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Table of Content

    21 April 2015, Volume 36 Issue 2
    Preparation and antitumor activity of a novel liposome of doxorubicin
    Li Feiyang, Cui Chunying, Wang Yuji, Wu Jianhui
    2015, 36(2):  157-160.  doi:10.3969/j.issn.1006-7795.2015.02.001
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    Objective To develop a novel preparation of liposomal doxorubicin. Methods The liposome was prepared by dispersion film assay, loaded doxorubicin by ammonium sulfate gradient method, the anti-proliferation activities of doxorubicin liposome and doxorubicin against cancer cells evaluated by MTT assay and the antitumor activities of doxorubicin liposome and doxorubicin on S180 mouse model were measured. Results The doxorubicin liposome encapsulation efficiency was more than 96.3% at different time points. Doxorubicin liposome effectively inhibited the proliferation of carcinoma cells. The in vivo anti-tumor activity and toxicity of doxorubicin liposome were significantly higher and lower than that of doxorubicin, respectively. Conclusion The method is practicable to prepare doxorubicin liposome having good antitumor potency in vivo.
    6-Mercaptopurine/verapamil-mesoporous silica and reversing multidrug resistance
    Xing Lu, Cui Chunying, Wang Yuji, Fang Chenjie, Wu Jianhui
    2015, 36(2):  161-165.  doi:10.3969/j.issn.1006-7795.2015.02.002
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    Objective To prepare 6-mercaptopurine/verapamil-loaded mesoporous silica, to examine its in vitro antitumor activity and to evaluate if it can reverse drug-resistance. Methods The mesoporous silica nanoparticles(MSNS) were synthesized with copolycondensation method. Separately loaded 6-mercaptopurine and verapamil onto MSNS to construct 6-mercaptopurine/verapamil-mesoporous silica. Dynamic dialysis was used to analyze the in vitro drug release. The reversal of drug resistance of 6-mercaptopurine/verapamil-mesoporous silica was evaluated on MCF-7 and MCF-7/ADM cells using MTT assay. Results There are spherical microstructure, uniform distribution and regular mesoporous channels in the prepared carrier system with a grain diameter ranging from 170 nm to 250 nm and the Zeta potential was in the range of -30 mV to -40 mV. The in vitro drug release showed that no 6-mercaptopurine be released before the addition of DTT but 6-mercaptopurine was suddenly and then slowly released with the addition of DTT. The results of MTT assays showed that 6-mercaptopurine/verapamil-mesoporous silica carrier system had a good effect on the reversal of drug-resistance. Conclusion 6-mercaptopurine/verapamil-mesoporous silica carrier system has a good antitumor activity and can reverse drug resistance.
    Preparation and evaluation of a liposome of PEG and epirubicin
    Huang Ping, Cui Chunying, Wang Yuji, Wu Jianhui
    2015, 36(2):  166-172.  doi:10.3969/j.issn.1006-7795.2015.02.003
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    Objective To prepare a novel lipsome of polyethylene glycol packaged with epirubicin(EPI) liposomes, to explore the properties of the liposome and evaluate its antitumor activities. Methods Emulsion solvent evaporation method was used to prepare the liposome. The morphology of the liposome was observed by transmission electron microscope(TEM) and scanning electron microscope(SEM). The characteristics such as particle size, zeta potential, drug entrapment rate, stability and releasing property in vitro were studied. The anti-proliferation activities of EPI liposome and EPI against cancer cells were evaluated by MTT assay. The antitumor activity in vivo was assayed on S180 mouse model. Results The average particle size and the zeta potential of the liposome were (231.4±2.0) nm and (-20~30) mV, respectively, and the average entrapment efficiency was (53.14±4.85)%. This novel liposome improved the stability, slowed the releasing rate, enhanced the activity, increased the tolerance and extended dosing interval. Conclusion PEG and EPI formed a novel liposome capable of long-acting, plasma-stabile, pH-dependent release, and highly inhibitory effect against tumor growth.
    Preparation and anti-tumor activity of a novel liposome-loaded drug
    Lu Xuemei, Wang Yuji, Wu Jianhui, Cui Chunying
    2015, 36(2):  172-177.  doi:10.3969/j.issn.1006-7795.2015.02.004
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    Objective To prepare an Arg-Gly-Asp-Phe-fatty alcohol(RGDFOC12) liposomes-loaded 17-allylamino-17-demethoxygeldanamycin(17-AAG). Methods RGDFOC12 liposomes-loaded 17-AAG(RLAs) was prepared by film dispersion method and evaluated by particle size analysis, Zeta potential, encapsulation efficiency, the release in vitro, plasma stability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and the anti-tumor activity in vivo. Results The RLAs was stable colloidal dispersion system in spherical shape of (130.6±0.6)nm in diameter and the Zeta potential was (-28.37±1.67)mV. The release of RLAs in vitro showed that the released percentage of RLAs in pH 5.4 is more than that in pH 7.4. The MTT assay proved that RLAs inhibited the proliferation of cancer cells. The anti-tumor assay showed that RLAs inhibited tumor growth and reduced the toxicity. Conclusion The RLAs were prepared by film dispersion method. RLAs showed anti-tumor activity in vivo and good potential in cancer therapy.
    Studies on anticancer activity of 17-AAG poly-butylcyanoacrylate nanoparticles
    Yan Yan, Wang Yuji, Wu Jianhui, Cui Chunying
    2015, 36(2):  178-184.  doi:10.3969/j.issn.1006-7795.2015.02.005
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    Objective To prepare 17-allylamino-17-demethoxygeldana-mycin poly-butylcyanoacrylate nanoparticles(17-AAG-PBCA-NPs) and study the anticancer activity of 17-AAG poly-butylcyanoacrylate nanoparticles. Methods 17-AAG was encapsulated in PBCA-NPs by interfacial polymerization method. Using single factor analysis combined with orthogonal design to compare the effect factors on 17-AAG-PBCA-NPs, optimizing the preparation method of 17-AAG-PBCA-NPs. Nanoparticle size analyzer, TEM and SEM were used to identify and analyze the characteristics of 17-AAG-PBCA-NPs. The dynamic dialysis method was used to determine the in vitro release of 17-AAG-PBCA-NPs. Results Optimal dosage of drug mass ratio was 1∶10. The conditions of the reaction system were: pH 2, 1 200 r/min, F-68, 3% Dextran70. The time of polymerization was 3 hours. The optimal encapsulation efficiency was more than 90%. The morphology of 17-AAG-PBCA-NPs was spherical shape with (180.5±12.0) nm in diameter and the Zeta potential ranged from -20 to -30 mV. The release of 17-AAG-PBCA-NPs in vitro was determined by dynamic dialysis method and it has been shown that the release of drug from the PBCA-NPs exhibited a rapid burst release followed by a sustained release. 17-AAG-PBCA-NPs showed good stability in plasma. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay of the 17-AAG-PBCA-NPs and 17-AAG using HepG2, A375, HeLa, MCF-7 and SHSY5Y cell line showed that the 17-AAG-PBCA-NPs and 17-AAG inhibited the growth of HepG2, A375, HeLa, MCF-7 and SHSY5Y cells, showing a time-dependent manner, respectively. Furthermore, in vivo anti-tumor activity of 17-AAG-PBCA-NPs was evaluated in sarcoma bearing mice following intraperitoneal injection. Compared with 17-AAG, 17-AAG-PBCA-NPs achieved superior sustained-release effect, and extended the dosing interval further, increased tolerated dose, reduced the side effect of the drug, and improved the compliance to medication, safety and medication. Conclusion 17-AAG-PBCA-NPs were prepared by using interfacial polymerization method. Further optimization of the preparation method is both beneficial to the characteristic and sustained release effect of 17-AAG-PBCA-NPs.
    Evaluation of anti-tumor activity of docetaxel loaded nanostructured lipid carriers with seal oil
    Shi Fan, Cui Chunying, Wu Jianhui, Wang Yuji
    2015, 36(2):  185-191.  doi:10.3969/j.issn.1006-7795.2015.04.006
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    Objective To design and prepare the nanostructured lipid carriers(NLC) and the lipid emulsion(LE) containing seal oil, and the anti-cancer drug docetaxel(DTX) as a model drug. Methods The DTX-NLC was obtained by high pressure homogenization. The release of DTX-NLC and DTX-LE in vitro were determined by dynamic dialysis method in PBS with 0.5% DMSO(pH7.4 and pH5.4), and in plasma. In vitro anti-tumor studies of DTX-NLC and DTX-LE were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. While, in vivo anti-tumor activities of DTX-NLC and DTX-LE were determined in S180 bearing mice. Results The morphology of DTX-NLC was in spherical shape with approximately 200 nm in diameter and the zeta potential ranged from -30 to -50 mV, and the encapsulation efficiency is about 95%.The release percentage of DTX-NLC is(4.30±0.32)% and (8.12±0.32)% in PBS with 0.5% DMSO at pH 7.4 and pH 5.4, and the releasing of the DTX from the DTX-NLC was more sustained from the DTX-LE. The DTX-NLC and DTX-LE showed a good stability in plasma. MTT assays showed that the DTX-NLC, DTX-LE and DTX could inhibit the growth of the cells. Conclusion Vivo anti-tumor studies in S180 bearing mice showed thatthe NLC had better sustained-release effect than that of LE, and extend the dosing interval further, increase tolerated dose, reduce the side effects of drugs, and improve the compliance of medication safety and medication.
    Preparation and anti-tumor bioassays of lipid emulsion composed of seal oil as a drug delivery system for docetaxel
    Wei Liyan, Cui Chunying, Wu Jianhui, Wang Yuji
    2015, 36(2):  192-198.  doi:10.3969/j.issn.1006-7795.2015.02.007
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    Objective To design arginine-glycine-aspartate-phenylalanine-dodecyl alcohols mediated docetaxel lipid emulsion[RGDFOC12-DTX-LE] and investigate its anti-tumor activity. Methods DTX a broadly used anti-cancer drug, was tested as the model drug. The RGDFOC12-DTX-LE was made of seal oil, lecithin, RGDFOC12 and glycerol. The formulation was obtained by high pressured homogenization. Results The physicochemical property of RGDFOC12-DTX-LE was evaluated by measuring mean particle size(190 nm~250 nm), Zeta potential(-30 mV to -40 mV), eccentricity constants(Ke<0.7), pH(6.5~8.0). Drug entrapment efficiency was greater than 90%. Transmission electron microscopy(TEM) and scanning electron microscopy(SEM) indicated that RGDFOC12-DTX-LEs appeared to be homogeneous and spherical particles. The result of dynamic dialysis method demonstrated that the drug was released from the emulsion slowly. Cytotoxicity studies in HepG2, A375, SH-sy5y, HeLa, MCF-7 cell lines were explored and DTX mixed suspension is the positive control. RGDFOC12-DTX-LE for each cell line had obvious time-dependent effect and presents the characteristics of slow releasing. The in vivo anti-tumor activity experiments showed that RGDFOC12-DTX-LE exhibited better anti-tumor activity. Conclusion In this work RGDF-mediated docetaxel lipid emulsion preparation, had a good in vitro stability, lower injection irritation, enhanced compliance, sustained release property and better anti-tumor effect.
    Prognostic impact of lymphovascular space invasion in women with early-stage cervical carcinoma
    Wang Man, Wang Shuzhen
    2015, 36(2):  199-204.  doi:10.3969/j.issn.1006-7795.2015.02.008
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    To determine whether lymphovascular space involvement (LVSI) in women with early cervical carcinoma is an independent prognostic factor,and how to deal with it. The literature was reviewed using Medline/Pubmed and known literature to determine if LVSI is an independent risk factor in patients with early-stage cervical carcinoma whether undergoing surgery or taking conservative treatment.A total of 34 articles were identified that satisfied the evaluation criteria; in ten LVSI was confirmed not to be an independent risk factor, and cannot be used as a criterion for reserving fertility in early cervical cancer. In 8 cases LVSI was identified to be related to the prognosis. Clinical observation showed that in LVSI-positive patients, conservative treatment need to be chosen with caution. Using LVSI as the independent determining factor for consideration of post radical hysterectomy radiotherapy appears questionable, and it may be related to lymph metastasis.
    An analysis of high-risk human papillomavirus DNA-negative cervical intraepithelial neoplasia in atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesion smears: a retrospective study
    He Xin, Tao Huicheng, Wang Shuzhen, Zhu Zhiqing, Wang Yue, Diao Xiaoli, Qu Jiuxin
    2015, 36(2):  205-211.  doi:10.3969/j.issn.1006-7795.2015.02.009
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    Objective To evaluate the cervical pathology among high-risk human papillomavirus(HR-HPV)-negative women with atypical squamous cells of undetermined significance(ASCUS) cells and low-grade squamous intraepithelial lesion(LSIL), and to investigate the reasonable management strategies for HR-HPV-negative patients with ASCUS and LSIL. Methods In this retrospective study, 356 cases who visited the Department of Gynecology, Beijing Chaoyang Hospital Affiliated to Capital Medical University during January 1, 2010 to 31 December 2012, and were HR-HPV-negative/ASCUS and LSIL on Pap smear were included; 306 patients were followed up. All the patients were examined by colposcopy and cervical biopsy if necessary. HR-HPV was detected by hybridization assay II, hybird capture Ⅱ(HC-Ⅱ). SPSS18.0 statistical software was used. Results Of the 202 cases, 16(7.9%) and 14 of 154(9.1%) HR-HPV negative ASCUS and LSIL women developed CIN-Ⅲ or worse compared with 32 of 140(22.9%) and 40 of 223(17.9%) HPV-positive ASCUS and LSIL women(P=0.000). The negative predictive value of HR-HPV testing for CIN-Ⅲ or worse in ASCUS and LSIL women were 87.13% and 84.42%, respectively. When cervical pathological data were further evaluated by age groups in HR-HPV-negative women with ASCUS and LSIL, the risk of ≥ CIN-Ⅱ were significantly different in various age groups(P<0.05), the highest group was 30-39 years old patients(18.2%). Conclusion HR-HPV-negative patients with an ASCUS or LSIL Pap test result have a chance of being cervical high grade lesions, 7.9% and 9.1% of HR-HPV-negative patients with ASCUS or LSIL had CIN-Ⅲ or worse, and 14.3% of these patients had CIN-Ⅱ or worse lesions, the negative predictive value of HR-HPV testing for CIN-Ⅲ or worse were lower. Colposcopy should be done for HR-HPV-negative patients with ASCUS or LSIL.
    Persistence and clearance high risk human papillomavirus types in the population of primary infection: a follow-up study
    Xu Shasha, He Xin, Liu Yingqiao, Liu Jun, Wang Shenzhen, Wang Yue, Diao Xiaoli, Qu Jiuxin
    2015, 36(2):  212-218.  doi:10.3969/j.issn.1006-7795.2015.02.010
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    Objective To investigate the persistence and clearance data regarding the 13 types of high risk human papillomavirus(HR-HPV) in the population of primary infection. Methods A total of 969 cases of HR-HPV positive patients were followed up between January 1, 2013 and December 31, 2013. The epidemiological characteristics of HR-HPV persistent infection were analyzed. The age, viral genotyping, HR-HPV multiple genotypes and initial viral load were evaluated. Statistical analyses were performed using SPSS software for Windows (version 18.0). The Kaplan-Meier survival function was used for calculating the time taken for the clearance of HR-HPV. The role of a group of independent variables concerning the time taken until clearance was evaluated using a Cox proportional-hazards regression model. P<0.05 was considered statistically significant. Results The average time of viral clearance was 9.52 months for all of 969 cases(9.09-9.95 months 95% CI). Older women had longer clearance time than the younger women groups(χ2=7.63, P=0.45). It was found that HPV-56, 35, 58, 52 and 18 had the lower probability of being cleared, when that HPV-45, 51 and 68 had the shorter time to be cleared. It was also found that multiple viral infection had the lower probability of being cleared than the single ones(χ2=18.67, P=0.000). Lower viral loads had better clearance compared to those having higher viral loads(χ2=18.68, P=0.000)). It was shown that age (regression coefficient=-0.082, P=0.046)and single or multiple HR-HPV infection (regression coefficient=-0.231, P=0.037) were associated with time to clearance in the present sample, while the lower or higher viral loads (regression coefficient=-0.185, P=0.108) was not associated with time to clearance in the Cox proportional-hazards regression model. Conclusion HPV infection clearance in this study was related to factors such as age, infection type, viral load and the multiple or single viral infections.
    Epidemiologic characteristics of high risk human papillomavirus(HR-HPV) infection in population undergone hospital opportunistic health screening and the relationship between cervical cancer lesion and HR-HPV infection
    He Xin, Tao Huicheng, Liu Chen, Wang Shuzhen, Wang Yue, Diao Xiaoli, Qu Jiuxin
    2015, 36(2):  219-225.  doi:10.3969/j.issn.1006-7795.2015.02.011
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    Objective To study epidemiological characteristics of population who underwent opportunistic screening for high risk-human papillomavirus(HR-HPV) infection, and to explore the relationship between HR-HPV infection and cervical precancerous lesions. Methods Data from 2 305 patients, according the inclusion criteria, who underwent HR-HPV DNAs and cervical cytology tests at the outpatient department of OB & GYN, Beijing Chaoyang Hospital from January 1, 2012 to December 31, 2012, were collected. HR-HPV DNAs were detected by hybrid capture-Ⅱ(HC-Ⅱ) method, cervical exfoliated cells were collected and inspected. SPSS 18.0 software was used for statistical analysis. Results 1) The infection rate of HR-HPV of the screening crowd was 34.84%, of which the young women(20-24 years old) is the highest(50.0%). With aging, the rate decreased gradually, the lowest rate of women aged 45-49 years(24.18%). It increased again after the age of 55 years(47.02%), showing second peak of infection. The age distribution of positive rate of cervical cytology tests also had two peaks, consistent with characteristics of age distribution of HR-HPV infection rate. 2) HR-HPV infection rates were respectively 23.98%, 66.47%, 87.21%, 98.04% and 100% in normal or inflammation, atypical squamous cells undetermined significance(ASCUS), low squamous intraepithelial lesion(LSIL), high squamous intraepithelial lesion(HSIL), squamous of cervical carcinoma(SCC), thin preparation test(TCT) results, respectively. HR-HPV infection rates of subjects with normal cervical biopsy or inflammation, cervical intraepithelial lesions(CIN)-Ⅰ, CIN-Ⅱ, CIN-Ⅲ, ASCUS, LSIL, HSIL and SCC were 67.44%, 86.96%, 88.24%, 94.57%, 90.48%, respectively. With the increasing level of cervical cytology lesions and cervical histological lesions, the rate of HR-HPV infection significantly increased. 3) In ASCUS patients, the incidence of high-grade cervical lesions was 31.73%, which in HR-HPV positive group was 36.93% and in negative group was 21.43%. The difference was statistically significant between the two groups(χ2=4.13, P=0.042). 4) In LSIL patients, the incidence of high-grade cervical lesions was 34.40%, which in HR-HPV positive group was 37.37% and in negative group was 14.29%. The difference was statistically significant between the two groups(χ2=5.76, P=0.016). Conclusion The infection rate of HR-HPV and the abnormal rate of cervical cytology are different with different age. Younger than 24 years and ≥ 55 years were peak of both of HR-HPV infection and abnormal cytology. HR-HPV infection and cervical lesion severity was positively correlated. Patients for ASCUS and LSIL of cervical cytology, if HR-HPV positive, will increase the probability of CIN-Ⅱ and above lesions, but negative patients cannot be excluded from high-grade cervical lesions, the colpscopic examination is necessary.
    Relationship between carbonic anhydrase-IX and high risk human papilloma virus infection in cervical exfoliated cells
    Liu Yingqiao, He Xin, Liu Jun, Wang Shuzhen, Xu Shasha, Wang Yue, Diao Xiaoli
    2015, 36(2):  226-231.  doi:10.3969/j.issn.1006-7795.2015.02.012
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    Objective To explore the value of carbonic anhydrase-IX(CA-IX) expressed in cervical exfoliated cells for predicting the natural outcome of cervical intraepithelial neoplasia(CIN)and analysis the relationship between high risk human papillomavirus(HR-HPV) infection and CA-IX. Methods Totally 333 cases who underwent cervical exfoliated cells cytologic examination through thinprep cytologic test(TCT) in Chaoyang hospital from May 2012 to Dec 2013 were enrolled into this study. Immunocytochemistry and HPV DNA tests were applied to study the relationship between HR-HPV infection and CA-IX. Results There was a correlation between CA-IX and HR-HPV infection, the staining of CA-IX in different HPV-infected cell was 60.6% in persistent HR-HPV infection, 38.4% in transient HR-HPV-infected cells and 15.6% in non-HR-HPV-infected cells. Conclusion There was a correlation between CA-IX and HR-HPV infection, the expression of CA-IX in persistent HR-HPV infection was much higher than that in transient HR-HPV-infected and non-HR-HPV-infected cells, showing that CA-IX may be one of the biomarkers of persistent HR-HPV infection leading to CIN.
    Immunoscreening and identification of a novel full-length cDNA encoding membrane protein of Trichinella spiralis
    Bi Kuo, Qin Jiajia, Yang Jing, Zhu Xinping
    2015, 36(2):  232-238.  doi:10.3969/j.issn.1006-7795.2015.02.013
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    Objective In order to obtain vaccine or diagnostic candidate antigens for trichinellosis, the adult cDNA library of T. spiralis was immunoscreened. Methods The adult cDNA library of T. spiralis was screened using the swine sera infected with T. spiralis. The cDNA sequence of the positive clone was analyzed and the recombinant protein was expressed and purified. The antigenicity and immunogenicity of the recombinant protein was analyzed with ELISA and Western blotting. Results Forty-two positive clones were identified by immunscreened. A novel gene Tmp10 of T. spiralis was sequenced and recombinant plasmid pET-28a(+)/Tmp10 was successfully constructed. The recombinant protein(rTmp10) was expressed and the molecular weight was about 40 000. The results of Western blotting showed that the rTmp10 could be recognized by sera from the mice and swine infected with T. spiralis respectively, which showed that the rTmp10 had the specific antigenicity. The rTmp10 could induce a high level of specific anti-Tmp10 IgG antibodies, which showed that the rTmp10 had the immunogenicity. Conclusion The novel gene Tmp10 of T. spiralis was obtained by immunoscreening and predicted to be a membrane protein. The recombinant protein Tmp10 had the high specific antigenicity and immunogenicity.
    Influence of Qiwei granules on the expression of CD2AP in podocyte of rats with diabetic nephropathy
    Wang Xinyao, Geng Jianguo, Gao Yanbin, Li Jiaoyang, Zhou Shengnan, Zou Dawei, Liu Yingxin
    2015, 36(2):  239-243.  doi:10.3969/j.issn.1006-7795.2015.02.014
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    Objective To investigate the effects of Qiwei granules on the expression of CD2-associated protein(CD2AP) in podocytes of rats with diabetic nephropathy(DN). Methods Ten normal rats were chosen randomly as the normal group, and the others were abdominally administrated with a single injection of streptozotocin(60 mg/kg) to induce diabetic nephropathy rat models.The model rats were separated into 3 groups randomly: the model group, the western medicine group and Qiwei granules group, and successive administration for 24 weeks. Fasting blood glucose(FBG), body weight(BW), 24 h urinary albumin excretion(UAER) were examined every 4 weeks.After 24 weeks, all the rats were sacrificed,and CD2AP in podocyte was detected by immunohistochemistry and Western blotting. Results As compared with normal control group, the BW in model group are significantly decreased(P>0.05), meanwhile the FBG and 24 hUAER are significantly increased(P<0.05). Compared with model group, BW and FBG in administation group did not change(P>0.05), 24 h UAER is decreased(P<0.05), and the expression of CD2AP is significantly increased(P<0.05). Conclusion 24 h UAER is decreased in Qiwei granules group.The protective effects of Qiwei granules group on kidney are achieved by up-regulating the expression of CD2AP.
    Effect of silencing Notch1 and Notch2 on the expression of Hes family genes in HPAC pancreatic cancer cells
    Li Bingqiu, Zhang Yuxiang
    2015, 36(2):  244-250.  doi:10.3969/j.issn.1006-7795.2015.02.015
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    Objective To explore the effect of silencing Notch1 and Notch2 on the gene expression of Hes family target genes in HPAC pancreatic cancer cells. Methods Notch1-siRNA and Notch2-siRNA were transfected as a liposomal formulation into HPAC cells, respectively, the protein expression of Notch1 and Notch2 were detected by Western blotting, and the mRNA levels of Hes1, Hes2, Hes4 and Hes6 were detected by real-time PCR, and the protein expression level of Hes1 was detected by Western blotting. Results The protein level of Notch1 in Notch1-siRNA group (0.124±0.005) was significantly decreased compared to that of the blank group (1.000±0.019) and RNAi control group (0.908±0.039). Down-regulation of Notch1 expression decreased the mRNA expression of Hes1 and Hes6(Hes1 vs that of the blank group, P=0.025, Hes1 vs RNAi control group, P=0.024; Hes6 vs the blank group, P=0.018, Hes6 vs RNAi control group, P=0.012), but has no effect on the expression of Hes2 and Hes4 mRNA(Hes2 vs the blank group, P=0.064, Hes2 vs RNAi control group, P=0.059; Hes4 vs the blank group, P=0.069, Hes4 vs RNAi control group, P=0.071. The protein expression of Notch2 in Notch2-siRNA group(0.350±0.009) was significantly decreased, compared to the blank group (1.000±0.015) and RNAi control group(0.990±0.017). Down-regulation of Notch2 decreased the mRNA expression of Hes1(Hes1 vs the blank group, P=0.015, Hes1 vs RNAi control group, P=0.014), but has no effect on the expression of Hes2, Hes4 and Hes6(Hes2 vs the blank group, P=0.178; Hes2 vs RNAi control group, P=0.121; Hes4 vs the blank group, P=0.097, Hes6 vs RNAi control group, P=0.100; Hes6 vs the blank group, P=0.053, Hes6 vs RNAi control group, P=0.088. Neither of down-regulation of Notch1 and Notch2 down-regulate the protein expression of Hes1. Conclusion The downstream target gene of Notch1 is Hes1 and Hes6, and the downstream target gene of Notch2 is Hes1. Our results indicate that Notch1 and Notch2 have different regulatory effects on the expressions of Hes family genes.
    NHERF3 expression level and its significance in renal cell carcinoma
    Fa Pengyan, Liu Hua, Zheng Junfang, Wang Lei, Dai Yuanping, He Junqi
    2015, 36(2):  251-254.  doi:10.3969/j.issn.1006-7795.2015.02.016
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    Objective To analysis the difference of the expression level of NHERF3 between renal cell carcinoma and its adjacent tissues. Methods To detect the expression of NHERF3 protein by immunohistochemistry and Western blotting, mRNA by RT-PCR in renal cell carcinoma and their corresponding adjacent tissues. Results NHERF3 protein and mRNA level is significantly decreased in renal cell carcinoma when compared with that in adjacent tissues. Conclusion NHERF3 gene expression was significantly inhibited at the transcriptional level.
    Establishment and evaluation of the striatal 6-OHDA lesioned mice model of Parkinson's disease
    Yu Yan, Wang Ke, Jia Jun, Wang Xiaomin
    2015, 36(2):  255-261.  doi:10.3969/j.issn.1006-7795.2015.02.017
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    Objective To establish different 6-hydroxydopamine(6-OHDA) lesioned mice model, we evaluated the behavioral, neurochemical and histological effect of different dosage injection of 6-OHDA on motor dysfunction and dopamine denervation. Methods Different doses of 6-OHDA(4 μg, 6 μg, 8 μg, 10 μg) were injected into the right dorsal striatum under the facilitation of stereotactic equipment. The body weight after 6-OHDA injection was observed. Apomorphine-induced rotation and rota-rod test were used to estimate motor impairment. Tyrosine hydroxylase(TH) immunostaining was performed to determine the dopaminergic cell loss. Dopamine(DA) content was determined by HPLC. Results 10 μg 6-OHDA injection induced a temporary weight loss, while as other dose of 6-OHDA had no obvious effect on the body weight of mice. At the 1st week after the injection of 6-OHDA, the positive apomorphine induced rotation was observed and this rotational performance was stable for at least 5 weeks. The mice of 8 μg and 10 μg 6-OHDA injection decreased the latency time on the rota-rod. TH-immunolabeling indicated that the 4 μg and 6 μg 6-OHDA injections reduced the number of 60% DA neurons and 40% fibers, as well as 8 μg and 10 μg 6-OHDA injections reduced the number of 80% DA neurons and 70% fibers. Besides, 6-OHDA injection leads to the significant decrease of striatal DA and its metabolites. The decreased of DA content showed a positive correlation with the injection dosage of 6-OHDA, and a negative correlation with dopamine availability(DOPAC+HVA/DA). Conclusion 4 μg and 6 μg 6-OHDA injection induced partial lesions of the nigrostriatal DA system to mimic the early phases of Parkinson's disease(PD). 8 μg and 10 μg 6-OHDA injection induced completely lesions of the nigrostriatal DA system to mimic the late phases of PD.
    Establishment of transgenic mice over-expressing insulin-like growth factor binding protein-4 in the brain
    Gou Rongbin, Jiang Xiaohua, Ju Rongkai, Niu Hanxu, Duan Deyi, Xu Qunyuan
    2015, 36(2):  262-269.  doi:10.3969/j.issn.1006-7795.2015.04.018
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    Objective To establish a transgenic mouse which may over express insulin-like growth factor binding protein-4(IGFBP-4) in the brain, in order to explore the effect of IGFBP-4 on brain development. Methods The neuron-specific transgene expression cassette of IGFBP-4 was constructed and then microinjected into the fertilized mouse egg. The genotype of the transgenic mice was identified using PCR. The expression of IGFBP-4 protein in the brain was detected by Western blotting, and the spatial expression of IGFBP-4 was detected by immunohistochemistry. Both weights of body and brain of mice were measured. Results Exogenous IGFBP-4 cDNA was successfully integrated into the mouse genome and stably passaged. The expression of IGFBP-4 protein in the adult brain was increased by 267% in the transgenic mouse, while its spatial expression pattern was similar to that in the wild type mouse. The body weight was insignificantly changeable, but the whole brain weight was found an 8.4% increase in transgenic mice(P<0.05). Especially, the weight of brain stem increased by 16.5%(P<0.05), with no significant changes in other areas of the brain. Conclusion The exogenous IGFBP-4 cDNA was stably integrated into the transgenic mouse genome, and the over-expression of IGFBP-4 could affect development of certain part of the brain. The newly established IGFBP-4 transgenic mouse may therefore provide a useful animal model for investigating brain development.
    Development of customized PDMS apparatus and dynamic assays for cancer cell migratory invasion
    Xing Tianyu, Niu Jing, Ding Wei
    2015, 36(2):  270-275.  doi:10.3969/j.issn.1006-7795.2015.02.019
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    Objective To develop assays for the dynamic analyses for the migratory invasion of tumor through biocompatible polydimethylsiloxane(PDMS) materials to produce tissue culture apparatus, thus to allow simultaneous morphological and biochemical measurements. Methods An adaptive PDMS mini-culture apparatus with a separate blade was designed and manufactured. U87MG glioblastoma cells were analyzed for time-dependent migration in correlation with the transforming growth factor-beta(TGF-β) luciferase reporter activities. The cell behavior in response to hypoxia treatment was investigated and compared with the results from classic TransWell assays. Results The dynamic monitoring and determination of cell invasiveness and migration could be conveniently achieved with flexibility using the customized PDMS apparatus. By separation and collection of the subpopulations of high-mobility cells, the involved signal transduction pathways could be detected with ease and precision. Conclusion The dynamic characterization and analyses using PDMS-based customized cell culture devices can be an attractive approach to investigate the process and underlining mechanisms of cancer cell migration. The exemplary apparatus demonstrated in this study showed promising potentials for the coordinated investigation from both morphological and biochemical measurements during the tumor cell migration process.
    Ceramide induces autophagic cell death in glioma cell through JNK-c-Jun pathway
    Zhang Luyong, Luo Feiya, Hu Peili, Shan Chun, Zhang Miao
    2015, 36(2):  276-281.  doi:10.3969/j.issn.1006-7795.2015.02.020
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    Objective To observe the autophagy of glioma cell induced by ceramide and explore the possible mechanism. Methods The viability of C6 cells was measured by MTT assay. The cell apoptosis was assayed by flow cytometry. Autophagic-related protein expressions of LC3B/LC3A and Beclin-1 were determined by Western blotting. The level of JNK-c-Jun induced by ceramide was measured by Western blotting with or without the treatment of JNK specific inhibitor SP600125. Results After treatment of ceramide for 24 hours, the growth of C6 cells were significantly inhibited in dose-dependent manner(P<0.05); and ceramide increased autophagic cell death also in dose-dependent manner(P<0.05). Compared with the control group, the levels of LC3B/LC3A and Beclin-1 were significantly increased in ceramide treatment group(P<0.05). JNK was activated in the C6 cells exposed to ceramide and the phosphorylation of c-Jun also increased. This activation of autophagy could be reversed by the pre-treatment of SP600125. Conclusion Ceramide may induce autophagy in glioma cell and the mechanism may be related to the activation of JNK-c-Jun signaling pathway.

    Clinical study of double needle puncture ozone injection combined with coblation in the treatment of discogenic low back pain
    Ge Weipeng, Zheng Guanrong, Du Meiqing, Xu Weimin, Wang Dalong, Zhang Haishan, He Mingwei
    2015, 36(2):  282-286.  doi:10.3969/j.issn.1006-7795.2015.02.021
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    Objective To observe the efficacy and safety of intervertebral disc double needle injection of ozone combined with ablation for the treatment of discogenic low back pain of coblation target through different minimally invasive methods. Methods Totally 120 patients received minimally invasive interventional treatment for lumbar intervertebral disc were randomly divided into 3 groups, Group A received coblation, Group B received double needle injection of ozone, and Group C received both ablation and double needle injection of ozone. Visual analogue scale(VAS), treatment effect, Oswestry disability index(ODI) and complications were evaluated before the treatment, 3 days, 30 days and 90 days after the treatment for all patients. Results The VAS values after treatment in each group significantly decreased, and the difference was statistically significant(P<0.05), and the VAS value reduction and the therapeutic effect of Group C 30 days and 90 days and the ODI of Group C 90 days after the treatment were much more significant than that of Group A and B(P<0.05). No significant complication happened during treatment. Conclusion The therapeutic effect of the combination of double needle injection of ozone and coblation was more obvious and lasted longer.

    Analysis of low density lipoprotein subclass in young patients with corneal arcus and normal serum lipids
    Gao Xinxiao, Guo Jia, Lin Jie, Meng Xin, Wang Jun
    2015, 36(2):  287-290.  doi:10.3969/j.issn.1006-7795.2015.02.022
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    Objective To exactly analyze low density lipoprotein(LDL) subclass and examine the association between small dense LDL(sdLDL) and corneal arcus in young patients. Methods We selected 12 cases of young patients with corneal arcus and normal serum lipids. Another 12 control cases with matched age, gender, smoking were recruited from Beijing Anzhen Hospital between March 2014 and October 2014. LDL particle separation was performed by Lipoprint System: LDL subfractions were obtained and sdLDL proportion(% of sdLDL particles) was calculated. sdLDL% was compared with the Student t test between two groups. Logistic regression analysis was performed to evaluate the correlation between corneal arcus and its related factors. Results SdLDL% was increased in corneal arcus group compared with controls[(19.64±6.52)% vs (13.16±6.29)%]. The difference between the two groups was significant(P=0.020). SdLDL% was significantly associated with corneal arcus(P=0.040) after controlling for age and LDL. SdLDL% played a role in the development of corneal arcus(OR=1.49, 95% CI: 1.01-2.19, P<0.05). Conclusion These findings suggested that young patients with corneal arcus and normal serum lipids had increased level of sdLDL proportion, indicating the abnormality of LDL subclass in such cases.

    Surgery management of implantable left ventricular assist device in sheep model
    Wu Guanghui, Lin Changyan, Hou Xiaotong, Li Haiyang, Chen Chen, Liu Xiujian, Xu Chuangye, Wang Jing, Yang Peng, Qu Wenbo
    2015, 36(2):  291-298.  doi:10.3969/j.issn.1006-7795.2015.04.023
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    Objective We performed this study to explore particular management requirement on sheep experimental implantation of left ventricular assist device(LVAD). Methods In accordance with the Beijing Laboratory Animal Management Regulations, the criteria of American Association for Laboratory Animal Science(AALAS) and the experiences of the other in vivo animal experiments of the LVAD, we implanted the LVAD in the ovine model. The LVADs were implanted in healthy sheep on beating heart. Considering with the characteristics of the animals and in accordance with the requirements of the LVAD, the managements were formulated on aspects of the perioperative, the perioperative and the postoperative. Results In total 6 sheep survived the surgical procedure, one case was terminated because of lung infection 3 days after the surgery, one case was terminated because of the inflow cannula obstruction associated with a thrombus and cellular fibrous tissue growths at the LV apex 25 days after the surgery, and the remaining four cases survived in good condition after implantation and reached the 30 d expectations. Conclusion Careful preparation before the surgery, development of strict surgical procedure, rigorous early postoperative care and a reasonable choice of anticoagulant strategies can effectively improve the success rate of implantable experiments.

    Clinical outcomes of postdilatation with non-compliant balloons following implantation of drug-eluting stents
    Hu Bin, Jia Dean, Fang Zhe, Yang Shiwei, Han Hongya, Xu Xiaohan, Yu Miao, Ma Qian, Shen Hua, Shi Dongmei, Zhou Yujie
    2015, 36(2):  299-302.  doi:10.3969/j.issn.1006-7795.2015.02.024
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    Objective To assess the clinical effects of postdilatation with non-compliant balloons following implantation of drug-eluting stents(DES) in patients with coronary artery disease. Methods From January 2011 to July 2011, 356 consecutive patients underwent postdilatation with non-compliant balloons following implantation of DES at Beijing Anzhen Hospital, Capital Medical University. Via propensity score methodology, these patients were matched with 1 group of 356 patients who underwent DES without postdilatation during the same period. Results The two groups were similar for age, sex, clinical presentation, main risk factors and the features of coronary artery lesions. At 18-month follow-up incidence of major adverse cardiac events(MACE) was 10.1% in the postdilatation treatment group and 15.7% in routine un-postdilatation group(P=0.020), with a significant difference in target vessel revascularization and stent thrombosis(6.2% vs 10.7%, P=0.010; and 1.1% vs 3.7%, P=0.030, respectively).There were no statistical differences in myocardial infarction(AMI) and death rates in the two groups(3.9% vs 4.6%, P=0.700; 1.7% vs 1.3%, P=0.229; and 1.1% vs 1.7%, P=0.760, respectively). Conclusion Postdilatation after DES implantation in patients with coronary artery disease is safe and associated improved clinical outcomes.

    Analysis on relationship between blood uric acid increasing rate and the related risk factors in the physical examination's population in Beijing
    Han Yumei, Chen Shuo, Ding Ran, Tang Jian, Yao Lang, Zhang Jingbo
    2015, 36(2):  303-307.  doi:10.3969/j.issn.1006-7795.2015.02.025
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    Objective To analyse blood uric acid increasing rate and related risk factors of physical examination population Beijing. Methods The research was performed by analyzing blood uric acid increasing distribution from 258 452 health check-up participants in 2008-2012. Then we chose blood uric acid, body mass index(BMI), lipid, blood pressure and fasting blood glucose subjects in 66 333 physical examinationparticipants in 2012. According to the blood uric acid values the experiments were divided into increased uric acidgroup and uric acid in normal ones. To compare with those two group's abnormal detection rate on BMI, lipid, blood pressure, blood glucose by different age groups. Results The prevalence of uric acid increased rates were 21.8%-27.2% in male and 5.5%-7.9% in female. The comparative difference has statistical significance(P<0.05). The overweight, obesity, dyslipidemia, blood pressure and blood glucose abnormal detection rate of the uric acid group are all higher than those of the normal uric acid group(P<0.05). Multivariate logistic regression analysis shows overweight obesity,dyslipidemia, hyperuricemia and hyperglycemia are the risk factors for hyperuricemia. Conclusion In all of the participants, the blood uric acid increasing detected rate in male is higher. Then younger female's uric acid detected rate annually increased. Therefore the health management for the population with blood uric acid increasing should be strengthened for the early detection, diagnosis and treatment of the disease.

    Main surgery procedures and mechanisms for treating type 2 diabetes
    Zhang Zhen, Kong Ke
    2015, 36(2):  318-323.  doi:10.3969/j.issn.1006-7795.2015.02.029
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    Bariatric surgery leads to glycemia normalization in most type 2 diabetes patients. There are a variety of surgery procedures and mechanisms for treating type 2 diabetes. Moreover, the effectiveness is different. These make it difficult for clinician to choose the appropriate surgery procedure. The objective of this study is to review the main surgery procedures and mechanisms for treating type 2 diabetes, contributing to making appropriate choice for clinician.

    Explorating on constructing a digitalized publishing platform of Chinese university sci-tech periodicals
    Zhang Junmin, Wang Xiaomin
    2015, 36(2):  324-327.  doi:10.3969/j.issn.1006-7795.2015.02.030
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    The practice of the science and technology(sci-tech) periodical industry at home and abroad proved that the clustering and digital publishing are an important way to improve the core competitiveness and academic influence of sci-tech periodicals. Chinese University Sci-tech Periodicals is an important part of Chinese Sci-tech Periodicals, it should apply the same way to make them more powerful and, expanded in scale and Chinese University Sci-tech Periodicals should also have a digital publishing platform for themselves development. Because of differences in managements, sponsors and places of the Chinese University Sci-tech Periodicals, they should base on the same or near scientific fields of periodicals to integrate and cluster, then to make a digital publishing platform together. Such a way will provide new insights for the digital development for Chinese University Sci-tech Periodicals.

    Comprehensive application and construction strategy of digital platform used in university academic journals
    Mu Meng
    2015, 36(2):  328-330.  doi:10.3969/j.issn.1006-7795.2015.02.031
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    In this study, measures that university academic journals were taken to response to the digital platform, had been discussed on the basis of understand their actual needs. By understanding, using and exploring digital platform, the journals can do more quickly spread on their contents, more effectively on their periodical office and more effective access to information and resources, as a result, overall academic level and publication quality of academic journals of universities could be improved.