1.Expression of bcl-2 mRNA and protein suppressed by antisense oligodexyonucleotide with phosphorothiote-modification in human melanoma A375 cells line
Shi QIU ; Yanli SHENG ; Liwei RAN
Chinese Journal of Medical Aesthetics and Cosmetology 2011;17(1):41-43
Objective To observe the effect of bcl-2 antisense oligodexyonucleotide (ASODN) by phosphorothiote-modification on the expression of bcl-2 mRNA and protein in human melanoma A375 cells line. Methods With the phosphorothiote-modification and liposome-encapsulation of ASODN, A375 cells were divided into ASODN group, nonsense oligodexyonucleotide (SODN) group and control group. Bcl-2 mRNA and protein were detected by RT-PCR and immunohistochemistry, respectively. Results The expression of bcl-2 protein was remarkably decreased in ASODN group than that in SODN group and control group (53.14 ±4.26 vs 138.22 ± 8.45, 53.14 ± 4.26 vs 141.08 ± 7.83, both P < 0.01 ). The level of bcl-2 mRNA was significantly lower than that in SODN group and control group (0.38 ± 0. 11 vs 0.96 ±0.13, 0. 38 ± 0.11 vs 0. 97 ± 0. 14, both P < 0. 01 ). Conclusion Bcl-2 antisense oligodexyonucleotide could down-regulate the bcl-2 level and block its protein expression.
2.Effects of miRNA-181a on proliferation and migration of multiple myelo-ma cells
Xiao YAN ; Yanli ZHANG ; Guifang OUYANG ; Qitian MU ; Lixia SHENG
Chinese Journal of Pathophysiology 2017;33(1):33-37
AIM:To investigate the effect of miRNA-181a inhibition on the proliferation, migration and cell cycle of the human multiple myeloma cell line RPMI 8226.METHODS:Real-time PCR was used to detect miRNA-181a expression in serum samples from multiple myeloma or healthy subjects .After transfection with miRNA-181a inhibitor, the cell viability was examined by CCK-8 assay and colony formation assay .The cell migration ability was analyzed by wound healing assay .The cell cycle was detected by flow cytometry .Moreover , the protein level of cyclin D 1 and the phosphoryla-tion of PI3K and Akt were determined by Western blot .RESULTS:The expression of miRNA-181a was significantly in-creased in the serum from multiple myeloma patients as compared with healthy group .Inhibition of miRNA-181a expression by transfection with miRNA-181a inhibitor remarkably decreased the cell viability , migratory ability, the population of G0/G1 phase and cyclin D1 protein expression in the RPMI8226 cells.However, the population of S phase and the phosphory-lation of PI3K and Akt were reduced .CONCLUSION:Down-regulation of miRNA-181a inhibits the viability and migra-tory ability in the RPMI8226 cells via inhibition of cell cycle and PI 3K/Akt signaling pathway .
3.Regulating effects of miR-144 on Beclin-1 gene expression in ovarian cancer cells SKOV-3
Hongna SHENG ; Yanli WANG ; Fan WU ; Caiyun ZHAO ; Jing LI
Basic & Clinical Medicine 2017;37(7):1021-1025
Objective To detect the influence of rapamycin on the expression of 4 kinds of miRNAs and the effect cell autophagy.To study the relationship of miR-144 and Beclin-1 gene.Methods SKOV-3 cells were treated with 50 ng/mL rapamycin 2 hours and 10 nmol/L 3-methyl adenine 12 hours,the expression of miR-17,miR-20a,miR-144 and miR-155 was detected by RT-qPCR in SKOV-3 cell of different groups,the protein expression of Beclin-1 was detected by Western blot.The targeting effect of miR-144 on Beclin-1 gene was verified by the dual-luciferase reporter assay,Western blot and RT-qPCR.Results The expression of miR-17,miR-144 and miR-155 were in creased compared with NC groups in rapamycin group (P<0.05);miR-17,miR-20a and miR-144 were down regulated compared with NC group in 3-MA group(P<0.05);the protein of Beclin-1 was down expression compared with NC group in rapamycin group.miR-144 could suppress Beclin-1 expression by targeting the specific 3'untranslated region sequence of Beclin-1 gene.Conclusions miR-144 can inhibit the autophagy-related gene Beclin-1 expression and regulate the autophagy process in SKOV-3 cells.
4.Analysis of the management staff and funds allocation in the new rural cooperative medical system in Guangxi
Yanli ZUO ; Sheng WANG ; Yusha GUO ; Fang SU ; Caiyuan WU ; Shu LI
Chinese Journal of Hospital Administration 2014;30(8):629-631
Objective To learn the management staff and funds allocation of the NRCMS in Guangxi,and come up with solution accordingly.Methods Collection of the annual report data of NRCMS in Guangxi from 2009 to 2012,and calculation of the management personnel and funds expenditure.Results The management personnel of NRCMS in Guangxi is found with a 1000-person gap between the actual staffing and approved staffing quota; the management funds rise year by year which come mostly from fiscal appropriation.Yet it accounts for less than 2.00% and with a high rate of surplus; per-capita funds for management personnel rise significantly,along with their per-capital salaries,yet the highest fall below 25000 yuan per year per person.Conclusion Staffing quota should be fixed more rationally to ensure the number and competence of NRCMS management staff; more funds and better use of management funds are required.
5.Cloning, expression, purification of spinach carboxyl-terminal processing protease of D1 protein with hydrolysis activity and preparation of polyclonal antibody.
Hui LI ; Wei ZHANG ; Mingxia SHENG ; Weiguo LI ; Yanli LIU ; Sufang LIU ; Chao QI
Chinese Journal of Biotechnology 2010;26(4):495-502
Carboxyl-terminal processing protease of D1 protein (CtpA) catalyzes carboxyl terminal processing of the D1 protein of photosystem II, which is essential for the assembly of a manganese cluster and consequent light-mediated water oxidation. It is a target for the discovery of wide-spectrum herbicide. We amplified the CtpA gene from spinach cDNA with standard PCR method and constructed it into pET-28a vector to generate a recombinant expression plasmid. Recombinant CtpA fusion protein with His-tag was expressed as soluble protein in Escherichia coli BL21(DE3) after induction with 0.1 mmol/L IPTG at 8 degrees C for 72 h. We purified the CtpA protein with the Ni-NTA affinity chromatography and Superdex 75 gel filtration chromatography respectively, and verified the protein by SDS-PAGE and Western blotting with anti-his antibody. Hydrolysis activity of CtpA was assayed by HPLC method with a synthetic 24-mer oligopeptide corresponding to carboxyl terminal of precursor D1 protein, and gave a total activity of 1.10 nmol/(mg x min). We used the purified CtpA protein as antigen to immune rabbit for the production of polyclonal antibody, and prepared antibody with high specificity and sensitivity. The results obtained in this paper provided the feasibility of high-throughput screening of lead compounds for the protease as inhibitors and mechanism analysis of CtpA enzyme.
Algal Proteins
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Antibodies
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metabolism
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Carboxypeptidases
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biosynthesis
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chemistry
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genetics
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immunology
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Cloning, Molecular
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DNA, Complementary
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genetics
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Escherichia coli
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genetics
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metabolism
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Hydrolysis
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Proprotein Convertases
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biosynthesis
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chemistry
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genetics
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immunology
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RNA, Plant
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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immunology
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Spinacia oleracea
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enzymology
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genetics
6.A phase II trial of cytoreductive surgery combined with niraparib maintenance in platinum-sensitive, secondary recurrent ovarian cancer: SGOG SOC-3 study
Tingyan SHI ; Sheng YIN ; Jianqing ZHU ; Ping ZHANG ; Jihong LIU ; Libing XIANG ; Yaping ZHU ; Sufang WU ; Xiaojun CHEN ; Xipeng WANG ; Yincheng TENG ; Tao ZHU ; Aijun YU ; Yingli ZHANG ; Yanling FENG ; He HUANG ; Wei BAO ; Yanli LI ; Wei JIANG ; Ping ZHANG ; Jiarui LI ; Zhihong AI ; Wei ZHANG ; Huixun JIA ; Yuqin ZHANG ; Rong JIANG ; Jiejie ZHANG ; Wen GAO ; Yuting LUAN ; Rongyu ZANG
Journal of Gynecologic Oncology 2020;31(3):e61-
Background:
In China, secondary cytoreductive surgery (SCR) has been widely used in ovarian cancer (OC) over the past two decades. Although Gynecologic Oncology Group-0213 trial did not show its overall survival benefit in first relapsed patients, the questions on patient selection and effect of subsequent targeting therapy are still open. The preliminary data from our pre-SOC1 phase II study showed that selected patients with second relapse who never received SCR at recurrence may still benefit from surgery. Moreover, poly(ADP-ribose) polymerase inhibitors (PARPi) maintenance now has been a standard care for platinum sensitive relapsed OC. To our knowledge, no published or ongoing trial is trying to answer the question if patient can benefit from a potentially complete resection combined with PARPi maintenance in OC patients with secondary recurrence.
Methods
SOC-3 is a multi-center, open, randomized, controlled, phase II trial of SCR followed by chemotherapy and niraparib maintenance vs chemotherapy and niraparib maintenance in patients with platinum-sensitive second relapsed OC who never received SCR at recurrence. To guarantee surgical quality, if the sites had no experience of participating in any OC-related surgical trials, the number of recurrent lesions evaluated by central-reviewed positron emission tomography–computed tomography image shouldn't be more than 3. Eligible patients are randomly assigned in a 1:1 ratio to receive either SCR followed by 6 cyclesof platinum-based chemotherapy and niraparib maintenance or 6 cycles of platinum-based chemotherapy and niraparib maintenance alone. Patients who undergo at least 4 cycles of chemotherapy and must be, in the opinion of the investigator, without disease progression, will be assigned niraparib maintenance. Major inclusion criteria are secondary relapsed OC with a platinum-free interval of no less than 6 months and a possibly complete resection. Major exclusion criteria are borderline tumors and non-epithelial ovarian malignancies, received debulking surgery at recurrence and impossible to complete resection. The sample size is 96 patients. Primary endpoint is 12-month non-progression rate.
7.Characterization of the antigens in inactivated porcine circovirus type 2 vaccines and virus-like particle vaccines by high-performance size-exclusion chromatography coupled with multi-angle laser light scattering.
Yuan XU ; Yanli YANG ; Xingqi ZOU ; Cui LI ; Yuanyuan ZHU ; Yixian QIN ; Yan LI ; Ya Nan SHENG ; Yebing LIU ; Guorui PENG ; Xiaoai XU ; Songping ZHANG ; Qizu ZHAO
Chinese Journal of Biotechnology 2022;38(8):2948-2958
This paper aims to detect the antigens in porcine circovirus type 2 (PCV2) vaccines by high-performance size-exclusion chromatography (HPSEC) coupled with multi-angle laser light scattering (MALLS). With purified inactivated PCV2 and PCV2 virus-like particles (VLP) as references, two inactivated vaccines (a and b) and two VLP vaccines (c and d) for PCV2 from four manufacturers were analyzed by HPSEC-MALLS after demulsification. The antigen peaks in HPSEC-MALLS were identified by PCV2 antigen test strips, Western blotting and transmission electron microscope (TEM). The repeatability and linearity of the method were investigated. The results showed the virus antigens in the two inactivated vaccines were eluted at about 13.3 min in HPSEC. The molecular weight of these antigens was 2.61×106 (±4.34%) Da and 2.40×106 (±2.51%) Da, respectively, as calculated by MALLS. The antigen peaks of the two VLP vaccines also appeared at 13.3 min and the molecular weight was 2.09×106 (±2.94%) Da and 2.88×106 (±11.85%) Da, respectively, which was close to the theoretical molecular weight of PCV2. Moreover, an antigen peak of VLP vaccine c was observed at 11.4 min and the molecular weight was 4.37×106 (±0.42%) Da. The antigen was verified to be the dimer of VLP by TEM. Vaccine d and purified Cap VLP antigens were tested repeatedly, and the RSD of the peak area (n=3) was all < 1.5%, indicating that the method was repeatable. The purified VLP were diluted in serial and tested for linearity. The result suggested good linear relationship between the peak area of VLP or VLP aggregates and the protein concentration of the sample with R2 of 0.999 and 0.997, respectively. Thus, the method met the requirement for quantification and aggregate analysis. This method is accurate and efficient in in vitro quality evaluation and improvement of PCV2 vaccine.
Animals
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Antibodies, Viral
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Capsid Proteins
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Chromatography, Gel
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Circoviridae Infections/prevention & control*
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Circovirus
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Lasers
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Swine
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Vaccines, Inactivated
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Vaccines, Virus-Like Particle
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Viral Vaccines
8.Comparison of the classification of pulmonary hypertension due to left heart disease according to transpulmonary pressure gradient or diastolic pressure difference methods.
Hao ZHANG ; Haifeng ZHANG ; Email: HAIFENG_ZHANG@163.COM. ; Wei SUN ; Yanhui SHENG ; Rong YANG ; Dongjie XU ; Fang ZHOU ; Ying XU ; Yanli ZHOU ; Xiangqing KONG ; Xinli LI
Chinese Journal of Cardiology 2015;43(9):769-773
OBJECTIVETo compare the features of patients with pulmonary hypertension due to left heart disease classified according to transpulmonary gradient (TGP) or diastolic pressure difference (DPD).
METHODSThirty-three patients with pulmonary hypertension due to left heart disease diagnosed by right heart catheterization were enrolled. Patients were divided into two groups according to TPG: 17 patients with TPG ≤ 12 mmHg (1 mmHg = 0.133 kPa) and 16 patients with TPG > 12 mmHg; or divided into two groups according to DPD: 23 patients with DPD < 7 mmHg and 10 patients with DPD ≥ 7 mmHg. McNemar's method was used to test the agreement of the two classification methods.
RESULTSBelow are the patients features according to the classification by TPG: central venous pressure ((9.0 ± 2.5) vs. (12.7 ± 5.4) mmHg), mean right atria pressure ((9.1 ± 2.4) vs. (12.8 ± 5.2) mmHg), right heart systolic pressure ((45.5 ± 9.8) vs. (66.8 ± 15.4) mmHg), right heart mean pressure ((22.6 ± 5.2) vs. (33.1 ± 7.5) mmHg), pulmonary systolic pressure ((44.2 ± 10.3) vs. (64.8 ± 14.2) mmHg), pulmonary diastolic pressure ((24.2 ± 4.5) vs. (33.1 ± 8.3) mmHg), pulmonary mean pressure ((32.3 ± 5.7) vs. (45.8 ± 8.6) mmHg), cardiac index ((2.6 ± 1.0) vs. (1.9 ± 0.9) L · min(-1) · m(-2)), right heart EF ((31.2 ± 12.6)% vs. (22.6 ± 7.1) %) and pulmonary vascular resistance ((2.3 ± 0.8) vs. (6.3 ± 2.6) Wood) were significantly different between the two groups (all P < 0.05). According to the classification of DPD, only right heart diastolic pressure ((7.4 ± 3.7) vs. (11.5 ± 5.7) mmHg), pulmonary diastolic pressure ((25.9 ± 6.4) vs. (34.7 ± 8.0) mmHg) and pulmonary vascular resistance ((3.3 ± 2.0) vs. (6.2 ± 3.4) Wood) were significantly different between the two groups (all P < 0.05). These was a weak agreement (κ = 0.386 6, 95% CI: 0.092 2-0.681 0) between the two classification methods.
CONCLUSIONTPG classification is superior to DPD classification for pulmonary hypertension patients due to left heart disease on identifying the hemodynamic differences.
Blood Pressure ; Cardiac Catheterization ; Diastole ; Heart ; Heart Failure ; Hemodynamics ; Humans ; Hypertension, Pulmonary ; Vascular Resistance
9.A review of deep learning methods for the detection and classification of pulmonary nodules.
Qingyi ZHAO ; Ping KONG ; Jianzhong MIN ; Yanli ZHOU ; Zhuangzhuang LIANG ; Sheng CHEN ; Maoju LI
Journal of Biomedical Engineering 2019;36(6):1060-1068
Lung cancer has the highest mortality rate among all malignant tumors. The key to reducing lung cancer mortality is the accurate diagnosis of pulmonary nodules in early-stage lung cancer. Computer-aided diagnostic techniques are considered to have potential beyond human experts for accurate diagnosis of early pulmonary nodules. The detection and classification of pulmonary nodules based on deep learning technology can continuously improve the accuracy of diagnosis through self-learning, and is an important means to achieve computer-aided diagnosis. First, we systematically introduced the application of two dimension convolutional neural network (2D-CNN), three dimension convolutional neural network (3D-CNN) and faster regions convolutional neural network (Faster R-CNN) techniques in the detection of pulmonary nodules. Then we introduced the application of 2D-CNN, 3D-CNN, multi-stream multi-scale convolutional neural network (MMCNN), deep convolutional generative adversarial networks (DCGAN) and transfer learning technology in classification of pulmonary nodules. Finally, we conducted a comprehensive comparative analysis of different deep learning methods in the detection and classification of pulmonary nodules.
Deep Learning
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Humans
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Multiple Pulmonary Nodules
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Neural Networks, Computer
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Solitary Pulmonary Nodule
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Tomography, X-Ray Computed