1.Construction of siRNA expression vector targeting IGF-1R and its ability to induce cell apoptosls in hmnan lung cancer cells
Ai-Qiang DONG ; Zhi-Yuan MA ; Min-Jian KONG ;
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(04):-
Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.
2.Relationship between Change of Cerebral Hemodynamic and Brain Injury in Neonates
xiao-li, MA ; jin-zhi, SONG ; jian-ming, LI
Journal of Applied Clinical Pediatrics 2004;0(08):-
The cerebral blood flow of infant is effected by physiological and pathological factors.As the cerebrovascular autoregulation of neonates is poor,in pathological cases,especially when hypoxemia and hypercapnia impaired regulation of its own,lead to changes in cerebral blood flow,then resulting in severe brain injury.It has made enormous progress in the research on the changes of cerebral blood flow in newborns in recent years.In normal infants,cerebral blood flow velocities is positively correlated to gestational age and body weight,and increase gradually with day-age in the first week after birth.The cerebral blood flow of newborn with brain injury can present as insufficiency,over-perfusion or low speed high-resistance earlier and high speed low-resistance later.Different results may be related to the duration and severity of asphyxia,but all of those are signs of damage of self-regulatory function of cerebral blood flow.Cerebral hemodynamic change is the important pathogenesis mechanism of brain injury.
3.Low testosterone levels are inversely correlated with carotid artery plaque formation in elderly women.
Qiang MA ; Qing-Li CHENG ; Jian MA ; Qiang-Guo AO ; Guo-Juan TAN ; Guang ZHI
Chinese Journal of Applied Physiology 2013;29(6):554-558
OBJECTIVETo study the relationship between serum testosterone levels and the plaque formation of the carotid artery in a population-based cohort of independently living healthy women above 60 years of age.
METHODSAnalysis of the healthy elders from a population-based cohort study in 9 communities of Beijing. Carotid intima-media thickness and atherosclerotic plaques were determined ultrasonographically. Serum testosterone levels were measured by immunoassay. The data were analyzed with ANOVA and logistic regression analysis.
RESULTSThere was an inverse correlation between testosterone and plaque formation in old females (P < 0.01), while no association was found in males. Female with testosterone levels in the lowest quartile (< 0.49 nmol/L) had more risk of plaque formation (OR = 3.805, P < 0.01) after adjusted with age and other traditional factors of atherosclerosis.
CONCLUSIONTestosterone concentrations are negatively associated with carotid artery atherosclerosis in old women in Beijing, experimental and prospective studies are needed to determine the possible therapeutic role of testosterone in atherosclerosis.
Aged ; Atherosclerosis ; blood ; Carotid Arteries ; pathology ; Carotid Intima-Media Thickness ; Carotid Stenosis ; blood ; Female ; Humans ; Male ; Prospective Studies ; Testosterone ; blood
4.Dynamic changes and clinical significance of serum Thl/Th2 cytokines in hemorrhagic fever with renal syndrome
Zhi-jun, MA ; Zhi-jian, SUN ; Hong, ZHAO ; Yu-hua, WANG ; Fen-yong, ZHU ; Dong-jun, ZHANG ; Suo-rong, WANG
Chinese Journal of Endemiology 2008;27(4):461-463
Objective To investigate the changes and pathogenic significance of serum interleukin-12p70 (IL-12), intefferon-γ,(IFN-γ) and IL-4 in the course of hemorrhagic fever with renal syndrome(HFRS). Methods Twenty five eases were divided into mild group (14 eases) and severe group (11 cases) according to the severity of illness. Blood samples were collected in various stages(fever, hypotensian and oliguria,diuresis stage). Serum IL-12 and IFN-γ levels were determined by enzyme-linked immunoserbent assay(ELISA), IL-4 by radioimmunoassay (RIA), blood urea nitrogen (BUN) and platelet by automatic biochemical analyzer and blood analyzer. Results Serum IL-12 levels in mild and severe groups were significantly different during various stages of HFRS (F=5.765, P<0.01). The IL-12 level of both patient groups significantly increased(P<0.01) in fever[ (0.87±0.38), (1.08± 0.77)μg/L], hypotension and oliguria [ (0.77±0.21), (2.11±2.13)μg/L] ,and diuresis stage [ (1.42±1.10), (1.20±0.88)μg/L], compared with control group [(0.56±0.10)μg/L]. In various stages, IFN-γ levels of both case groups were respectively (8.04±13.05), (5.94±8.24), (15.95±18.05), (4.41±4.10), (1.09±1.24), (1.38±1.74), (1.12±1.26), (0.19±1.29)μg/L, and the difference was statistically significant compared with control [ (0.27±0.15)rig/L]. K,-4 levels did not change significantly in the stages(F=0.682, P0.05), while the ratios of IFN-γ and IL-4 contents in mild and severe cases were significantly higher than control [(0.36±0.26) μg/L] in fever[ (2.46±3.52), (16.92±22.77)p.g/L], hypotension and oliguria[(2.52±2.72), (1.77±2.06) μg/L],diuresis stage [(1.45±2.28), (2.32±3.98)μg/L], the difference had statically significant (P<0.05 or 0.01).The curve of IL-12 was similar to that of BUN, but was contrary to blood platelet count. Conclusions The elevated levels of IL-12 and IFN-γ, with the imbalance of Th1/Th2 might be the main cause of systemic inflammatoryresponse and involved in the pathogenesis of HFRS.
5.Inhibitory effect of 8-prenylnaringenin on osteoclastogensis of bone marrow cells and bone resorption activity.
Xiang Lü ; Ying ZHOU ; Keming CHEN ; Zhi ZHAO ; Jian ZHOU ; Xiaoni MA
Acta Pharmaceutica Sinica 2013;48(3):347-51
This study is to investigate the effect of 8-prenylnaringenin (8-PNG) on osteoclastogensis of bone marrow cells and bone resorption activity of osteoclasts. Osteoclasts were separated from long bone marrow of newborn rabbits and cultured in alpha-MEM containing 10% FBS. 8-PNG was added into culture media at 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) mol xL(-1), separately. 17beta-Estradiol (E2, 1 x 10(-7) mol x L(-7)) was used as positive control. T RAP staining and TRAP activity measurement were performed after 5 days, and the bone resorption pits were analyzed after 7 days. Annexin V staining for the detection of apoptotic osteoclasts was performed after 2, 4, 8, 12, 24, 36 and 48 h separately. The mRNA expression level of TRAP and cathepsin K (CTSK) was measured by real-time RT-PCR. 8-PNG significantly reduced the number of osteoclasts which was TRAP staining positive and with more than three nucleus, the area and number of bone resorption pits decreased obviously in 8-PNG-supplemented groups. The apoptosis rate peaked earlier in the 8-PNG-supplemented groups and the mRNA expression level of TRAP and CTSK decreased significantly. All these inhibitory effects were in a dose dependent manner, the highest effect was obtained by 1 x 10(-5) mol x L(-1) 8-PNG. 8-PNG inhibits bone resorption activity of osteoclasts by inducing osteoclast apoptosis and inhibiting the gene expression and enzyme activity including TRAP and CTSK, and restrains bone marrow cells to osteoclast differentiation.
6.Change of JNK and c-Jun in lung injury associated with paraquat poisoning of rats.
Jian-Hui LIU ; Zhi-Ping SUN ; Yu-Teng MA
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(7):406-409
OBJECTIVETo investigate the change of JNK and c-Jun in lung injury associated with paraquat poisoning of rats.
METHODS46 Rats were randomly divided into four groups: PQ group (n = 12), control group (n = 10), PQ + ZnPP group (n = 12) and PQ + Hm group (n = 12). The rats were injected with 2% PQ (25 mg/kg, ip) in PQ group. ZnPP and Hemin (10 mg/kg, 10 mg/ml) were injected through inguinal vein before intraperitoneal administration of 2% paraquat in PQ + ZnPP group and PQ + Hm group respectively. The rats were injected NS (1 ml/kg, ip) in control group. HE dyeing of lung tissue and MDA content of plasma were used for estimating the injury of lung tissue. The content of CO in the lung tissue was determined. The expression of HO-1 mRNA of the lung tissue was detected by the reverse transcription-polymerase chain reaction. The phosphorylation of JNK and c-Jun was evaluated by Western blot analysis.
RESULTSThe degree of lung injury in PQ group and PQ + ZnPP group was higher than that in control group and PQ + Hm group. But in PQ + Hm group the degree of lung injury was lower. The content of MDA in PQ group and PQ + ZnPP group was higher than that in control group and PQ + Hm group (P < 0.01). The content of MDA in PQ + Hm group was higher than that in control group (P < 0.05). The content of CO in lung tissue in PQ group, PQ + ZnPP group and PQ + Hm group was and (1.08 +/- 0.15 mg/L) respectively, and higher than that in control group (P < 0.01). The content of CO in lung tissue in PQ + Hm group was significantly higher than that in PQ + ZnPP group (P < 0.01). The expression of HO-1 and the phosphorylation of JNK (55.24 +/- 9.34, 38.15 +/- 10.71, 128.55 +/- 19.43) and c-Jun (23.16 +/- 4.85, 15.49 +/- 3.13, 44.89 +/- 10.37) were increased remarkably in PQ group, PQ + ZnPP group and PQ + Hm group. Those in PQ + Hm group were higher significantly than PQ group and PQ + ZnPP group (P < 0.01). Those in PQ + ZnPP group were lower than PQ group (P < 0.05).
CONCLUSIONThe increase of CO of lung tissue in rats at the lung injury associated with paraquat poisoning reduces the acute lung injury of rats. The level of JNK and c-Jun phosphorylation increases obviously, especially after Hemin is utilized.
Acute Lung Injury ; chemically induced ; metabolism ; Animals ; Disease Models, Animal ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; metabolism ; Male ; Paraquat ; poisoning ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Sprague-Dawley
8.Comparison of adenosine and exercise stress 201Tl myocardial perfusion imaging for diagnosing coronary heart disease in women
Jiang-jin, LI ; Shu-ren, MA ; Tao, MENG ; Zhi, BAO ; Jian-he, CUI
Chinese Journal of Nuclear Medicine 2011;31(1):39-41
Objective To compare the diagnostic value of adenosine and exercise stress myocardial perfusion imaging (MPI) for detecting coronary heart disease (CHD) in women. Methods One hundred and thirty-eight patients with CHD were randomly divided into two groups: adenosine stress group (n = 69)and exercise stress group (n = 69). All patients underwent myocardial SPECT evaluation. Coronary angiography (CAG), referred as "gold standard" , was performed in each patient within 1 week before or after MPI. The diagnostic value of the two stress MPI was compared with χ2 test or Fisher's exact test. Results In adenosine stress group, the sensitivity, negative predictive value and accuracy were 88.2% (45/51),72.7% (16/22), 88.4% (61/69), respectively, which were not significantly different from those of the exercise stress group (91.7% (44/48), 66.7% (8/12), 81.2% (52/64); χ2 =0. 571, 0. 714, 0.249, P >0.05). However, the false positive rate of adenosine stress (11.1%, 2/18) was significantly lower than that of exercise stress (50.0%, 8/16), P = 0.023. Conclusions Adenosine and exercise stress MPI have similar value for CHD diagnosis in women, however, adenosine stress MPI may have an advantage of low false positive rate.
9.Relationship between diopter and refractive factors of high school students
Jian-Zhou, MA ; Guo-Hua, YANG ; Jia-Zhi, LI ; Xiao-Ping, ZHANG
International Eye Science 2015;(2):375-377
AlM: To explore a formula between the diopter , cornea diopter ( CD ) , anteriou chamber depth ( ACD ) , lens thickness ( LENS ) , vitreous depth ( VlTR ) , and ocular axis longitude ( AL ) of high school students in certain middle school, and to predict the diopter with the biometry data of refractive factors on the students. METHODS: The related ocular examinations were performed to 364 eyes of 182 high school students in certain middle school by cluster sampling method. Optometry was for diopter and A-ultrasonic biometry was for ACD, LENS, VlTR, and AL. The data were analyzed with SPSS to sum up a poly-variation liner regression formula that can express their quantitative relationship.RESULTS: A poly-variation liner regression was got. The formula was D=69. 750-0. 724×CD-0. 630×ACD-2. 207× LENS-1. 728×VlTR (r=0. 87).CONCLUSlON: The diopter of high school students seems able to be predicted through biometric data of CD, ACD, LENS and VlTR.
10.Cut-off value of rose bengal plate agglutination test in rapid diagnosis of brucellosis
Zhi-dong, MA ; Yun-xia, LIU ; Yan-hong, LI ; Jian-yun, LI
Chinese Journal of Endemiology 2013;32(5):493-495
Objective To find out the cut-off value of rose bengal plate agglutination test(RBPT) in rapid diagnosis of brucellosis.Methods From May to June 2009,398 people who came to the outpatient department of Inner Mongolia Center for Endemic Disease Control and Research were diagnosed brucellosis by RBPT and tube agglutination test (SAT).Tube agglutination test as a gold standard,rose bengal plate agglutination test,its authenticity and reliability were evaluated.Results Taking positive predictive value 100.0% as the selection standard,the cut-off value of RBPT was ++ which could be used to diagnose brucellosis.The sensitivity was 83.3%; the specificity was 100.0%; the Youden index was 0.832; and the compliance rote was 89.9%.Conclusion The cut-off value ++ of RBPT to diagnosis brucellosis is worthy of clinical promotion.