1.Constructing the dual luciferase reporter vector containing human DRD1 promoter region
Chunhong WANG ; Zhe LI ; Minghan WANG ; Lili DENG ; Huan WANG
Chinese Journal of Tissue Engineering Research 2016;20(40):6060-6066
BACKGROUND:The polymorphisms of dopamine receptor in promoter region wil affect the expression of the receptor, thereby affecting the dopaminergic neurotransmitter, final y lead to related diseases.
OBJECTIVE:To construct the dual luciferase reporter vector containing human DRD1 promoter region and determine its activity, which could provide the basic tool for studying the transcriptional regulation of DRD1 gene.
METHODS:DRD1 promoter sequence was amplified by PCR using the human blood genomic DNA and cloned into pGM-T vector. After sequencing, the correctly constructed vectors were ligated to the firefly luciferase reporter plasmid pGL3-Basic. The cloned pGL3-Basic vectors were transfected into HEK293 using cationic liposome method. In the meanwhile, PGL3-Basic vector with no promoter was co-transfected with pGL3-TK plasmid as negative control group. The relative fluorescence intensity was measured by chemiluminescence.
RESULTS AND CONCLUSION:(1) Recombinant luciferase reporter gene vectors were confirmed by restriction analysis and sequencing. (2) Compared with the negative control group, the HEK293 cel s transfected by recombinant vectors presented transcriptional activity. (3) In conclusion, luciferase reporter gene vectors containing DRD1 promoter region are successful y constructed and can provide the basic tool for further study on the transcriptional regulation of DRD1.
2.Expression and significance of aryl hydrocarbon receptor in atrial tissues of patients with atrial fibrillation
Jianming LEI ; Hua XIAO ; Qiang LI ; Xiao WEI ; Jingwen GUO ; Minghan XIAO
Chinese Journal of Pathophysiology 2017;33(5):826-831
AIM:To investigate the expression of aryl hydrocarbon receptor (AhR) in atrial tissues of the patients with rheumatic heart disease (RHD), and the effects of AhR on rheumatic atrial fibrosis.METHODS:Right atrial specimens obtained from the patients with RHD requiring valve replacement surgery were divided into chronic atrial fibrillation (RHD+cAF, n=11) group and sinus rhythm (RHD+sinus rhythm, n=25) group.The patients with congenital heart disease (CHD) and sinus rhythm (CHD+sinus rhythm, n=12) who underwent heart surgery served as controls.The collagen volume fraction in the atrial specimens was examined by Masson`s trichrome staining.The protein expression and distribution of AhR, AhR nuclear translocator (ARNT) and CYP1A1 were detected by the methods of immunohistochemistry and Western blot.The mRNA expression of AhR, ARNT and CYP1A1 was detected by real-time fluorescence quantitative PCR.RESULTS:Compared with CHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+sinus rhythm group and RHD+cAF group.Compared with RHD+sinus rhythm group, the collagen content and the expression of AhR, ARNT and CYP1A1 were significantly increased in RHD+cAF group (P<0.05).CONCLUSION:The expression of AhR is correlated with the degree of fibrosis.The expression of AhR/ARNT/CYP1A1 is increased in atrial tissues of patients with RHD, suggesting that AhR/ARNT/CYP1A1 should be involved in atrial fibrosis of the patient with RHD.
3.Expression of caspase-4 in Hodgkin's lymphoma and anaplastic large cell lymphoma
Zihua ZENG ; Jiwu WEI ; Peie ZHENG ; Guangyu JIANG ; Huanzhen YANG ; Wenqing LIU ; Hong LI ; Minghan XIA
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: This study is based on the result of the study in HL and ALCL employing gene chip technique, in which writer found that there was distinctly different expression of caspase-4 between HL and ALCL cell lines at the level of mRNA. From the point of view, we try to identify at the level of protein whether there is different expression of this gene in HL and ALCL tissues as well. METHODS: HE staining, the monoclonal antibodies CD30 (BerH2), CD15 (C3D-1), CD20 (L26) and CD45RO (UCHL1) were used for selecting the cases of HL and ALCL. Specific high affinitive anti-caspase-4 polyclonal antibody was used by immunohistochemical staining to analyze the expression of caspase-4 in 18 cases of HL and 15 cases of ALCL. RESULTS: The expression of caspase-4 demonstrated a strong positive staining in all ALCL cases (15/15,100%), whereas negative in 16 HL cases (88 8%), while other two cases were weakly stained (11 2%), showing a distinct difference (P
4.Clinical Research on Reversion of Precancerous Lesions of Gastric Carcinoma by Compound Gastritis Mixture
Minghan HUANG ; Jian HUANG ; Sihan LI ; Ping LIN ; Jianlong LIN ; Xin WANG ; Guodong ZHONG ; Hengqing HUANG
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(4):20-23
Objective To discuss the clinical efficacy of compound gastritis mixture (CGM) in treating precancerous lesions of gastric carcinoma (PLGC).Methods Totally 85 PLGC patients were randomly divided into treatment group and control group. The treatment group took CGM and the control group took Vitacoenzyme tablets. One therapeutic course was three months, and the treatment lasted for two courses. The clinical symptoms, electronic gastroscopy presentation, and pathological tissues before and after treatment were observed, the clinical efficacy was evaluated.Results There was statistical significance in TCM syndrome between the two groups (P<0.05), and the effective rate in the treatment group was more obvious than that in the control group (P<0.01). The symptoms of the two groups were significantly improved (P<0.05,P<0.01), but the improvements of the main symptoms in the treatment group were superior to those in the control group (P<0.05,P<0.01). The total effective rate of electronic gastroscopy presentation was 80.0% (36/45) in the treatment group, which was better than that of the control group (P<0.05). Pathological curative effects of chronic atrophic gastritis, intestinal metaplasia, and dysplasia in the treatment group were also better than those in the control group (P<0.05).Conclusion CGM has definite clinical efficacy in treating PLGC.
5.Effects of Jianpi Qinghua Chinese herbal compound on TLR4-MyD88-dependent pathway protein expression and TNF-αin animal model of chronic atrophic gastritis(CAG) in rats
Minghan HUANG ; Jian HUANG ; Qing CHEN ; Sihan LI ; Jianlong LIN ; Guodong ZHONG ; Hengqing HUANG ; Ping LIN
Chinese Pharmacological Bulletin 2016;32(9):1321-1325
Aim To investigate Jianpi Qinghua Chinese herbal compound( JQCC) on the expressions of the rel-evant proteins of TLR4 and its downstream MyD88-de-pendent pathways, and on the inflammatory factor TNF-α in the animal model of chronic atrophic gastritis ( CAG) in rats, so as to discuss the molecular mecha-nism of JQCC in the treatment of CAG. Methods 53 Wistar rats were randomly divided into the blank con-trol group(n=8) and the CAG model group(n=45), and the animal model of CAG in rats was replicated by the “ammonia + sodium deoxycholic acid + ethanol”method. After the successful modeling was confirmed, the rest of the 40 CAG rats in the CAG model group were divided into the model group, the vitacoenzyme-tablet group, the low dose of JQCC group, the medium dose of JQCC group, the high dose of JQCC group ( each group n =8 ) . The experimental animals of all the groups were given intragastric administration of medication continuously for 30 days. Then the patho-logical histological changes were observed by HE stai-ning. The protein expressions of TLR4, MyD88, NF-КB and COX-2 were tested by Western-blot assay. And the serum TNF-α level was measured by ELISA. Results The protein expressions of TLR4 , MyD88 , NF-КB and COX-2 and the serum TNF-α level in the rats in the model group were increased evidently ( P<0. 01). Compared with the model group, the gastric mucosa lesions were improved in the low dose of JQCC group, the medium dose of JQCC group, the high dose of JQCC group, together with significant decreases of the protein expressions of TLR4 , MyD88 , NF-κB and COX-2 and the serum TNF-α level ( P <0. 05 or P <0. 01 ) . Conclusion JQCC could effectively improve the pathological and histological changes in the gastric mucosa in CAG rats, and the therapeutic mechanism might be related to the expressions of the relevant pro-teins of TLR4-MyD88-dependent pathways and the ex-pressions of anti-inflammatory cytokines.
6.Evaluation of community intervention on cardiocerebrovascular disease in Guzhen town, Zhongshan city, Guangdong province
Wenli CHEN ; Chenghuo CAI ; Tingbiao HUANG ; Zhenshan YAN ; Baohui LI ; Bilin ZHU ; Xigou LI ; Minghan KUANG ; En XU ; Xuefen LU
Chinese Journal of Tissue Engineering Research 2005;9(37):124-126
BACKGROUND: It has been explained in many big sample experiments that community comprehensive prevention from the risks of cardiocerebrovascular disease can reduce its morbidity and mortality. But, it is required more samples to verify the evaluations of the process and effect of intervention.OBJECTIVE: To understand the effect of community comprehensive prevention on cardiocerebrovascular disease and hygienic costs demanded and summarizes the suitable patterns of comprehensive prevention on cardiocerebrovascular disease in countryside.DESIGN: Healthy people in community were taken as the objects and community intervention was designed.SETTING: Department of Community Hygienic Service of People's Hospital in Guzheng Town, Zhongshan City, Guangdong Province and Neurological Institute of Guangzhou Medical College.PARTICIPANTS: Totally 32 000 inhabitants were collected from 8 villages in Guzheng Town, Zhongshan City, Guangdong Province from March 1992 to March 2002, aged varied from 20 to 74 years, of which, 14 600inhabitants were male and 17 400 inhabitants were female.METHODS: The intervention of community-based prevention was applied on cardiocerebrovascular disease, including managementof hypertension,smoking control, rational nutrient, nationwide body building and diabetic management.MAIN OUTCOME MEASURES: [1] Awareness, attitude and behavior related to cardiocerebrovascular disease. [2] Management rate and control rate of hypertension. [3] Morbidity and mortality of cerebral apoplexy.RESULTS: [1] By 10-year intervention, awareness rate of hypertension in whole group was 70.29%. [2] Management rate and control rate of hypertension were increased yearly, in which, the management rate was increased from 25.66% in 1997 to 80.50% in 2002, the total control rate was increased from 4.43% in 1997 to 45.80% in 2002, the control rate of blood pressure <160/95 mm HG (1 mm Hg=0.133 kPa) was 58.80%and that <140/90 mm Hg was 32.80%. [3] The systolic and diastolic pressures in intervention community were decreased averagely, in which the D-values are 7.23 mm Hg and 3.92 mm Hg respectively. [4] The awareness, attitude and behavior related to cardiocerebrovascular disease were enhanced remarkably. [5] Morbidity and mortality of cerebral apoplexy were in tendency of decreasing yearly, from 146.90/100 000 and 108.63/100 000 in 1997 to 105.83/100 000 and 69.90/100 000 in 2002 respectively, by 41.07/100 000 and 38.73/100 000 respectively in 6 years and the age of incidence was postponed. [6] It was indicated in economic analysis that the average ratio of profit to cost between 1997 and 2001 was 2.32.CONCLUSION: Community-based comprehensive prevention on cardiocerebrovascular disease improves the awareness, attitude and behavior in community group and reduces morbidity and mortality of cerebral apoplexy.
7.Imaging etiology of pathological myopia results in visual impairment
Yanru CHEN ; Xiaoxin LI ; Minghan LI
Chinese Journal of Experimental Ophthalmology 2021;39(6):528-533
Objective:To analyze the imaging etiology of patients having vision loss with pathological myopia.Methods:A cross-sectional study was conducted.The clinical data of 110 cases (138 eyes) who had vision loss with pathological myopia diagnosed in Xiamen Eye Center of Xiamen University from June 1st, 2016 to May 31st, 2017 was collected and analyzed.Fundus photography was used to observe lacquer cracks; spectral domain optical coherence tomography (SD-OCT), fundus fluorescein angiography, indocyanine green angiography and optical coherence tomography angiography (OCTA) were employed to evaluate the choroidal neovascularization (CNV) and punctate inner choroidopathy (PIC). The macular retinoschisis (MRS), macular atrophy, macular hole and epiretinal membranes were assessed by SD-OCT.The proportion and age distribution of different fundus lesions of pathological myopia complicated with vision loss were analyzed.The study protocol was approved by an Ethics Committee of Xiamen Eye Center of Xiamen University (No.XMYKZX-2016-KY-010). Written informed consent was obtained from each patient before study.Results:Among the imaging causes of visual impairment caused by pathological myopia, there were 87 (63.0%) eyes of myopic CNV (MCNV) with the highest proportion, followed by 53 (38.4%) eyes of lacquer cracks, 48 (34.8%) eyes of MRS, 44 (31.9%) eyes of macular atrophy, 42 (30.4%) eyes of epiretinal membranes, 14 (10.1%) eyes of macular lamellar hole, 19 (13.8%) eyes of full-thickness macular hole (FTMH), and 3 (2.2%) eyes of PIC.The average age was (53.00±1.51) years of MCNV, (53.00±1.77) years of lacquer cracks, (58.00±1.64) years of MRS, (57.00±1.76) years of macular atrophy, (59.00±1.48) years of epiretinal membranes, (61.00±3.90) years of macular lamellar hole, (59.00±3.39) years of FTMH with retinal detachment (RD), and (67.00±0.50) years of FTMH without RD.The average age of PIC patients was (31.00±8.50) years, which was significantly smaller than that of the other groups (all at P<0.05). Conclusions:The main cause of visual impairment resulted from pathological myopia is the obvious abnormality of macular structure, and MCNV is the most common type.
8.The expression of E-cadherin, vimentin, β-catenin, transforming growth factor-β1 in oral squamous cell carcinomas.
Li ZHANG ; Zhiyuan LU ; Cao YIN ; Minghan XIA ; Siming XIE
Chinese Journal of Stomatology 2015;50(4):228-234
OBJECTIVETo investigate the expression of E-cadherin, vimentin, β-catenin and transforming growth factor-β1 (TGF-β1) in oral squamous cell carcinomas (OSCC).
METHODSEighty-nine cases of OSCC and 20 cases of normal oral mucosa were collected. Then the 89 cases of OSCC were classified as grade I, II, III. The semiquantitative method was used to calculated the positive intensity and positive rate. The relationship between the OSCC differentiation and the four biomarkers was analyzed.
RESULTSThe median of E-cadherin was 9.00 in the normal tissue, 9.00, 6.00 and 6.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-4.211, P=0.000). The median of vimentin was 0.00 in the normal tissue, 0.00, 0.00 and 4.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-3.675, P=0.000). The median of β-catenin was 9.00 in the normal tissue, 3.00, 4.00 and 3.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-6.300, respectively. There was significant difference between the normal group and OSCC group (Z=-3.329, P=0.000). E-cadherin expression was positively correlated to β-catenin expression (r=0.327, P=0.002), negtively correlated to vimentin expression (r=-0.386, P=0.001) and positively correlated to TGF-β1 expression (r=-0.304, P=0.004). Vimentin expression was positively correlated to TGF-β1 expression (r=0.401, P=0.000).
CONCLUSIONSE-cadherin and β-catenin in OSCC had a down-regulated expression, while the vimentin has an up-regulated expression.
Cadherins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Humans ; Mouth Mucosa ; cytology ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Transforming Growth Factors ; metabolism ; Vimentin ; metabolism ; beta Catenin ; metabolism
9.Study on rabbit mesenchymal stem cells differentiation to the adipogenic or osteogenic lineage in vitro.
Shengfu LI ; Dingqiang HUANG ; Xiaofeng LU ; Jin LIU ; Minghan SUN ; Youping LI ; Jingqiu CHENG ; Hong BU ; Chuanyu LIANG
Journal of Biomedical Engineering 2003;20(2):209-213
Rabbit bone marrow-derived mesenchymal stem cells(MSCs) are multipotent. We studied the adipogenic and osteogenic differentiation potent using adipogenic supplement (AS) or osteogenic supplement (OS) in vitro. Specific markers of this induced adipogenic and osteogenic lineage were identified. The findings showed that the rabbit MSCs are capable of differentiating into adipogenic and osteogenic lineages spontaneously. On the 21st day, approximately 75% rabbit MSCs were induced to adipogenic or osteogenic cells in medium containing AS or OS, respectively. These results demonstrated that the differentiation of MSCs could be regulated in vitro. The underlying molecular mechanisms of adipogenic or osteogenic differentiation await elucidation.
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10.Application of Circulating Tumor DNA in Precision Diagnosis and Treatment of Colorectal Cancer
Wentao YANG ; Minghan LI ; Jidong CAI ; Cong LI ; Wang SONG ; Ye XU
Cancer Research on Prevention and Treatment 2024;51(3):157-162
Colorectal cancer (CRC) is one of the most common malignant tumors recorded worldwide. This condition has high morbidity and mortality and seriously endangers people's health. Traditional diagnostic models fail to meet people's current needs for real-time monitoring of tumors. Compared with traditional detection methods, ctDNA detection is not only noninvasive but can also attain real-time detection of comprehensive genomic information of tumors. The advancement of detection technology has gradually highlighted the potential of ctDNA detection in the clinical treatment of CRC. This article reviews the advancements on the clinical application of ctDNA in early screening, minimal residual disease detection, and guidance on individualized treatment of CRC patients.