1.Influence of testing system and sample diluents on cyclosporine analysis
Zhidong GU ; Xinming SHI ; Qiuya LU ; Qishi FAN
Chinese Journal of Laboratory Medicine 2003;0(08):-
Objective To obtain conversion formulae of cyclosporine test results between TDX and AXSYM system and evaluate the influence of different sample diluents in over-range samples on AXSYM system.Methods One hundred samples with different concentration were analyzed by TDX and AXSYM system, respectively. The results were compared. Fifty over-range samples were diluted with prepared diluents such as whole blood, phosphate buffer, self-prepared diluents and zero point calibrator and then analyzed on AXSYM system.Results There was a significant difference in the results between TDX and AXSYM system. Reasonable formulae were concluded based on different concentration ranges. Results obtained by using whole blood and phosphate buffer as diluents differed from that got by using zero point calibrator. In contrast, there was no significant difference between self-prepared diluents and zero point calibrator.Conclusion The conversion formulae are helpful to evaluate the variation of the results while the analysis system was changed. Self-prepared diluents could be used to replace zero point calibrator as sample diluents.
2.Expression of EGFR mutation-specific antibody and its significance in lung adenocarcinoma
Xia GU ; Jieyu WU ; Xinming HE ; Yunen LIN ; Ping HE ; Qinian WU ; Guangqiu LI
Chinese Journal of Clinical and Experimental Pathology 2015;(6):652-656
Purpose To investigate the expression and significance of epiderma1 growth factor receptor( EGFR)mutation-specific anti-bodies in 1ung adenocarcinoma. Methods Immunohistochemica1( IHC)technique was used to detect the expression of EGFR muta-tion-specific antibodies(EGFR-19,EGFR-21)and EGFR tota1 protein antibody(EGFR-P)in 171 cases of 1ung adenocarcinoma with resected specimens,and EGFR gene mutation was a1so performed by amp1ification refractory mutation aystem-PCR( ARMS-PCR). The expression of EGFR-19,EGFR-21 and EGFR-P mutant proteins was compared with EGFR gene mutation and their re1ationship with histo1ogica1 c1assification and c1inica1 characteristics were ana1yzed. Results The expression of EGFR mutant protein was corre1ated with the poor differentiation group inc1uding micropapi11ary and so1id predominant types( P=0. 021 ). EGFR-21 high expression was re1ated to p1eura1 invasion(P=0. 005). The coherence of IHC(with EGFR-19/21 antibodies)and ARMS-PCR was existed(Kappa>0. 4 ). Taking ARMS-PCR as a standard,the sensitivity and specificity of EGFR-19 and EGFR-21 were 65. 0% and 89. 4%, 70. 0% and 97. 6%,respective1y. Conclusions Expression of EGFR mutation-specific antibodies is associated with poor differentia-tion and p1eura1 invasion. It suggests a worse prognosis indicator in 1ung adenocarcinoma. Because of the coherence with ARMS-PCR, using IHC with mutation-specific antibodies to detect the mutant proteins of EGFR-19/21 may act as a pre1iminary screening method of EGFR gene mutation.
3.Innovative practice and popularization analysis of problem-based learning in the teaching of clinical molecular biology testing
Ying CHEN ; Xinming XU ; Changbao XIE ; Rui HU ; Xin WANG ; Qian WANG ; Yong JIAO ; Ping MA ; Bing GU
Chinese Journal of Medical Education Research 2018;17(12):1205-1210
Objective To carry out innovative practice as well as questionnaire survey and conduct promotion analysis of clinical molecular biology testing based on problem-based learning (PBL). Methods 137 Students of medical laboratory technology discipline were chosen as subjects to conduct the innovative practice of PBL method. The teaching results were shared and promoted by video online through WeChat. Micro-questionnaire was conducted in 335 students, and the results of the survey were statistically analyzed. Result The improved PBL teaching based on speech contest can boost students to integrate into classroom teaching actively in which they had a better understanding of the knowledge so that the expected teaching quality of clinical molecular biology testing was achieved effectively. Questionnaire showed that the impact of PBL teaching method on students was not extensive currently, indicating that there was still more space to promoting such teaching methods. Although traditional teaching methods still held a place in students' minds (43.9%, 147 cases), the innovative teaching of PBL, which is novel, interesting and colorful, is more in line with the expectations of medical laboratory students in the new era and has the highest acceptance (49.6%, 166 cases). Conclusion The innovative practice of PBL teaching based on speech contest can achieve the purpose of strengthening students' knowledge acquisition and flexibility in the practical use of knowledge, which is worthy of promoting. Teachers could gradually integrate the new PBL teaching model into the tradi-tional teaching methods in the future, making students study efficiently in a relaxed and active atmosphere
4. Influencing factors for keratinized differentiation of keratinocytes
Xinxin DING ; Xue LI ; Jia WANG ; Xinming GU ; Anqian WU ; Yanmin ZHOU
Chinese Journal of Stomatology 2018;53(4):284-288
Keratinized mucosa in oral cavity plays an important role in periodontal health. The defect of keratinized mucosa may increase the risks of complication of oral implant surgery and restoration. Graft of keratinized tissue and connective tissue are still the gold standard for treating keratinized mucosa defect now. The current research focus on how to modulate non-keratinized mucosa to highly-efficient and minimally-invasive keratinized mucosa. Keratinocytes are critical components of oral mucosa and its final differentiation into keratinized mucosa is controlled by the connective tissue microenvironment involving a variety of molecules and ions. To fully understand keratinized differentiation of keratinocyte, this review focuses on its influence factors and possible mechanisms under the differentiation.
5. Research progress in the pathogenic mechanisms of Porphyromonas gingivalis fimbriae
Liuyi DU ; Xiaolin SUN ; Weixian YU ; Jingyi REN ; Xinming GU ; Yanmin ZHOU
Chinese Journal of Stomatology 2018;53(10):703-707
FimA has been characterized as an important virulence factor for Porphyromonas gingivalis (Pg). These structures play a major role in the mechanisms of adhesion and invasion of Pg to host cells, and can induce cellular activation and cytokines release. FimA can also promote biofilm formation and induce immuno-inflammatory response of host cells. Many studies have characterized FimA to be associated with periodontitis and cardiovascular disease. Pg strains are classified into six types based on divergent nucleotide sequences of the fimA gene (types Ⅰ、Ⅰb、Ⅱ、Ⅲ、Ⅳ andⅤ). The expression of fimbriae is regulated by the fimA gene, which may be the key factor that leads to virulence diversities of Pg, At present, the research on the pathogenesis of FimA mainly focuses on periodontitis and atherosclerosis, which is of great significance for the prevention and treatment of diseases. This paper reviewed the pathogenic effect of FimA in the development of above mentioned two diseases and its application in the prevention.
6.Expression characteristics of glutamine synthetase of wheat in Escherichia coli.
Mingxin GU ; Yihao WEI ; Xiting JIA ; Shuping XIONG ; Xinming MA ; Xiaochun WANG
Chinese Journal of Biotechnology 2018;34(2):264-274
Glutamine synthetase is a key enzyme in plant nitrogen assimilation. To study the structure of wheat glutamine synthetase isoenzymes, GS1, GSr, GSe, GS2 and GS2p of wheat were cloned into pET-21a, and the expression condition was optimized. Although wheat glutamine synthetase isoenzymes had 70%-80% amino acid sequence homology, the isoforms expressed with different characteristics. Induced at 30 °C, the most expression level of GSr, GSe and GS2 was after 3 h, and of GS1 was at the 7 h whereas no GS2p was expressed, and the GS isoenzymes showed different expression level, with the order of GS1 (22%)>GSr (15%)>GS2 (12%)>GSe (5%). GSe expressed as soluble protein, and GS1 expressed mainly as soluble protein whereas GSr and GS2 expressed as insoluble proteins. Induced at 30 °C for 3 h, mRNA transcript levels of GS isoforms were different, with the order of GSr (7.59)>GS2 (1.84)>GS2p (1.66)>GSe (1.46)>GS1 (1.00). The levels of mRNA transcription were not consistent with the level of the protein translation. The analysis of mRNA secondary structure showed the free energy of translation initiation region of glutamine synthetase isoforms was different, with the order of GS1 (14.4)
7.Heterozygous CARD9 mutation favors the development of allergic bronchopulmonary aspergillosis.
Xia XU ; Haiwen LU ; Jianxiong LI ; Jielin DUAN ; Zhongwei WANG ; Jiawei YANG ; Shuyi GU ; Rongguang LUO ; Shuo LIANG ; Wei TANG ; Fengying ZHANG ; Jingqing HANG ; Juan GE ; Xin LIN ; Jieming QU ; Xinming JIA ; Jinfu XU
Chinese Medical Journal 2023;136(16):1949-1958
BACKGROUND:
Previous research demonstrated that a homozygous mutation of g.136372044G>A (S12N) in caspase recruitment domain family member 9 ( CARD9 ) is critical for producing Aspergillus fumigatus -induced ( Af -induced) T helper 2 (T H 2)-mediated responses in allergic bronchopulmonary aspergillosis (ABPA). However, it remains unclear whether the CARD9S12N mutation, especially the heterozygous occurrence, predisposes the host to ABPA.
METHODS:
A total of 61 ABPA patients and 264 controls (including 156 healthy controls and 108 asthma patients) were recruited for sequencing the CARD9 locus to clarify whether patients with this heterozygous single-nucleotide polymorphisms are predisposed to the development of ABPA. A series of in vivo and in vitro experiments, such as quantitative real-time polymerase chain reaction, flow cytometry, and RNA isolation and quantification, were used to illuminate the involved mechanism of the disease.
RESULTS:
The presence of the p.S12N mutation was associated with a significant risk of ABPA in ABPA patients when compared with healthy controls and asthma patients, regardless of Aspergillus sensitivity. Relative to healthy controls without relevant allergies, the mutation of p.S12N was associated with a significant risk of ABPA (OR: 2.69 and 4.17 for GA and AA genotypes, P = 0.003 and 0.029, respectively). Compared with patients with asthma, ABPA patients had a significantly higher heterozygous mutation (GA genotype), indicating that p.S12N might be a significant ABPA-susceptibility locus ( aspergillus sensitized asthma: OR: 3.02, P = 0.009; aspergillus unsensitized asthma: OR: 2.94, P = 0.005). The mutant allele was preferentially expressed in ABPA patients with heterozygous CARD9S12N , which contributes to its functional alterations to facilitate Af -induced T H 2-mediated ABPA development. In terms of mechanism, Card9 wild-type ( Card9WT ) expression levels decreased significantly due to Af -induced decay of its messenger RNA compared to the heterozygous Card9S12N . In addition, ABPA patients with heterozygous CARD9S12N had increased Af -induced interleukin-5 production.
CONCLUSION
Our study provides the genetic evidence showing that the heterozygous mutation of CARD9S12N , followed by allele expression imbalance of CARD9S12N , facilitates the development of ABPA.
Humans
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Aspergillosis, Allergic Bronchopulmonary/complications*
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Aspergillus fumigatus/genetics*
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Asthma/genetics*
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Aspergillus
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Mutation/genetics*
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CARD Signaling Adaptor Proteins/genetics*
8.Expert consensus on clinical application of 177Lu-prostate specific membrane antigen radio-ligand therapy in prostate cancer
Guobing LIU ; Weihai ZHUO ; Yushen GU ; Zhi YANG ; Yue CHEN ; Wei FAN ; Jianming GUO ; Jian TAN ; Xiaohua ZHU ; Li HUO ; Xiaoli LAN ; Biao LI ; Weibing MIAO ; Shaoli SONG ; Hao XU ; Rong TIAN ; Quanyong LUO ; Feng WANG ; Xuemei WANG ; Aimin YANG ; Dong DAI ; Zhiyong DENG ; Jinhua ZHAO ; Xiaoliang CHEN ; Yan FAN ; Zairong GAO ; Xingmin HAN ; Ningyi JIANG ; Anren KUANG ; Yansong LIN ; Fugeng LIU ; Cen LOU ; Xinhui SU ; Lijun TANG ; Hui WANG ; Xinlu WANG ; Fuzhou YANG ; Hui YANG ; Xinming ZHAO ; Bo YANG ; Xiaodong HUANG ; Jiliang CHEN ; Sijin LI ; Jing WANG ; Yaming LI ; Hongcheng SHI
Chinese Journal of Clinical Medicine 2024;31(5):844-850
177Lu- prostate specific membrane antigen (PSMA) radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China. Based on domestic clinical practice and experimental data and referred to international experience and viewpoints, the expert group forms a consensus on the clinical application of 177Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.