1.Expressions and significance of miRNA - 101,enhancer of ZESTE homolog 2 and transforming growth factor-β1 in renal obstruction models in young rats
Jing CHANG ; Zhan GUO ; Lingang CUI ; Chuanchuan REN ; Xinmei GAO ; Shuman HUANG ; Bing BAI ; Jianguo WEN
Chinese Journal of Applied Clinical Pediatrics 2015;(21):1656-1659
Objective To explore the expressions and significance of miRNA - 101,enhancer of ZESTE homo-log 2(EZH2)and transforming growth factor(TGF)- β1 in the kidneys with complete uniateral ureteral obstruction (CUUO). Methods Thirty male SD rats,(6 ± 1)weeks old,weighted(150 ± 10)g,were divided into sham group, 7 - day group with CUUO and 14 - day group with CUUO by using random number table method,10 rats in each group. The obstructed kidney samples were collected in 7 and 14 days,respectively,for detecting the expression of miRNA -101 by real time - polymerase chain reaction(RT - PCR)and TGF - β1 and EZH2 protein by Western blot,immuno-histochemistry and hematoxylin - eosin staining. Their correlated expressions were analyzed. Results RT - PCR results showed that the expressions of miRNA - 101 in sham group and 7 - day group with CUUO were(12. 69 ± 1. 60)times and(3. 74 ± 1. 24)times which were higher than those of 14 - day group with CUUO,respectively,there was a signifi-cant difference among these 3 groups(P ﹤ 0. 05). The expressions of TGF - β1 and EZH2 proteins were 1. 14 ± 0. 12, 1. 04 ± 0. 14,0. 76 ± 0. 18 and 1. 04 ± 0. 04,0. 89 ± 0. 03,0. 73 ± 0. 02 in 14 - day group with CUUO,7 - day group with CUUO and sham group,respectively. There was a negative correlation between miRNA - 101 with EZH2( r =- 0. 92,P ﹤ 0. 05),and negative correlation with TGF - β1(r = - 0. 63,P ﹤ 0. 05),and positive correlation between EZH2 and TGF - β1(r = 0. 67,P ﹤ 0. 05);the expressions of miRNA - 101,EZH2 and TGF - β1 were associated with each other in obstruction renal in different time periods. Conclusions With the extension of obstruction time, miRNA - 101 expression decreased,EZH2 and TGF - β1 expression increased,evidently,which indicates that the de-velopment of renal interstitial fibrosis may be affected through regulating EZH2 expression of renal obstruction through miRNA - 101 in the young rats.
2.Analysis of nuclear magnetic resonance-based metabonomics of pancreatic cancer
Xianchao LIN ; Bohan ZHAN ; Shi WEN ; Zhishui LI ; Jianghua FENG ; Heguang HUANG
Chinese Journal of Digestive Surgery 2016;15(6):574-578
Objective To investigate the clinical value of serum metabonomic profile of pancreatic cancer using nuclear magnetic resonance (NMR)-based metabonomics.Methods The retrospective case-control study was adopted.The clinical data of 23 patients with pancreatic cancer (PC group) and 16 healthy volunteers (control group) who were admitted to the Fujian Medical University Union Hospital between December 2013 and December 2014 were collected.The serum of the 2 groups was measured by 1H NMR spectroscopy.Multivariate statistical analyses were performed to identify the characteristic metabolites in the 2 groups,including principal component analysis (PCA),partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA).Observation indicators included:(1) multivariate statistical analysis of serum metabonomic profile,results of PCA,PLS-DA and OPLS-DA,(2) screening of metabolites.Measurement data with normal distribution were presented as x ± s.The comparison between groups was evaluated with the t test.The count data were analyzed using the chi-square test.Results (1) The multivariate statistical analysis of serum metabonomic profile:results of PCA showed that expression rates of principal component 1 (PC1) and principal component 2 (PC2) to original data were 54.9% and 23.5%,with both cumulative contribution rate of 78.4%.Results of PLS-DA showed that the separative trend between PC group and control group was appeared,and variance of X and Y matrixes and predictive value were 0.254,0.816 and 0.385.Results of OPLS-DA showed that the differences of samples between the 2 groups were further increased,and differential metabolites were screened according to the distinction of scores between the 2 groups,value of R2X,R2Y and Q2 was 0.254,0.816 and 0.433.(2) Screening of metabolites:35 serum metabolites were detected in the 2 groups.Compared with the control group,levels of 3-hydroxybuyarate,citrate,formate,glutamate,isoleucine,methionine and phenylalanine in the PC group were elevated (r =0.524,0.511,0.656,0.566,0.503,0.498,0.648,P <0.05),and levels of 3-methylhistidine,alanine,glutamine,LDL and VLDL in the PC group were decreased (r =-0.607,-0.508,-0.560,-0.568,-0.559,P < 0.05).Conclusions Compared with healthy controls,several amino acids,citrate and lipoproteins demonstrate the metabolic differences in the serum of patients with pancreatic cancer.NMR based metabonomic profile technology can distinguish the difference of serum metabolites between patients with pancreatic cancer and healthy controls.NMR based metabonomic technology may be a promising method for the diagnosis of pancreatic cancer.
3.Generation of monoclonal antibodies against complexed prostate specific antigen and development of an antibody-based chemiluminescence immune quantifica-tion assay
Luting ZHAN ; Guiping WEN ; Min ZHAO ; Han YI ; Jiangwu LIU ; Xiaoyi GUO ; Haijun LIN ; Liunü HUANG ; Zizheng ZHENG
Chinese Journal of Immunology 2016;32(8):1171-1174,1178
Objective:To construct a chemiluminescense immune quantification assay based one paired mAbs against complexed prostate specific antigen ( c-PSA).Methods:Six week-old female BALB/c mice were immunized with the commercial c-PSA antigen.After serum titer reaching a platform stage ,the spleen was immunized and fused with mouse myeloma cell lines ( Sp2/0 ) .The hybridoma were screened by indirect ELISA ,and eight generated antibodies were paired to obtain a quantitative analysis of the chemical luminescence.Results:7D6 specifically recognized c-PSA,while 1A10 recognized total PSA(t-PSA).And the paired antibody 1A10/7D6 were determined to successfully construct a chemiluminescense immune response quantitative detection method through the detection of c-PSA standard and clinical serum samples .had,positive samples have statistically significant difference ( P<0.000 1 ) with negative samples.And the correlation coefficient R 2 was 0.97 between our c-PSA quantitative results and that of the Siemens c-PSA chemiluminescense immunoassay kit .The detection linear range was 0.1-100 ng/ml,and the sensitivity was 0.005 ng/ml.Conclusion:The paired monoclonal antibodies specifically detecting c-PSA were generated and a c-PSA chemiluminescense immunoassay were developed in this study .The detection capability of our method was comparable with that of the international commercial kit .
4.DNA microarray screening analysis in children with profound hearing impairment in Hubei province.
Yue ZHAN ; Xia WU ; Yujuan HU ; Xiang HUANG ; Jiade DUAN ; Haihua CHEN ; Jing JIN ; Dan LI ; Wen XIE ; Weijia KONG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(10):680-684
OBJECTIVE:
To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling.
METHOD:
Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan.
RESULT:
One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan.
CONCLUSION
Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.
Adolescent
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Child
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Child, Preschool
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China
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Connexin 26
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Connexins
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genetics
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DNA Mutational Analysis
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Deafness
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genetics
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Female
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Genetic Testing
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Humans
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Infant
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Male
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Membrane Transport Proteins
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genetics
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Mutation
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Oligonucleotide Array Sequence Analysis
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Sulfate Transporters
5.Synthesis and biodistribution of a bi-functional agent 99Tcm-Gd-DTPA-DG in tumor bearing nude mice
Wei, ZHANG ; Liang, CAI ; Yue, CHEN ; Zhan-wen, HUANG ; Zhi-ling, DING ; Feng, CAO ; Li, ZHANG ; Ling, HE
Chinese Journal of Nuclear Medicine 2011;31(2):117-120
Objective To evaluate the stability and biodistribution of a novel SPECT-MRI bi-functional agem 99Tcm-Gd-DTPA-DG in tumor-bearing nude mice. Methods DTPA-DG was synthesized and then conjugated with Gd2O3 to generate Gd-DTPA-DG. The tumor-bearing nude mice were scanned by MRI to evaluate the tumor targeting ability of Gd-DTPA-DG. The orthogonal experiment was applied to optimize pH value of reaction medium and reaction temperature. The radiolabeling efficiency was measured by thin layer chromatography. The distribution of 99Tcm-Gd-DTPA-DG in nude mice was evaluated by scintigrapy in vivo. The % ID/g was measured at different time after intravenous injection of 99Tcm-Gd-DTPA-DG. Results The tumor was significantly enhanced by Gd-DTPA-DG with MRI. The radiochemical purity of 99Tcm-Gd-DTPADG was about 98.5% and remained 96.2% at room temperature for 6 h. The tumor was well visualized by 99TcmGd-DTPA-DG SPECT at 2 h after injection. The tumor uptake was (1.48 ±0.12) %ID/g, and the rumor to muscle radioactivity ratio was 2.91. Conclusions MRI contrast of Gd-DTPA-DG may enhance tumor detection. 99Tcm-labeled Gd-DTPA-DG may be useful for tumor imaging and might have a potential role as a SPECT-MRI bi-functional agent.
6.Anterior expansion of sacral foramen and decompression of sacral plexus via lateral-rectus approach for sacral fractures complicated with sacral plexus injury
Qiubao ZHENG ; Shicai FAN ; Zhiyong HOU ; Chengla YI ; Qiguang MAI ; Tao LI ; Xiaorui ZHAN ; Fuming HUANG ; Xiangyuan WEN ; Yuancheng LIU
Chinese Journal of Orthopaedic Trauma 2021;23(1):47-54
Objective:To evaluate the anterior expansion of sacral foramen and decompression of sacral plexus via the lateral-rectus approach (LRA) in the surgical treatment of sacral fractures complicated with sacral plexus injury.Methods:From January 2013 to June 2018, 11 patients were treated at Department of Orthopaedics, The Third Hospital Affiliated to Southern Medical University for obsolete sacral fractures complicated with sacral plexus injury. They were 8 males and 3 females, aged from 17 to 54 years (average, 38 years). According to the Denis classification, all the sacral fractures belonged to Denis Zone Ⅱ. According to British Medical Research Council (BMRC) grading system, the nerve injury was complete damage in 2 cases and partial damage in 9. The mean time from injury to surgery was 6 months (range, from 0.7 to 12.0 months). After the sacroiliac joint was exposed via the LRA, the lumbosacral trunk was exposed and released between iliac vessels and the iliopsoas. Next, the S1 foramen was expanded and the S1 nerve root was released after separation of the median sacral artery and the internal iliac artery. Reduction and fixation of the sacroiliac joint was carried out for patients with unstable sacral fracture. X-ray and CT examinations of the pelvis were performed to evaluate fracture healing and neurological function recovery postoperatively.Results:Of this cohort of 11 cases, operation succeeded in 10 but failed in one whose sacral fracture was found to have completely healed with the S1 foramina totally occluded. The surgical time averaged 110 min (range, from 70 to 220 min) and the blood loss 1, 100 mL (range, from 450 to 2, 800 mL). Postoperative X-ray and CT examinations showed that the sacral foramens were expanded significantly without any complications. The follow-up time averaged 18 months (range, from 12 months to 4 years). By the BMRC grading system at the last follow-up, the neural function was completely recovered in 5 cases, partially recovered in 4 cases and not recovered in one.Conclusion:Significant anterior expansion of sacral foramen and decompression of sacral plexus via the LRA is a viable and effective alternative for treatment of sacral fractures complicated with sacral plexus injury.
7.The expression and clinical implication of plasma miR-328 in patients with atrial fibrillation.
Xi WANG ; Chun-guang QIU ; Zhen-wen HUANG ; Zhan-ying HAN ; Wen-jie LU ; Xiao-jie CHEN
Chinese Journal of Cardiology 2013;41(2):126-129
OBJECTIVETo observe the expression and clinical implication of plasma miR-328 in patients with atrial fibrillation (AF).
METHODSFifty-eight patients with AF (AF group: 17 paroxysmal AF, 21 persistent AF, and 20 permanent AF) and 15 healthy volunteers (Control group) were included. General clinical data and related biochemical parameters were collected. Plasma miR-328 levels were detected with quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The correlation between plasma miR-328 and AF risk factors was analyzed.
RESULTS(1) Compared with the control group, the expression level of plasma miR-328 was significantly elevated in AF group (fold 7.72 ± 9.32) (P < 0.05). (2) In AF group, the expression of plasma miR-328 was significantly different in different type of AF[paroxysmal AF with (1.98 ± 0.81), persistent AF with (6.57 ± 5.82) and permanent AF with (13.47 ± 12.29)] (P < 0.05), and which was increased in proportion to the duration of AF. (3) There was a positive correlation between plasma miR-328 level and left atrial diameter in the AF group (r = 0.310, P < 0.05).
CONCLUSIONmiR-328 expression is significantly increased in patients with AF, which may be involved in the atrial remodeling process of AF.
Aged ; Atrial Fibrillation ; blood ; Case-Control Studies ; Female ; Humans ; Male ; MicroRNAs ; blood ; Middle Aged
8.Rapid detection of influenza A/H1N1 virus by polymerase spiral reaction
Wen MA ; Derong DONG ; Dayang ZOU ; Ningwei LIU ; Xiaoming HE ; Da AO ; Zhan YANG ; Simo HUANG ; Yaqing XU ; Wei LIU ; Liuyu HUANG
Military Medical Sciences 2017;41(6):449-452
Objective To establish a polymerase spiral reaction (PSR) method for rapid detection of influenza A/H1N1 virus.Methods Six sets of primers were designed for influenza A/H1N1 virus HA gene, and the results were determined with real time kinetic turbidimetric assay and colorimetry method.Results and Conclusion The best primers were selected from six sets of primers, and the best temperature was determined as 65 degrees Celsius.Further experiments showed that the best primer had good specificity for detection of influenza A/H1N1 virus,without cross reactions with 14 other respiratory tract pathogenic nucleic acids.The sensitivity was up to 100 copies,and consistent with that of PCR.So a PSR method is established for rapid detection of the influenza A/H1N1 virus, which is simple, quick, highly specific and sensitive,and especially applicable to field and grass-roots units.
9.Cephalometry of Li people through 3-dimensional CT.
Wen-Jian YOU ; Lü-Ping HUANG ; Jian-Jun LI ; Nian WU ; Wang ZHAN ; Yun LIU
Chinese Journal of Plastic Surgery 2008;24(5):374-376
OBJECTIVETo investigate the skull characteristics of the Li people in Hainan through 3-D CT.
METHODSCT scan and 3-D reconstruction are very helpful for the cephalometry including the distance and angle measurement. The image can also be enlarged to make the measurement more precisely. 80 Li volunteers underwent the cephalometry through 3-D CT. The data were analyzed and compared with those an people.
RESULTSThe results showed difference between the genders of Li people. Compared with Han people, Li people has their own facial characteristics, such as wider face and wider orbital distance.
CONCLUSIONSCephalometry through 3-D CT can show the skull characteristics precisely. The data in this study has great significance in craniomaxillofacial surgery and ethnology.
Adolescent ; Adult ; Asian Continental Ancestry Group ; Cephalometry ; methods ; Female ; Humans ; Imaging, Three-Dimensional ; Male ; Skull ; diagnostic imaging ; Tomography, Spiral Computed ; Young Adult
10.Establishment of a rat model of mycoplasma pneumonia and the treatment.
Xiao-Hong HUANG ; Ming GONG ; Tao LI ; Wen-Ling JIANG ; Jing LIU ; Zhan-Qiu YANG
Journal of Southern Medical University 2009;29(11):2219-2221
OBJECTIVETo establish a rat model of mycoplasma pneumonia (MP) for investigating the pathogenesis of MP and its therapy with drugs.
METHODSThirty Wistar rats were randomly divided into 6 groups (n=5), including a control group, a MP model group, a erythromycin lactobionate group and 3 erythromycin microspheres groups (high, middle, and low dose groups). With the exception of those in the control group, all the rats received intranasal MP administration followed by corresponding treatments administered via tail vein injection. At different time points after inoculation of the pathogen, the lungs of the rats were taken for histopathological scoring.
RESULTSIn the MP model group, the lung pathology was characterized by patchy interstitial pneumonitis with predominantly lymphocyte infiltration and mucosal edema. The bronchiolar walls became thickened and the lumens narrowed. In erythromycin lactobionate and erythromycin microspheres treatment (high and middle dose) groups, clear cell boundaries were observed in the lungs where no obvious pathological changes were found. RT-PCR amplification showed positive results of MP RNA in the model group, erythromycin lactobionate group and erythromycin microsphere groups.
CONCLUSIONThe approach described is practicable to establish rat models of MP. Erythromycin microspheres can effectively relieve the lung inflammations and has therapeutic effect on MP.
Animals ; Disease Models, Animal ; Erythromycin ; therapeutic use ; Female ; Male ; Microspheres ; Pneumonia, Mycoplasma ; drug therapy ; Random Allocation ; Rats ; Rats, Wistar