1.Gas chromatography-mass spectrometry (GC-MS) and its application in metabonomics.
Juan LI ; Lujing REN ; Guannan SUN ; He HUANG
Chinese Journal of Biotechnology 2013;29(4):434-446
Metabonomics involves the unbiased quantitative and qualitative analysis of the complete set of metabolites present in cells, body fluids and tissues (the metabolome) based on modern analytic technique with high throughput, high sensitivity, and high resolution. Gas chromatography-mass spectrometry (GC-MS) is used to gain qualitative results of detected metabolites for biological samples as it provides superior distinguishability, detection sensitivity and integrated standard mass spectrometry library. In this article, the historic developments of GC-MS and its application in metabonomics in the past several years were reviewed. Firstly, the classification and the derivative methods of GC-MS were introduced. Subsequently, sample pretreatment process, qualitative and quantitative analysis and data analysis during detecting metabolites by GC-MS were introduced, then its application in microorganism, plant and disease diagnosis was systematically summarized. Finally, the problems in metabonomics study based on GC-MS and the research prospect in the future were discussed.
Gas Chromatography-Mass Spectrometry
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methods
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Metabolomics
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methods
2.Effect of Tangshenping Medicated Serum on Proliferationand RhoA / ROCK Signaling Pathway of High Glucose-induced Epithelial Cells of Renal Tubules
Yonghui MIAO ; Zongjiang ZHAO ; Guannan YANG ; Ting WANG ; Yawei HUANG ; Ziyan ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(6):1050-1056
To study the effect of the Tangshenping containing serum on the proliferation of the high glucose-induced epithelial cells of renal tubules.To prepare durg-contained serum from rats to enter into in vitor reaction system,the cellscultured via 10% FBS-RPMI 1640 were randomly divided into 7groups:the normal group,themodel group,the Y27632 group,theirbesartangroup,the small dose Tangshenping group,the medium dose Tangshenping group and the high dose Tangshenping group and The cells were cultured in 3000 cells/well and grown in 96-well plates.Each group had 8 wells,then detect the effects of all serum sections on the proliferation of high glucose-induced epithelial cells of kidney tubules by the MTT colorimetric method after cultured for 12 h,24 h,48 h,and 60 h.Based on the results above,cell protein were extracted from each group at 24 h,and the expression of RhoA,ROCK1,α-SMA and E-cadherin in each group were detected by Western blotting.After high glucose stimulation,the shape of cell was shuttle-like or irregular triangle,the way it grew was radial;after the intervention of the corresponding serum,the shape of the cell was fiat and irregular polygonal.Started with 12h,compared with the normal group,OD value of other groups increased;at the 24h、48hand 60h,compared with the normal group,OD value of high glucose groupincreased significantly (P<0.01);compared with the high glucose group,OD value of treatment groups decreased (P<0.05);and 48 h,compared with the Y27632group,irbesartan groupand Tangshenping high dose group,OD value of Tangshenping low and medium dose groups decreased (P<0.05);60 h,compared with Y27632 group,OD value of Tangshenping medium dose groups decreased;compared with irbesartan group,OD value of o Tangshenpinggroupsdecreased (P<0.05);compared with Tangshenping high dose group,OD value of Tangshenpinglow groupsdecreased (P<0.05) Western blotting analysis showed that compared with normal group,the expression of E-Cadherin protein in high glucose group reduced,and the expression of RhoA,ROCK1 and α-SMA protein increased;compared with high glucose group,the expression of E-Cadherin protein in each treating group increased,and the Tangshenping large dose group wassignificantly different (P<0.01);the expression of RhoA,ROCK1 and or-SMA protein reduced,Tangshenping,the large dose group was significantlydifferent (P<0.01);Compared with the Y27632 group,the expression of E-cadherin,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference,while the expression of RhoAproteinreduced (P <0.01).Compared with theirbesartan group,the expression of E-cadherin,RhoA,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference (P>0.05).The Tangshenping containing serum is abletoinhibit the proliferation of high glucose-induced epithelial cells of kidney tubules,and can reverse renal tubular-epithelial cell transdifferentiation via regulating RhoA/ROCK signaling pathway,and restrain renal interstitial fibrosis,thereby delaying the pathogenesis of diabetic kidney disease.
3.Identification and analysis of two Mycobacterium abscess with different morphology
Xiaojun ZHOU ; Duanduan XUAN ; Guannan ZHANG ; Tao HUANG ; Xuming WANG ; Lina NIU ; Mingguan LIN ; Chuiwen DENG
Chinese Journal of Laboratory Medicine 2021;44(9):860-863
Two types of Mycobacterium abscesses (Mab) were found in sputum from a patient with severe pneumonia in May 2020. One was Mycobacteriumabscessus with smooth morphology (Mab S), the other was Mycobacteriumabscessus with rough morphology (Mab R). Both of them were compared and drug susceptibility testing were performed to provide clinical scientific diagnosis and treatment. Morphology, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA were used to analyze Mab S and Mab R, phylogenetic evolution tree was constructed by gene sequence alignment for homology, proportion and broth drug test were used for in vitro drug sensitivity test. There were morphological differences between Mab S and Mab R. MALDI-TOF MS analysis showed that there were 223 protein peaks in Mab R and 147 protein peaks in Mab S. Mab S contained 1 397 bp and Mab R contained 1 402 bp as 16s rRNA gene sequencing revealed. Drug susceptibility testing showed that both of them were almost resistant to all antituberculosis drugs, but sensitive to most of antibiotics. Mab S and Mab R were not only different in manifestations, but also in protein and gene comparison. Both of them were generally resistant to antituberculosis drugs. Antibiotic combined therapy has been confirmed to be an effective treatment in clinic.
4.High grade B cell lymphoma with concurrent MYC rearrangement and 11q aberrations:a clinicopathological analysis
Susu LU ; Guannan WANG ; Wugan ZHAO ; Dandan ZHANG ; Yanping ZHANG ; Sixia HUANG ; Enjie LIU ; Weiwei WANG ; Lan ZHANG ; Wencai LI
Chinese Journal of Clinical and Experimental Pathology 2024;40(1):24-28
Purpose To investigate the clinicopathological features,molecular genetics and prognosis of high grade B cell lymphoma with concurrent MYC rearrangement and 11q aberra-tions(HGBCL-MYC-11q).MethodsThree cases of HGBCL-MYC-11q were reviewed and analyzed using hematoxylin-eosin staining,immunohistochemistry,EBER in situ hybridization and fluorescence in situ hybridization.Clinical data were collected with follow-up.Results All three patients were male,age was 10,61,and 74 years,respectively.All patients had Ann Arbor stage Ⅳ disease.All three cases were biopsies occurring in the nasopharynx,upper pharynx and ileocecus,respectively.Three cases were morphologically similar to diffuse infiltrative growth of tumor cells,moderate or moderately large cells,round to slightly irregular nuclei and easily visible mitotic figures.Focal necrosis was noted in one case.One case exhibited the distinct"starry sky"pattern.All cases expressed CD20,BCL6 and MUM1 and high Ki67 index,two cases expressed CD10 and two cases ex-pressed BCL2.CD3,CD30 and TDT were all negative.EBER in situ hybridization was all negative.FISH analyses using C-MYC break-apart probes were all positive and all cases had 11q aberrations.One case only had the 11q23.3 amplification;and one case only had the 11q24.3 loss.After a follow-up for 1-18 months,one patient died and two patients survived with disease.ConclusionHGBCL-MYC-11q is rare,morphologically similar to BL/HGBCL,with MYC rearrangement and 11q abnormali-ties.We should enhance awareness of the disease and improve more accurate diagnosis and differential diagnosis of the disease.
5.Application value of ultrasound in evaluating the morphological and hemodynamic changes of aorta in mice model with aortic dissection induced by angiotensinogen Ⅱcombined with β-aminopropionitrile
Guannan HUANG ; Huiping NIU ; Tianjiao WU ; Yanan LI ; Yanqin ZHAO ; Lili XUE ; Yingxin XIE ; Dong MA ; Yapei ZHAO
Chinese Journal of Ultrasonography 2022;31(12):1084-1091
Objective:To observe the morphological and hemodynamics changes of aortic segments in mice with angiotensinogen Ⅱ(Ang II) combined with β-aminopropionitrile(BAPN) induced-aortic dissection by color Doppler ultrasound(CDUS).Methods:Twenty male mice of 6-8 weeks old C57BL/6 were randomly divided into two groups: the model group( n=10) was induced by intraperitoneal injection of Ang Ⅱ combined with BAPN to establish mice model with aortic dissection; the control group( n=10) was intraperitoneally injected with normal saline.The body weight, systolic and diastolic blood pressure of the mice were routinely recorded. On the 42th day, CDUS was used to measure the indexes of ascending aorta(AoA), descending thoracic aorta(DAo) and suprarenal aorta(SAo) in both groups, including the inner diameter of the cross section, peak systolic velocity(PSV), the end diastolic velocity(EDV), the resistance index(RI), the pulsatility index(PI), time average mean velocity(TAMV), the heart rate(HR) and the maximal shear rate(SR). Then, the aortas were harvested from the root to the bifurcation of the renal artery. The pathological changes of the aortic wall were observed using hematoxylin-eosin(HE) staining. Results:①There were statistically significant differences in body weight, systolic blood pressure, diastolic blood pressure and heart rate between the model group and the control group(all P<0.05). Compared with the control group(0/10), the incidence of the AoA dissection(8/10) in the model group was obviously higher, the difference was statistically significant( P<0.05); while the incidence of the DAo dissection(4/10) and the SAo dissection(3/10) in the model group was slightly higher, the differences were not statistically significant (all P>0.05). ②Compared with the ascending aorta of the control group, the inner diameter, PSV, EDV, TAMV, PI and SR in the model group were significantly higher(all P<0.05), while RI showed no significant difference between the two groups ( P>0.05). For the descending thoracic aorta, PSV, EDV, TAMV, PI and SR in model group were higher than those of the control group(all P<0.05), however the inner diameter and RI were not significantly different between the two groups (all P>0.05). And for the superior renal aorta, PSV, TAMV, RI, PI and SR in the model group were obviously higher than the control group(all P<0.05), whereas the inner diameter and EDV were not significantly different between the two groups (all P>0.05). ③The HE of the tissue section in the model group showed, the aortas were obviously dilated, irregular, with inhomogeneously thickening wall; the endothelial cell nuclei were slightly stained, and some intima and middle layer ruptured and protruded outward to form dissecting aneurysms. The adventitias were markedly infiltrated with inflammatory cells. Conclusions:Ultrasonography could primarily evaluate the hemodynamic changes of aorta in hypertension with aortic dissection, and the PSV, TAMV, PI and SR of aorta may be important indicators for early predicting the occurrence of aortic dissection in hypertension.