2.Inhibitory effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3 and its mechanisms
Xilin LIU ; Qiang JIAN ; Ye MIAO ; Min HUANG ; Chengxin LI
Chinese Journal of Dermatology 2014;47(7):499-502
Objective To evaluate the effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3,and to investigate its mechanisms.Methods Cultured Sa3 cells were classified into four groups:normal control group transfected with green fluorescent protein (GFP),Rab23-overexpressing group transfected with a GFP-labelled Rab23-overexpressing plasmid,Rab23-silencing group transfected with a plasmid carrying a Rab23-targeting shRNA,empty vector group transfected with an empty vector.After additional culture for different durations,plate colony formation assay and flow cytometry were performed to evaluate the proliferative activity of Sa3 cells,and Western blot was conducted to detect the expression of Erl/phosphorylated-Erk in Sa3 cells.Statistical analysis was carried out by t test,one-way analysis of variance and Bonferroni's multiple comparison test.Results Stable Sa3 cell lines with overexpression or silencing of Rab23 were established by plasmid construction and lentivirus-mediated transfection.The plate colony formation assay showed that the colony formation rate was significantly lower in the Rab23-overexpressing group than in the normal control group (2.3% ± 0.2% vs.3.6% ± 0.3%,P < 0.05),but higher in the Rab23-silencing group than in the empty vector group (4.1% ± 0.2% vs.1.8% ± 0.03%,P < 0.01).Rab23 overexpression induced G1 phase arrest in Sa3 cells.The proliferation index was significantly decreased in the Rab23-overexpressing group compared with the normal control group (0.581 ± 0.035 vs.0.698 ± 0.018,P < 0.05),but increased in the Rab23-silencing group compared with the empty vector group (0.567 ± 0.015 vs.0.444 ± 0.014,P < 0.01).As Western blot showed,there were no significant changes in the expression of Erk in the Rab23-silencing or-overexpressing group compared with the normal control group,whereas the expression of p-Erk was attenuated in the Rab23-overexpressing group compared with the normal control group,but enhanced in the Rab23-silencing group compared with the empty vector group.Conclusions Rab23 could inhibit the proliferation of Sa3 cells,which may be associated with the Erk pathway.
3.Influence of injection rate of isobaric ropivacaine on spinal anesthesia
ting-ting, WANG ; shao-qiang, HUANG ; xiao-min, ZHOU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
0.05).Conclusion There may be no significant difference in sensory block,motor block and prevalence of adverse effects between injection rates of 0.27 mL/s and 0.04 mL/s in spinal anesthesia with 20 mg isobaric ropivacaine.
5.Down-regulation of kallikrein in prostate cancer
Qiang XUAN ; Xiaoli YANG ; Linjian MO ; Fengyu HUANG ; Min QIN ; Min HE ; Youhong PANG ; Zengnan MO
Chinese Journal of Urology 2008;29(5):346-349
Objective To study the expression of kallikrein 7 (KLK7) in different prostate tissues and its clinical significance. Methods KLK7 mRNA levels in normal prostate epithelia (5 cases), benign prostat(ic) hyperplasia (BPH) epithelia (13 cases), prostate cancer and prostate cancer cell lines (8 cases) were analyzed by using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Western blot was used to analyze the protein levels of human kallikrein 7 (hK7) in benign prostate epithelia and prostate cancer cell lines, hK7 expressions were examined in 20 normal prostate tissue specimens, 50 BPH specimens and 103 prostate cancer specimens by immunohistochemical staining.Results The mRNA levels of KLK7 in normal prostate, BPH and prostate cancer were 0.59, 0.52 and 0.02 respectively, mRNA levels of KLK7 were significantly different among the three groups (F=13.03, P<0.01). mRNA levels of KLK7 were decreased in prostate cancers compared with that in benign hyperplastic prostate epithelial cells (P<0.01) and in normal prostate epithelial cells (P<0.01). No significant difference of KLK7 mRNA levels was found between normal prostate and BPH. The protein levels of KLK7 in normal prostate, BPH, DU145, LNCaP, PC3,22RV1 and BPH1 was 0.22, O. 40, 0.01, 0.05, 0, 0.03 and 0.14 respectively, hK7 protein level was down-regulated in prostate cancer cell lines compared to benign prostate epithelial cells. The expression of bK7 was observed in benign prostate epithelial cells, whereas little or no staining was observed in prostate cancer cells in immunohistochemical study, hK7 protein was detected in 13 of 20 (65%)normal prostate specimens, 38 of 50 (76%) BPH specimens and 18 of 103 (17.5%) prostate cancer specimens. The difference between the normal prostate and prostate cancer was significant (Z=-4.43, P<0.01). The difference between BPH and prostate cancer was significant (Z=-7.77,P<0.01) as well. However, no significant difference of hK7 protein level was found between normal prostate and BPH (Z=-1. 52, P>0.05). Conclusions KLK7 expression level is down regulated in prostate cancer. KLK7 may play an important role in prostate cancer progression.
6.Expression and significance of BCL-6 and ZEB2 in breast cancer
Li ZHENG ; Min ZHAO ; Lin ANG ; Jin HUANG ; Jin WANG ; Qiang ZOU ; Zhengsheng WU
Chinese Journal of Clinical and Experimental Pathology 2017;33(1):50-54
Purpose To explore the biological significance of BCL-6 and ZEB2 in invasion,metastasis and prognosis of breast cancer.Methods The expressions of BCL-6,ZEB2 protein and mRNA were detected respectively in 228 cases of breast cancer and 80 cases of breast benign lesions by immunohistochemical SP two-step staining and situ hybridization.Result The expression levels of BCL-6,ZEB2 protein and mRNA in breast cancer tissues were significantly higher than in breast benign lesions (P < 0.05).The expressions of BCL-6 was positively correlated with tumor size,lymphatic metastasis,histological grade,TNM staging and HER-2 expression (P < 0.05).The expressions of ZEB2 was positively correlated with tumor size,lymphatic metastasis,TNM staging and HER-2 expression (P < 0.05).The overall survival and relapse-free survival of BCL-6 and ZEB2 positive expression were significantly less than the negative expression (P < 0.01).Conclusion The BCL-6 and ZEB2 are closely correlated with the evolution process of breast cancer,which may become important means for monitoring and warning the metastasis,invasion,and prognosis of breast cancer.
7.Clinically predictive factors of Gleason score upgrading in patients after radical pros-tatectomy
Qiang ZUO ; Fan ZHANG ; Yi HUANG ; Lulin MA ; Min LU ; Jian LU
Journal of Peking University(Health Sciences) 2016;48(4):603-606
Objective:To assess the discrepancy between preoperative needle biopsy (NB)Gleason score and pathological specimen Gleason score (GS)after radical prostatectomy,and to explore the risk factors of postoperative upgrading of GS.Methods:We retrospectively evaluated 160 patients who suf-fered from biopsy proved prostatic carcinoma and performed radical prostatectomy.Age of the patients was 57 -82 years,with the average age of 71.6;prebiopsy prostate specific antigen (PSA)was 0.31 -40.32 μg/L,with the average PSA of 11.29 μg/L;body mass index (BMI)was 16.41 -32.04 kg/m2 , with the average BMI of 23.63 kg/m2;prostate volume (PV)was 9.52 -148.46 mL,with the average PV of 40.19 mL.All the patients included in the study had complete information for clinical variables, including age,BMI,prebiopsy PSA level,PV,number of biopsy cores obtained,percentage,clinical stage,and biopsy GS.Grading of NB Gleason score was compared with their corresponding radical pros-tatectomy specimens,and the discrepancy between the NB and prostatectomy specimens GS assessed. Upgrading was defined as any increase in the pathological GS over that of the biopsy GS as a total sum of primary and secondary grades or a change in the order of primary and secondary grades towards higher ones.Univariable and multivariable Logistic regression analyses were used to identify predictors of patho-logical grading changes.Results:Of the 160 patients,the specimen GS was upgraded in 49 (30.6%) patients and remained with no change in 82 (51.3%)patients.Univariate and multivariate regression analysis showed that prostate volume and biopsy GS were independent predictors with postoperative upgra-ding of GS.Age,BMI,PSA before needle biopsy,clinical stage and needle number showed no statistical significance (P >0.05).Conclusion:Lower biopsy GS and smaller prostate volume are increased risks for clinically upgrading of GS after radical prostatectomy.This fact should be kept in mind when deciding on therapy decisions for patients with prostate cancer.
8.Effect of nervous growth factor on the proliferation of the limbal stem cells in vitro
Jie GAO ; Qiang LIU ; Rong HU ; Yue HUANG ; Min SU ; Hong LI
Journal of Regional Anatomy and Operative Surgery 2017;26(6):391-394
Objective To investgate the effect of nervous growth factor(NGF) on the proliferation of the limbal stem cells(LSCs) in vitro,and the relationship bewteen expression of its receptors and cell proliferation.Methods After primary cultured,LSCs were divided into the control group and the NGF group.Selected cells cultured of 1 d,3 d and 5 d in the two groups and examined the expression of p63,TrkA,p75 with immunohistochemistry.Results The average gray scale values of expression of p63,TrkA and p75 at 1 d,3 d and 5 d in NGF group were significant decreased compared with the corresponding data in the control group(P<0.05).Pearson's correlations analysis showed that the average gray scale values of expression of TrkA and p63 were of statistically significant differences(P<0.05).Conclusion These results highlight that NGF could maintain the stem cell properties of LSCs.LSCs could exepress the NGF receptors of TrkA and p75,and the expression of TrkA showed a correlation with LSCs proliferation.
9.Investigation of the influence of different etiologies on serum level of carbohydrate antigen 125 in elderly patients with heart failure
Qi HUANG ; Mingbin XIE ; Min WANG ; Zhengming XU ; Dean PEI ; Qiang YAO ; Ronghua LUO ; Shien HUANG ; Shisheng WANG
Chinese Journal of Geriatrics 2011;30(6):464-466
Objective To investigate the effect of different etiology on the serum level of carbohydrate antigen 125 (CA125) in elderly patients with chronic congestive heart failure (CHF), and to assess any correlation of CA125 with serum level of B-type natriuretic peptide (BNP). Methods The 155 aged patients with New York Heart Association (NYHA) class Ⅲ or Ⅳ were enrolled and grouped into four reasons of hypertension, coronary heart disease (CHD), cardiomyopathy and other reasons, and 25 healthy old persons as control.CA125 and BNP levels were measured by automatic chemiluminescent immunoassay and enzyme immunoradiometric assay, respectively. Results CA125 level in patients with CHF was (83.4±6.6)U/L for hypertension, (36.8±1.4)U/L for CHD, (38.1±1.6)U/L for cardiomyopathy and (38.4±1.4)U/L for other reasons, which significantly higher than for healthy controls [(14.3±1.15) U/L, P<0.05].Especially, CA125 level in hypertension group was notable higher than in other reasons of group (P<0.05), and was positively related to BNP level (r=0.67,P<0.05). Conclusions Serum CA125 level is a predictor for clinical pathogen of CHF.Therefore, it may be a useful additional marker for the evaluation of clinical treatment of these patients
10.Synthesis and antitumor activity of fluoroquinolon-3-yl-s-triazole sulfide ketones and their derivatives from ciprofloxacin.
Li-li NI ; Qiang YAN ; Shu-min WU ; Yu-suo XIE ; Liu-zhou GAO ; Ying-jie LIU ; Wen-long HUANG ; Guo-qiang HU
Acta Pharmaceutica Sinica 2015;50(10):1258-1262
To discover an efficient strategy for the conversion of the antibacterial activity of fluoroquinolones into the antitumor activity, the three series of C-3 s-triazole-based derivatives including sulfide ketones (6a-6g), thiosemicarbazones (7a-7g) and fused heterocyclic thiazolotriazoles (8a-8g) were synthesized from ciprofloxacin (1), respectively. The structures were characterized by elemental analysis and spectral data. The antitumor activity was tested against three tumor cell lines (Hep-3B, Capan-1 and HL60) using the MTT assay. The three types of compounds all exhibited stronger anti-proliferative activities than ciprofloxacin in the test. The order of their activities was in compounds 7>8>6, and the order of selectivity against cancer cell lines was Capan-1, Hep-3B and HL60. Meanwhile, the SAR revealed that some compounds with electron-drawing group substituted such as fluoro- and nitro-phenyl compounds (6f, 7f, 8f) and (6g, 7g, 8g) displayed more significant activity than the control compounds, especially the IC50 values of thiosemicarbazone compounds 7f and 7g against Capan-1 was comparable to doxorubicin. Thus, a five-membered triazole as the C-3 bioisostere modified with the functionalized side-chain of sulfide-ketone thiosemicarbazone warrants special attention and further investigation.
Anti-Bacterial Agents
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chemistry
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Antineoplastic Agents
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pharmacology
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Cell Line, Tumor
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Ciprofloxacin
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chemistry
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Doxorubicin
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pharmacology
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HL-60 Cells
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Humans
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Ketones
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pharmacology
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Sulfides
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pharmacology
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Triazoles
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pharmacology