1.The mechanism of cardiac protection of Hsp27 against cardiac dysfunction during endotoxema in mice
Hongmei ZHOU ; Wenjun YOU ; Xiaojin ZHANG ; Zhengnian DING ; Yunlin CHENG ; Li LIU
Chinese Journal of Emergency Medicine 2010;19(5):482-487
Objective To investigate the cardiac protection of Hsp27 against endotoxic cardiac depression mediated by activation of PI3K/Akt pathway and the suppression of NFκB-mediated inflammatory response in mice. Method (1) Transgenic mice with cardiac specific overexpression of Hsp27 (Hsp27 Tg) and wild littermate controls (WT) were given 10 mg/kg LPS injected intraperitoneally to induce endotoxemia, (2) The cardiac function measurement in mice was performed by using echocardiography 6 hours after LPS treatment (n = 6), (3) The activity of PBK/Akt pathway was evaluated by Western blot for [hosphor-Akt (p-Akt) and phosphor-Gsk-3β (p-Gsk-3β) one hour after LPS administration ( n = 4)], (4) Activity of inflammatory response was evaluated by protein degradation of IκBα (n = 4), (5) The apoptosis of myocardial cells was determined by TUNEL assay on the paraffin section of cardiac tissue 24 hours after LPS exposure (n = 4). Results (1) Hsp27 attenuated cardiac dysfunction significantly following LPS treatment. Compared with the primary value, LPS induced the depression of cardiac function both in WT rats and Hsp27Tg rats. However, the cardiac dysfunction was attenuated significantly in Hsp27Tg rats compared with that in WT rats ( P < 0.01 or 0.05) . (2) Hsp27 attenuated IκBα degradation after LPS administration. Compared with the primary value, LPS led to LκBα degradation by (72.92 + 9.20) % in WT rats and by (41.43 + 24.10) % in Hsp27Tg rats. The overexpression of Hsp27 lessened the IκBα degradation significantly (P < 0.05). The similar results were obtained in rat myocardial cell culture of experiments. (3) Hsp27 enhanced the activation of PI3K/Akt signaling following LPS exposure. One hour after LPS administration, the relative levels of p-Akt and p-GSK-30 were (3.11 + 0.83) and (3.19 + 1.04), respectively in WT rats, and (5.13 + 0.73) and (5.71 + 1.20) in Hsp27Tg rats, respectively. Compared with WT rats, the levels of p-Akt and p-GSK-3β were significantly higher in Hsp27Tg rats (P < 0.05). (4) The Hsp27 lessened LPS-induced the apopto-sis of myocardial cells. Twenty-four hours after LPS treatment, the percentages of myocardial cell apoptosis were (6.46+ 1.74)% in WT rats and (2.88 + 0.91)% in Hsp27Tg rats. Compared with WT rats, LPS-induced apoptosis in myocardial cells was significantly decreased in Hsp27Tg rats (P < 0.01). Conclusions The overexpression ofHsp27 attenuates cardiac dysfunction significantly during endotoxemia, and the mechanisms may be attributed to the activation of PDK/Akt signaling pathway.
2.Diatoms in drowned and postmortem immersed rabbits' lungs.
Xiang-Yang LI ; Jian ZHAO ; Chao LIU ; Sun-Lin HU ; You-Chuan ZHANG ; Jin-Feng WEN ; Jian-Ding CHENG
Journal of Forensic Medicine 2014;30(2):81-87
OBJECTIVE:
To investigate the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs.
METHODS:
Sixty-two rabbits were randomly divided into drowning group (n = 30), postmortem immersion group (n = 30) and land death group (n=2), and the diatoms in each lung lobe were analyzed quantitatively and qualitatively by microwave digestion and scanning electron microscopy.
RESULTS:
In the drowning group, the diatoms were detected in each lung lobe with Cyclotella and Melosira in the majority. In the postmortem immersion group, Cyclotella was in the majority. And the diatoms weren't detected in some lung lobes in postmortem immersion. There were significant differences in the detection rates of upper lobe of left lung, middle lobe and cardiac lobe of right lung in two groups (P < 0.05).
CONCLUSION
Based on the microwave digestion and scanning electron microscopy, the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs can be analyzed and used as references for testing theory.
Animals
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Autopsy
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Diatoms/isolation & purification*
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Drowning
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Lung/microbiology*
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Microscopy, Electron, Scanning
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Microwaves
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Rabbits
3.Effect of heat shock protein 27 on the cardiac dysfunction induced by endotoxemia
Li LIU ; Wenjun YOU ; Xiaoyan MIN ; Xiaojin ZHANG ; Bo QIAN ; Juncheng DAI ; Zhengnian DING ; Xiang GAO ; Yunlin CHENG
Chinese Journal of Emergency Medicine 2009;18(4):386-391
Objective To study the effects of heat shock proetin 27(Hsp27)on the cardiac dysfunction induced by endotoxemia.Method All experiments were performed in the geriatric lab of the First Affiliated Hospital Of Nanjing Medical University,and in the Animal Model Center of Nanjing University.The genotyping of the transgenic mice with cardiac-specific overexpression of Hsp27(Hsp27 Tg)was assayed by PCR and the expression of Hsp27 was determined by western blot.Hsp27 Tg and its wild type littermates(WT)were intraperitoneally injected with LPS(10 mg/kg),and 24 hours later,cardiac function was measured by echocardiography(n=6/group).The accumulated mice mortality was recorded within 70 hous after intraperitoneal injection of LPS(20mg/kg)(n=37/WT,n=27/Hsp27Tg).The NF-kB activity for cardiac-tissue samples was analyzed by electrophoretic mobility shift assay(ENSA)and for cell culture samples by dual-reporter gene assay(n=4/group).The comparison of multiple groups was performed by one-way analysis of variance(ANOVA),and comparison of two groups was performed by Scheffe-test.Survival curves were analyzed by the log-rank test.P<0.05 wns considered to be significant.Results The high expression of Hsp27 exhibited in myocardium of Hsp27 Tg,whereas not in myocardium of WT.LPS significantly reduced the cardiac function both in Hsp27 Tg and WT.However,compaled with LPS-treated WT,cardiac function was more significantly improved as evidenced by the increases of EF by 27.33%and FS by 37.09%(P<0.01 or P<0.05).Seventy hours after LPS injection,the mortality was 11.11% in Hsp27 Tg and 37.84% in WT.Compared with WT,the survival rate of Hsp27 Tg significantly increased(P<0.05).The NF-kB activation was significantly inhibited by Hsp27(P<0.01 or P<0.05).Conclusions The high cardiac-specific expression of Hsp27 significantly inhibits cardiac dysfunction induced by endotoxemia,and at the same time improve the survival rate.The mechanism may be connected with Hsp27 downregulating NF-kB-activation induced by LPS.
4.A study on the long-term outcome of hepatitis B e antigen-negative chronic hepatitis B compared with that of hepatitis B e antigen-positive chronic hepatitis B
Gui-Cheng WU ; Wei-Ping ZHOU ; You-Tong ZHAO ; Shu-Hua GUO ; Ai-Long HUANG ; Hong REN ; Ding-Feng ZHANG ;
Chinese Journal of Infectious Diseases 2001;0(03):-
0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.
5.Down-regulation of APLP1 mRNA expression in hippocampus of pilocarpine-induced epileptic rats.
Cheng WANG ; Zi-Li YOU ; Ding-Ding ZHANG
Neuroscience Bulletin 2009;25(3):109-114
OBJECTIVETo investigate the expression of amyloid beta precursor-like protein 1(APLP1) gene on the transcription level in hippocampus of pilocarpine-induced epileptic rats.
METHODSEpileptic rats were developed by LiC1 (3 mmol/kg, i.p.) approximately 20 h prior to pilocarpine (30 mg/kg, i.p.) administration. The 3' end partial sequence of rat APLP1 gene was cloned, and the expression levels of APLP1 mRNA in hippocampus of epileptic rats at 6 h, 30 h, 7 d and 15 d were determined by semi-quantitative RT-PCR.
RESULTSThe 3'end partial sequence of rat APLP1 gene shared a 97% homology with that of mice, and 90% with that of human. The APLP1 amino acid sequence of rat was identical with that of mouse, but was different from that of human in 3 residues. Moreover, pilocarpine induced a significant down-regulation of APLP1 mRNA expression at 6 h after epilepsy initiation (P< 0.05), and at 30 h, this down-regulation became more dramatic (P< 0.01), which lasted till day 15 (P< 0.01).
CONCLUSIONThe 3' end of APLP1 gene is highly conserved, and APLP1 mRNA expression is kept at low level in hippocampus of pilocarpine-induced epileptic rats.
Amino Acid Sequence ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Base Sequence ; Disease Models, Animal ; Down-Regulation ; physiology ; Epilepsy ; chemically induced ; pathology ; Hippocampus ; metabolism ; Humans ; Male ; Mice ; Molecular Sequence Data ; Pilocarpine ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sequence Alignment ; Time Factors
6.DNA extraction from bones and teeth using AutoMate Express forensic DNA extraction system.
Lin-Lin GAO ; Nian-Lai XU ; Wei XIE ; Shao-Cheng DING ; Dong-Jing WANG ; Li-Qin MA ; You-Ying LI
Journal of Forensic Medicine 2013;29(2):127-129
OBJECTIVE:
To explore a new method in order to extract DNA from bones and teeth automatically.
METHODS:
Samples of 33 bones and 15 teeth were acquired by freeze-mill method and manual method, respectively. DNA materials were extracted and quantified from the triturated samples by AutoMate Express forensic DNA extraction system.
RESULTS:
DNA extraction from bones and teeth were completed in 3 hours using the AutoMate Express forensic DNA extraction system. There was no statistical difference between the two methods in the DNA concentration of bones. Both bones and teeth got the good STR typing by freeze-mill method, and the DNA concentration of teeth was higher than those by manual method.
CONCLUSION
AutoMate Express forensic DNA extraction system is a new method to extract DNA from bones and teeth, which can be applied in forensic practice.
Automation
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Bone and Bones/chemistry*
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DNA/isolation & purification*
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DNA Fingerprinting/methods*
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Forensic Medicine/methods*
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Humans
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Microsatellite Repeats
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Polymerase Chain Reaction
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Specimen Handling/methods*
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Tooth/chemistry*
7.Long-term health-related quality of life in chronic hepatitis B patients.
Gui-cheng WU ; Wei-ping ZHOU ; You-rong ZHAO ; Shu-hua GUO ; Zhi-yi WANG ; Shu-bi ZOU ; Quan-hai ZHANG ; Hong REN ; Ai-long HUANG ; Ding-feng ZHANG
Chinese Journal of Hepatology 2003;11(5):275-277
OBJECTIVETo evaluate their long-term outcome and the efficacy and economic significance of antiviral drugs by investigating the long-term health-related quality of life (HQL) in chronic hepatitis B (CHB) patients.
METHODSThe HQL of 101 CHB patients with biopsy-proven 6 to 18 years ago and 105 persons of general population as control was studied with revised SF-36 questionnaire.
RESULTSThe HQL in CHB patients was lower than that in general population in physical functioning, role physical, general health, mental health, and specific symptoms (mu > or = 2.10, P<0.05).
CONCLUSIONSThe long-term HQL in chronic hepatitis B patients is poor.
Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Cost of Illness ; Female ; Follow-Up Studies ; Hepatitis B, Chronic ; drug therapy ; economics ; Humans ; Male ; Middle Aged ; Quality of Life ; Surveys and Questionnaires
8.Relationship between urokinase-type plasminogen activator expression and peritoneal metastatic potency in different gastric cancer cell lines.
You-cheng DING ; Zheng-gang ZHU ; Bing-ya LIU ; Yu-bao JI ; Xue-hua CHEN ; Yi ZHANG ; Ying-ya YU ; Yan-zhen LIN
Chinese Journal of Gastrointestinal Surgery 2005;8(3):249-251
OBJECTIVETo compare the expression and activities of urokinase type plasminogen activator (uPA) among different gastric cancer cell lines and investigate their relations with peritoneal metastatic potency.
METHODSThe uPA expression in 4 gastric cancer cell lines (AGS, SGC7901, MKN45, MKMN28) was detected using ELISA and Western blot methods. uPA activity was detected simultaneously using uPA activity kit. The gastric cancer cells were cultured with confluent mesothelial cells in 24-well plates or Boyden chambers for different times. The adhesive cells were counted directly under a microscope. The motility and invasion of gastric cancer cells were determined by MTT assay.
RESULTSAmong four gastric cancer lines,the highest expression of uPA was found in SGC7901 and the highest uPA activity in MKN45, while the lowest expression and activity of uPA in AGS. Compared with the other three lines, MKN45 had stronger adhesion than MKMN28 (P< 0.05), SGC7901 (P< 0.05), and AGS (P< 0.01), but there were no significant differences in motility and invasion among MKN45, MKN28 and SGC7901. The adhesion,motility and invasion of AGS were weaker compared with those of the other three cell lines.
CONCLUSIONThe uPA expression and activity are significantly different among 4 gastric cancer cell lines, and positively correlated with their peritoneal metastatic potency.
Blotting, Western ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasm Metastasis ; Peritoneal Neoplasms ; metabolism ; secondary ; Stomach Neoplasms ; classification ; metabolism ; pathology ; Urokinase-Type Plasminogen Activator ; metabolism
9.uPA expression of gastric cancer cell lines and its correlation with peritoneal seeding.
You-Cheng DING ; Zheng-Gang ZHU ; Bing-Ya LIU ; Yu-Bao JI ; Xue-Hua CHEN ; Yi ZHANG ; Ying-Yan YU ; Yan-Zhen LIN
Chinese Journal of Oncology 2005;27(1):13-15
OBJECTIVETo study the correlation between expression of urokinase-type plasminogen activator (uPA) and capability of tumor cell seeding to the peritoneal membrane by different gastric cancer lines.
METHODSExpression of uPA in 4 human gastric cancer cell lines was examined by semi-quantitative RT-PCR, ELISA and Western blot. uPA activity was determined by an assay kit. After ip inoculation of cancer cells to nude mice, tumors on peritoneal membrane was grossly examined for tumor cell seedings.
RESULTSSGC7901 was the highest in uPA expression among human gastric cancer cell lines AGS, SGC7901, MKN45, and MKN28. MKN45 had the strongest uPA activity, while AGS was lowest in both uPA expression and activity. Peritoneal seeding tumors of various sizes were observed in mice inoculated with SGC7901 and MKN45 cells. In addition to peritoneal seedings, bloody ascites was present in mice inoculated with MKN28. The MKN45-inoculated mice took the least time to develop tumors and had the shortest surviving period. No peritoneal seeding was seen in mice inoculated with AGS cells.
CONCLUSIONThree of 4 human gastric cancer cell lines studied express uPA mRNA and activity, which correlate with their peritoneal seeding potentials.
Adenocarcinoma ; enzymology ; secondary ; Animals ; Cell Line, Tumor ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Seeding ; Peritoneal Neoplasms ; secondary ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; enzymology ; pathology ; Urokinase-Type Plasminogen Activator ; biosynthesis ; genetics
10.Prevalence on overweight and obesity in Han, Uygur and Hazakh in adults from Xinjiang
Cheng LIU ; Xiang MA ; Yi-Tong MA ; Fen LIU ; Yi-Ning YANG ; Ding HUANG ; Xiao-Mei LI ; Ying HUANG ; You CHEN ; Bang-Dang CHEN ; Xiang XIE
Chinese Journal of Epidemiology 2010;31(10):1139-1143
Objective To investigate the prevalence and distributing feature of overweight and obesity in Han, Uygur and Hazakh population in adults from Xinjiang. Methods Four-stage selected random samples with maternal age at 35 or over were used to analyze the prevalence and distributing feature of self-reported congestive heart failure in different nationalities, age, sex. The sampled adult population were collected from 6 localities(Urumqi, Kelamayi, Fukang, the Turfan Basin locality, Hetian locality, Yili Hazakh autonomous prefecture), 23 municipalities and 7 locality and 5 autonomous counties in Xinjiang. Results 16 460 people were surveyed. The prevalence rates of overweight and obesity were 36.1% and 26.9% in Han, Uygur and Hazakh population in Xinjiang,respectively from February, 2007. The prevalence rates of overweight and obesity were 41.4% and18.4% in Han population, 34.9% and 28.9% in Uygur population, but 32.8% and 40.1% in Hazakh population. The prevalence rate of overweight and obesity was higher in males(x2= 135.00, P<0.05).The prevalence rates of overweight and obesity were different between different ethnic groups(x2=338.232, P<0.05). The prevalence of overweight was highest in Han population, with the highest seen in Hazakh population. The prevalence rates of overweight and obesity were increasing with age (x2=246.80,P<0.05). The overweight rate in 45-54 year olds and the obesity rate in 55-64 year olds reached their peak values. Results from logistic regression model analyses indicated that the prevalence of overweight and obesity in Xinjiang were statistically associated with age, educational level, jobs, smoking and alcohol consumption. Conclusion The prevalence rates of overweight and obesity were much higher in the population of Xinjiang but different among ethnicities. The prevalence of overweight was the highest in Han male population and the rate of obesity in Hazakh male population was the highest.