2.Preparation and characterization of the antibodies against human myofibrillogenesis regulator 1 and application in neonatal rat cardiomyocytes
Xiaoreng WANG ; Xiuhua LIU ; Quansheng SONG ; Ting LI ; Wenling HAN
Chinese Journal of Pathophysiology 2010;26(1):42-47
AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.
3.Current situation and influential factors of cognitive frailty among elderly patients in Urumqi, China
Yongxiu LIU ; Li YU ; Ting HAN ; Ping LI
Chinese Journal of Practical Nursing 2021;37(6):424-430
Objective:To investigate the current status of cognitive frailty among elderly patients in Urumqi and to identify its influencing factors.Methods:From March to December 2019, the elderly from 3 tertiary hospitals′ geriatrics centers in Urumqi were recruited using the general information questionnaire, FRAIL Scale, Montreal Cognitive Assessment and Clinical Dementia Rating.Results:A total of 1 006 elderly patients were surveyed, among which, 131(13.0%) cases were deemed to have developed cognitive frailty. The results of multivariate logistic regression analysis showed that age, depression, Instrumental Activities of Daily Living Scale score and diabetes were influencing factors of cognitive frailty ( P<0.001). Conclusion:The prevalence rate of cognitive frailty in elderly patients is relatively high. Medical staff should attach great importance to the assessment of cognitive frailty in elderly patients and take targeted intervention in time to prevent, slow down or reverse the onset and development of cognitive frailty.
4.Experimental study on participation of interstitial cells of Cajal and connexin 43 in the pathogenesis of diabetic gastroparesis
Han-Ni WU ; Han TIAN ; Xi-Ting ZHANG ; Wei QIAN ; Jing-Song LIU ; Xiao-Hua HOU ;
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
The changes of interstitial cells of Cajal(ICC)distribution and connexin 43(Cx43)expression in gastric muscle layers were assayed in gastroparesis models of streptozotocin-induced diabetic rats.The results showed that gastric emptying was significantly delayed,the contraction frequency and amplitude of gastric muscle segments were greatly decreased,Cx43 gray values were significantly increased and Cx43 distributed homogeneously with ICC immunopositivity in the model rats.These changes appear to be related to the pathogenesis of diabetic gastroparesis.
5.Effect of different cult ure conditions on differentiation of Treg and Th17 in PB-MC
Hongwei SHAO ; Hailin YANG ; Chunmei LIANG ; Xiaomin LU ; Ting LIU ; Han SHEN ; Fenglin WU ; Shulin HUANG
Chinese Journal of Immunology 2015;(7):874-878
Objectiv e:To investigate the effect of different culture conditions on the differentiation of Treg and Th17 to lay a foundation for exploring the methods to reverse the immune tolerance induced by tumor microenvironment.Methods:The IL-6 gene was cloned and stablely transferred into the tumor cell line expressing TGF-β.The conditioned mediums ( CM) were prepared by collecting the culture supernatants of tumor cell lines with or without IL-6 expression and used in the in vitro culture of peripheral blood mononuclear cells ( PBMC ) .The changes of Treg and Th17 in PBMC treated with different CM were detected with flow cytometry ( FCM) .Results:The expression of TGF-βin BEL-7402 was higher than that in HepG2.Thus the BEL-7402 was selected for preparation of cell line stablely transfected with IL -6 gene.ELISA detection confirmed the effective expression of IL -6 by the identified cell lines.It was showed that the Treg increased in PBMC treated with culture supernatants of tumor cells .However,the presence of IL-6 reversed the increase of Treg and promoted the differentiation of Th 17.Conclusion: The culture supernatants of tumor cells increases the proportion of Treg.However,the presence of IL-6 in this CM can reverse the increase of Treg and raise the proportion of Th 17.
6.Multi-slice spiral CT findings of Castleman disease
Peiling LI ; Miao CHANG ; Ting LIU ; Yuchen HAN ; Qiang WANG ; Li ZHAO ; Ke XU
Chinese Journal of Radiology 2013;(1):64-67
Objective To analyze the CT imaging features of Castleman disease and enhance our knowledge of Castleman disease.Methods Twenty two patients with lymph node biopsy-proved or surgeryproved Castleman disease were retrospectively reviewed in this study.Of the 22 patients,18 had localized lesion and 4 patients had multicentric lesions.Correlation was made between CT and pathologic findings.Results Eighteen patients with localized Castleman disease had the hyaline-vascular type and showed well-circumscribed masses with soft-tissue density [mean CT value,(45 ± 16) HU],punctate or bifurcate calcification and linear low-density areas on non-enhanced CT images.All localized masses showed significant enhancementwith an increase of(56 ± 22)HU on arterial phase and showed residual enhancement and some low-density areas on delayed phase.Enhancing patterns were variable,including homogeneous enhancement,gradual enhancement from the edge to the center of mass and heterogeneous enhancement.Four patients with localized lesion demonstrated enhancing vessels around masses.Four patients with muhicentric CD belonged to the plasma cell type and had multiple enlarged lymph nodes.Plasma cell type masses with homogeneous density also showed enhancement after injection of contrast media but appeared to reveal a less increase of (32 ± 10) HU than the hyaline vascular type.Conclusions The localized Castleman disease showed certain characteristics on CT imaging includingcalcification and contrast enhancing patterns,which could help in the differential diagnosis of this disease.The muhicentric Castleman disease did not reveal any useful imaging features.
7.Total flavonoids of Epimedium attenuate aging-related inflammation in rat brain by inhibiting MAPK/NF-κB signaling pathway
Laixin SONG ; Changcheng ZHANG ; Ting WANG ; Zhencai LIU ; Guifang HAN ; Ding YUAN ; Haixia ZHAO
Chinese Pharmacological Bulletin 2017;33(1):84-89,90
Aim To investigate the effect of the total flavonoids of Epimedium (TFE)on MAPK/NF-κB sig-naling pathway and the inflammatory reaction in the hippocampus of natural aging male rats.Methods The morphological changes of the hippocampus com-posed of three areas (CA1 ,CA3 and DG)were ob-served using haematoxylin-eoin (HE ) staining.The protein expression levels of senescence-associated pro-tein p21,apoptosis-related proteins Bax and Bcl-2, nuclear transcription factor-κB p65 (NF-κB p65 )and its downstream inflammatory factors TNF-α,IL-1βand COX-2,and MAPK signaling pathway-related proteins (ERK1/2,p-ERK1/2,JNK,p-JNK,p38MAPK,p-p38MAPK)in hippocampal were detected by Western blot.Results Compared with natural aging group, TFE obviously improved the morphology and structure of hippocampal neurons,and the nerve cells arranged neatly and closely. Furthermore, TFE significantly downregulated the protein expression levels of p2 1 and Bax,upregulated the protein expression levels of Bcl-2 and the ratio of Bcl-2/Bax,and reduced the expression of NF-κB p65 and of its downstream inflammatory fac-tors TNF-α, IL-1β, COX-2, and MAPK signaling pathway-related proteins (p-ERK1/2,p-JNK and p-p38 MAPK ) in hippocampus of natural aging rats. Conclusions TFE effectively protects against inflam-matory reaction in brain aging of SD male rats.The mechanism is related with inhibition of NF-κB nuclear translocation and reduction of its downstream inflamma-tory cytokines expression by inhibiting MAPK signaling pathway activation.
8.Determination of the concentration of very long chain fatty acids in serum by liquid chromatography-tandem mass spectrometry
Liu HAN ; Junjun NI ; Ting XIANG ; Huiyuan GAO ; Wei LI ; Lijun WU
Chinese Journal of Laboratory Medicine 2011;34(1):30-35
Objective To establish a method for very long chain fatty acids( VLCFA )with liquid chromatography-tandem mass spectrometry( LC-MS/MS ). Methods One hundred and one healthy cases and 35 suspected ALD patients collected from April to June in 2009 were enrolled into this study. Quantitative analyzed the concentrations of VLCFA in serum was performed using liquid chromatography-tandem mass spectrometry. The precision, accuracy and recovery were analyzed, and the stability of VLCFA concentration of sample under room temperature and repeated freeze-thawing were also investigated. Serum levels of VLCFA in 101 normal cases were determined and analyzed statistically. The results for the 35 randomly chosen serum samples were compared with those from MDI in Germany. Results Serum VLCFA were separated well under these gradient condition with small interference. The linear range of C22:0 was from 2 mg/L to 64 mg/L, the recovery was 99. 92% -102. 05%, and the relative standard deviation ( RSD ) of intra-day and inter-day was less than 6% and 9% respectively. For C24:0 they were 2-64 mg/L. 95. 12%-100. 44%. ≤6%, ≤7%,respectively. For C26:0, they were 0-8 mg/L, 92.21%-103.71%, ≤7%, ≤8%, respectively. The accuracy of C22: 0,C24:0 and C26:0 were among 85% to 115%. The samples could be stable within 12 h at room temperature and repeated 10 times freeze-thawing. The values of VLCFA in 101 normal cases followed a normal distribution and the measured values were C22:0 =( 19. 43 ±4.43 ) mg/L,C24:0 =( 19. 10 ±4. 58 )mg/L, C26:0 = ( 0. 21 ± 0. 11 ) mg/L, the ratio of C24: 0/C22:0 and C26:0/C22: 0 were ( 0. 99 ± 0. 13 )and ( 0. 01 ±0. 01 ) respectively. The statistical analysis showed the concentration of C26:0 in adults ( 0. 18±0. 10 ) mg/L and children ( 0. 21 ± 0. 08 ) mg/L, C24: 0/C22:0 in adults ( 1.01 ± 0. 10 ) and children ( 0. 99 ±0. 14 ) has no significant( t values were 1. 439,0. 806, respectively, all P > 0. 05 ); the ratio of C24:0/C22:0 in male (1.05 ± 0. 10 ) and female (0.97 ± 0. 10 ) has significant difference ( t =3. 394,P =0. 001 ). Compared the values determined by MDI laboratory, the results of C22: 0( 16. 93 ±4. 30 ) mg/L,C24: 0( 19. 57 ± 6. 40 ) mg/L by this method and C22:0 ( 13.85 ± 3. 17 ) mg/L, C24:0( 16. 10 ±5.84 ) mg/L by MDI have significant differences( t = 8. 401 ,P =0. 000;t =9. 914,P =0. 000 ),but C26:0( 0.68 ±0.48 ) mg/L, C24:0/C22:0( 1.20 ±0.40 ), C26: 0/C22:0 ( 0.04 ±0.04 )by this method and C26: 0( 0. 65 ± 0. 67 ) mg/L, C24:0/C22: 0( 1.19 ± 0. 43 ), C26:0/C22: 0 ( 0. 05 ± 0. 05 )by MDI have no differences( t values were 0. 372,0. 317,0. 945 ,respectively ,all P >0. 05 ). Conclusions The quantitative analysis method for serum very long chain fatty acid using LC-MS/MS is accurate, sensitive,specific and stable. It could provide important biochemistry information for diagnosis in clinic.
9.Research of 24-hour dynamic sleep monitoring and melatonin changes in patients with delirium in intensive care unit
Ting SUN ; Fang HAN ; Yunliang SUN ; Jianghua LIU ; Jiabin YANG ; Xiaozhi WANG
Chinese Critical Care Medicine 2014;(10):726-729
Objective To dynamic monitor and analyze the characteristic of polysomnography (PSG)and melatonin levels of delirium patients in intensive care unit (ICU). Methods A prospective observational study was performed from December 2013 to April 2014. The patients admitted to ICU of Affiliated Hospital of Binzhou Medical College for more than 72 hours were evaluated with confusion assessment method for the ICU (CAM-ICU),and were divided into delirium group and non-delirium group. Sleep patterns of all the patients underwent continuous PSG for up to 24 hours were evaluated. Melatonin levels were determined every 4 hours with enzyme linked immunosorbent assay (ELISA)duration sleep monitoring. Results Eighteen patients were enrolled,and 9 were delirium patients. All the patients had sleep disorders:a decrease in rapid eye movement (REM)sleep 〔(5.91±5.26)%〕,an increase in the sleep fragmentations 〔arousal index was (15.40 ±12.79)times/h〕,and the N3 sleep stage was on the lower limit of normal 〔(14.67 ±11.10)%〕. Compared with non-delirium group,the REM sleep was significantly decreased in delirium group〔(0.10±0.20)%vs.(8.83±3.81)%,t=4.782,P=0.001〕. Melatonin levels lost rhythm between day and night,and there was no difference in melatonin between delirium group and non-delirium group (time effect:F=1.370,P=0.287;between-group effect:F=1.646,P=0.250;interaction effect:F=1.558,P=0.247). The peak of melatonin levels of delirium group appeared on 06:00 〔(137.84±62.21)ng/L〕and 14:00 〔(148.24±58.8)ng/L〕, the minimum value on 22:00 〔(64.47 ±26.97) ng/L〕. But in non-delirium group,the peak of melatonin levels appeared on 02:00 〔(63.52 ±39.75)ng/L〕,the minimum value on 10:00 〔(44.87 ±11.19)ng/L〕. Conclusions ICU patients have sleep disorders,and the delirium patients have less REM stage. Normal rhythmic melatonin secretion changes of ICU patients were lost. The delirium peak of patients appears in the daytime.
10.Rapid Identification of Traditional Chinese Medicine Using Electronic Nose Based on RBF-RF Cascade Classifier
Huiqin ZOU ; Shuo LI ; Yonghong YAN ; Yong LIU ; Ting ZHAO ; Yu HAN ; Yuzhen SU ; Lian PENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(9):1876-1881
This study was aimed to apply the electronic nose (E-nose) in the research of traditional Chinese medicine (TCM). The discussion was made on difficulties of using E-nose. The solution plan was proposed and the discrimination model was established. It provided a simple, rapid and effective analysi method in the identification of TCM. It also provided new ideas for the research and application of gas sensor arrays. E-nose was used in the ex-traction of TCM scent characteristics. Based on ion mobility spectrometry of MOS sensor, the fingerprint of TCM scent was established. The maximum response value of the sensor was used as analysis index. According to the diffi-culties of identification, two solution plans were proposed. Firstly, different detectors were employed to complete the classification. Secondly, radial basis function (RBF) and random forests (RF) were combined and then a cascade classifier was constructed in order to achieve the maximum of information obtained in conditions where the number of measurements, metal oxide semiconductor sensors in E-nose was limited. The results showed that both plans were accurate and practical with relatively high upper correct judge rate and better cross-validation (The highest upper correct judge rates were 95% and 100%, 96% and 80%, respectively). It was concluded that this study firstly ap-plied cascade classifier in the establishment of TCM identification by E-nose. With limited amount of sensors, the maximum information was received through data mining. Using E-nose in the identification of TCM was rapid and accurate. The established pattern recognition method was maneuverable with accurate identification rate and stability compared to conventional sensory identification method. It provided a simple and rapid analysis method for the iden-tification of TCM.