1.Preparation of Monospecific Polyclonal Antibodies Against Glechoma hederacea Agglutinin
Progress in Biochemistry and Biophysics 2006;33(11):1113-1119
Glechoma hederocea agglutinin (Gleheda) is a novel glycosylated lectin isolated from the leaves of G. hederacea. Like other glycosylated proteins, the detection of Gleheda by immunological methods is often hampered by the cross-reactivity of the polyclonal antibodies with unrelated glycoproteins. Hence a protocol to purify monospecific polyclonal antibodies from a crude antiserum raised against Gleheda was developed. After selective ammonium sulfate precipitation and successive affinity chromatography on columns of Sepharose 4B with immobilized Gleheda and Robinia pseudoacacia agglutinin (RPA), respectively, ion-exchange chromatography on a column of Q Fast Flow was used for further purification. The specificity of the antibody fractions from each step was tested by double immunodiffusion assay and analyzed by Western blot. Results revealed that affinity chromatography of the immunoglobulin fraction on the immobilized Gleheda antigen yielded an antibody preparation that still cross-reacted with many proteins in leaf extracts. Depletion of nonspecific cross-reacting antibodies directed against the glycan part of the glycoprotein by affinity chromatography on immobilized RPA removed most but not all nonspecifically reacting antibodies. Only upon further purification by ion exchange chromatography an IgG fraction of monospecific antibodies that reacted exclusively with Gleheda could be obtained and accordingly was suitable for immunodetection studies. This antibody purification procedure promises simplicity and efficiency. In addition, this method does not require expensive facilities.
2.Application value of CT- based three-dimensional digital navigation technology in interventional microwave ablation of lung cancer
Feng WANG ; Hu SONG ; Ruidong LI ; Ning DING ; Kongyuan ZHANG
Chinese Journal of Radiological Health 2022;31(4):477-481
Objective To explore the application value of computed tomography (CT)-based three-dimensional digital navigation technology in microwave ablation of lung cancer. Methods A total of 92 patients with lung cancer in our hospital were retrospectively analyzed, and randomized to receive microwave ablation with the three-dimensional digital navigation technology or traditional CT-guided microwave ablation (three-dimensional navigation group and traditional group). The two groups were paired according to tumor location, size (the maximum diameter difference of less than or equal to 2 mm), and microwave ablation conditions, for a total of 46 pairs. The operation time, times of microwave ablation needle puncture, CT dose index, incidence rates of intraoperative complications, and postoperative lesion control were compared between the two groups. Results The operation time [(30.07 ± 6.36) min vs (47.20 ± 9.65) min], times of puncture [(1.72 ± 0.69) times vs (7.13 ± 3.00) times], CT dose index [(11.16 ± 2.20) mGy vs (26.67 ± 8.72) mGy], and incidence of intraoperative complications (10.87% vs 34.78%) in the three-dimensional navigation group were lower than those in the traditional group, and the response rate was higher in the three-dimensional navigation group than in the traditional group (93.48% vs 71.74%) (P < 0.05). Conclusion CT-guided three-dimensional digital navigation technology for microwave ablation of lung cancer makes the operation of interventional puncture more accurate and safe.
3.Comment on “Prognostic Role of Claudin-1 Immunohistochemistry in Malignant Solid Tumors: A Meta-Analysis”
Journal of Pathology and Translational Medicine 2019;53(6):411-411
No abstract available.
Claudin-1
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Immunohistochemistry
4.Value Rational Return of Medical Humanities Education under the Concept of People First
Chinese Medical Ethics 2023;36(8):909-914
The concept of people first has important inspirations for the value rational return of medical humanities education in the new era. It is consistent with the educational value, curriculum attribute, and practical attribute of medical humanistic education, and puts forward new requirements for the value rational return of medical humanities education in terms of educational supply content, practical curriculum setting, educational development form, and educational effect evaluation. Medical colleges and universities should further adhere to the medical humanities education concept centered on people’s health, optimize the practice curriculum of medical humanistic education, innovate the practice platform of medical humanistic education, and construct a value evaluation mechanism of medical humanistic education, so as to promote the value rational return of medical humanities education under the concept of people first.
5.Molecular genetic analysis and identification of novel alleles of ABO subtypes
Jun SU ; Xigang WANG ; Hongxia YANG ; Lingling CHE ; Tiantian REN ; Chunqing YANG ; Ling ZHAO ; Sheng WANG
Chinese Journal of Blood Transfusion 2024;37(2):145-150
【Objective】 To study the molecular mechanism of 95 samples of serological ABO subtypes. 【Methods】 A total of 95 samples with discrepancy between forward and reverse blood grouping were subjected to serological confirmation, and genotyped by polymerase chain reaction with sequence-specific primers (PCR-SSP). For those subtype alleles could not be detected by PCR-SSP, ABO gene exon 1-7 sequencing and gene single strand sequencing were performed successively to determine the mutation site and the gene location. 【Results】 A total of 34 ABO alleles were detected in 95 samples. Five common ABO alleles (ABO*A1.01, ABO*A1.02, ABO*B.01, ABO*O.01.01 and ABO*O.01.02) and 29 rare ABO alleles were identified, including 16 named alleles by ISBT (ABO*A2.01, ABO*A2.05, ABO*A2.13, ABO*A3.07, ABO*AW.37, ABO*AEL.05, ABO*B3.01, ABO*B3.05, ABO*BW.03, ABO*BW.07, ABO*BW.27, ABO*BEL.03, ABO*cisAB.01, ABO*cisAB.05, ABO*BA.02, ABO*BA.04) and 5 named alleles by dbRBC(A223, B309, Bw37, Bel09, Bw40)and eight unnamed alleles [ABO*B.01+ 978C>A, ABO*A1.02+ 248A>T, ABO*B.01+ 125dupT, ABO*B.01+ (98+ 1G>A), ABO*A1.02/ABO*B.01+ 1A>G, ABO*A1.02/ABO*O.01.01+ 28G>T, ABO*A1.02/ABO*B.01+ 538C>T, ABO*A1.02/ABO*O.01.01+ 797insT] .The last four samples could not be verified by single strand because of insufficient samples. In 95 samples, 76 samples (21 named alleles of ISBT and dbRBC) were identified by PCR-SSP, and the remaining 19 samples were identified by exon 1-7 sequencing of ABO gene, of which 8 were identified as unnamed alleles, and the remaining 11 samples were not identified as subtype alleles. 【Conclusion】 The molecular genetic mechanism of 95 serological ABO subtypes was revealed, and 8 rare novel alleles were identified. The detection of ambiguous blood groups is influenced by factors such as patient pathology and physiology, therefore the combination of serological testing and genetic testing is suggested for the identification of ABO subtype.
6.Research Progress of Pharmacological Therapy and Nutritional Support for Cachexia in Lung Cancer Patients.
Jiemin WANG ; Weihui JIA ; Danyang LI ; Yanmei SONG ; Ningxin SUN ; Ke YANG ; Hongli LI ; Chonggao YIN
Chinese Journal of Lung Cancer 2022;25(6):420-424
Cachexia is a common complication in patients with lung cancer. It aggravates the toxic and side effects of chemotherapy, hinders the treatment plan, weakens the responsiveness of chemotherapy, reduces the quality of life, increases complications and mortality, and seriously endangers the physical and mental health of patients with lung cancer. The causes and pathogenesis of tumor cachexia are extremely complex, which makes its treatment difficult and complex. Controlling cachexia in lung cancer patients requires many means such as anti-tumor therapy, inhibition of inflammatory response, nutritional support, physical exercise, and relief of symptoms to exert the synergistic effect of multimodal therapy against multiple mechanisms of tumor cachexia. To date, there has been a consensus within the discipline that no single therapy can control the development of cachexia. Some therapies have made some progress, but they need to be implemented in combination with multimodal therapy after fully assessing the individual characteristics of lung cancer patients. This article reviews the application of drug therapy and nutritional support in lung cancer patients, and looks forward to the research direction of cachexia control in lung cancer patients.
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Cachexia/therapy*
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Combined Modality Therapy
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Humans
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Lung Neoplasms/drug therapy*
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Neoplasms/complications*
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Nutritional Support/adverse effects*
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Quality of Life
7.Qualitative Study on the Psychological Experience of Cancer Patients after Withdrawing from Drug Clinical Trials
Yihong JIANG ; Jinhong YANG ; Yanfen LI ; Tingting WANG ; Shujuan CHEN ; Yuxiu LIU
Chinese Medical Ethics 2023;36(6):624-629
【Objective:】 To understand the real psychological feelings of cancer patients after withdrawing from drug clinical trials, and provide reference for clinical healthcare professionals to serve personalized care for patients in the transitional period after withdrawing from drug clinical trials. 【Methods:】 With the descriptive phenomenology method, 11 subjects who withdrew from drug clinical trials in the oncology department of a tertiary hospital in Shandong Province were selected by intentional sampling method to conduct semi-structured in-depth interviews. Colaizzi seven-step analysis method was used to analyze and sort out records, and refine themes. 【Results:】 The psychological experiences of cancer patients after withdrawing from drug clinical trials were summarized into four themes: benefits of participating in clinical trials, losses of withdrawing from clinical trials, attitudes towards future treatment, and expectations of receiving help. 【Conclusion:】 Cancer patients face great psychological pressure after withdrawing from drug clinical trials and require deep emotional support and humanistic care. Nursing staff should take personalized care measures to meet the special problems of patients’ psychological needs, so as to help patients smoothly pass through the withdrawal stage and enter subsequent treatment.
8.Study of IFN-α in combination with all-trans retinoic acid on the proliferation and differentiation of acute promyelocytic leukemia cell lines NB4 and NB4-R1 cells.
Gongai WANG ; Haiying WANG ; Zhanju WANG ; Kewei XUE ; Chuanxiang MA ; Anhua FENG ; Yuqing TIAN
Chinese Journal of Hematology 2015;36(2):166-167
9.Mechanism of DERL3 Affecting the Proliferation, Invasion and Metastasis of Lung Adenocarcinoma A549 Cells.
Dandan ZHOU ; Jiemin WANG ; Ke YANG ; Liping ZHANG ; Quan ZHENG ; Jun BAI ; Yaqiong HU ; Qingjie MU ; Chonggao YIN ; Hongli LI
Chinese Journal of Lung Cancer 2020;23(8):638-645
BACKGROUND:
Derlin 3 (DERL3) is downregulated in colorectal cancer (CRC) samples. Its level is closely linked to lymphatic metastasis or distant metastasis rate in CRC patients. However, its biological behavior in lung adenocarcinoma were rarely reported. The aim of this study is to investigate the ectopic expression of DERL3 in lung adenocarcinoma tissues and its effect on the invasion and metastasis of lung adenocarcinoma A549 cell line to reveal the possible mechanism of invasion and metastasis of lung adenocarcinoma.
METHODS:
Lung adenocarcinoma microarray gene chip data included 3 cases of lymph node metastasis and 3 cases of lung adenocarcinoma tissue without lymph node metastasis. The GEDS and Kaplan-Meier plot queries the survival curve and expression level of DERL3. Western blot was used to detect the expression of DERL3 in lung adenocarcinoma cells. The efficiency of knockdown DERL3 gene was detected by Western blot assay. Transwell detected the number of cells passing through the basement membrane of the transwell. EDU assay detected cell proliferation ability. Western blot detected the expression of epithelial-mesenchymal transition related proteins E-cadherin and Vimentin.
RESULTS:
The microarray gene chip results showed that compared with lung adenocarcinoma tissues without lymph node metastasis, 1,314 mRNAs in lung adenocarcinoma tissues with lymph node metastasis were up-regulated, 400 mRNAs were down (P<0.05). The expression of DERL3 increased in lung adenocarcinoma (P<0.05). The results of survival curve showed that the lung cancer patients with high expression of DERL3 with poor prognosis (P<0.05). Western blot results indicated that plasmid transfection was successful. Knockdown of DERL3 suppressed the ability of proliferation, invasion and migration in A549 cells (P<0.05). After knockdown of DERL3, the expression level of Vimentin was decreased, while E-cadherin expression increased (P<0.05).
CONCLUSIONS
Knockdown of DERL3 inhibited the proliferation, invasion and metastasis of A549 cells.
10.Mechanism of miR-186-5p Regulating PRKAA2 to Promote Ferroptosis in Lung Adenocarcinoma Cells.
Lu LIU ; Xin GUAN ; Yanqiao ZHAO ; Xiaona WANG ; Chonggao YIN ; Qinghua LIU ; Hongli LI
Chinese Journal of Lung Cancer 2023;26(11):813-821
BACKGROUND:
Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer, and any change of miRNAs expression will affect the degree of target regulation, thus affecting intracellular homeostasis. This study verified that miR-186-5p could inhibit the proliferation, migration and invasion of LUAD cells by regulating PRKAA2.
METHODS:
Previous investigations found that the expression of miR-186-5p was markedly suppressed in LUAD. Bioinformatics method is used to predict the target protein related to ferroptosis downstream and inquire about its expression level in LUAD and its influence on the survival of patients. Double luciferase verified the binding site of PRKAA2 and miR-186-5p. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PRKAA2. The effects of miR-186-5p of LUAD cells as well as the mechanism by which miR-186-5p inhibits Fer-1's sensitivity to ferroptosis were confirmed by EdU, Transwell, and scratch assays. The effect of miR-186-5p on the amount of reactive oxygen species (ROS) in LUAD cells was discovered using ROS experiment. Malondialdehyde (MDA) and glutathione (GSH) experiments were used to detect the effects of miR-186-5p and PRKAA2 on ferroptosis index of LUAD cells. The concentration of lipid ROS (L-ROS) in LUAD cells were measured using the L-ROS tests to determine the effects of miR-186-5p and PRKAA2.
RESULTS:
The expression of PRKAA2 is up-regulated, and a high level of PRKAA2 expression was associated with a poor prognosis for patients with LUAD. Overexpression of miR-186-5p decreased the gene and protein expression of PRKAA2. By promoting ferroptosis, miR-186-5p overexpression prevented lung cancer cells from proliferating, invading, and migrating. ROS could be produced in higher amounts in LUAD cells due to miR-186-5p. Overexpression of miR-186-5p and knockdown PRKAA2 up-regulated MDA content and reduced GSH content in LUAD cells, respectively. miR-186-5p could increase the content of L-ROS and promote the ferroptosis sensitivity of LUAD cells by targeting PRKAA2.
CONCLUSIONS
miR-186-5p promotes ferroptosis of LUAD cells through targeted regulation of PRKAA2, thus inhibiting the proliferation, invasion and migration of LUAD.
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Humans
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Lung Neoplasms/genetics*
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Carcinoma, Non-Small-Cell Lung
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Ferroptosis/genetics*
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Reactive Oxygen Species
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Adenocarcinoma of Lung/genetics*
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MicroRNAs/genetics*
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3,4-Methylenedioxyamphetamine
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Cell Proliferation/genetics*
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Cell Movement/genetics*
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Gene Expression Regulation, Neoplastic
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Cell Line, Tumor
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AMP-Activated Protein Kinases