1.Modified method of chromosome G-banding in human peripheral blood
Zizhao WU ; Liqing CHENG ; Lingmin MU ; Zhengyue CHEN
Chinese Journal of Tissue Engineering Research 2007;11(11):2185-2186,2193
BACKGROUND: Routine methods of chromosome G-bending are complicated, time-consuming and worse in effect, which is not suitable for clinical examination and teaching investigation of chromosomal disorder.OBJECTIVE: To seek for proper method of improving the effect of chromosome G-banding.DESIGN: Observation experiment.SETTING: Xinxiang Medical University.MATERIALS: The experiment was conducted in the Laboratory of Morphous, Xinxiang Medical University between January 2001 and January 2005. 376 blood samples were obtained from patients with infertility and sterility or those had abnormal childbearing history, who came from the Clinic of Gynecology and Obstetrics, Third Hospital Affiliated to Xinxiang Medical College between 2001 and 2004. Main agents: RPMI1640 culture fluid (GIBCO Co., Ltd), 20% calf serum (CS), colchicines, 0.075 mol/L KCL, methanol, glacial acetic acid, trypase (SIGMA Co., Ltd) and Giemsa staining solution.METHODS: Modified method of human peripheral chromosomal preparation and G-banding: The procedures were the same as routine methods, while partial influential factors were regulated, for example, the action time of colchicines was set at 2 hours before the ending; 1 mL of fixation fluid (the ratio of methanol and glacial acetic acid was 2:1) was added for pre-fixation, and samples were mixed and centrifuged at 1 500 r/minute for 10 minutes. After re-centrifugalization,fresh fixation fluid was added to prepare for cell suspension and glass slide by according to the amount of cells.Above-mentioned glass samples were baked at 50 ℃ for 1-2 hours, naturally cooled to 37 ℃, digested for 3-5 minutes by immersing into trypase, rapidly stained for 10 minutes with Giemsa staining solution, and counted by the test under microscope to observe 3 000 metaphases, and the percentages; The percentage of 400-600 metaphases and well-dispersion rate were calculated. Well-dispersion rate of chromosome referred to the overlapping after dispersion with complete numbers. The trabant and kinetic body were obvious and in bright color with clear shape. Moreover, all chromosomes were on the same plane. The percentage of metaphase = the number of metaphase cells (400-600)/number of cells under observation×100%.MAIN OUTCOME MEASURES: The percentage of 400-600 pieces of metaphases in samples and good integration chromosome.RESULTS: 400-600 metaphases obtained with routine methods accounted for 52% with a well-dispersion rate of 68%,while 400-600 metaphases obtained with modified methods accounted for 75% with a well-dispersion rate of 86%, and there were significant differences between the two groups (P < 0.05).the stripes are chear.CONCLUSION:Chromoseme metaphases obtained with modified methods are more in number and good in dispersion with proper size. Giemsa shows that the stripes art chear.
2.Effects of Liuwei Shunji Capsule on Visceral Hypersensitivity and Content of 5-HT of Rats with Liver Depression and Spleen Deficiency-type Irritable Bowel Syndrome
Guiqiu DENG ; Bei ZHANG ; Zhe ZHANG ; Feiyan CHEN ; Zizhao LAO ; Geng LI ; Qingguang WU ; Honghui CHENG
Chinese Journal of Information on Traditional Chinese Medicine 2014;(8):46-48
Objective To observe the therapeutic effects of Liuwei Shunji Capsule on survival state, visceral hypersensitivity, the content of 5-HT of model rats with liver depression and spleen deficiency-type irritable bowel syndrome (IBS), and explore its mechanism.Methods SD rats were randomly divided into normal group, model group, pinaverium bromide group and Liuwei Shunji Capsule of high, medium and low dose groups. Senna with restraint stress was used in duplicating liver depression and spleen deficiency-type IBS model. From the second day after the model was established, each drug treatment group was administered with corresponding drugs once a day for two weeks. The effects of Liuwei Shunji Capsule on survival state, visceral hypersensitivity, the content of 5-HT in serum and hypothalamus were observed.Results Compared with model group, Liuwei Shunji Capsule could raise weight growth rate of model rats with liver depression and spleen deficiency-type IBS (P<0.05), improve their diarrhea, fatigue demeanor, fur quality, irritability and other symptoms, significantly decrease their visceral hypersensitivity, the content of 5-HT in serum and hypothalamus (P<0.01).Conclusion Liuwei Shunji Capsule can effectively improve survival state and intestinal disorders of rats with liver depression and spleen deficiency-type IBS, and the mechanism of treatment could be realized through regulating 5-HT level.
3.Conventional cytogenetics and fluorescence in situ hybridization as methods for detecting MLL gene rearrangements in leukemia.
Xu-Ping LIU ; Cheng-Wen LI ; Shuang QIN ; Yun DAI ; Ji-Gang XIAO ; Qi HUANG ; Fang-Yun XU ; Jin-Ying GONG ; Shi-He LIU
Journal of Experimental Hematology 2005;13(5):798-803
This study was aimed to compare the values of conventional cytogenetics (CC), interphase FISH and sequential R-banding and FISH analysis as methods for detecting MLL gene rearrangements. 37 acute leukemia patients were studied by CC and interphase FISH. The results showed that among them, 10 cases were 11q23(+)/MLL(+), 2 cases were 11q23(-)/MLL(+) (5.4%), 3 cases were 111q23(+)/MLL(-) (8.1%) and 22 cases were 11q23(-)/MLL(-). For some patients, different results were obtained by using CC and interphase FISH for detecting 11q23/MLL gene rearrangements. After sequential R-banding and FISH analysis for 6 patients, the chromosome related to MLL gene translocation was seen clearly in karyotypes and FISH image. It is concluded that for accurate diagnosis both CC and FISH are needed for detecting 11q23/MLL gene rearrangements, and evaluation is needed in combination of these two results. When necessary, it needs to do sequential R-banding and FISH or molecular analysis.
Adolescent
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Adult
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Aged
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Child
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Child, Preschool
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Chromosome Aberrations
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Chromosome Banding
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Chromosomes, Human, Pair 11
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genetics
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Female
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Gene Rearrangement
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Histone-Lysine N-Methyltransferase
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Humans
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In Situ Hybridization, Fluorescence
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methods
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Infant
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Karyotyping
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Leukemia
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genetics
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Male
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Middle Aged
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Myeloid-Lymphoid Leukemia Protein
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genetics
4. Bruton tyrosine kinase inhibitors and refractory mantle cell lymphoma
Anqi LOU ; Qiang SU ; Anqi LOU ; Junxian YU ; Zizhao CHENG
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(6):680-686
Bruton tyrosine kinase (BTK) is a key mediator of B-cell receptor signalling cascade and an effective target for treating mantle cell lymphoma (MCL). BTK inhibitors play a critical role in the treatment of MCL. Here we introduced the mechanism of action of BTKI in the treatment of MCL. Though generally well prescribed, Ibrutinib, as the first BTKI, still has limitations of toxicity and resistance. New BTK inhibitors, such as zanubrutinib, acalabrutinib and orelabrutinib, are designed to improve on the safety and efficacy as first-generation BTK inhibitors. Comparing the similarities and differences of the two generations of BTKI in structure and function provides a basis for better clinical application of BTKI. On November 15, 2019, FDA approved zanubrutinib for marketing for patients with adult mantle cell lymphoma. Compared with Ibrutinib, zanubrutinib was found with higher target selectivity, longer-lasting inhibition, fewer adverse reactions, and better patient benefit. Zanubrutinib provides a viable treatment option for patients with r/r MCL. At the same time, it is also actively carrying out clinical researches on the treatment of other B-cell lymphomas. It is a very promising targeted drug.