT1 (1.26 ± 0.36 vs.1.22 ± 0.33)and bcl-2 (1.14 ± 0.38 vs.1.03 ± 0.27)in the HPAECs of AAT1 +CoCl2group did not change (all P>0. 05).Moreover,no difference was observed in the HPAECs apoptosis assessed by TUNEL and the ratio of cleaved Caspase-3/Caspase-3 (0.51 ± 0.17 vs.0.50 ± 0.11)between the two AAT1 -overexpressed groups (all P >0. 05).Conclusion Endo-genous SO2inhibited the hypoxic HPAECs apoptosis stimulated by the treatment of CoCl2.
		                        		
		                        		
		                        		
		                        	
9.Influence of endothelin-1 on nitric oxide and hydrogen sulfide pathways in vascular smooth muscle cells in rats
Xiaoyu TIAN ; Qingyou ZHANG ; Yaqian HUANG ; Da ZHANG ; Xinjing TANG ; Hongfang JIN ; Junbao DU ; Chaoshu TANG ; Yan SUN
Chinese Journal of Applied Clinical Pediatrics 2018;33(13):1013-1017
		                        		
		                        			
		                        			Objective To explore the effect of different concentrations of endothelin-1 (ET-1)on the en-dogenous nitric oxide (NO)and hydrogen sulfide (H2S)pathways of vascular smooth muscle cells (A7r5 cell lines)in rats.Methods A7r5 cell lines were divided into the control group and the experimental group.ET-1 at a concentra-tion of 10 -8-10 -6 mol/L was added into the experimental group,and as for the control group,the same volume of sterile phosphate buffered saline (PBS)buffer solution was added.The content of NO and H2S in A7r5 cell lines was detected by fluorescent NO probe and H2S probe after ET-1 stimulation for 48 h,respectively.The content of NO in the supernatant was measured by NO assay kit at 48 h of the incubation.The content of H2S in the supernatant was measured by polarographic H2S sensor at 48 h of the incubation. The expressions of inducible nitric oxide synthase (NOS2),endothelial nitric oxide synthase (NOS3),cystathionine -γ -lyase (CSE),cystathionine -β -synthase (CBS)and proliferating cell nuclear antigen (PCNA)were detected by the Western blot method.Results The rela-tive fluorescence intensity of the content of NO in the A7r5 cell lines of ET-1 10 -8,10 -7 and 10 -6mol/L groups (0. 078 ± 0. 080,0.075 ± 0.002,0.056 ± 0.009)was markedly lower than that in the control group(0.094 ± 0. 061), and the differences were statistically significant(F=15.248,P<0.05);Compared with the control group[(2. 131 ± 0. 484)μmol/L],the content of NO in the supernatant of the experimental groups [(1.391 ± 0.134 )μmol/L, (1.219 ± 0. 280)μmol/L,(1.116 ± 0.181)μmol/L]was significantly decreased,and the differences were statistically significant(F=20.833,P<0.01);NOS2 protein expression(0.457 ± 0.097,0.462 ± 0.116,0.438 ± 0.180)was decreased markedly compared with that of the control group(0.721 ± 0.222),and the differences were statistically sig-nificant(F=6.196,P<0.01),but the expression of NOS3 showed no significant differences(F=2.669,P>0.05). The relative fluorescence intensity of the content of H2S in the A7r5 cell lines of ET-1 10 -8,10 -7 and 10 -6mol/L groups (0.063 ± 0.002,0.056 ± 0.008,0.042 ± 0.009)was markedly lower than that in the control group (0.082 ± 0. 006),and the differences were statistically significant(F =16.297,P<0.01);Compared with the control group [(29.439 ±4.236)μmol/L],the content of H2S in the supernatant of the experimental groups [(17.516 ±5.144) μmol/L,(14.481 ± 4.885)μmol/L]was significantly decreased,and the differences were statistically significant (F=12.518,P <0.01).CBS protein expression(0.359 ± 0.096,0.270 ± 0.038,0.174 ± 0.051)was decreased markedly compared with that of the control group(0.707 ± 0.107),and the differences were statistically significant (F=20.833,P<0.01),and the expression of CSE showed no significant differences(F=0.708,P>0.05).The data showed that PCNA protein expression in the 10 -7mol/L ET-1 group(0.686 ± 0.180)significantly increased com-pared with that of the control group(0.437 ± 0.191),and the difference was statistically significant (t= -2.840,P<0.01).Conclusion ET-1 stimulation can lead to the proliferation of vascular smooth muscle cells and down-regu-late its endogenous NO and H2S pathways.
		                        		
		                        		
		                        		
		                        	
10.Effects of different modes of mild treadmill exercise on cognitive function of Alzheimer's disease mice
Wenying SONG ; Jiawei ZHANG ; Fang PENG ; Junbao LIU
Chongqing Medicine 2018;47(5):614-617
		                        		
		                        			
		                        			Objective To investigate the effects and possible mechanisms of different modes of mild treadmill exercise on cognitive function in Alzheimer's disease(AD) APP transgenic mice.Methods Twenty-four 6-month-old APP+ mice were randomly and equally divided into the four groups:control group(Con),regular exercise group(RE),irregular time-of-day exercise group (ITE) and irregular different duration exercise group(IDDE).After treadmill exercise,the spatial cognitive ability was tested.Soluble and insoluble Aβ40,42 levels,activity of β-secretase and γ-secretase in hippocampal tissue were compared among 4 groups.The mRNA levels of NEP,IDE and MMP-9 in the hippocampal tissue were detected.The number of hippocampal survival neurons and protein levels of BDNF,TrkB and p-TrkB were detected.Results Compared with the control group,the spatial cognitive ability in the RE group was significantly increased(P<0.05),the levels of soluble and insoluble Aβ40,42 and activity of β-secretase and γ-secreatase were significantly reduced in the hippocampal tissue of th RE group(P< 0.05).However,the mRNA levels of NEP,IDE and MMP-9 had no statistically significant difference among the four groups(P>0.05).Compared with the control group,the number of survival neurons and protein levels of BDNF and p-TrkB in the hippocampal of the RE group tissue were significantly increased(P<0.05).The cognitive function,levels of Aβ40,42,BDNF and p-TrkB,number of survival neurons in the RE group had statistically significant differences when compared with the ITE and IDDE groups(P<0.05).Conclusion Mild and regular treadmill exercise can effectively increase the cognitive function of AD mice,which might be related with the regulation of BDNF/TrkB pathway.
		                        		
		                        		
		                        		
		                        	
            
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