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10 October 2018
Volume 56 Issue 10
Research progress of exosomes biomarkers in tumor development
WANG Chuanxin
2018, 56(10):  18-23.  doi:10.6040/j.issn.1671-7554.0.2018.1026
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Exosomes, a double-layer lipid inclusion with a diameter of 50-200 nm, can carry a variety of proteins, lipids and nucleic acids, playing an important role in intercellular information transfer. As a novel biomarker, exosomes has become a hot topic in clinical reasearches in recent years. Studies have found that exosomes secreted by tumor cells play an important role in the development of tumors such as tumor immunity, angiogenesis, and tumor microenvironment 山 东 大 学 学 报 (医 学 版)56卷10期 -王传新.外泌体生物标志物与肿瘤发生发展的研究进展 \=-regulation, and may become the natural carrier of drugs for tumor treatment. With the development of proteomics, genomics and other technologies, the research on the biological characteristics and functions of exosomes has been further deepened, and the extraction and detection of exosomes have been continuously improved, providing new ideas and opportunities for the clinical diagnosis and treatment of tumors.
Cancer biomarkers of intrahepatic cholangiocarcinoma and the clinical application
GAO Chunfang, HUANG Chenjun
2018, 56(10):  24-30.  doi:10.6040/j.issn.1671-7554.0.2018.1011
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Intrahepatic cholangiocarcinoma(ICC)is the second most common primary liver tumor with increasing incidence worldwide. The early symptoms are insidious. With rapid progression and high malignancy, the tumor shows poor 山 东 大 学 学 报 (医 学 版)56卷10期 -高春芳,等.肝内胆管细胞癌肿瘤标志物研究与临床应用进展 \=-outcomes. Early diagnosis and treatment can effectively improve patients’ survival. Detection of the cancer biomarkers is conducive not only for related studies on the genesis, development and recurrence of ICC, but also for its diagnosis and treatment.
Research advances in markers for the evaluation of lung cancer efficacy
PAN Shiyang
2018, 56(10):  31-39.  doi:10.6040/j.issn.1671-7554.0.2018.1030
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Lung cancer is one of the most common malignant tumors worldwide with the highest mortality rate. Chemotherapy is the most common treatment for advanced lung cancer. The Lack of sufficient specificity and sensitivity makes it difficult for individualized treatment based on imaging examination. Therefore, it is particularly important to find high-sensitivity evaluation indicators. This article briefly reviewed the latest research advances in imaging and biological markers for the clinical evaluation of lung cancer treatment.
Detection of circulating tumor cell: actualities and challenges
CUI Wei, ZHANG Yujuan
2018, 56(10):  40-45.  doi:10.6040/j.issn.1671-7554.0.2018.1013
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Circulating tumor cell(CTC), as a non-invasive and real-time means of monitoring curative effects, has a potential clinical application. However, due to its distinctive characteristics, the detection of circulating tumer cell is still challenging. This paper will review the actualities and challenges of CTC detection, in order that it may eventually be widely applied in clinical practice.
Applications of digital PCR in cancer detection and treatment
GUAN Ming, ZHANG Xinju
2018, 56(10):  46-50.  doi:10.6040/j.issn.1671-7554.0.2018.1035
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Digital PCR is a novel technique that facilitates the absolute quantitation of nucleic acid targets with high sensitivity. Digital PCR combined with liquid biopsies can be used in the detection of infinitesimal nucleic acid samples and rare mutations. It is particularly suitable for cancer patients who need repeated sampling. Digital PCR can provide adequate gene information in cancer diagnosis and relapse monitoring.
Construction and screening of CA50 targeted antibody library and establishment of immunoassay system
DU Lutao, ZHAO Qiaohui, TIAN Xiaoping, WU Xuewei, FU Guangyu, QU Hai, MIAO Yongjun, WANG Lili, ZHENG Guixi, GUO Haiyang, LI Guilin, WANG Chuanxin
2018, 56(10):  51-57.  doi:10.6040/j.issn.1671-7554.0.2018.1027
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Objective To obtain antibodies against CA50 antigen with high affinity and specificity, and to establish a diagnostic kit so as to enhance the specificity and sensitivity of the diagnosis of tumor related diseases. Methods The mouse single chain antibody(ScFv)genes were obtained by PCR and cloned into pcomb3XSS phagemids. ScFv phage antibody library was obtained by electrotransformation. CA50 antigen containing epitope LST a(sialic acid lactose-N-four sugar a)was used for the elutriation screening. Random clone sequencing results showed that the antibody library 山 东 大 学 学 报 (医 学 版)56卷10期 -杜鲁涛,等. CA50靶向抗体文库的构建筛选及应用评价 \=-had good diversity. The human mouse chimeric antibody was constructed by overlap-PCR. The double antibody sandwich method CA50 kit was established after antibody expression purification and matching screening. Meanwhile, 54 clinical positive samples and 146 negative control samples were collected. The CA50 kit established was used for comparation with Roches CA50 kit to evaluate the detection correlation. Results The phage antibody library had a capacity of 2.5×108, and had a good diversity. The positive clones of 24 strains were identified by PCR. The sequencing results showed that 20 strains were correct and the sequences were different. After 3 rounds of panning, 5 positive clones were screened out, and the correlation between the results of CA50 kit and Roches CA50 kit was 0.98. Conclusion The antibodies with high specificity and affinity against CA50 were successfully screened by phage library technology, which made the application of CA50 possible in tumor diagnosis.
A study of serum PIVKA-II in patients with liver metastasis of colorectal cancer
ZHU Yu, WANG Hongjie, HAN Binbin, FENG Shana, ZHANG Chun, LI Xuexiang, HAN Xiaohong, CUI Wei
2018, 56(10):  58-63.  doi:10.6040/j.issn.1671-7554.0.2018.1019
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Objective To evaluate the diagnostic value of serum prothrombin induced by vitamin K absence or antagonist-II(PIVKA-II)in patients with liver metastasis of colorectal cancer(CRC). Methods A total of 120 CRC patients(including 80 without organ metastasis and 40 with liver metastasis), 80 patients with early-stage hepatocellular carcinoma(HCC)and 80 healthy controls were involved in the study. The levels of serum PIVKA-II and carcinoembryonic antigen(CEA)were determined to evaluate the optimal cut-off values. The diagnostic sensitivity and specificity of PIVKA-II and CEA for the liver metastasis of CRC were analyzed. Results The median concentrations of serum PIVKA-II in healthy controls, CRC patients without organ metastasis, CRC patients with liver metastasis and early-stage HCC patients were 18(15-20.8)mAU/mL, 20(17-24)mAU/mL, 32(22.3-41.8)mAU/mL and 345(31.8-3 787)mAU/mL, respectively. The median PIVKA-II level was significantly greater in CRC patients with liver metastasis 山 东 大 学 学 报 (医 学 版)56卷10期 -朱宇,等.人异常凝血酶原(PIVKA-II)在监测结直肠癌肝转移中的应用 \=-than that in healthy controls (P<0.001)and CRC patients without organ metastasis(P<0.001), but significantly lower than that in early-stage HCC patients(P<0.001). There was no significant difference between PIVKA-II and CEA in the area under the receiver operating characteristic curve(AUC)[0.818(95%CI: 0.736-0.901)vs 0.799(95%CI: 0.710-0.888), Z=0.332, P=0.74]. The optimal cut-off value of PIVKA-II for the detection of CRC liver metastasis was 28.5 mAU/mL, with sensitivity of 60% and specificity of 90%. A combination of PIVKA-II and CEA increased the sensitivity(85%)in detecting CRC liver metastasis. Conclusion PIVKA-II could serve as a marker for CRC liver metastasis screening.
Umbilical cord blood NK cells improved liver injury in sub-acute aging rats
LAN Jing, LI Dong, SHI Qing, LIU Zilin, ZHOU Panpan, JU Xiuli
2018, 56(10):  64-71.  doi:10.6040/j.issn.1671-7554.0.2017.795
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Objective To investigate the feasibility and molecular mechanism of umbilical cord blood NK cells in improving liver injury in sub-acute aging rat models. Methods NK cells in cord blood(CB)were amplified from fresh CB mononuclear cells using genetically engineered K562 cells. Male Wistar rats aged 8-10 weeks were randomly divided into four groups: Normal group, Model group, Mod-Con group and Mod-NK group. Body weight and liver index, superoxide dismutase(SOD)and malondialdehyde(MDA)levels in serum and liver, and genes related to apoptosis were tested one week after the experiment. The morphology of liver sections was observed. Results Sub-acute aging rat models could be constructed successfully by subcutaneous injection of D-galactose. The transfusion of high-purity and high-concentration NK cells could significantly promote the recovery of liver injury and body weight(P<0.01), decrease the liver index(P<0.05), enhance the SOD activity(P<0.01), and reduce the level of MDA in serum and liver tissues(P<0.01). The liver sections showed that hepatic edema relieved. Masson staining indicted that fibrosis of liver decreased. Expression of apoptosis promoting genes decreased(P<0.01), while anti-apoptotic gene and fibroblast growth factor increased(P<0.01). Conclusion Transfusion of umbilical cord blood NK cells can effectively improve the liver injury in sub-acute aging rats.
Effects of Notch1 on neural stem cell differentiation induced by all-trans retinoic acid
XU Jiaming, MA Aihua, CHEN Na, HAN Bo
2018, 56(10):  72-78.  doi:10.6040/j.issn.1671-7554.0.2017.1194
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Objective To investigate the role of Notch1 in neural stem cell differentiation induced by all-trans retinoic acid and to reveal its effects on neural tube development. Methods The expressions of Notch1 in nestin, neurofilament(NF), glial fibrillary acidic protein(GFAP)and galactocerebroside(GALC)were identified with immunofluorescence. The neural stem cells were cultured in normal differentiation medium and the all-trans retinoic acid(ATRA)medium. The expressions of nestin, NF, GFAP and GALC on day 1, 3, 5 and 7 were detected with Western blotting. The neural stem cells were divided into 4 groups and treated with normal differentiation medium(group A), normal differentiation medium + DMSO(group B), normal differentiation media + DMSO + 25 μmol/L γ-secretase inhibitor(group C), and normal differentiation media + DMSO + 50 μmol/L γ-secretase inhibitors(group D). The expressions of Notch1 in these groups were detected with Western blotting. Results Nestin, NF, GFAP and GALC were positively expressed, and Notch1 distributed in the cell membrane and cytoplasm. In the normal differentiation medium, Notch1 山 东 大 学 学 报 (医 学 版)56卷10期 -徐加明,等.Notch1在全反式维甲酸诱导的神经干细胞分化中的作用 \=-significantly increased on day 5 compared with day 1, NF decreased significantly on day 5 compared with day 3, GFAP and GALC significantly decreased on day 5 compared with day 1. In the ATRA medium, Notch1 significantly reduced on day 7 compared with day 1, whereas NF, GFAP, and GALC significantly increased on day 7. In after adoling the γSI, the intracellular domain(NICD)of Notch1 decreased significantly. Conclusion Notch1 signal pathway inhibits neural stem cell differentiation, while ATRA promotes neural stem cell differentiation by inhibiting Notch1.
Effects of TAZ knockdown on the proliferation and apoptosis of TSCC cell line CAL27 and the molecular mechanism
CHEN Xiyan, WANG Qi, GU Weiting, WEN Yong
2018, 56(10):  79-85.  doi:10.6040/j.issn.1671-7554.0.2017.985
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Objective To explore the effects of TAZ knockdown on the proliferation and apoptosis of tongue squamous cell carcinoma(TSCC)cell line CAL27, and the possible molecular mechanism. Methods CAL27 cells were divided into the interference group and control group. The former was transfected with targeting TAZ shRNA, and the latter with non-specific targeting sequence. The interference efficiency was detected with real-time fluorescent quantitative PCR(RT-PCR)and Western blotting. The changes of proliferation were detected with CCK-8 assay and EdU staining assay. The cell cycle distribution and cell apoptosis were tested with flowcytometry. The expressions of proteins related to cell cycle and apoptosis were determined with Western blotting. Results The results of RT-PCR and Western blotting showed 山 东 大 学 学 报 (医 学 版)56卷10期 -陈希彦,等.干扰TAZ基因对舌鳞癌细胞CAL27增殖凋亡的影响及其机制 \=-that the mRNA and protein expressions of the interference group decreased sufficiently. The CCK-8 and EdU staining assay indicated that knockdown of TAZ led to the inhibition of proliferation activity. Flowcytometry results showed that cell cycle arrested in G0/G1 phase and cell apoptosis increased after TAZ knockdown. Meanwhile, knockdown of TAZ decreased the expressions of CYCLIN D1, CDK4, and CDK6. By contrast, the expressions of proapoptotic proteins like Bax and cleaved Caspase9 increased, while the expression of anti-apoptotic protein like Bcl-2 decreased. Conclusion Knockdown of TAZ can affect the proliferation and apoptosis of TSCC cell line CAL27 by modulating the expression of proteins related to cell cycle and apoptosis.
Effects of mucin1-C on the biological behaviors of esophageal squamous cell carcinoma cells
GONGSUN Xin, XIN Zhongwei, SHI Mo, QIN Qiming, JIANG Bin, SONG Xuemin, LIU Xiangyan
2018, 56(10):  86-92.  doi:10.6040/j.issn.1671-7554.0.2017.925
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Objective To explore the expression of mucin1-C(MUC1-C)in esophageal squamous cell carcinoma(ESCC)and its role in the malignant biological behaviors of ESCC cells. Methods The expressions of MUC1-C in the ESCC tissues and normal tissues were detected with Western blotting. After MUC1-C entrance into the nucleus was inhibited with Go-201, the activity, growth and proliferation of ESCC cells were assessed with colony-forming assay and CCK-8 test; the cell migration and invasion abilities were measured with wound healing and Transwell assay, respectively; the apoptosis rate was determined with flow cytometry. Results The expression of MUC1-C in ESCC tissues was higher than that in normal tissues. The activity, growth, proliferation, migration and invasion of ESCC cells decreased while the apoptosis rate increased after MUC1-C entrance into the nucleus was inhibited with Go-201. Conclusion MUC1-C can promote the growth, proliferation, migration and invasion while inhibit the apoptosis of ESCC cells.
Consumption of soybean products and gastric cancer risk: a systematic review and meta-analysis
NI Yingchun, YANG Xiaorong, DU Jinge, LÜ Ming
2018, 56(10):  93-102.  doi:10.6040/j.issn.1671-7554.0.2017.1017
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Objective To investigate the association between consumption of soybean products and risk of gastric cancer(GC). Methods Relevant literature was searched in PubMed, Web of Science, Embase, CNKI and Wanfang Database from the inception to March 2017. The association between soybean product consumption and GC risk was analyzed with pooled odds ratio(OR)and 95% confidence interval(CI). Subgroup analyses were further performed according to different types of design, genders and geographic regions. Results A total of 53 studies were obtained, including 18 cohort studies with 5 153 cases and 890 145 participants, and 35 case-control studies with 10 692 cases and 25 324 controls. Finally 3 studies with low quality were excluded and 50 studies were included in the systematic review and meta-analysis. The results indicated that higher soybean, tofu and soybean milk consumptions were associated with a decreased risk of GC(soybean, OR=0.75, 95%CI: 0.66-0.85; tofu, OR=0.77, 95%CI: 0.68-0.88; soybean milk, OR=0.78, 95%CI: 0.65-0.94). Subgroup analyses suggested that higher soybean and tofu consumptions were negatively associated with GC risk(soybean, OR=0.80, 95%CI: 0.74-0.86; tofu, OR=0.77, 95%CI: 0.68-0.88)among Asians. Conclusion High intake of soybean, tofu and soybean milk can reduce the risk of GC.
Effect of IQ motif-containing GTPase activating protein 3 on the proliferation and migration of cervical cancer cells
WANG Yong, JIANG Chen, ZHOU Shiying, YANG Xiaomei
2018, 56(10):  103-109.  doi:10.6040/j.issn.1671-7554.0.2018.590
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Objective To investigate the expression of IQ motif-containing GTPase activating protein 3(IQGAP3)in the cervical cancer tissues and cell lines, and its subsequent influences on cell proliferation, apoptosis and migration. Methods The expression of IQGAP3 was analyzed by immunohistochemistry and real-time PCR in cervical cancer and adjacent normal tissues. The siRNA targeting IQGAP3 was used to knockdown the expression of IQGAP3. The influences of IQGAP3 silencing on cancer cell proliferation, apoptosis and migration were determined by MTT methods, flow cytometry and transwell migration assay, respectively. Results The expression of IQGAP3 was up-regulated in the cervical cancer tissues and cervical cancer cell lines. The up-regulation of IQGAP3 was associated with the poor prognosis of patients with cervical cancer(χ2=6.15, P=0.01). Down-regulation of IQGAP3 decreased proliferation and promoted apoptosis in CaSki and HeLa cells. Moreover, as compared with siScramble transfected cells, the number of migrating cell was significantly decreased in CaSki and HeLa cells due to si-IQGAP3 transfection. In addition, Western blotting analysis showed that knockdown of IQGAP3 increased the expression of epithelial marker E-cadherin and decreased the 山 东 大 学 学 报 (医 学 版)56卷10期 -王永,等.IQ结构域GTP酶激活蛋白3表达对宫颈癌细胞增殖和转移的影响 \=-expression of mesenchymal marker N-cadherin(all P<0.001). Conclusion IQGAP3 is up-regulated in cervical cancer tissues and cells, and its ver-expression promots cell proliferation and the migration capacity in cervical cancer cells. IQGAP3 plays a key role in cervical cancer progression and could be a potential therapeutic target for cervical cancer.
Clinical research on liquid-solid medium culture method in the detection of genitourinary tract mycoplasma infection
JIA Zhonglan, XU Lifeng
2018, 56(10):  110-113.  doi:10.6040/j.issn.1671-7554.0.2018.456
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Objective To compare the liquid medium culture, solid medium culture and liquid-solid medium culture of mycoplasma detection methods, and to optimize culture methods for mycoplasma. Methods A total of 1 642 samples from female genital tract were cultured by the liquid medium culture, solid medium culture and liquid-solid medium culture parallelly from October 2016 to August 2017. The liquid culture results of ureaplasma urealyticum(Uu)and mycoplasma hominis(Mh)were interpretated at 24 and 48 h, respectively; the solid culture result was interpretated at 48 h. The positive rate and false positive rate were calculated and statistical data were analyzed by χ2 test. Results The total positive rate of liquid culture method was 40.38%, among which the positive rates of Uu, Mh and mixed infection were 30.09%, 5.42%, and 4.87%, respectively, and the false positive rate was 5.67%; the total positive rate of solid culture method was 39.46%, among which the positive rates of Uu, Mh and mixed infection were 32.2%, 2.80%, and 4.44%, respectively, and the false positive rate was 0.37%; the total positive rate of the liquid-solid culture method was 32.70%, among which the positive rates of Uu, Mh and mixed infection were 25.70%, 2.74%, and 4.26%, respectively, and the false positive rate was 0.12%. The positive rate exhibited a statistical difference between the liquid-solid medium culture method and liquid medium culture method/solid medium culture method(χ2=16.267, P<0.001; χ2=20.848, 山 东 大 学 学 报 (医 学 版)56卷10期 -贾忠兰,等.液体-固体联合培养法检测泌尿生殖道支原体的临床研究 \=-P<0.001). The false positive rate exhibited a statistical difference between the liquid medium culture method and solid medium culture method(χ2=5.192, P=0.019)/ liquid-solid medium culture method(χ2=4.851, P=0.033). Conclusion The combination of liquid culture and solid culture can accurately identify Uu and Mh. It can eliminate mycoplasma colonizationn in female genital tract and significantly reduce the false positive result caused by the liquid culture method due to bacterial or fungal contamination.
Impacts of leptin resistance developed in boyhood on the health condition of young adults
WANG Min, WANG Chunhua, LIU Hongli, HE Zhiliang, CUI Weiwei, WANG Lei, LIU Jiejun
2018, 56(10):  114-118.  doi:10.6040/j.issn.1671-7554.0.2018.789
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Objective To investigate the impacts of leptin resistance(LR)developed in boyhood on the health condition of young adults. Methods Subjects of a previous study conducted by us 19 years ago on the relationship between serum leptin level and early onset obesity were followed up. LR group included 9 subjects who had LR in their childhood. The normal control(NC)group consisted of 28 adults randomly selected from people with normal body mass index who came for health examination in our hospital during the same period. The control group was of the same age as LR group. Height, weight, waist circumference, blood pressure, fasting blood insulin(FINS), glucose, blood lipids, liver and kidney functions were examined in both groups. The visceral fat was determined and parents’ histories of diseases were collected in LR group. The incidence of metabolic diseases such as obesity and hypertension was compared between the two groups. Results In the LR group, the incidence of central obesity(waist circumference ≥90 cm)was 100%, visceral fat area was(209.79±55.46)cm2, index of insulin resistance(HOMA IR)was 9.42±8.00 and first-degree relatives incidence of type 2 diabetes was 100%. The systolic and diastolic pressures were significantly higher in LR group than in NC group(P=0.001 1, P=0.000 5, respectively). The levels of FBG, triglyceride(TG)and uric 山 东 大 学 学 报 (医 学 版)56卷10期 -王敏,等.男性儿童瘦素抵抗对成年后健康状况的影响 \=-acid(UA)were significantly higher in LR group than in NC group(P=0.005 0, P=0.029 8, P<0.001). The levels of glutamic-pyruvic transaminase(ALT), glutamic oxalacetic transaminase(AST)and glutamyltranspeptidase(GGT)were significantly higher in LR group than in NC group (P=0.003 1, P=0.027 1, P<0.001). The incidence of fatty liver was significantly higher in LR group than in NC group(P<0.001). Conclusion Boys with early onset obesity and LR tend to develop severe obesity and insulin resistance when they grow up. Besides, they have high risks of type 2 diabetes, dyslipidemia, cancer and liver dysfunction. LR in children can predict the risk of type 2 diabetes in the first-degree relatives.