1.Expression of miR-7850 in renal cell carcinoma and its effect on proliferation and migration of renal cell carcinoma
Fang XIE ; Geng HUANG ; Zhihua YE ; Xiaoying WANG ; Dingwen GUI
Journal of Chinese Physician 2021;23(5):668-673
Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.
2.Molecular mechanisms of TRP channels in mechano-sensory transduction.
Wen-juan ZOU ; Gui-fang HUANG ; Li-jun KANG
Journal of Zhejiang University. Medical sciences 2012;41(2):222-228
Channels from the TRP superfamily have essential roles in a wide variety of sensory transductions, especially in mechano-sensation, such as hearing, touch and mechanical pain. TRP channels are also implicated in major channelopathies, including deafness, chronic pain, autosomal dominant polycystic kidney disease (ADPKD) and ventricular hypertrophy. As the leading candidates for mechano-sensitive channels, some TRP channels appear to be mechano-receptor, which can be activated by mechanical forces directly, such as C. elegans TRPN homolog TRP-4; whereas others may act as signal modulators, receiving and amplifying signals indirectly. This review is to introduce the function of TRPs in mechano-sensory transduction and to discuss the underlying molecular mechanisms.
Animals
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Humans
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Neural Conduction
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Sensation
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physiology
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Signal Transduction
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Transient Receptor Potential Channels
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metabolism
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physiology
3.Effect of advanced glycosylation end products on oxidative stress and MCP-1 in human renal mesangial cells.
Min FENG ; Cheng-Bo XU ; Jun-Ping WEN ; Gui-Fang LIN ; Qi LV ; Guo-Liang HUANG
Chinese Journal of Applied Physiology 2014;30(4):306-313
OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).
METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.
RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.
CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.
Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology
4.Effect of lentivirus-mediated interference with long non-coding RNA LINC00630 expression in vitro on proliferation and migration of bladder cancer cells
Geng HUANG ; Dingwen GUI ; Xiaoying WANG ; Wei PENG ; Yunfei ZHAO ; Jinghua WAN ; Fang XIE
Cancer Research and Clinic 2021;33(4):254-258
Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.
5.Antibacterial activity of turmeric (Curcuma longa L ) extract and effect on bacterial cell membranes
Lu HAN ; Chao ZHOU ; Xiu-fang BI ; Mei-gui HUANG ; Gang HAO
Acta Pharmaceutica Sinica 2024;59(8):2265-2272
In the present study, the antibacterial spectrum of turmeric extract was analyzed by measuring the minimum inhibitory concentration (MIC), and the antibacterial mechanism of turmeric extract was elaborated by determining its effects on the permeability and integrity of the cytoplasmic membrane, energy metabolism, and the morphology of the tested bacteria (
6.Determination of 27 elements in Maca nationality's medicine by microwave digestion ICP-MS.
Gui-fang YU ; Hai-jie ZHONG ; Jun-hua HU ; Jing WANG ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(23):4545-4551
An analysis method has been established to test 27 elements (Li, Be, B, Mg, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Sr, Mo, Cd, Sn, Sb, Ba, La, Hg, Pb, Bi) in Maca nationality's medicine with microwave digestion-ICP-MS. Sample solutions were analyzed by ICP-MS after microwave digestion, and the contents of elements were calculated according to their calibration curves, and internal standard method was adopted to reduce matrix effect and other interference effects. The experimental results showed that the linear relations of all the elements were very good; the correlation coefficient (r) was 0.9994-1.0000 (Hg was 0.9982) ; the limits of detection were 0.003-2.662 microg x L(-1); the relative standard deviations for all elements of reproducibility were lower than 5% (except the individual elements); the recovery rate were 78.5%-123.7% with RSD lower than 5% ( except the individual elements). The analytical results of standard material showed acceptable agreement with the certified values. This method was applicable to determinate the contents of multi-elements in Maca which had a high sensitivity, good specificity and good repeatability, and provide basis for the quality control of Maca.
Lepidium
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chemistry
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Mass Spectrometry
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methods
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Microwaves
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Reproducibility of Results
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Trace Elements
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chemistry
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isolation & purification
7.Study on the causes and risk factors on vertigo and balance disorders in 118 elderly patients.
Wei-ning HUANG ; Jin XU ; Bo GAO ; Jin-mei ZHOU ; Gui-fang LIU
Chinese Journal of Epidemiology 2005;26(9):720-722
OBJECTIVETo understand the various factors causing vertigo and balance disorders in the elderly.
METHODS118 elderly patients (aged equal or older than 60 years of age) with vertigo or balance disorders were retrospectively analyzed through clinical symptoms, audio-vestibular function tests, X-ray, CT scan or MRI in cervical vertebras, brain and inner ears, ultrasonography, transcranial doppler (TCD) or magnetic resonance angiography (MRA) in blood vessels on head and neck.
RESULTSOf 118 patients, 70 (23%) of them suffered perip heral vestibular disorders while 29 (58%) having cerebral vertigo or dizzness, leaving 19 cases (16%) as unclassified.
CONCLUSIONFor elderly patients, vertigo and balance disorders were commonly caused by many kinds of peripheral and cerebral vestibular pathological disfunctions while the functional weakness of vestibular organs and systems affected by the physiological process of ageing and different concommitant diseases as well as environmental, psychogenic factors should also be considered.
Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Postural Balance ; Retrospective Studies ; Risk Factors ; Vertigo ; etiology ; pathology ; physiopathology
8.Observation on therapeutic effect of acupuncture and moxibustion on disorders of myometrial gland.
Hong YAN ; Xiao-Hui HUANG ; Gui-Fang DENG
Chinese Acupuncture & Moxibustion 2008;28(8):579-581
OBJECTIVETo explore the therapeutic effect of acupuncture on disorders of myometrial gland and the mechanism.
METHODSSixty-six cases were randomly divided into an acupuncture group and a medication group, 33 cases in each group. The acupuncture group were treated with acupuncture at Zhongji (CV 3), Shuidao (ST 28), Tainshu (ST 25), Qugu (CV 2), Zigong (EX-CA 1) as main; the medication group were treated with oral administration of Danazol. Changes of estradiol (E2) level, hemoglobin (Hb) and blood platelet counter (BPC) were observed in the acupuncture group, and the therapeutic effects of the two group were compared.
RESULTSThe total effective rate was 97.0% in the acupuncture group and 72.7% in the medication group, the former being better than the latter (P<0.05). After treatment, E2 level decreased and Hb and BPC increased in the acupuncture group.
CONCLUSIONAcupuncture has obvious therapeutic effect, which is better than that of simple western medicine. Acupuncture can decrease E2 level.
Acupuncture Therapy ; Adult ; Danazol ; therapeutic use ; Endometriosis ; therapy ; Female ; Humans ; Middle Aged ; Moxibustion ; Myometrium ; Uterine Diseases ; therapy
9.The expression of interferon-regulatory factor genes in patients with systemic lupus erythematosus
Jie QIAN ; Nan SHEN ; Gui-Mei GUO ; Nian-Hong WAN ; Yan LIN ; Xin-Fang HUANG ; Hui WU ; Shun-Le CHEN ;
Chinese Journal of Rheumatology 2003;0(09):-
Objective To observe whether the expression of interferon-regulatory factor genes are re- lated to systemic lupus erythematosus (SLE).Methods The clinical data of 45 SLE patients and 37 normal controls were collected.Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression (indicated as-??Ct value) of IRFI,IRF4,IRF8 in patients with SLE and those in the controls.Results The levels of IRF1,IRF4 and IRF8 mRNA were-3.90?0.19,-8.04?0.25 and 3.60?0.15 respectively in normal controls.In SLE patients, IRF4 mRNA expression was -8.82?0.18,higher than that in normal (P=0.011).But IRF8 mRNA expression was 3.09?0.13,lower than that in normal (P=0.012).Conclusion Abnormal IRF mRNA expression is found in the peripheral blood of SLE patients.IRFs may play roles in the pathogenesis of SLE by affecting the differen- tiation of Th cells.
10.Prenatal nicotine exposure aggravated abnormal adrenal function and its gender differences in adult offspring rats with high-fat diet
fang Fang DUAN ; Lian LIU ; gui He HUANG ; yan Chun ZHU ; Zheng HE ; Hui WANG
Chinese Pharmacological Bulletin 2017;33(11):1530-1534
Aim To explore the alterations of blood corticosterone (CORT) level and adrenal steroidogenic function,as well as its sex specificity in intrauterine growth retardation (IUGR) rats induced by prenatal nicotine exposure (PNE) with high-fat diet (HFD) after birth,and to make clear its mechanism through insulin-like growth factor-1 (IGF-1) signaling pathway.Methods IUGR model was established by PNE (2.0mL · kg-1 · d-1),and the offspring rats were administered with HFD until postnatal week (PW) 24 after weaning.Blood CORT concentration,adrenal steroidogenesis enzymes,expressions of IGF-1 signaling pathway and 11β-HSDs/CR system were tested.Results Compared with HFD control group,the CORT concentration in male offspring of PNE group represented a decrease trend,while an increase trend in female;the expressions of adrenal steroidogenesis enzymes (such as StAR,3β-HSD and P450cll) in male offspring decreased,while increased in female offspring (such as SF-1 and P450c21);the expressions of IGF-1 signalling pathway (IGF-1 and IGF-1R) in male offspring increased,and they significantly increased in female offspring;the expression levels of 11 β-HSD2 and GR decreased,but 11β-HSD1/11β-HSD2 ratio was enhanced in male PNE group,while in female PNE group,the corresponding gene expressions increased.Conclusions PNE could induce abnormal alterations of adrenal steroidogenic function,and exhibit apparent gender differences.The potential mechanism is related to low adrenal steroidogenesis function programming induced by nicotine and catch-up growth mediated by IGF-1 after birth.