1.Expression and biological role of Matrix metalloproteinases 16 in esophagus squamous cell carcinoma
Qiuli WU ; Biyun LIN ; Mengjie WU ; Ming LIU
China Oncology 2014;(6):423-432
Background and purpose:Esophageal carcinoma is one of main malignancies with rapid course and a poor prognosis in China. The reasons of poor overall survival are the invasion and metastasis of the tumor. Matrix metalloproteinase (MMPs) play essential roles in promoting tumor invasion and metastasis. In this study, we aimed to investigate the expression and functional signiifcance of matrix metalloproteinase 16(MMP-16) in esophageal squamous cell carcinoma (ESCC). We expect to ifnd a lead molecule for the beneift of early detecting tumor and the development of novel treatment of ESCC. Methods:The expression levels of MMP-16 protein and mRNA in human ESCC and the matched normal tissues were determined by immunohistochemistry, Western blot and Real-Time PCR (RT-PCR). The stable Ec109 cell line with MMP-16 knockdown and negative controls were established by RNA interference technology. The cell migration, invasion, proliferation and cell apoptosis of MMP-16 in stable interfered Ec109 cell line was examined by cell counting, scratch test, Transwell test and lfow cytometry assays. The data were analyzed by t test. Results:MMP-16 protein was downregulated in cancerous group compared with the matched normal tissue and correlated with the clinical features of histological differentiation (P<0.05) and tumor stage (P<0.05). The levels of MMP-16 mRNA and protein in Ec109 were signiifcantly decreased by RNA intetrence (P<0.05). We demonstrated that MMP-16 silencing signiifcantly promoted cell invasion and migration (P<0.05), and inhibited cell apoptosis (P<0.05), while no significant effect was observed on cell proliferation (P>0.05). Conclusion: MMP-16 is downregulated in human ESCC tissues. The cell migration and invasion is promoted by interference of MMP-16 in Ec109, while the cell apoptosis is inhibited. MMP-16 may be considered as a target gene for therapy of ESCC.
2.A case of fructose-1, 6-bisphosphatase deficiency with delayed diagnosis for 7 years and review of literature
Mengjie YANG ; Yili LU ; Huiping WU ; Xiaoou SHAN
Chinese Journal of Endocrinology and Metabolism 2021;37(5):457-461
To report a case of fructose-1, 6-bisphosphatase deficiency child with delayed diagnosis for seven years, and review the literature. A 13-year-old boy presented with recurrent episodes of hypoglycemia after infection since 6 years old, he also had convulsion and short stature. The laboratory finding revealed ketotic hypoglycemia, lactic acidosis, transient elevated ALT and UA. The genetic analysis showed compound heterozygous mutations of c. 960-961insG and c. 355G> A in FBP1 gene. Among eighteen patients reported in China, 88.9% had convulsion, 16.7% had growth retardation, the average delayed diagnosis spent 2.8 years, which could result in permanent brain damage and high death risk. The most common mutation was c. 960-961insG, followed by c. 355G>A and c. 490G>A, these mutations maybe hot spot sites of FBPl gene in China.
3.Therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis in rats
Xiaohong WANG ; Mengjie WU ; Wenyuan MA ; Ronglong WEI ; Qing MIAO ; Xun ZHUANG ; Xianglong XUE
Chinese Journal of Digestion 2016;36(10):670-675
Objective To investigate the therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis (SAP) in rats.Methods A total of 80 healthy Sprague-Dawley rats were divided into eight groups:SAP group (three hours、48 hours),Saussurea involucrate treated group (three hours、48 hours),ulinastatin control group (three hours、48 hours) and sham operation group (three hours、48 hours),10 rats in each group.After modeling,the rats of SAP group were regularly feeded and the rats of other three group were treated with Saussurea involucrate injection (1.04 mL/kg) intraperitoneal injection,ulinastatin 10 000 U/L tail vein injection,and saline femoral vein injection,respectively and injected every 12 hours.At three hours and 48 hours after treated,blood and pancreatic tissue samples were obtained.The mortality rate,serum amylase level and pathological changes of the pancreas of each group were observed.Serum tumor necrosis factor alpha (TNF-α),interleukin (IL)-6 and IL-10 levels were detected by enzyme linked immunosorbent assay (ELISA).The content of malondialdehyde (MDA) in pancreatic tissues was determined by chemical colorimetry.The level of TNF-α mRNA,IL 6 mRNA and IL-10 mRNA in pancreatic tissues were measured with reverse trascription-polymerase chain reaction (RT-PCR).The activity of nuclear factor kappa B p65 (NF-κB p65) in the pancreatic tissue was tested by immunohistochemistry.Single factor analysis of variance was used to compare multiple groups,and the least significant difference (LSD) method was used in the multiple comparisons between groups.Fisher's exact probability method was performed for rates comparison.Results At 48 hours,there was no statistically significant difference in mortality rate among Saussurea involucrate treated group,SAP group and ulinastatin groups (all P>0.05).At 48 hours,the histopathology score (8.13 ± 0.64),levels of serum amylase ((2 597.0±214.0) U/L),TNF-α ((254.4±11.6) ng/L),IL-6 ((441.4±14.6) ng/L),levels of pancreatic tissues MDA ((311.0±10.6) mmol/L),TNF-α mRNA(2.04±0.08),IL-6 mRNA (1.77±0.04)and activity of NF-κB p65 ((25.90±2.90)%) of Saussurea involucrate treated group were all lower than those of SAP group (11.40±0.89,(4 780.0±101.0) U/L,(396.0±7.4) ng/L,(664.4± 7.6) ng/L,(418.0± 10.6) mmol/L,2.94±0.03,2.63±0.08 and (51.60±5.27) %;however level of serum IL-10 ((133.5±6.9) ng/L vs (95.1±5.2) ng/L) and IL-10 mRNA of the pancreatic tissue (1.38±0.06 vs 0.85±0.03) significantly increased (F=253.07、441.63、489.40、2 465.00、196.65、477.89、562.79、131.70、560.18、570.04,all P<0.01).There was no significant differences in all above parameters between Saussurea involucrate treated group and ulinastatin groups (7.56±0.88,(2 607.0±239.0) U/L,(252.2 ±9.2) ng/L,(443.4±9.6) ng/L,(308.4±9.2) mmol/L,2.10±0.12,1.74±0.04,(26.00±3.67)%,(134.5±7.8) ng/L and 1.42±0.06) at 48 hours (all P>0.05).Conclusion Saussurea involucrate injection can eliminate oxygen free radicals and prevent to xidation,inhibit NF-κB activation,regulate synthesis and release of cytokines,and alleviate pancreatic injury in SAP rats,but it can not decrease mortality.
4.Analysis of nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cy-tomegalovirus strains in Guangzhou
Lijun CAI ; Jingjing HU ; Bo WANG ; Haihao SU ; Yuanbin WU ; Yingge MI ; Mengjie GUO
Chinese Journal of Microbiology and Immunology 2015;(6):407-413
Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.
5. A study of establishment of non-standard methods in qualification accreditation of virological laboratories
Jie LI ; Xiaozhou HE ; Mengjie YANG ; Xuejun MA ; Guizhen WU
Chinese Journal of Experimental and Clinical Virology 2018;32(2):211-213
The laboratories of National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, have developed a series of experimental method . These method have unique advantages over the national standard method and industry standard method . How to make these method become public products through legal procedures to serve disease prevention and control more extensively is an obligatory task for national virological laboratories. This article explores the establishment, verification, validation, and examination of virological non-standard method under laboratory certification and accreditation conditions through empirical research on the " RT-RAA method for detecting EV71/CA16 and other viruses" .
6.Stratified random sampling survey on senile chronic kidney diseases among elderly people aged over a certain Crops
Xue SONG ; Wenli WU ; Jiang CHENG ; Min GE ; Xinhong LU ; Zhaohui DENG ; Mengjie LIANG ; Yingbo SONG ; Ye ZHANG ; Weidong YI ; Xueli LIANG ; Shuang LIU ; Lei WANG ; Yan LI ; Xin ZHANG
Chongqing Medicine 2016;45(10):1360-1363,1366
Objective To understand the prevalence rate and related factors of chronic kidney disease (CKD) among elderly people aged more than 65 years old in the 66th regiment of the fourth division of A Crops in Xinjiang .Methods A total of 2 030 elderly people aged more than 65 years old in the 66th regiment of the fourth division of XPCC were distributed in 6 communities . Totally 334 permanent residents aged more than 65 years old were chosen from 2 communities by the stratified random sampling method .The renal injury indicators and related factors were detected .Results Among 329 residents with intact data ,after the age correction ,the prevalence rate of albuminuria ,hematuria and renal function decrease were 22 .2% ,14 .2% ,4 .9% ,respectively .The prevalence rate of CKD in this group was 32 .8% ,CKD stage 1―3 were dominated .The awareness rate was 15 .1% .The multiva‐riate Logistic regression analysis showed that gender and hypertension were independently associated with CKD .Conclusion The prevalence rate of CKD among elderly people aged over 65 years old in the 66th regiment of the fourth division of this Crops is high‐er .The related factors are gender and hypertension .
7.Schisantherin B Improves the Pathological Manifestations of Mice Caused by Behavior Desperation in Different Ages-Depression with Cognitive Impairment
Mengjie XU ; Feng XIAO ; Mengshi WANG ; Tingxu YAN ; Huilin YANG ; Bo WU ; Kaishun BI ; Ying JIA
Biomolecules & Therapeutics 2019;27(2):160-167
Depression is a major mood disorder. Abnormal expression of glial glutamate transporter-1 (GLT-1) is associated with depression. Schisantherin B (STB) is one bioactive of lignans isolated from Schisandra chinensis (Turcz.) Baill which has been commonly used as a traditional herbal medicine for thousands of years. This paper was designed to investigate the effects of STB on depressive mice induced by forced swimming test (FST). Additionally, we also assessed the impairment of FST on cognitive function in mice with different ages. FST and open field test (OFT) were used for assessing depressive symptoms, and Y-maze was used for evaluating cognition processes. Our study showed that STB acting as an antidepressant, which increased GLT-1 levels by promoting PI3K/AKT/mTOR pathway. Although the damage is reversible, short-term learning and memory impairment caused by FST test is more serious in the aged mice, and STB also exerts cognition improvement ability in the meanwhile. Our findings suggested that STB might be a promising therapeutic agent of depression by regulating the GLT-1 restoration as well as activating PI3K/AKT/mTOR pathway.
Animals
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Cognition
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Cognition Disorders
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Depression
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Glutamic Acid
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Herbal Medicine
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Learning
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Lignans
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Memory
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Mice
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Mood Disorders
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Physical Exertion
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Schisandra
8.In vitro expression of human cytomegalovirus UL148 RNA and prediction of its functional sites
Jingjing HU ; Yuanbin WU ; Qiqi TAN ; Haihao SU ; Juncai DING ; Yuanyuan GUO ; Binhua XIE ; Lijun CAI ; Mengjie GUO ; Bo WANG
Chinese Journal of Microbiology and Immunology 2018;38(2):94-97
Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.
9.Temporal and spatial pattern of RhoA expression in injured spinal cord of adult mice.
Xiaoning KANG ; Jingkun WEN ; Xianghai WANG ; Mengjie PAN ; Weiwei ZHANG ; Xiaoduo ZHAN ; Zhongying LIU ; Wutian WU ; Jiasong GUO
Journal of Southern Medical University 2013;33(4):463-468
OBJECTIVETo quantitatively analyze the temporal and spatial pattern of RhoA expression in injured spinal cord of adult mice.
METHODSA spinal cord transection model was established in adult mice. At 1, 3, 7, 14, 28, 56 and 112 days after the surgery, the spinal cords were dissected and cryosectioned for RhoA/NF200, RhoA/GFAP, RhoA/CNPase or RhoA/IBA1 double fluorescent immunohistochemistry to visualize RhoA expressions in the neurons, astrocytes, oligodendrocytes and microglia. The percentages as well as the immunostaining intensities of RhoA-positive cells in the parenchymal cells were quantitatively analyzed.
RESULTSRhoA was weakly expressed in a few neurons and oligodendrocytes in normal spinal cord. After spinal cord injury, the percentage of RhoA-positive cells and RhoA expression intensity in the spinal cord increased and peaked at 7 days post injury (dpi) in neurons, oligodendrocytes and astrocytes, followed by a gradual decrease till reaching a low level at 112 dpi. In the microglia, both the RhoA-positive cells and RhoA expression intensity reached the maximum at 14 dpi and maintained a high level till 112 dpi.
CONCLUSIONTraumatic spinal cord injury can upregulate RhoA expression in the neurons as well as all the glial cells in the spinal cord. RhoA expression patterns vary with post-injury time, location and among different parenchymal cells in the injured spinal cord.
Animals ; Astrocytes ; metabolism ; Female ; Mice ; Mice, Inbred Strains ; Microglia ; metabolism ; Neuroglia ; metabolism ; Neurons ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; rho GTP-Binding Proteins ; metabolism
10.An integrated segmentation method for 3D ultrasound carotid artery.
Xin YANG ; Huihui WU ; Yang LIU ; Hongwei XU ; Huageng LIANG ; Wenjuan CAI ; Mengjie FANG ; Yujie WANG
Chinese Journal of Medical Instrumentation 2013;37(4):235-239
An integrated segmentation method for 3D ultrasound carotid artery was proposed. 3D ultrasound image was sliced into transverse, coronal and sagittal 2D images on the carotid bifurcation point. Then, the three images were processed respectively, and the carotid artery contours and thickness were obtained finally. This paper tries to overcome the disadvantages of current computer aided diagnosis method, such as high computational complexity, easily introduced subjective errors et al. The proposed method could get the carotid artery overall information rapidly, accurately and completely. It could be transplanted into clinical usage for atherosclerosis diagnosis and prevention.
Algorithms
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Angiography
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methods
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Carotid Arteries
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diagnostic imaging
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Humans
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Imaging, Three-Dimensional
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methods
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Ultrasonography