2.PLCE1 modulates p53 expression and apoptosis in esophageal cancer cells
Yun LI ; Junhang ZHANG ; Jun AN ; Jinyuan HE ; Shaohong HUANG
Chinese Pharmacological Bulletin 2015;(1):82-86
Aim To investigate the role of phospho-lipase C epsilon 1 ( PLCE1 ) in modulating the apoptot-ic mechanism in esophageal cancer ( Eca ) cells. Methods Eca cell lines, OE33 and CP-C cells were cultured to assess the expression of PLCE1 . siRNA suppress expression of PLCE1 . The expression of PLCE1 and p53 was evaluated by quantitative real time PCR and Western blot. Methylation analyses of p53 were performed by bisulfite conversion of genomic DNA. Apoptosis was assessed by flow cytometry. Results OE33 and CP-C cells expressed high levels of PLCE1 . Knockdown of PLCE1 markedly increased the expression and hypomethylation of p53 , and in-creased the frequency of apoptosis. Conclusion PLCE1 suppresses apoptosis of Eca cells via promoting p53 promoter methylation and inhibiting expression of p53 .
3.Activation of protease-activated receptor 2 inhibits apoptosis of lung cancer cells
Shaohong HUANG ; Jun AN ; Yun LI ; Junhang ZHANG ; Jian RONG
Chinese Pharmacological Bulletin 2014;(5):684-687,688
Aim To investigate the activation of prote-ase-activated receptor 2 ( PAR2 ) in regulation of the expression of epidermal growth factor receptor ( EGFR) and apoptosis of lung cancer ( LC) cells. Methods LC cells A549 and its EGFR-silenced counterpart were incubated in the medium added with tryptase. Quanti-tative RT-PCR and Western blotting were used to de-tect the expression of EGFR in A 5 4 9 cells . The apop-tosis and Bax/Bcl-xL of cells were also recorded. Re-sults Treating A549 cells with tryptase in the culture for 48 hrs resulted in a marked increase in the expres-sion of EGFR in A549 cells. Marked decreases in a tryptase dose-dependent manner in apoptotic A549 cells were detected in the presence of tryptase. Expo-sure to tryptase markedly decreased the levels of Bax and increased the levels of Bcl-xL in A549 cells, which were not shown in EGFR-deficient A549 cells. Conclusion Tryptase can increase the expression of EGFR in LC cell line, A549 cells, which protects A549 cells from apoptosis, increases Bcl-xL, and sup-presses Bax in A549 cells.
4.Determination of Hydroxycamptothecin Concentration in Rabbit Tissues by HPLC
Yun FANG ; Jun CHEN ; Lili HUANG ; Jian MA ; Liping ZHAO
China Pharmacy 1991;0(04):-
OBJECTIVE: To establish a HPLC - UV method for determination of hydroxycamptothecin(HCPT) in rabbit tissues (liver, kidney, stomach, lung, spleen, heart, intestine) .METHODS: After homogenization, tissue samples and internal standard, eamptothecine, were precipitated with CH3OH-CH3CN( 1 : 1) and then centrifugalized.20?l of supernatant was injected and measured by HPLC - UV method.The chromatographic column was Lichrospher C18 column: (250mm ? 4.6mm, 5?m), CH3CN - 0.075mol/L NH4AC buffer(pH6.4) (30 : 70, contain 5mol/L tyiethylamine) served as mobile phase with flow rate of 1.0ml/ min .Detection wavelength was 384nm .RESULTS: The retention time of hydroxyeamptothecin was 4.5min.A good linearity was shown in the concentration range of (40- 1 600)ng/ml .The recovery was between 95.24% and 107.58% . The intra day RSD was less than 7.70% and the inter - day RSD was less than 6.69% .CONCLUSION: This method is simple,pratical and accurate.It could be applied to pharmacokinetic study of hydroxyeamptothecin.
5.Immobilized Lipases Cooperates to Catalyze Transesterification Reaction of Lard
Ying HUANG ; Huan GAO ; Hai ZHEN ; Yun-Jun YAN ;
China Biotechnology 2006;0(01):-
The transesterification reaction conditions of lard with methyl acetate with combined use of immobilized lipases as catalysts were conducted. Initially, according to single factorial experiments, the studies on Lipozyme TL IM and Novozym 435 respectively catalyzed transesterification of lard showed that the optimal parameters of transesterification reaction were: the molar ratio of methyl acetate to oil of 14∶1, 40% enzyme added based on oil weight, temperature 50℃. Combined use of Lipozyme TL IM and Novozym 435 was proposed further to improve the catalytic performance by the response surface method (RSM). Herein, a 5-level-3-factor central composite rotated design was employed to evaluate the effects of lipase loading, the proportion of the two lipases and amount of methyl acetate. The optimum conditions were as followings: 40% lipase loading based on oil weight, 50%/50% the proportion of lipases (Novozym 435/Lipozyme TL IM), and the molar ratio of methyl acetate to oil of 14∶1. And under the optimal conditions, the highest biodiesel yield of 97.6% could be attained, which was higher than the biodiesel yield with each single one of the two lipases. The results suggested that the technics of combined use of certain immobilized lipases catalyzed transesterification reaction of lard for biodiesel production with methyl acetate as the acyl acceptor could raise the FAME yield and save the production cost.
6.Outcomes of unexpected pathologic N2 disease after total video-assisted thoracic surgery lobectomy for non-small cell lung cancer
Yingtai CHEN ; Yuqing HUANG ; Yun LI ; Luming JIN ; Jian CUI ; Jun LIU ; Jun WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;28(2):86-89
Objective To assess early and medium outcomes of pathologic N2 disease unexpectedly detected in patients undergoing total video-assisted thoracic surgery lobectomy for non-small cell lung cancer.Methods Between Sep.2006 and Dec.2010,348 patients with Non-small cell lung cancer underwent total video-assisted thoracic surgery lobectomy,and within them,35( 10.1% ) were found to have pathologic N2 disease after operation.We retrospectively reviewed the clinical and pathologic features of patients with unexpected N2 disease after video-assisted thoracic surgery lobectomy and their early and medium outcomes,including survival and recurrence pattern.Results No perioperative mortality was noted.26 patients received a lobectomy directly,and the other 9 patients after a wedge resection.All the patients had R0 resection.The medium operation time was 190 minutes and medium blood loss was 200ml.The medium stations and numbers of dissected N2 lymph nodes in operation were 4 and 10,respectively.And the medium stations and numbers of metastatic N2 Lymph nodes were 1 and 2,respectively.Among patients with pathologic N2 disease,18 (51.4%) had single-station involvement.The median duration of chest tube placement was 8 days.The median length of hospital stay was 11 days.15 complications occurred in 12 (34.3%) patients.All of the patients underwent adjuvant chemotherapy with platinum postoperatively.The median follow-up time was 23 months.The 1 - and 2-year overall survival (OS) was 80.9% and 67.9%,and the medium OS was not reached.During follow-up,16 (45.7%) patients had a recurrence.The pattern of recurrence was locoregional in 5,distant in 11.The 1 - and 2-year disease-free survival (DFS) was 71.9% and 44.2%,and the medium DFS was 20 months (95%,8.1 to 31.9 months).Divided the patients with pathologic N2 disease into two groups considering single-station involvement or not,the 1-and 2-year OS and DFS for the single-station group and for the multiple-station group were 87.7%,78.9% ; 88.9%,49.4%and 67.6%,59.1% ; 55.3%,39.5%.The medium DFS for both the two groups was 23 and 16 months respectively.Conclusion For non-small cell lung cancer with N0 disease confirmed by an exactly preoperative staging workups,if it is feasible in technology,a total video-assisted thoracic surgery lobectomy should be recommended.Even if N2 lymph node metastasis is unexpectedly detected postoperatively,the metastasis was mostly micro- or single-station involved,and a similar outcome with conventional thoracotomy can be achieved.
7.Intralesional bleomycin injection treatment for 44 cases of pharyngolaryngeal haemangioma.
Guo-jun LIU ; Qi-jun FAN ; Xue-jun LIU ; Li-yan NI ; Jin-jian GAO ; Sai-yu HUANG ; Bo-bei CHEN ; Jia-yun HUANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2013;48(10):843-845
Adolescent
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Adult
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Bleomycin
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therapeutic use
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Child
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Child, Preschool
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Female
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Hemangioma
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drug therapy
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Humans
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Laryngeal Neoplasms
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drug therapy
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Male
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Middle Aged
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Pharyngeal Neoplasms
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drug therapy
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Young Adult
8.Inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide-stimulated human mast cells.
Yun-jiang ZHOU ; Hu WANG ; Li LI ; He-huan SUI ; Jia-jun HUANG
Acta Pharmaceutica Sinica 2015;50(6):702-707
This study is to investigate the inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide(LPS)-stimulated HMC-1 mast cells. The cytotoxicity of kaempferol to HMC-1 mast cells were analyzed by using MTT assay and then the administration concentrations of kaempferol were established. Histamine, IL-6, IL-8, IL-1β and TNF-α were measured using ELISA assay in activated HMC-1 mast cells after incubation with various concentrations of kaempferol (10, 20 and 40 µmol.L-1). Western blot was used to test the protein expression of p-IKKβ, IκBα, p-IκBα and nucleus NF-κB of LPS-induced HMC-1 mast cells after incubation with different concentrations of kaempferol. The optimal concentrations of kaempferol were defined as the range from 5 µmol.L-1 to 40 µmol.L-1. Kaempferol significantly decreased the release of histamine, IL-6, IL-8, IL-1β and TNF-α of activated HMC-1 mast cells (P<0.01). After incubation with kaempferol, the protein expression of p-IKKβ, p-IKBa and nucleus NF-κB (p65) markedly reduced in LPS-stimulated HMC-1 mast cells (P<0.01). Taken together, we concluded that kaempferol markedly inhibit mast cell-mediated inflammatory response. At the same time, kaempferol can inhibit the activation of IKKβ, block the phosphorylation of IκBα, prevent NF-KB entering into the nucleus, and then decrease the release of inflammatory mediators.
Cells, Cultured
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Histamine
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metabolism
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Humans
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I-kappa B Kinase
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metabolism
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I-kappa B Proteins
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metabolism
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Inflammation
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metabolism
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Interleukin-1beta
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metabolism
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Interleukin-6
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metabolism
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Interleukin-8
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metabolism
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Kaempferols
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pharmacology
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Lipopolysaccharides
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Mast Cells
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drug effects
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NF-KappaB Inhibitor alpha
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NF-kappa B
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metabolism
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Tumor Necrosis Factor-alpha
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metabolism
9.Airway epithelial cell-derived insulin-like growth factor-1 triggers skewed CD8 +T cell polarization
Shaohong HUANG ; Yun LI ; Jie QIN ; Jun AN ; Junhang ZHANG ; Jian RONG
Chinese Pharmacological Bulletin 2015;(2):204-209,210
Aim To investigate the effects of airway epithelial cell-derived insulin-like growth factor-1 (IGF1) on CD8 +T cell polarization. Methods Hu-man airway epithelial cell line, RPMI2650 cells, was cultured in the presence of a mice allergen, Der p1, for 72 h. IGF1 expression was checked with quantita-tive RT-PCR and Western blot. Der p1-primed RP-MI2650 cells, recombinant IGF1 and anti-IGF1 anti-body was cocultured respectively with CD8 + T cells, which were activated by anti-CD3/CD8 Ab. Apoptotic cells frequency was calculated with flow cytometry. The alteration of p53 gene hypermethylation in CD8 + T cells elicited by Der p1-primed airway epithelial cell and IGF1 was plotted. Results Both mRNA(23. 1%± 5. 2% vs 5. 2% ± 2. 3%, P < 0. 01 ) and protein (33. 4 ± 6. 4 vs 9. 2 ± 4. 6, P <0. 01 ) expression of IGF1 in RPMI2650 cells markedly increased after ex-posure to Der p1 . The increase of apoptotic CD3/CD28 Ab-activated CD8 + T cells was abolished by the pres-ence of Derp1-primed epithelial cells ( 41. 7% ± 8. 2%vs 5. 2% ± 1. 8%, P <0. 01 ) . The results were con-firmed by the addition of recombinant IGF1 . Anti-IGF1 antibody abolished the effect of the epithelial cells. Derp1-primed epithelial cells inhibited p53 gene mR-NA( 29. 1% ± 5. 9% vs 16. 2% ± 4. 3%, P <0. 01 ) and protein ( 63. 3 ± 8. 9 vs 26. 9 ± 5. 6 , P <0. 01 ) ex-pression. Anti-IGF1 antibody abolished the effect. Re-combinant IGF1 promoted CD8 + T cells′p53 gene hy-permethylation. Conclusion Der p1 induces RP-MI2650 cells to produce IGF1 , and this factor prevents CD8 + T cell apoptosis by inducing p53 gene hyperm-ethylation.
10.Multicentric prospective randomized controlled study of efficacy of mannitol,furosemide and albumin in reducing intracranial pressure in patients with severe brain injury
Guodong HUANG ; Jun JIA ; Yun ZHEN ; Jiangong WEI ; Richu LIANG ; Weiping LI ; Yongzhong GAO
Chinese Journal of Trauma 2008;24(9):680-683
Objective To compare the effect of difierent combinatio of mannitol, furesemide and albumin in reducing intracranial pressure in 451 patients with severe traumatic brain injury (sTBI). Methods A total of 451 patients with an admissiou Glasgow Coma Scale of or less from 5 medical centers were randomly divided into 5 groups, ie, Group A(250 ml 20% mannitol each time as control), Group B(125 ml 20% mannitol each time), Group C(alternate use of 250 ml 20% mannitol each time or 40 mg furosemide), Group D(alternate use of 125 ml 20% mannitol each time and 20 mg furosemide)and Group E(alternate use of 125 ml 20% mannitol and moderate or large dose of albumin). We monitored intracraniai pressure continuously and observed the changes of intracranial pressure, electrolytes, hemato-crit and renal function after use of 5 combinations of mannitol. Furosemide and albumin. Results Man-nitol and furosemide could independently reduce intracranial pressure after 1-3 hours (P<0. 05). Semis mannitol plus furosemide or albumin could more signifieantly reduce intracranial pressure, with statistical difference compared with full dose of mannitol. Semis mannitol and alternate use of mannitol and furose-mide in aspect of intracranial pressure reduction and persistence time(P<0. 05). Alternate use of man-nitol and furosemide begot higher incidence rate of electrolyte abnormality, compared with the other com-binations (P<0. 05). Rebound rate of intracranial pressure was higher in full dose of mannitol than other combinations (P<0. 05). Incidence of renal function abnormality was higher in combination involved al-bumin than alternative use of mannitol and furosemide as well as combination of semis mannitol and furo-semide (P<0. 05). Abnormality of electrolyte and renal function wag reversible. Conclusion The use of 125 ml 20% mannitol each time plus 20 mg furesemide is more reasonable than other combina-tions. Meanwhile, semis mannitol combined with moderate or large dose of albumin has certain advantages too.