1.Immunocytochemistry analysis of human neutrophil defensins
Qi WU ; Luo-Ling XU ; Bo-Yao WANG ;
Chinese Journal of Immunology 1985;0(06):-
Our laboratory isolated three defensin molecules from the circulating neutrophils of healthydonors.The three molecule mixture was conjugated with BSA and immunized New Zealand rab-bit.ELISA showed that the rabbit antisera was of a very high titters,which were specific for hu-man defensins.Immunoperoxidase staining with this rabbit antisera indicated that normal humanneutrophils,acute myeloid leukemia cell lines KG1 and HL-60 were highly positive.In con-trast,normal mononuclear leukocytes,erythrocytes,erythroleukemia cell line K562 were all neg-ative,even rabbit and guinea pig neutrophils had no response to this rabbit anti-human de-fensins antisera.Our preliminary experiments suggested that the rabbit antisera was highly spe-cific for human defensins.Probably,defensin is a differentiation antigen of granunocyte lineage,and antibody against human defensins would be a candidate immunological reagent specific for a-cute myeloid leukemia.Further work for this should be done.
2.Clinical Analysis of Cardiac Involvement in Children with Mitochondriopathies
jian-guang, QI ; ying, ZHANG ; yu, QI ; yan-ling, YANG ; ye, WU ; yu-wu, JIANG ; jiong, QIN ; jun-bao, DU
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the clinical characteristics of cardiac involvement in children with mitochondriopathies.Methods The clinical data of 23 children with mitochondriopathies were reviewed.The changes of electrocardiography,echocardiography and heart enzymes were analyzed.Results In 15 cases of mitochondrial encephalomyopathy,lactic acidosis,and stroke-like episode(MELAS syndrome),electrocardiography was performed on 9 cases,6 of them showed abnormal electrocardiographic findings,including right bundle branch block,ST-T change,Wolff-Parkinson-White syndrome,et al.On echocardiographic examination in 9 MELAS syndrome ca-ses,only 1 case showed hypertrophy cardiomyopathy.Six cases had increased plasma creatine kinaseMB(CK-MB) mass and only one of 12 MELAS syndrome cases had increased cardiac troponin I(cTnI) level.In 8 cases of subacute necrotizing encephalomyopathy(Leigh syndrome),electrocardiography was performed on 5 cases,4 of them showed abnormal electrocardiographic findings,including sinus tachycardia,ST-T change and low voltage.Two cases showed normal electrocardiography.Three out of 6 cases with Leigh syndrome showed increased plasma CK-MB mass.The molecular genetic examinations were performed in 13 cases of MELAS syndrome and 6 cases of Leigh syndrome.The mitochondrial DNA nt 3243 A→G mutation was found in white blood cells of 9 MELAS syndrome cases,the mutation rate being 37%-60%.The mitochondrial DNA nt 8993 T→C mutation was found in white blood cells of 2 Leigh syndrome cases.Conclusion In children with mitochondriopathies,myocardiac involvement is comparatively common,and even cardiomyopathy can occur.
3.A High-throughput and Quantitative Assay Based on Fluorescence Intensity for Detection of Apoptosis
Ling-Ling YE ; Hong LIU ; Xing-Mao LIU ; Shi-Chong LI ; Ben-Chuan WU ; Qi-Wei WANG ; Zhao-Lie CHEN ;
China Biotechnology 2006;0(06):-
Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P
4.Nicotine- and tar-free cigarette smoke extract reduces the penile erectile function of rats.
Shu-chao LIU ; Tian-peng WU ; Qi-xin DUAN ; Ling-qi LIU ; Chao SONG ; Wen-biao LIAO ; Si-xing YANG
National Journal of Andrology 2015;21(12):1071-1076
OBJECTIVETo evaluate the impact of nicotine- and tar-free cigarette smoke extract (fCSE) on the serum testosterone (T) level and erectile function of male rats.
METHODSWe randomized 30 male SD rats to three groups of equal number to receive subcutaneous injection of PBS (1.0 ml / 300 g body weight per day), fCSE (1.0 ml/300 g body weight per day), and reduced glutathione hormone (GSH, 200 mg per kg body weight per day) in addition to fCSE (fCSE + GSH), respectively, all for 8 weeks. Then we evaluated the erectile function of the rats by measuring the maximal intracavernous pressure (MICP), mean arterial pressure (MAP), ICP/MAP ratio, time of stimulation to MICP (Tmax), and cavernosal filling fate (CFR). We determined the serum T level, the activities of superoxide dismutase (SOD) , malondialdehyde (MDA), and nitric oxide synthase (NOS) in the cavernosal tissue, and also observed the morphological changes of the corpus cavernosum.
RESULTSCompared with the controls, the rats of the fCSE group showed obvious decreases in the levels of serum T ([5.37 ± 1.43] vs [3.22 ± 1.11] μg/L), NOS ([2.90 ± 0.27] vs [1.67 ± 0.18] U/mg) , and SOD ([18.41 ± 1.09] vs [13.36 ± 1.18] U/mg prot) and erectile function-related indexes MICP ([85.92 ± 6.36] vs [58.99 ± 10.76] mmHg), MICP/MAP (0.86 ± 0.09 vs [0.56 ± 0.08]), and CFR (2.14 ± 0.44 vs 0.89 ± 0.44), but markedly increased Tmax ([29.90 ± 5.78] vs [42.90 ± 8.56]s), with a positive correlation between the serum T level and CFR (r = 0. 364, P < 0.05). Masson staining revealed a lower ratio of the corpus cavernosum smooth muscle tissue to collagen fiber in the fCSE group (0.27 ± 0.04) than in the control (0.98 ± 0.12). Compared with the fCSE group, the fCSE + GSH group exhibited significantly improved MICP ([58.99 ± 10.76 ] vs [77.95 ± 7.71] mmHg), MICP/MAP (0.56 ± 0.08 vs 0.77 ± 0.09), and CFR (0.89 ± 0.44] vs 1.76 ± 0.42) and shortened Tmax ([42.90 ± 8.56 ] vs [32.10 ± 5.84 ] s). The ratio of the corpus cavernosum smooth muscle tissue to collagen fiber was higher in the fCSE + GSH than in the fCSE group (0.77 ± 0.09 vs 0.27 ± 0.04) but still lower than in the control (0.98 ± 0.12).
CONCLUSIONNicotine- and tar-free cigarette smoke extract reduces the serum T level and erectile function of rats, which is related to oxidative stress. Antioxidant therapy can improve erectile function but has a limited value for morphological protection of the penile tissue.
Animals ; Erectile Dysfunction ; chemically induced ; Male ; Malondialdehyde ; metabolism ; Muscle, Smooth ; pathology ; Nicotine ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Superoxide Dismutase ; metabolism ; Tars ; Tobacco ; adverse effects
5.Study on mechanism of total flavonoids from hemerocallis fulva on oxidative stress and hepatocyte apoptosis in alcoholic liver injury
Bo XU ; Yan LI ; Pengyan JI ; Ling QI ; Qian LU ; Weinan WU ; Nan SHEN
Chongqing Medicine 2017;46(10):1304-1307
Objective To study the influence of total flavonoids of hemerocallis fulva(TFHF) on hepatocyte apoptosis and related protein expression in mice with alcoholic hepatic injury.Methods A total of 40 mice were randomly divided into four groups:blank control,model control andsmall and high dose TFHF groups,10 cases in each group.The mice were given the continuous gavage administration for 7 d.Then the model group was given once gavage by 50% ethanol 12.0 mL/kg after 1 h of the last administration.The blank control group was given the equal volume of distilled water.The activity levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum as well as the superoxide dismutase(SOD) activity and malondialdehyde (MDA) content in liver tissue hemogenate were detected.Hematoxylin and Eosin(HE) staining was performed for observing the pathological changes of the liver tissue.The flow cytometer was used to test the apoptosis ratio in hepatocyte suspension.The expressions of caspase-3,Bcl-2 and Bax protein were detected by Western blot.Results The various TFHF groups could decrease the activities of ALT and AST in serum (P<0.05),while could decrease the MDA content in liver tissue hemogenate (P<0.01) and increased the SOD activity;the liver tissue pathological examination showed that the high dose TFHF group could make the liver cell degeneration,alleviated the necrosis degree and relieved the pathological change of hepatic tissue;compared with the model group,the hepatocyte apoptosis rate in each TFHF group was decreased significantly;Western blotting results showed that the caspase-3 protein level in each TFHF group was decreased,expression of Bcl-2 protein was increased,whereas which of Bax protein was decreased and Bax/Bcl-2 ratio was reduced.Conclnsion TFHF has obvious protective effect on mice acute hepatic injury induced by ethanol,and can inhibit the hepatocyte apptosis,its action mechanism may be related to its antioxidation and regulation of caspase-3,Bcl-2 and Bax expression.
6.Comparative Study on the Effect of Seroquel and Chlorpromazine on Schizophrenia
Hui TIAN ; Qi HE ; Ling DU ; Renhui WU ; Shuguo HUI ; Qiulian ZHENG ; Yinghong BING
China Pharmacy 2001;0(09):-
0.05).Seroquel showed high incidence rate of drowsiness,dizziness and weight gain(26.1%,23.9%,17.3%),chlorpromazine showed high incidence rate of extrapyramidal side effect,pyknocardia and erect collapse(60.9%,39.1%,32.6%).CONCLUSIONS:There was no significant difference of curative effect for the treat-ment of schizophrenia between seroquel-treated group and chlorpromazine-treated group,but seroque showed low side effect and high safety.
7.Early posttransplant hepatorenal functions in the prediction of liver graft recipient's prognosis
Xiao XU ; Jian WU ; Qi LING ; Feng GAO ; Lin ZHOU ; Shusen ZHENG
Chinese Journal of General Surgery 2008;23(10):781-783
Objective To monitor the dynamic changes of liver and kidney functions in the first week after liver transplantation and to assess the value of liver and kidney functions in predicting patient's survival. Methods clinical data of 161 recipients with benign liver diseases were retrospectively reviewed.Total bilirubin ( TB ), aminoleucine transferase ( ALT), aspartate aminotransferase ( AST), prothrombin time (PT) and serum creatinine (SCr) were recorded and analyzed during the first post-transplant week.The data collected at dl were analyzed in the multivariate COX regression. Results From d1 to d7 post-transplant, the median value changed from 116. 2 mmol/L to 66. 7 mmol/L ( Z = 5.901, P < 0. 01 ) for TB,19.4 s to 15.0 s (Z = 11. 657, P <0.01 ) for PT, 285 U/L to 100 U/L (Z = 12.619, P<0. 01 ) for ALT,264 U/L to 50 U/L (Z =9. 776, P <0. 01 ) for AST, 103.4 mmol/L to 86. 6 mmol/L (Z = 1. 353, P 0. 05 ) for SCr. Post-transplant SCr ( RR = 3. 477, P < 0. 001 ) and TB ( RR = 2. 088, P < 0. 05) levels wereindependent factors influencing patient survival. A prognostic score formula was then established as 1. 276 ×InSCr (mg/dL) + 0. 730 × InTB (mg/dL). Conclusion In a successful liver transplantation,transplanted liver function recovers promtly within one week. SCr and TB levels on post-transplant d1 have good prognostic value.
8.The activated pathways by hypoxia in patients with Obstructive sleep apnea hypopnea syndrome
Ling LUO ; Xinglin GAO ; Dongfeng LI ; Shaohua LUO ; Jian WU ; Qi LIN ; Qiong OU ; Ruijin CEN
The Journal of Practical Medicine 2014;(24):3902-3905
Objective To investigate the activation of nuclear factor-B (NF-B)and hypoxia-inducible factor-1 (HIF-1)in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Subjects were classified into mild-to-moderate OSHAS group (n = 16), severe OSAHS group (n = 14) and the matched control group (n=30). Gene and protein expressions of NF-B and HIF-1 were measured by RT-PCR, Western blot in peripheral blood mononuclear cells (PBMC). Results NF-κB p65 mRNA and NF-κB p65 protein,HIF-1α mRNA and HIF-1α protein in PBMC were significantly higher in severe OSAHS group than those in mild–to-moderate group and control group (P<0.01). But there were no significant difference in NF-κB p65 mRNA and NF-κB p65 protein between mild-to-moderate group and control group (P=0.068, P=0.254 respectively). Only gene (P<0.05) not the protein (P=0.777) of HIF-1αwas higher in mild-to-moderate as compared with control group. Both NF-κB p65 mRNA and HIF-1αmRNA were positively correlated with AHI (r=0.493, P=0.006, r=0.508, P=0.004), while negatively correlated with nighttime lowest blood oxygen saturation (LSaO2)(r=-0.488, P=0.006, r=-0.46, P=0.011). There was a positive correlation between NF-κB p65 protein level and AHI (r=0.669, P<0.001). HIF-1αprotein level was positively correlated with AHI, ODI (r=0.628, P=0.001;r=0.480, P=0.018). There were positive correlations between NF-κBp65mRNA and HIF-1αmRNA (r=0.543, P=0.002), NF-κBp65 and HIF-1αprotein respectively (r=0.716, P<0.001). Conclusions As the gene and protein of NF-κB and HIF-1 were up-regulated in patients with OSAHS, and also positively correlated with the severity of sickness. We conclude that both NF-κB and HIF-1 were involed in the pathogenesis of OSAHS.
9.Optimization of combined CT pulmonary angiography and renal venography for the detection of venous thromboembolism in patients with nephrotic syndrome
Ling ZHENG ; Xinsheng WU ; Changsheng ZHOU ; Li QI ; Longjiang ZHANG ; Guangming LU
Chinese Journal of Radiology 2013;47(12):1124-1127
Objective To optimize combined CT pulmonary angiography (CTPA) and renal venography (rCTV) and to evaluate its value for the detection of venous thromboembolism in patients with nephrotic syndrome (NS).Methods Ninety NS patients suspected of venous thromboembolism because of abnormal D-dimer value were included in this prospective study.The first 45 patients were defined as group 1 (protocol 1).These patients underwent CTPA,then rCTV 25 s after CTPA,last inferior vena cava scanning after another 25 s.The following 45 patients as group 2 (protocol 2).These patients underwent CTPA and then inferior vena cava scanning 50 s after CTPA,3 patients in group 2 were excluded because of unavailable CTPA or rCTV.Vessel enhancement of CTPA and rCTV,and radiation doses for two protocols were compared with independent sample student t test.Incidence and distribution of pulmonary embolism and renal vein thrombosis were recorded and compared by using Chi-square test.Results Enhancement values of pulmonary trunk for groups 1 and 2 were (335.5 ± 111.3) and (335.0 ± 76.0) HU,right renal vein were (142.7 ±33.3) and (140.7 ±25.9) HU,left renal vein were (141.6 ±26.4) and (138.8 ±33.6) HU respectively,without any statistical difference (t values were 0.026,0.322 and 0.452,P >0.05).Radiation dose to the patients receiving protocol 2 were lower than that of patients receiving protocol 1 [volume CT index,(19.7 ± 4.3) vs (13.6 ± 3.0) mGy; dose length product,(1019.9 ± 878.5) vs (532.0 ± 132.9) mGy · cm; both P <0.01].Of 87 NS patients,44 venous thromboembolism events were detected,resulting in the incidence of 50.6% (44/87),including pulmonary embolism in 19 patients,renal vein thrombosis and pulmonary embolism in 17,renal vein thrombosis in 7,and inferior vena cava in one patient.The detection rate were 48.9% (22/45) and 52.4% (22/42),there was no significant difference (x2 =0.106,P =0.745).Conclusion Combined CTPA and rCTV with protocol 2 (CTPA and inferior vena cava phase scanning 50 s after CTPA) is suitable to clinical application and pulmonary embolism is the most common thromboembolism complication in NS patients.
10.Protective effect and mechanism of MST1 inhibition on kidney tissue in diabetic rats induced by streptozotocin
Weihua WU ; Ling XUE ; Santao OU ; Ying LI ; Fan ZHANG ; Qi LIU ; Jian LIU
Chinese Journal of Nephrology 2015;31(11):835-841
Objective To investigate the protective effect and mechanism of MST1 inhibition on kidney tissue in diabetic rats,and to find a new therapeutic target for diabetic nephropathy.Methods Total of 54 male SD rats enrolled in this study were divided into 3 groups including normal control (group A,n=18),MST1 inhibition group (Group B,n=18) and diabetes group (group C,n=18).Diabetes was induced by a single streptozotocin (STZ,50 mg/kg) injection in group B and group C.rats in group B received lentiviral vector contain Mst1 interference RNA (shRNA) and the rats in group C received empty vector.The end of 4th,8th and 12th week after modeling were considered as time points in this study.At each time point,the level of 24 hours urine protein (24-HUP),blood glucose and serum creatinine were examined.Pathological changes were observed with HE stain; Injury of podocyte and glomerular basement membrane (GBM) were examined with transmission electron microscope (TEM).The intensity and location of MST1 in kidney tissue were detected by immunohistochemistry.The level of MST1,Phosphorylated-MST1,nephrin,Caspase-3 and FasL were detected by western bloting.Results (1) At the starting point,there were no significant differences among groups in terms of weight,activity,eating and drinking.Since the end of 72nd hour after modeling,the levels of glucose in both group B and group C,compared to those in group A,significantly increased (P < 0.05).There was no significant difference between group B and group C for glucose level at each time point (P > 0.05); the level of 24-HUP increased significantly since the end of 4th week after modeling,and the level in group C was higher than its counterpart in group B at the same point (P < 0.05); (2) There was no significant pathological lesion observed in group A.Without obvious K-W nodular changes,mesangial proliferation was observed in group B and group C.It was shown by TEM that podocyte fusion and thickening of the GBM could be found in group B and group C.The pathological change in group B was better than that in group C; (3) Compared to group A,it was shown by western blot that the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B and group C were significantly higher (P < 0.05),and the levels of nephrin in group B and group C were significantly lower (P < 0.05) since the end of 4th week after modeling.Meanwhile,the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B were significantly lower than that in group C at each time point (P < 0.05),the level of nephrin in group B was significantly higher than the one in group C; (4) It was shown by immunohistochemistry that there was low MST1 expression in normal condition,especially in cytoplasm of tubular epithelial cells.The level of MST1 in group B and group C significantly increased after modeling,and the change could be the same as Western blot shown.Conclusions MST1 pathway could be involved in kidney injury induced by diabetes.MST1 inhibition could alleviate the kidney injury in STZ-induced diabetes animal model.