1.Effects of traditional Chinese medicine Yiqi Huoxue Tongluo Jiedu prescription on pneumocyte apoptosis and c-Jun N-terminal protein kinase pathway in mice after lung ischemia/reperfusion injury
Lu SHI ; Dong SONG ; Xuguang JIA ; Ziyin LUO ; Bingqian XIANG ; Yongyue DAI ; Min LUO ; Wantie WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(1):73-77
Objective To observe the effects of Yiqi Huoxue Tongluo Jiedu fang (YHTJF) on pneumocyte apoptosis after lung ischemia/reperfusion (I/R) injury (LIRI) in mice and to investigate whether c-Jun N-terminal protein kinase (JNK) is involved in the mechanism of apoptosis.Methods Seventy C57BL/6J male mice were randomly divided into seven groups:normal control group (C group),carboxyl methyl cellulose-Na+normal control group (CMC-Na+C group),CMC-Na+sham group (CMC-Na+S group),CMC-Na+I/R group (CMC-Na+I/R group) and CMC-Na+YHTJF-low,-middle,-high dose groups (CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups).C group did not undergo any processing;in CMC-Na+S group,only was chest opened without clipping the lung hilum;in the rest of the four groups,they all underwent opening of the chest and clipping the lung hilum for 30 minutes,then the clipping of artery was relieved and left lung reperfusion was carried out for 3 hours.After operation,the mice were sacrificed,the lung tissues were harvested.Under light and electron microscopes,the lung morphological and ultra-structural changes were observed,and the changes of index of quantitative evaluation for alveolar damage (IQA) were determined.The terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was applied to evaluate the apoptosis index (AI) of the lung tissues.The protein and mRNA expressions of JNK and glucose regulating protein 78 (GRP78) in lung tissues were detected by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR);the correlations between lung AI and the expressions of mRNA and protein of JNK and GRP78,IQA were analyzed.Results Compared with CMC-Na+S group,IQA,AI and mRNA and the protein expressions of JNK and GRP78 in CMC-Na+I/R group were obviously higher [IQA:(74.00 ± 7.31)% vs.(7.00 ± 1.23)%,AI:(64.40 ± 11.97)% vs.(5.60 ± 1.14)%,JNK mRNA (gray value):1.143 ± 0.284 vs.0.152 ± 0.128,GRP78 mRNA (gray value):0.897 ± 0.129 vs.0.284 ± 0.044,JNK protein (A value):0.428 ± 0.074 vs.0.073 ± 0.052,GRP78 protein (A value):1.075 ± 0.145 vs.0.589 ± 0.060].Compared with CMC-Na+I/R group,the IQA,AI,protein and mRNA expressions of JNK and GRP78 in CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups were all lower,and the degree of reduction in group CMC-Na+YM was the most remarkable,greater than that in CMC-Na+YL or CMC-Na+YH group [IQA:(26.20 ± 3.35)% vs.(34.00±5.34)%,(41.20±9.18)%,AI:(29.40±3.05)% vs.(48.20±3.83)%,(39.20±6.14)%,JNK mRNA (gray value):0.681 ± 0.130 vs.0.804 ± 0.153,0.938 ± 0.11,GRP78 mRNA (gray value):0.450 ± 0.105 vs.0.747 ± 0.231,0.566 ± 0.115,JNK protein (A value):0.188 ± 0.049 vs.0.261 ± 0.065,0.209 ± 0.063,all P < 0.01],compared with the CMC-Na+I/R group,the expression of GRP78 protein was obviously higher in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkably high was in CMC-Na+YH group (A value:1.429 ±0.226 vs.1.130±0.169,1.128 ±0.177,all P < 0.01).The apoptosis of each group was mainly in the pulmonary vascular endothelial cells and alveolar epithelial cells,and brown particles were positive cells under light microscope.Under transmission electron microscope:nuclear pyknosis and margination under the nuclear membrane,cytoplasm condensed,lamellar bodies decreased and emptying increased,cell membrane microvilli decreased or disappeared,mitochondria swelling,inflammatory cells increased in alveolar septum and adhering onto the capillary walls could be seen in CMC-Na+I/R group.Compared with CMC-Na+I/R group,the lung tissue ultrastructural damage alleviated,ultrastructure of alveoli clearly seen,nuclear chromatin relatively uniform,cytoplasm increased,type Ⅱ alveolar epithelial cell surface microvilli relatively plenty,lamellar corpuscle number increased,mitochondria swelling ameliorated in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkable one was CMC-Na+YM group.AI was significantly positive correlated with the mRNA and protein expressions of JNK,GRP78 and IQA (r =0.907,0.928,0.880,0.712,0.911,all P < 0.01).Conclusions YHTJF may effectively alleviate the cell apoptosis in mice LIRI,and its mechanism may be related to the inhibition of JNK pathway.
2.Construction of a eukaryotic expression plasmid of Humanin.
Ben-yan LUO ; Xiang-ming CHEN ; Min TANG ; Feng CHEN ; Zhi CHEN
Journal of Zhejiang University. Science. B 2005;6(1):11-13
OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.
METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.
RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.
CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.
Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry
3.CTU diagnosis of 85 cases with repeat kidney and ureter duplication and its comparison with B-mode ultrasonography and IVU
Shanxi CHEN ; Yuantong GAO ; Xiaochan XIANG ; Min LUO ; Yang LI ; Xiaoyang WANG
The Journal of Practical Medicine 2014;(3):397-399
Objective To investigate the application values of B-mode ultrasonography , IVU and spiral CT urography (CTU) in duplication of kidney and ureter. Methods Data of the results from B-mode ultrasonography , IVU and spiral CTU on 85 cases with duplication of renal pelvis and ureter were retrospectively analyzed. Results The diagnostic accordance rates of B-mode ultrasonography , IVU and spiral CTU for duplication of kidney and ureter were 44.7%, 60.0% and 100%, respectively. CTU examination and image post-processing could clearly display the locations and degrees of dilatation or obstruction in duplication of kidney and ureter, and could also display the ectopic location and assess renal function. Conclusion The diagnostic accuracy rate of CTU examination and image post-processing for duplication of kidney and ureter is higher than those of B-mode ultrasonography and IVU.
5.Cellular expression profile of RhoA in rats with spinal cord injury.
Wen-Jie, WEI ; Zhi-Yuan, YU ; Huai-Jie, YANG ; Min-Jie, XIE ; Wei, WANG ; Xiang, LUO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(5):657-62
RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.
6.Effects of indomethacin on human choroidal melanoma OCM-1 xenografts in nude mouse
Xin-rui, LUO ; Hao, CHEN ; Qin-xiang, ZHENG ; Ling, QIN ; Min, LI ; Wen-sheng, LI
Chinese Journal of Experimental Ophthalmology 2011;29(1):8-12
Background Choroidal melanoma(CM)is a common form of primary ocular cancer in adults.It is reported that indomethacin has inhibitory effect on many tumor in vitro and in vivo,but whether it can inhibit the growth of CM has not been published. Objective This study was to investigate the anti-tumor activity of indomethacin on the growth of human CM OCM-1 cell xenografts in nude mice. Methods OCM-1 cells were subcutaneously implanted on 24 SPF female BALB/C.nu/nu nude mice to establish ectopic models of human CM.The nude mice with the tumor 5 mm were randomly divided into 4 groups:untreated group,normal saline solution(NS) group,indomethacin 1 ms/kg group,indomethacin 2 ms/kg group.The 1 mS/kg or 2 ms/kg indomethacin was intraperitoneally injected for 14 consecutive days in indomethacin 1 ms/kg group and indomethacin 2 me/kg group respectively.and 0.2 ml of 2%NS-DMSO was used at a same way in the NS group.No any agent was used as the untreated group.The volume and weight of implanted tumor as well as inhibitory rates of indomethaein on tumor were calculated.The expression of ki67 and survivin proteins were measured with immunohistochemistry,and the expression of survivin mRNA in CM was assessed by RT-PCR. ResuIts The tumor of indomethacin treatment group was reduced in volume and weight with a significant difference between treatment group and control group as well as indomethacin 1 ms/ks group and indomethacin 2 ms/kg group(P<0.05).The inhibitory rate of indomethacin 1 ms/kg and 2 ms/kg for tumor was 22.86%,48.00%respectively.The prolifiration index (PI)of ki67 in these 4 groups were (76.73±3.34)%,(73.30±2.95)%,(55.97±2.24)%,(32.87±2.91)%respectively,and significant difference was found in PI between indomethacin 2 mg/kg group and untreated group or NS group(P<0.05),but there was not significant difference between indomethacin 1 mg/kg and 2 ms/kg group(P>0.05).Compared with the control group,the indomethacin treatment groups showed the decreased expression of survivin protein and mRNA,and significant difference was found between indomethaein 2 ms/kg group and untreated group or NS group(P<0.05),however,no significant difference was found between indomethacin 1 mg/kg and 2 mg/kg group(P>0.05). Conclusion Indomethacin inhibits the growth of CM in nude mice through inhibiting the expression of survivin in the tumor and accelerating cell apoptosis and inhibiting tumor cell proliferation.
7.Application of MSCT and post-processing images to fractures of nasal bone in forensic identification.
Xiang-min CHEN ; Shi-guang LUO ; Zi-xuan WANG
Journal of Forensic Medicine 2005;21(3):183-187
OBJECTIVE:
To evaluate the application of MSCT and post-processing images to fractures of nasal bone in forensic identification.
METHODS:
134 cases were examined by thin slice scanning with MSCT and all of the data were sent to workstation for MPR and SSD. The result of MSCT was compared with that of X-ray.
RESULTS:
There are 55 (41.04%) cases of linear fracture, 46 (34.33%) cases of comminuted fracture, 27 (20.15%) cases of depressed fracture and 6 (4.48%) cases of no fracture in this sample. With X-ray or CR, 48 (35.82%) cases were misdiagnosed or underdiagnosed. 133 (99.25%) cases were confirmed by MSCT. Significance difference was found between X-ray and MSCT (chi2= 45.0816, P<0.001).
CONCLUSION
MSCT and post-processing images might be the chief evidence for nasal fractures in forensic identification.
Adolescent
;
Adult
;
Chromatography, Thin Layer
;
Female
;
Forensic Medicine/methods*
;
Fractures, Comminuted/diagnostic imaging*
;
Humans
;
Imaging, Three-Dimensional
;
Male
;
Middle Aged
;
Nasal Bone/injuries*
;
Skull Fractures/diagnostic imaging*
;
Tomography, Spiral Computed/methods*
;
Young Adult
8.Receptor-specific Ca2+ signaling in polarized cells.
Dong Min SHIN ; Min Goo LEE ; Xiang LUO ; Shmuel MUALLEM
Journal of Korean Medical Science 2000;15(Suppl):S46-S48
No abstract available.
Calcium Signaling/physiology*
;
Cell Polarity/physiology*
;
Epithelial Cells/physiology*
;
Epithelial Cells/cytology*
9.Risk factors for adverse neonatal outcomes in 254 twins.
Xiang CAO ; Xin-Hua QIAN ; Ying LUO ; Wei-Min HUANG
Chinese Journal of Contemporary Pediatrics 2010;12(10):777-780
OBJECTIVEThe aim of this study was to identify the risk factors for adverse neonatal outcome in twins in order to provide a basis for the improvement of the survival and neonatal outcomes of twins.
METHODSData from 254 twins admitted to Nanfang Hospital of Southern Medical University From January 2005 to December 2009 were retrospectively studied. Risk factors for adverse neonatal outcomes were assessed by logistic regression analysis.
RESULTSOf the 254 twins, 84 (33.1%) had an adverse outcome, including 10 (3.9%) neonatal deaths. Logistic regression analysis demonstrated that gestational age (≤34 weeks), cord abnormalities, meconium-stained amniotic fluid and 5-min Apgar scores (≤7) were independent risk factors for adverse neonatal outcomes (OR=4.434, 4.731, 3.424, 18.958, respectively; P=0.021, 0.001, 0.037, 0.011, respectively). Conception by assisted reproductive technology was shown as a protective factor for adverse neonatal outcomes (OR=0.389, P=0.037).
CONCLUSIONSThe twins with gestational age ≤34 weeks, cord abnormalities, meconium-stained amniotic fluid or 5-min Apgar scores (≤7) are subject to adverse neonatal outcome.
Apgar Score ; Female ; Gestational Age ; Humans ; Infant Mortality ; Infant, Newborn ; Logistic Models ; Male ; Risk Factors ; Twins