1.Expression and significance of mucosal β-defensin-2,TNFα and IL-1β in ulcerative colitis CHANG
Chinese Journal of Internal Medicine 2008;47(1):11-14
Objective To investigate the expression and significance of human β-defensin-2 (HBD2),TNFα and IL-1βin ulcerative colitis(UC).Methods Thirty-five patients with active UC diagnosed by the department of gastroenterology in West China Hospital were included in this study.Ulcerative colitis disease activity index(UCAI)was assessed and the pathological grades of UC were classified.Immunohistochemistry assay and real-time quantitative PCR were used for the expression of HBD2,TNFα,IL-1β in colonic mucosa of UC.Results Among the 35 patients with UC,10 cases were mild.13 moderate and 12 severe.Of the 35 cases.there were 11 with grade Ⅰ.13 grade Ⅱ and 11 grade Ⅲ lesion according to Truelove criteria.The score of UCAI had positive correlation with pathological grading (r=0.890,P<0.01).The expressions of HBD2,TNFα,IL-1β in colonic mucosa of UC with immunohistochemistry and real-time quantitative PCR were significantly higher than those in healthy control (P<0.05);the expressions increased gradually with the severity of pathological grade and there was a higher expression of them in inflamed area than in non-inflamed(P<0.05).A good positive correlation was also found between HBD2 and other inflammatory cytokines.Conclusions It is shown that there is a higher expression of HBD2 in colonic mucosa as compared with healthy control.a higher expression of it in inflamed area than in non-inflamed area and a positive correlation of expression between HBD2 and pro-inflammatory cytokines such as TNFα and IL-1 β,implying that HBD2 and pro-inflammatory cytokines are interdependent and interactive playing an important role in magnifying and aggravating inflammatory injury in UC.
2.Anti-migratory and anti-invasive effect of somatostatin receptor type2 gene in human pancreatic carcinoma cell
Yanping FENG ; Jun GAO ; Tao HUANG ; Qin CHANG ; Renyi QIN
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the anti-migratory and anti-invasive effect of somatostatin receptor type 2(SSTR2) gene transfection mediated by adenovirus in human pancreatic carcinoma cell and the mechanisms involved in this effect.Methods The full length human SSTR2 cDNA was introduced into pancreatic cancer cell line BXPC-3 by adenovirus-mediated transfection,and stable expression of RNA and protein of SSTR2 were detected by RT-PCR and Westen-blot.The Matrigel coated Transwell was used to detect the migratory and invasive ability of SSTR2expressing cells,Adv-GFP control cells and mock control cells.Furthermore,the expressions of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) were detected by RT-PCR method in these cells.Results The stable expression of SSTR2 was detected in BXPC-3 cells transfected by Adv-GFP-SSTR2.A dramatic decrease of BXPC-3 expressing SSTR2 cell(migrated) through a Matrigel-coated filter was observed,as compared with Adv-GFP control cells and mock control cells(P
3.HSP12.1, A Small Heat Shock Protein in C.elegans, Has Chaperone-like Activity
Yan QIN ; Hui WANG ; Zengyi CHANG
Progress in Biochemistry and Biophysics 2006;0(06):-
Many kinds of small heat shock proteins (sHSPs) are able to prevent protein aggregation in stress, which show the ATP independent chaperone-like activity. The smallest protein HSP12.1 in sHSP family of the nematode Caenorhabditis elegans exhibits chaperone-like activities in vitro. It prevents protein aggregation in a certain extent when use insulin, ADH and lysozyme as the substrates, though it is not as efficient as the typical chaperones (such as HSP16.1 in C. elegans). By contrast, the other three sHSP12s (HSP12.2, HSP12.3 and HSP12.6), which have similar molecular masses and primary structure, appear devoid of in vitro chaperone-like activities. In addition, overexpressing HSP12.1 enhances cell thermotolerance of Escherichia coli. The survival rate of the HSP12.1 overexpressed cells is 4-fold higher than the control, yet whether it does the same function in C. elegans is still unknown. Results indicate that C-terminal region is not necessary for the chaperone-like activity of sHSPs, for HSP12.1 terminates a short C-terminal tail. N-terminal domain may play a relatively important role in the exhibition of chaperone-like activities, while ?-crystalline domain may also involve in this function.
4.Construction of Thymosin ?4 Shrna Lentivirus and Its Effects on the Expression of Thymosin ?4 in 293T Cells
Sijian HUANG ; Chang LIU ; Zelian QIN
Chinese Journal of Minimally Invasive Surgery 2005;0(07):-
Objective To construct a recombinant lentiviral vector for human thymosin ?4 (TMSB4) and to test the mRNA and protein expression of TMSB4 in 293T cells after being infected by shRNA lentivirus. Methods We designed a specific sequence of small hair RNA targeting TMSB4 gene; the complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was cloned into the pGCSIL-GFP vector to construct a lentiviral vector. The vector was converted into the competent DH5a coli,which confirmed by PCR and sequencing. Then the viral vector and the packed systemic vector were cotransfected 293T cells to get the lentivirus. The virus titer was determined. Afterwards,in the 293T cells infected with the lentivirus,the expression of TMSB4 was detected by real time PCR and immunocytochemistry. Similarly,a primary fibroblast was also infected with the lentivirus. Results Compared with negative control cells,the mRNA and protein levels of TMSB4 in 293T cells infected with the lentivirus were reduced by 44% (0.56?0.11 vs 1.00?0.06,F=89.673,P
5.An analysis of misdiagnosed Crohn's disease in China
Yuying CHANG ; Qin OUYANG ; Renwei HU
Chinese Journal of Digestive Endoscopy 1996;0(06):-
Objective In order to improve the diagnostic level of Crohn's disease( CD),the relevant literatures with CD in China were reviewed and the causes of misdiagnosis about CD were analyzed. Methods Papers on CD published in medical journals from 1989 to 2003 in CMCC and VIP are collected and reviewed one by one, and then the characteristics of the misdiagnosed and missed CD were recorded and the causes of these cases were analyzed. Results From 1989 to 2003, there are 428 papers about CD, of which 261 papers are about clinical studies(60.9%); In these papers, total of 3397 CD cases were reported. In these cases there are 2357 cases missed, clinical symptoms are various and complex including abdomen pain (62.5%),diarrhea(51.5%), abdomenal lumps(16.6%) and so on. The most commonly involved sites of CD are terminal ileum (62.5%), colon (26.9%) and ileum combining with colon (38.9%). Cranulomas were identified in 61.2% of operative specimens and in 27.6% of biopsy specimens. The most common missed diagnosed diseases are intestinal tuberculosis(32. 2% ), simplex intestinal block (23.2%) and intestinal block caused by tumor(6.4%) and acute appenditis(27. 9% ). There are 389 cases misdiagnosed as CD, the most common diseases are tuberculosis(53.5%), small intestinal lymphoma(14.4%) and colonal cancer (17.0%). Conclusion Papers on CD are increased gradually during the periods from 1989 to 2003 with rather high missed or misdiagnosis. As a result, it is necessary for us to improve the cognitzability on CD and to improve the accuracy of diagnosis.
6.Effect of 23 G minimally invasive vitrectomy without irrigation in cataract ultrasonic phacoemulsification and trabeculectomy of phakic malignant glaucoma
Zhan-Feng, WANG ; Chang-Qin, XU
International Eye Science 2016;16(10):1879-1882
AIM: To investigate the efficacy of 23G minimally vitrectomy without irrigation in cataract phacoemulsification and trabeculectomy of malignant glaucoma, and to analyze such compound operative procedures for phakic malignant glaucoma.
●METHODS:A total of 21 phakic malignant glaucoma patients (21 eyes) underwent anterior vitrectomy without irrigation by using 23G vitrectomy. During surgical course phacoemulsification with anterior and posterior continuous circular capsulorhexis, trabeculectomy combined with iridectomy would be completed. lntraocular pressure, anterior chamber depth changes and postoperative complications were observed after the operation.
●RESULTS:ln the three-month follow-up, intraocular pressures were reduced from ( 57. 18 ± 6. 18 ) mmHg to (16. 15 ± 2. 43 ) mmHg, there was statistical difference compared with pre - operation ( P < 0. 001 ). The preoperative anterior chamber depth (ACD) was (0. 88± 0. 25) mm, the postoperative ACD was (2. 44±0. 37) mm 3mo later, there were significant difference (P<0. 001). The best corrected visual acuity improved significantly, no serious postoperative complication appeared.
● CONCLUSION: The compound surgical method of anterior vitrectomy combined with phacoemulsifier and trabeculectomy can effectively treat phakic malignant glaucoma. Early diagnosis and early compound surgery may effectively reduce the intraocular pressure of malignant glaucoma.
7.Influence of Insoine on Expressions of Vascular Endothelial Growth Factor and Cyclooxygenase-2 in Newborn Rat with Hypoxic-Ischemic Encephalopathy
bin-chang, GAO ; qin, LI ; hong, LUAN
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To observe the expressions of vascular endothelial growth factor(VEGF) and cyclooxygenase-2(COX-2) in the cerebral tissue following hypoxic-ischemic encephalopathy(HIE) in newborn rats and explore the machanism of inosine in protecting against hypoxic-ischemic brain damage(HIBD).Methods Sixty-six newborn rats aged 7 days were divided into sham,control and experimental groups.HIE models were made by clamping the right cervical artery and making hypoxia for 2 hours.The rats in experimental group began injecting inosine,at 1 day before experiment,and the rats in the sham and control groups saline solution with same dose.The samples were made at the given time,and expressions of VEGF and COX-2 were investigated by immunohistochemical technique.Results The cerebral tissue had no expression of VEGF and COX-2 in sham group.From 2 hours on cortex and striatum after HIE in control and experimental groups,expressions of VEGF and COX-2 increased rapidly,peaking at 12-24 hours,and then decreased gradually.Expressions of VEGF and COX-2 were higher in experimental group(All P
8. Antibacterial mechanism of cryptotanshinone
Chinese Pharmaceutical Journal 2012;47(21):1706-1710
OBJECTIVE: To study the antimicrobial activity and antibacterial mechanisms of cryptotanshinone against SA, MRSA and ESBLS-SA. METHODS: MIC value was determined by disc diffusion method. IC50 value was determined by liquid culture. The antibacterial mechanisms of cryptotanshinone were investigated by determining the changes in electric conductivity, concentration of AKP, protein content, and the changes of protein electrophoretic bands in SDS-PAGE. RESULTS: The antibacterial rings, MIC and IC50 values showed that the antimicrobial activity against SA was better than that against MRSA and ESBLs-SA. Treated with cryptotanshinone, the electric conductivity, concentration of AKP, and protein content were increased, and protein bands in SDS-PAGE were changed obviously. CONCLUSION: Cryptotanshinone significantly inhibits SA, MRSA and ESBLs-SA and can damage the structures of cell wall and cell membrane, which results in the increase of permeability of cell membrane and release of cell components. Cryptotanshinone can influence the synthesis of bacteria protein, destroy the protein or reject the anabolism or expression of the protein, and finally leads to the loss of normal physiological function of bacterium.
9.Determination of furbenicillin sodium and its related substances by HPLC.
Yan CHANG ; Ye TIAN ; Yue MA ; Chang-qin HU
Acta Pharmaceutica Sinica 2015;50(12):1632-1636
Furbenicillin is a broad-spectrum semisynthetic penicillin with strong antibacterial activity against Gram-negative bacteria. Furbenicillin sodium is determined by volumetric method in current criteria. However, the criteria does not contain an assay of related substances of furbenicillin sodium. In this study, we established a method for detection and analysis of furbenicillin sodium and its related substances by HPLC. The analysis was performed with a C18 column under a gradient elution, the detection wavelength was 225 nm, and the column temperature was 35 degrees C. The reliability and accuracy of established method was validated in this study. Pure samples of furbenicillin sodium and its related substances were prepared. The structures, biological activities, and chromatographic retention behaviors of furbenicillin sodium and its related substances were identified using NMR, CLSI agar dilution method, and HPLC. All results in the current study provide ample evidence that this method is able to determine the reasonable limits in the quality-control protocol for furbenicillin sodium.
Anti-Bacterial Agents
;
chemistry
;
Chromatography, High Pressure Liquid
;
Magnetic Resonance Spectroscopy
;
Penicillins
;
Quality Control
;
Reproducibility of Results