3.Construction and application of a new rat-holding device
Jieru GUO ; Wen ZHU ; Chenghao LI ; Fei YIN ; Guangwei ZHANG ; Can TAO ; Yi ZHOU
Chinese Journal of Comparative Medicine 2015;(8):76-78
Objective To provide a practical device and protocol to hold conscious rats for subsequent operations which can overcome the disadvantages of existing methods .Users can complete the experiment more efficiently , with or without prior experience .Methods Using transparent plastic film , plastic sealing machine and sponge to make a simple device for holding rats , by taking advantage of their escaping nature .To compare the performance of the new method and existing methods for holding and injecting rats .Results Compared with existing methods , the new device and method can reduce the time-consuming to hold rats by 44.7%, from 18.13 seconds to 10.03 seconds.For holding and injecting , the new method can reduce the time-consuming by 55.3%, from 139.33 seconds to 52.26 seconds .Conclusions The new device and method is good for holding and injecting rats or drawing blood from the caudal veins .It can shorten the time of operation and reduce the stress reaction in the animals .It’ s especially helpful for inexperienced experimenters such as students in teaching and research tasks .
4.The Effect of p65 siRNA on the Expression of Pro-Inflammatory Factors under Conditions Mimicking Ischemia/Reperfusion in Human Umbilical Vein Endothelial Cells
Min ZENG ; Zhiyong WU ; Yin ZHENG ; Yangli HE ; Yi FEI ; Xiaojun LIU
Tianjin Medical Journal 2014;(2):97-100
Objective To investigate the effect and regulation mechanism of mimic ischemia/reperfusion (I/R) cul-ture of human umbilical vein endothelial cells (HUVECs) on levels of tumor necrosis factor(TNF)-αand intercellular adhe-sion molecule (ICAM)-1. Methods HUVECs were randomly divided into four groups:control group (normal media cell cul-ture+control siRNA transfection), mimic I/R+control siRNA transfection group (HUVECs were transfected with control siR-NA, for 48 h ,and then received mimic ischemic media culture for 30 min followed by normal media culture for 4 h), normal culture+p65 siRNA transfection group and mimic I/R+p65 siRNA transfection group. The expression levels of TNF-αand ICAM-1 mRNA and protein were determined by real-time PCR and enzyme linked immunosorbent assay (ELISA), respec-tively. Results The mRNA and supernatant protein levels of TNF-αand ICAM-1 were significantly increased in mimic I/R culture group (4.96±0.16 and 3.33±0.30)μg/L than those of other tree groups (P<0.05). The level of ICAM-1 mRNA was significantly higher in I/R+p65 siRNA transfection group (1.87±0.21)μg/L than that of control group (1.58±0.15) μg/L and normal culture+p65 siRNA transfection group [(1.69±0.21)μg/L, P<0.05]. The levels of TNF-αand ICAM-1proteins were (329.98 ± 12.18) μg/L and (654.74 ± 64.79) μg/L in mimic I/R+control siRNA transfection group, which were significantly higher than those of other three groups (P<0.05). The levels of TNF-αand ICAM-1 proteins were (129.65±22.42)μg/L and (185.76±11.27)μg/L in mimic I/R+p65 siRNA transfection group, which were significantly lower than those of contro group [(183.50±11.77)μg/L and (280.43±13.76)μg/L, P<0.05]. Conclusion The silencing of p65 through transfection of p65 siRNA in HUVECs inhibited mimic I/R-induced mRNA and protein expressions of TNF-αand ICAM-1.
6.Formaldehyde inhalation may damage olfactory bulb and hippocampus in rats.
Yi-qiao LI ; Hao-hao CHEN ; Yi-fei YIN ; Fei HAN ; Xue-song YE ; Shu-cai LING
Journal of Zhejiang University. Medical sciences 2010;39(3):272-277
OBJECTIVETo investigate the effects of formaldehyde inhalation on the morphological damage, and Glu, GABA and NOS contents in olfactory bulb and hippocampus of rats.
METHODSTwenty SD rats were equally divided into two groups: rats in the control group inhaled fresh air, while the animals in experimental group were exposed to the air containing formaldehyde (12.5 mg/m(3), 4 h/d) for 7 days. Then rats were sacrificed and frozen sections of olfactory bulb and hippocampus were prepared. The morphological changes were examined and the Glu, GABA and NOS contents were detected using Nissl-staining, immunohistochemistry and Western blot, respectively.
RESULTCompared with the control group, there was a significant confusion and shrink of neuron morphology in experimental group, the number and staining intensity of Glu and NOS positive cells and protein contents were reduced. The protein expression of GABA was also decreased in the formaldehyde group.
CONCLUSIONFormaldehyde inhalation can cause a severe morphological damage of olfactory bulb and hippocampus in SD rats,which may further impair memory and learning ability through the reduction of Glu, GABA and NOS expression.
Animals ; Formaldehyde ; toxicity ; Glutamic Acid ; metabolism ; Hippocampus ; drug effects ; metabolism ; pathology ; Inhalation Exposure ; Learning ; drug effects ; Neurons ; drug effects ; metabolism ; pathology ; Nitric Oxide Synthase ; metabolism ; Olfactory Bulb ; drug effects ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism
7.GAD67-GFP expression and co-localization with bNOS in main olfactory bulb of GAD67-GFP knock-in mouse.
Fei HAN ; Jing YANG ; Wen-fu YU ; Yi-fei YIN ; Sheng-xi WU ; Shu-cai LING
Journal of Zhejiang University. Medical sciences 2012;41(2):159-170
OBJECTIVETo investigate the distribution of GAD67 and the co-localization with bNOS in the main olfactory bulb of GAD67-GFP knock-in mouse.
METHODSPolymerase chain reaction was applied to identify the genotype of GAD67-GFP knock-in mouse, the animals were sacrificed and frozen sections of olfactory bulb were prepared. The Nissl-staining was performed to show an framework of the neuron in the olfactory bulb. The distribution of GAD67 and co-localization with bNOS were detected by immunofluorescence technique.
RESULTSThe proportion of GAD67-positive cells among DAPI-positive cells were (42.98 ± 0.92)% in glomerular layer, (23.64 ± 0.84)% in mitral cell layer and (77.75 ± 0.84)% in granule cell layer; the bNOS-positive cells mainly existed in glomerular layer and mitral cell layer, very few in granule cell layer. No co-localization of GAD67 and bNOS in granule cell layer and mitral cell layer was found, but there was dispersed distribution in glomerular layer.
CONCLUSIONGAD67-positive neurons mainly appear in glomerular layer and granule cell layer, and the bNOS is mostly expressed in glomerular layer and mitral cell layer; while the co-localization of GAD67 and bNOS only occurs in glomerular layer of olfactory bulb.
Animals ; Gene Knock-In Techniques ; Glutamate Decarboxylase ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Mice, Transgenic ; Neurons ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; Olfactory Bulb ; metabolism ; Tissue Distribution
8.Risk factors of the failure in digit replantation.
Fei YIN ; Jing-yi MI ; Yong-jun RUI ; Ya-jun XU ; Qun YAO ; Yang QIU ; Zun-shan KE ; Zhen-zhong SUN
China Journal of Orthopaedics and Traumatology 2015;28(5):429-432
OBJECTIVETo explore the relative factors on the failure in digit replantation in order to take preventions to control the risk factors.
METHODSFrom January 2013 to December 2013, 236 consecutive patients (311 fingers) underwent digit replantation were collected to analyze retrospectively, involving 183 males and 53 females with an average age of 34.5 years old ranging from 2 to 62 years old (6 cases under 6 years old and 230 cases elder than 6 years old). There were 51 thumbs, 87 index fingers, 78 middle fingers, 63 ring fings and 32 little thumbs. Forty cases(forty fings) who were failured as the observation group, the others as the control group. The factors of age, gender, finger, cause of injury, smoking history, ischemia duration, plane of division, condition of venous drainage and condition of arterial repair we assessed.
RESULTSAll 236 cases with 311 fingers were replanted, 40 fingers were failured after operation. The relative factors on the failure in digit replantation included smoking history, cause of injury, plane of division, condition of venous drainage and condition of arterial repair (P< 0.05). There were no significant correlation between the failure and age, gender, finger and ischemia duration (P>0.05).
CONCLUSIONSmoking history, causes of injury, plane of division, condition of venous drainage and condition of arterial repair are risks of failure in digit replantation. Before choosing the type of operation, it should be think about the patient's general conditions, injury status, grasp firmly the operative indications and actively carry out surgical treatment.
Adolescent ; Adult ; Child ; Female ; Finger Injuries ; surgery ; Fingers ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Replantation ; Retrospective Studies ; Risk Factors ; Thumb ; injuries ; surgery ; Treatment Failure ; Young Adult
10.Altered expression of PTEN gene and LOH of its epigenetic microsatellite in gastric carcinoma.
Jin-yi LI ; Hua-chuan ZHENG ; Lin YANG ; Lei XU ; Xue-Fei YANG ; Hong GAO ; Yin-chang ZHANG ; Yan XIN
Chinese Journal of Oncology 2004;26(7):389-392
OBJECTIVETo investigate the expression of PTEN and loss of heterozygosity (LOH) of its epigenetic microsatellite in gastric carcinoma and explore their roles in progression of gastric carcinoma.
METHODSLOH of epigenetic microsatellites of PTEN (D10S541, D10S583 and D10S1687) in advanced gastric cancer was detected by PCR-SSCP. Expression of PTEN mRNA and protein in normal gastric mucosa and gastric cancer was evaluated by RT-PCR and SABC immunohistochemistry, respectively. The relationship between expression of PTEN mRNA and protein and lymph node metastasis or LOH of microsatellites was discussed.
RESULTSLOH of D10S541, D10S583 and D10S1687 was found in 37.5% (21/56) of advanced gastric cancers. The positive rates of PTEN mRNA expression were 80.4% (45/56), 45.5% (5/11) and 32.1% (18/56) in normal mucosa, early and advanced gastric carcinomas, respectively, while 78.6% (44/56), 44.5% (5/11) and 28.6% (16/56) at the protein level. PTEN mRNA and protein were less frequently expressed in early and advanced gastric carcinomas than that in normal gastric mucosa (P < 0.05). There was positive correlation between PTEN mRNA expression and LOH of microsatellites in advanced gastric carcinomas. PTEN protein expression paralleled with its mRNA expression (P < 0.05). The expression of PTEN mRNA and protein was negatively correlated with lymph node metastasis of advanced gastric carcinomas (P < 0.05).
CONCLUSIONDown-regulated expression of PTEN gene is found in different stages of gastric carcinoma, and is closely correlated with LOH of its epigenetic microsatellites, which probably is its underlying molecular mechanisms. It suggests that altered PTEN gene contributes to tumorigenesis and progression of gastric carcinomas.
Gastric Mucosa ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Loss of Heterozygosity ; Lymphatic Metastasis ; genetics ; Microsatellite Repeats ; genetics ; Neoplasm Staging ; PTEN Phosphohydrolase ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; biosynthesis ; genetics