1.The status quo and future prospects of the next generation sequencing technologies in clinical diagnostics
Chinese Journal of Laboratory Medicine 2017;40(7):483-485
During the past decade, tremendous progress has been made in terms of speed, read length, and throughput, along with a sharp reduction in per-base cost.Together, these advances democratized next generation sequence (NGS) and paved the way for the development of a large number of novel NGS applications in clinical diagnostics, especially in the field of non-invasive prenatal detection, rare genetic disease and cancer companion diagnostics.As technology advances, long-read single molecule sequencing began to emerge.Single cell, long-reads, transcriptome, and low cost will be the NGS direction.Due to the special nature of clinical testing, the current NGS clinical application system,including genetic counseling, testing standards, quality control, supervision, database construction etc, does not match the national conditions well and still faces a few challenges, needs to be constantly improved through the routine clinical practice in the future.
2.Molecular diagnosis in China:present status and challenges ahead
Chinese Journal of Laboratory Medicine 2016;39(7):473-476
Molecular diagnosis is rapidly developed in recent years , mainly applicated in the diagnosis of hereditary disease , infectious pathogens, tumor susceptibility and molecular typing , companion diagnosis and prognosis assessment , playing more and more important role in many diseases diagnosis and treatment.Molecular diagnosis was developed from the eighties of the last century in our country .Nowadays, the mainly applied technologies in the clinical laboratory include fluorescence in situ hybridization , quantitative PCR, microarray and DNA sequencing. These molecular technologies make up for the insufficiency of routine testing and take up a central role in the development of modern laboratory medicine . With the continuous development in transformation research of molecular technology recent years , there will be more molecular diagnostic techniques applied in clinicaldiagnosis in the future .But it still exists some drawbacks in the performance of molecular diagnosis in our country according to the current situation , such as imbalanced regional development , mismatched policies, non-standardized laboratory construction , deficiency of quality control and supervision , etc., which requires the joint effort of the government , hospital, professional association and clinical laboratory itself to promote the healthy and orderly development of molecular diagnosis.
4.ATRA combined with IFN-α2a promotes iodine uptake of FTC-133 follicular thyroid carcinoma cells
Zhen ZHAO ; Guohua SHEN ; Huawei CAI
Journal of Medical Postgraduates 2016;29(9):910-912
Objective Some drugs can effectively improve the uptake of 131 I in dedifferentiated thyroid cancer .This study was to investigate the effect of all-trans retinoic acid (ATRA) combined with interferon-α2a (IFN-α2a) on the iodine uptake and re-tention rate of FTC-133 follicular thyroid carcinoma cells . Methods FTC-133 cells were cultured in the presence of 2 μmol/L AT-RA and/or 1500 IU/mL IFN-α2a for 72 hours.The the cells were collected for measurement of the uptake and retention rate of Na 125 I. Results The Na125 I uptake of the FTC-133 cells was significantly increased after 72-hour incubation with 2μmol/L ATRA and 1500 IU/mL IFN-α2a (2423.1 ±237.5) as compared with those of the control (1279.5 ±102.8), ATRA (1438.2 ±149.6), and IFN-α2a groups (1355.3 ±198.4) (P<0.05).Statistically significant differences were observed in the retention rate of Na 125I in the FTC-133 cells at different time points in the each of the blank control , 2μmol/L ATRA, 1500 IU/mL IFN-α2a, and combination groups (P<0.01), but not in that of Na125I among these groups (P<0.05). Conclusion ATRA in combination with IFN-α2a can promote the Na125 I uptake but cannot improve the Na 125 I retention rate in FTC-133 cells.
5.Research advancement of pro-apoptotic mechanisms of GRIM-19 and its related proteins
Guohua SHEN ; Huawei CAI ; Zhen ZHAO
Journal of Medical Postgraduates 2015;(3):303-307
Cancer development is a complex process that involves multiple genetic changes and multiple signaling pathways . Recent findings show that the GRIM-19 is a novel apoptosis regulation gene , and its gene mutations and loss of protein expression have been observed in many tumor types such as urinarysystem tumor , digestive system neoplasm , which are closely related to cancer devel-opment.Thus, GRIM-19 may be a potential target for gene therapy .Pro-apoptotic mechanisms of GRIM-19 and its related proteins such as STAT3,GW112,p16INK4aare overviewed in this article.
6.Expression and role of IL-17 in nasal polyposis.
Yang SHEN ; Suling HONG ; Guohua HU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(8):516-519
OBJECTIVE:
To study the expression of interleukin-17 (IL-17) in nasal polyps from both atopic and nonatopic patients, and its associations with histological features of polyps tissue.
METHOD:
Thirty patients with nasal polyps (NP) were included and divided into atopic and nonatopic groups according to the skin prick test. Histological characteristics were assessed by eosinophilic infiltration with HE staining. IL-17 expression in polyps tissue was detected by ELISA and RT-PCR.
RESULT:
Eosinophilic infiltration was significantly higher in atopic NP patients than in nonatopic NP patients (P < 0.01). IL-17 protein and IL-17 mRNA levels were significantly upregulated in both atopic (P < 0.01) and nonatopic (P < 0.05) patients compared with controls. Furthermore, IL-17 levels were significantly higher in the atopic group than in nonatopic group. Significantly positive correlations were found between IL-17 levels and eosinophilic infiltration in NP patients.
CONCLUSION
These results indicated that expression of IL-17 was significantly upregulated in NP patients and was especially higher in atopic NP patients, suggesting that IL-17 may play an important role in the pathogenesis of NP and atopy may contribute to NP by stimulating the production of IL-17.
Adult
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Aged
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Eosinophils
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cytology
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Female
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Humans
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Hypersensitivity, Immediate
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metabolism
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pathology
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Interleukin-17
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metabolism
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Male
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Middle Aged
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Nasal Polyps
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metabolism
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pathology
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Young Adult
7.Experimental study on the differentiation of SHI-1 cells induced by puerariae radix flavones in vitro
Guohua ZHU ; Qi ZHANG ; Haiping DAI ; Yunliang ZHAI ; Qun SHEN
Journal of Leukemia & Lymphoma 2013;22(10):582-585
Objective To explore the possible effects on differentiation of SHI-1 cells induced by puerariae radix flavones(PRF)in vitro.Methods SHI-1 cells were treated with PRF in various concertration,then the inhibitory effects of cell proliferation were detected by MTT assay,the cell cycles were analyzed by flow cytometry(FCM),the cells reduction rates were detected by NBT reduction test,and the expression of CD11b and CD14 were tested by FCM.Results 10-50 μg/ml PRF could inhibit the proliferation of SHI-1 cells in a time-and dose-dependent manner,and the cell cycles were arrested in S phase.When SHI-1 cells were treated with 10,30 and 50 μg/ml PRF in 48 houres respectively,the NBT reduction rates of cells were increased in a dose-dependent with PRF(P<0.05),and the expression of cells surface differentiation antigen CD14 was also increased along with the concentration of PRF.Conclusion The SHI-1 cells could be induced to differentiation partially after treated with 10,30 and 50 μg/ml PRF in vitro.
8.Research advances in quantitative detection of single cell protein
Yiyun SHEN ; Xiemin QI ; Qinxin SONG ; Guohua ZHOU
Journal of China Pharmaceutical University 2015;(5):521-531
Proteins presence and differences of the expression level can clarify the physiological or pathological changes in organisms;so the quantitative detection of proteins is vital for disease mechanism research;diagnosis and prognosis evaluation.Traditional protein quantitation methods at the tissue level reflected the average protein expression in cells;but ignore the differences between individual cells.In contrast;approaches for quantitative detection at single-cell level can better reflect the differences.Recently;a number of approaches for such detec-tion have been proposed;including microfluidics;microwell-based technology;optical fiber nanobiosensor;activity-based probe technology and mass spectrometry.The principles;advantages and drawbacks of these approaches are briefly introduced in this review.
9.Inhibitory effect of curcumin on the expressions of MAPKs and MMPs in RPMI8226 cells during their pro-liferation
Guohua ZHU ; Qi ZHANG ; Haiping DAI ; Qun SHEN
Journal of Medical Postgraduates 2014;(10):1016-1019
Objective curcumin can suppress the proliferation , induce apoptosis and partial differentiation , and inhibit the migration of many kinds of tumor cells .The aim of this study was to investigate the expressions of mitogen-activated protein kinase (MAPKs) and matrix metalloproteinases (MMPs) when the proliferation of human multiple myeloma RPMI8226 cells was inhibited by curcumin in vitro, and to reveal the antitumor molecular mechanism of curcumin . Methods RPMI8226 cells were treated with various concentrations of curcumin for different periods of times .The inhibitory rate of curcumin on cell proliferation was detected by MTT assay , the cell cycle analyzed by flow cytometry , the protein levels of MAPKs measured by Western blot , and the activity of MMPs analyzed by Gelatin zymography . Results Curcumin inhibited the proliferation of RPMI 8226 cells in a time-and dose-dependent manner , and the cell cycle was arrested in the G 2/M phase ([12.72 ±0.68]%vs [4.79 ±0.15]%).The expressions of JNK and p-JNK showed a con-centration-dependent increase in the RPMI8226 cells treated with curcumin at 6.25, 12.50, and 25.00 μmol/L, respectively (P<0.01) , but the expressions of ERK 1/2 and P38 MAPK did not change significantly compared with the control group (P>0.05).In addition, the activities of MMP-2 and MMP-9 were decreased in a dose-depend-ent manner in the supernatant of RPMI8226 cells ( P <0.01). Conclusion A certain concentration of curcumin could not only acti-vate the JNK signalling pathway of the MAPKs family and induce the apoptosis of RPMI8226 cells, but also inhibit the activity of MMPs and influence the invasion and metastasis of RPMI 8226 cells.
10.Combination of taurine and diazepam has neuroprotective effect on focal cerebral ischemia-reperfusion in rats
Guohua WANG ; Zhenglin JIANG ; Xia LI ; Xingjuan FAN ; Lihua SHEN
Chinese Journal of Pathophysiology 2010;26(4):737-741
AIM: To observe the neuroprotective effect of combined treatment with taurine and diazepam against focal cerebral ischemia-reperfusion in rats. METHODS: Sixty male Sprague-Dawley rats were randomly divided into five groups: sham-operation group, vehicle group, taurine group (200 mg/kg, ip), diazepam group (10 mg/kg, ip) and combined treatment group (taurine 100 mg/kg+diazepam 5 mg/kg). Focal cerebral ischemia was induced by the method of middle cerebral artery occlusion (MCAO) in rats, and reperfusion was emerged by removing the thread 2 h later. The drugs were administered respectively at the time of reperfusion, and subsequently repeated once 12 h later. The animals in vehicle group were intraperitoneally injected with isodose normal saline. The neurological deficit score, the brain water content and cerebral infarction were measured 48 h after MCAO. Other 5 group animals of focal cerebral ischemia-reperfusion (n=16 in each group) were set up as mentioned above and accepted treatments 10 h after reperfusion, likewise repeated once 12 h later. Twelve animals in each group were adopted the same management as the previous 5 groups at 48 h after MCAO. The remained 4 animals in each group were sacrificed until two weeks after MCAO to observe the histopathological changes by nissl staining. RESULTS: Compared to vehicle group, the animals in combined treatment group at 2 h or 12 h after MCAO both decreased the neurological deficit score, reduced the brain water content and infarct volume (P<0.01 or P<0.05). The combined treatment significantly alleviated the neurological necrosis as well. The neuroprotective effect of the combined treatment was superior to that of using taurine or diazepam alone. CONCLUSION: These results suggest that combination of taurine and diazepam treatment has a coordinate neuroprotective effect on both the acute and chronic brain damage of focal cerebral ischemia-reperfusion.