1.Gastrodin ameliorates oleic acid-induced fat accumulation through activation of AMPK pathway in HL-7702 cells
Yana GENG ; Bin YU ; Weijia KONG
Chinese Pharmacological Bulletin 2015;(1):39-44
Aim To study the inhibitory effect of gast-rodin (GSTD) on oleic acid (OA)-induced fat accu-mulation in HL-7702 cells and explore possible cellular signaling pathways. Methods The MTT method was used to study the impact of GSTD on cell viability in HL-7702 cells. Cellular steatosis was induced by 1 mmol·L-1 of OA administration for 24 h, and differ-ent concentrations of GSTD were added at the same time. Oil red O ( ORO) staining was used to determine fat accumulation in cells, and intracellular triglyceride ( TG) contents were assayed. Western blot was used to determine the phosphorylation levels of AMPKα and ACC in cells after GSTD administration. Compound C was used to treat the cells in order to study its influ-ence on the efficacies of GSTD. Results GSTD had no obvious toxicity in HL-7702 cells when its concen-tration was≤3 386. 5 μmol · L-1 . After 24 h of OA administration, there were large amounts of lipid drop-lets accumulated in HL-7702 cells, and intracellular TG contents greatly increased as well. However, when 169. 3 or 338. 7 μmol · L-1 of GSTD was added to-gether with OA, fat accumulation in cells was greatly inhibited, and intracellular TG contents were reduced averagely by 35% and 43 . 6%, respectively ( P<0. 01 vs OA alone ) . After administration, GSTD could in-crease the levels of p-AMPKα and p-ACC in HL-7702 cells time and dose dependently. Compound C could completely abolish the stimulating activity of GSTD on AMPK pathway and block its reducing effect on hepatic TG accumulation. Conclusions GSTD greatly inhibits OA-induced fat accumulation and reduces intracellular TG contents in HL-7702 cells;the efficacy of GSTD is dependent on the activation of cellular AMPK pathway.
2.Effect of transketolase-like protein 1 on proliferation of human nasopharyngeal carcinoma cells
Song ZHANG ; Juhong YANG ; Weijia KONG ; Yu SUN
Chinese Journal of Pathophysiology 1986;0(03):-
0.05). However,the expression level of TKTL1 gene was significantly downregulated in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector. Cancer cells were arrested in G0/G1 phase,and cancer cell proliferation was significantly inhibited in the CNE cells transfected with siRNA TKTL1 construct. CONCLUSION:TKTL1 plays an important role in the total transketolase activity and cell proliferation of human nasopharyngeal carcinoma. TKTL1 may be considered as a potential target for novel anti-cancer therapy.
3.Properties of ACh-sensitive BK channels in mice type Ⅱ vestibular hair cells and its modulation by calcium
Changkai GUO ; Song ZHANG ; Weijia KONG ; Qingsong YU ; Qingtian LI
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To explore the properties of the acetylcholine(ACh)-sensitive potassium channel in type Ⅱ vestibular hair cells(VHCs Ⅱ) in mice saccular macula and the modulation effect of calcium ions.METHODS: Under the whole-cell patch mode,the pharmacology properties of ACh-sensitive potassium channel and the modulation of calcium ions on ACh-sensitive potassium channel were investigated.RESULTS: Following extracellular perfusion of ACh,VHCs Ⅱ displayed a slow and sustained outward current,which was sensitive to tetraethylammonium(TEA,5 mmol/L) and charybdotoxin(CTX,100 nmol/L),but not sensitive to 4-aminopyride(4-AP,15 ?mol/L).ACh-sensitive potassium current was inhibited by intracellular application of ethylene glycol-bis(B-aminoethylether)-N,N,N',N'-tetraacetic-acid(EGTA,5 mmol/L) and extracellular perfusion of Cd2+ and Ni2+,respectively.Intracellular application of heparin(8 g/L) failed to inhibit ACh-sensitive potassium current.CONCLUSION: Extracellular application of ACh activates the big conductance,calcium-dependent potassium current(BK) in VHCs Ⅱ of mice,which is potently modulated by extracellular Ca2+ ions.However,intracellular IP3-dependent Ca2+ ions release mechanism is not involved in the activation of the ACh-sensitive BK channel.
4.Status and training of preoperative anesthetist visit in anesthesia residents
Yahong GONG ; Weijia WANG ; Chunhua YU ; Lijian PEI ; Yuguang HUANG
Basic & Clinical Medicine 2017;37(8):1202-1205
Objective To investigate the resident performance on preoperative anesthetist visit in Beijing hospitals,thereby providing introduction for further training.Methods A self-designed questionnaire was distributed among the anesthesia residents who were receiving anesthesia residency training in Beijing through WeChat.The questionnaire covers geographic data of the residents,information on preoperative visit and existing training program.Results160 questionnaires were reclaimed.History-taking and physical examination were not comprehensive in many residents,the nature of surgery was not evaluated by most residents.The capacity of risk assessment and risk informing were not competent in many residents.The most desired training methods for preoperative visit were scenario simulation and bedside observation.Conclusions Scenario simulation with standard patient may have a promising prospect in preoperative anesthetist visit training.
5.The effect of ΔNp63 knockdown on the growth of pancreatic cancer PANC1 cells
Qingrong MO ; Yaqun YU ; Shuqun LI ; Qian CHEN ; Weijia LIAO
Chinese Journal of Pancreatology 2016;16(5):310-313
Objective To investigate the effect of deltaNp63(ΔNp63) silencing on the proliferation of pancreatic cancer PANC1 cells.Methods ΔNp63 mRNA level in 23 pairs of pancreatic cancer and adjacent tissue specimen was detected by real-time PCR, andΔNp63 protein in human normal pancreatic ductal cell line HPDE6-C7 and pancreatic cancer cell line PANC1, CFPAC1 and BXPC3 was detected by Western blot. PANC1 cells were transfectedΔNp63 specific siRNA (ΔNp63-siRNA ) and scramble siRNA ( Con-siRNA ) using liposome, and untransfected cells served as control.ΔNp63 mRNA and protein was detected by real-time PCR and Western blot to validate the silencing ofΔNp63 expression.MTT assay and BrdU method were used to detect the proliferation and DNA synthesis of transfected PANC1 cells.Results TheΔNp63 mRNA expression in pancreatic cancer tissues and matched adjacent normal tissues was 0.99 ± 0.07 and 0.70 ±0.07, respectively.ΔNp63 mRNA expression in pancreatic cancer tissue was significantly up-regulated compared with that in the normal tissue (P=0.0034).TheΔNp63 protein expression in HPDE6-C7, PANC1, CFPAC-1 and BxPC3 cells was 0.97 ±0.09,3.06 ±0.16,2.57 ±0.11 and 2.45 ±0.08, respectively.TheΔNp63 protein level in pancreatic cancer cells were higher than that in HPDE6-C7 cells (P<0.001).ΔNp63 mRNA level in control, Con-siRNA and ΔNp63-siRNA group was 0.97 ±0.07,0.97 ±0.07 and 0.28 ±0.03, respectively, andΔNp63 protein expression level was 0.97 ±0.06,1.00 ±0.10 and 0.26 ±0.03.The expression ofΔNp63 mRNA and protein inΔNp63-siRNA group were significantly down-regulated comparing with those in Con-siRNA group (P<0.01).Significant inhibition on cell proliferation was observed in ΔNp63-siRNA group, which was statistically different from that in control and Con-siRNA group.The A490 value (DNA synthesis) of control, Con-siRNA andΔNp63-siRNA group was 0.55 ±0.04, 0.56 ±0.01 and 0.55 ±0.00 at 24 h after transfection, and 0.84 ±0.05,0.87 ±0.07 and 0.71 ±0.05 at 48 h after transfection.The DNA synthesis inΔNp63-siRNA group was significantly down-regulated compared with that in control and Con-siRNA group (P<0.05).Conclusions Knockdown ofΔNp63 could greatly inhibit the proliferation and DNA synthesis of pancreatic cancer PANC1 cells.
6.Primary nasopharyngeal tuberculosis presenting as durative aural fullness: one case report and literature review.
Xia WU ; Yu SUN ; Weijia KONG ; Maoli DUAN ; Yanjun WANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(11):824-826
OBJECTIVE:
To study the clinical features, diagnosis and treatments of primary nasopharyngeal tuberculosis.
METHOD:
A case report was presented, and meanwhile etiopathogenesis and differential diagnosis were also reviewed.
RESULT:
A biopsy was taken and the histopathological examination showed tuberculosis granuloma with caseous necrosis. After anti-tuberculosis therapy, the symptoms disappeared.
CONCLUSION
Not only otologic disorders but also nasopharyngeal diseases need to be considered when aural fullness exists. More importantly, primary nasopharyngeal tuberculosis should be taken as one of the differential diagnosis.
Adult
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Female
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Humans
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Nasopharyngeal Diseases
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diagnosis
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microbiology
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Tuberculosis
;
diagnosis
7.An oligonucleotide microarray for detection of cytochrome P450 enzyme gene polymorphisms related to paclitaxel metabolism
Weijia FANG ; Jiong QIAN ; Jing CHEN ; Lanfang YU ; Lin ZHONG ; Nong XU
Chinese Journal of Laboratory Medicine 2008;31(2):195-199
Objective To develop an accurate,rapid,high throughout genotyping method based on oligonucleotide microarray for cytochrome P450 gene polymorphisms related to paclitaxel metabolism.Methods The mutant points of 2C8 * 3,3A4 * 18 and 3A5 * 3C from cytochrome P450 gene were regarded as targets.Based on the sequences in the GenBank,the wild-type and mutant-type probes were specially designed for each mutant point.PCR primers were located in the both sides of mutant point,and furthermore the fragments of PCR products were less than 200 bp.Each type of standard plasmids was constructed.Thus,all the olignucleotide probes were modified with 3'amino-group,and the reverse primers were labeled with fluorescein (Cy3).The probes were immobilized onto certain glass slides.The specific fragments of three genes were amplified and then hybridized with oligonucleotide microarray.The results were analyzed by using certain software.Finally this assay was applied to detect 50 clinical blood specimens.Results When PCR products from standard plasmids were hybridized with DNA microarray,the corresponding probes produced positive signals.Meanwhile,the non-specific hybridization signals did not appear.The results of clinical specimens showed that the mutant rate of CYP2C8 * 3 was 2%.The point of CYP3A4 * 18 for all the clinical specimens was wild-type and the mutant rate of CYP3A5 * 3C was 62%. Meanwhile,the results from detecting 50 clinical blood specimens using oligonucleotide microarray were the same as sequencing analysis.Conclusions Oligonucleotide microarray is a reliable and accurate genotyping assay for cytochrome P450 2C8 * 3.3A4 * 18 and 3A5 * 3C polymorphisms related to paclitaxel simultaneously.This genotyping assay is a high-throughout method for guiding personalized therapy and analyzing metabolism of paclitaxel in vivo.
8.Vascular endothelial growth factor165-regulated nasopharyngeal carcinoma cell lines invasion and migration involve expression and activation of matrix metalloproteinase-2.
Yanjun, WANG ; Weijia, KONG ; Jianxin, YUE ; Dawei, SUN ; Wei, LI ; Qi, YAO ; Yu, SUN ; Jihua, DONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(5):621-4
The effect of vascular endothelial growth factor (VEGF) overexpression on matrix metalloproteinase-2 (MMP-2) in nasopharyngeal carcinoma (NPC) cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.
9.Dysregulation of E-cadherin in chronic rhinosinusitis with nasal polyps.
Weijia, KONG ; Junhua, WU ; Yanjun, WANG ; Jianxin, YUE ; Song, ZHANG ; Yanping, YU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):509-13
E-cadherin is a key epithelial protein and adhesive molecule. This study detected the E-cadherin expression in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and controls, and analyzed its possible role in the pathogenesis of CRSwNP. The expression of E-cadherin was detected by using Western blotting and immunohistochemistry in controls and patients with CRSwNP. Computed tomography (CT) scan findings were scored. The results showed that the E-cadherin expression was up-regulated in patients with CRSwNP as compared with controls (P=0.039) and the positive staining was predominantly localized on the epithelial cell membrane. E-cadherin level was correlated negatively with Lund-Mackay scores in patients with CRSwNP (r=-0.604, P=0.005). It is suggested that E-cadherin may be involved in the pathogenesis of CRSwNP and correlated with disease severity.
10.Protective roles of alpha-lipoic acid in rat model of mitochondrial DNA4834bp deletion in inner ear.
Wei, PENG ; Yujuan, HU ; Yi, ZHONG ; Bei, CHEN ; Yu, SUN ; Yang, YANG ; Weijia, KONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):514-8
The protective roles of alpha-lipoic acid in the rat model of mitochondrial DNA (mtDNA) 4834bp deletion in inner ear were investigated. Forty female Wistar rats at 4 weeks of age were divided into four groups: group A (D-galactose group, n=10), group B (D-galactose+alpha-lipoic acid group, n=10), group C (alpha-lipoic acid group, n=10), and group D (control group, n=10). Auditory brainstem response (ABR) was used to detect the hearing threshold. Colorimetry was used to analyze activity of superoxide dismutase (SOD) and concentration of malondialdehyde (MDA). The percentage of mtDNA4834bp deletion in inner ear was identified by real-time PCR. There was no significant difference in ABR threshold shift among all groups. The percentage of mtDNA4834bp deletion in group A was higher than that in other groups, but there was no significant difference in percentage of mtDNA4834bp deletion among groups B, C, and D. The activity of SOD in group A was lower than that in other groups. The concentration of MDA in group A was higher than that in other groups. It was concluded that there was no significant hearing loss when the percentage of mtDNA4834bp deletion was lower than 12.5%. alpha-Lipoic acid could prevent the reactive oxygen species (ROS)-induced mtDNA4834bp deletion in inner ear of rats.