1. Expression of MMP-9 in brain tissue of mouse cryptococcal meningitis model
Academic Journal of Second Military Medical University 2010;29(7):778-780
Objective: To explore the expression of MMP-9 in mouse brain tissue and its significance. Methods: Cryptococcosis ssuspension alone (fungal suspension group) or combined with aprotinin (aprotinin group) was injected in to female C57 BL/6 mice via femoral vein cannulation. The expression of MMP-9 was examined immunohistochemically 8 h later. Meanwhile, the mouse brain tissue homogenate colony count (CFU) was observed and the results were compared with that of the immunohistochemical examination. Results: The expression of MMP-9 was markedly increased in the fungal suspension group compared with the normal control group (P<0.01); the expression in the aprotinin group was not significantly different from that of the normal control group (P>0.05). The amount of cryptococcosis in the brain tissue was positively correlated with expression of MMP-9. Conclusion: MMP-9 is overexpressed in the brain of mice with Cryptococcal meningitis; MMP-9 might lead to increased permeability of blood brain barrier.
2. Activity detection of extracellular proteinase and serine protease secreted by Cryptococcus neo formans
Academic Journal of Second Military Medical University 2006;27(2):125-128
Objective: To evaluate the activities of extracellular proteinase and serine protease secreted by Cryptococcus neoformans (C. neoformans) of different sources and serotypes and incubated under different conditions. Methods: Thirty-six C. neoformans strains of different sources and serotypes were incubated in protein agar medium at 30°C and 37°C separately. Activities of extracellular proteinase and serine protease were analyzed by determining Clearance Halos(CHs) produced by C. neoformans. Activity changes of the 2 proteases were analyzed after treated with serine protease inhibitors. The activity and the molecular weight of extracellular proteinase in the concentrated supernatant were measured using zymography assay. Results: Thirty-three of 36 C. neoformans strains produced specific Clearance Halos around the colonies, and their mean CHs at 30°C and 37°C were 0.558±0.170 and 0.575±0.169, respectively (P>0.05). The mean CHs of serotype A(n=13), B(n=13), and D/AD (n=6) strains were 0.564±0.144, 0.515±0.078 and 0.482±0.072, respectively (P>0.05); the mean CHs of clinical(n=23), environmental(n=9),and uncapsuled strains(n=4) were 0.570±0.177, 0.513±0.069, and 0.942±0.075, respectively (P<0.05). The mean CHs of control group (without protease inhibitor) and groups treated with protease inhibitor (1.2 mU and 1.6 mU) were 0.459±0.188, 0.975±0.287 and 0.733±0.252, respectively(P<0.01). Rich extracellular protease was found in the concentrated supernatant. Conclusion: The incubation temperature (30°C and 37°C) has no effect on activities of extracellular proteinase and serine protease, and the activities of clinical strains are significantly stronger than those of other 2 strains; the activities in different serotypes groups have no significant difference and can be inhibited by serine protease inhibitors.
3. Cell microarray in detection of serine protease expression in Cryptococcus neo formans
Academic Journal of Second Military Medical University 2006;27(2):129-131
Objective: To determine the serine protease expression in Cryptococcus neoformans (C. neoformans) by cell microarray technique,so as to investigate the role of serine protease in the pathogenesis of C. neoformans. Methods: The cell microarray was constructed with tissue microarray. Thirty-six strains of C. neoformans of different sources and homologous serotypes were examined for their serine protease expression by cell microarray technique and immunohistochemistry. Results: Strong expressi on of serine protease was found in 25(67.0%) strains. The rates of strong serine protease expression in serotype A, B and D/AD strains were 46.2%(6/13), 92.3%(12/13) and 66.7%(4/6), and in environment-isolated, clinically isolated and uncapsuled strains were 55.6%(5/9),82.6%(19/23) and 25%(1/4), respectively. Serine protease expressions in serotype B and clinically-isolated strains were significantly higher than that in other group (P<0.05). Conclusion: Mic roarray of strain cells is a new method for identifying pathogenic fungus. Higher expression of serine protease in clinically-isolated strains is associated with strong virulence of clinical isolates strains, suggesting that serine protease plays a major role in the pathogenesis of C. neoformans.
4.Research progress in HIV auxiliary proteins counteracting host restriction factors.
Qian-Qian CHEN ; Qing-Gang XU ; Chi-Yu ZHANG
Chinese Journal of Virology 2014;30(1):84-90
Identification and functional analyses of antiviral restriction factors in hosts have become hot research topics. Four HIV restriction factors, APOBEC3G, Trim5alpha, Tetherin, and SAMHD1, have been identified in recent years. By encoding auxiliary proteins, lentiviruses can counteract host restriction factors. For example, the auxiliary proteins Vif, Vpu, and Vpx of HIV antagonize APOBEC3G, Tetherin, and SAMHD1, respectively. Furthermore, these auxiliary proteins enable the entry of HIV into host cells and influence the replication and pathogenicity of HIV. In this paper, we review the research progress in the functions of the three HIV auxiliary proteins that can antagonize the host restriction factors.
Animals
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HIV
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metabolism
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physiology
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Host-Pathogen Interactions
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Humans
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Viral Proteins
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metabolism
5.Effect of kidney-supplementing and blood-activating method on expression of Malb and RBP in senile patients with isolated systolic hypertension
Yelin SONG ; Weijun XU ; Guangyu YU ; Lingbo WEI ; Weifeng CHI ; Yingying NIE
International Journal of Traditional Chinese Medicine 2013;35(4):313-315
Objective Kidney-supplementing and blood-activating method was adopted in treating senile patients with isolated systolic hypertension to observe its decompression effects and influences on microalbunminuria (Malb),retinol binding protein (RBP) level in 24 hours.Methods 90 patients with simple systolic hypertension were randomly recurited into two groups.52 cases in the treatment group were administered with kidney-supplementing and blood-activating decoction,including 1 case falling off and 51 cases entering statistical analysis; 38 cases in the control group were administered with oral placebo,among them 2 cases were fallen offand 36 cases were entered statistical analysis.Both groups were treated for 8 weeks.Results () Blood pressure:systolic blood pressure at 4 and 8 weeks after the treatment in the treatment group [(144.03±12.33)mmHg (1 mmHg=0.133kPa) and (132.27±13.15)mmHg] wassignificantlyimproved than before the treatment [(156.32±12.05)mm Hg] (P<0.05),and also significantly better than the control group at 4,8 weeks after the treatment [(151.19± 13.83)mm Hg,(152.74± 12.03)mm Hg] (P<0.05).②The Malb,RBP level:Malb,RBP level [(40.80±13.51)mg/L,(150.43±23.62)mg/L] after the treatment in the treatment group was reduced than before the treatment [(50.14± 15.61)mg/L,(220.04±30.20) mg/L] (P<0.05),and was significantly different to the control group after treatment [(52.12±14.69)mg/L,(219.34±34.37)mg/L] (P<0.05).Conclusion Kidney-supplementing and blood-activating method can improve kidney function,and thus to reduce the effect of systolic blood pressure.
6.Clinical and pathological analysis of primary biliary cirrhosis-autoimmune hepatitis overlap syndrome
Chi-Hong WU ; Xiao-Yuan XU ; Hai-Ying LU ; Yan-Yan YU ; Gui-Qiang WANG ;
Journal of Peking University(Health Sciences) 2003;0(06):-
Objective:To explore clinical and histopathological characteristics of primary biliary cirrho-sis-autoimmune hepatitis overlap syndrome.Methods:Clinical data and pathological findings of 10 pa-tients were reviewed.Results:Serum glutamine transpeptidase,alkaline phosphatase levels,alaninetransaminase,aspartate transaminase,serum IgG and IgM were elevated in all the patients.They were allpositive for anti-mitochondrial antibody and AMA-M2.Nine patients were positive for anti-nuclear anti-body and one patient was positive for anti liver-kidney microsome antibody.Liver biopsies in these pa-tients revealed:ten patients had bile duct lesion,hepatitis activities ranged from moderate to severe,andfibrosis ranged from S1 to S3.Conclusion:PBC-AIH overlap syndrome is mostly found in middle-agedwomen.It has the clinical and histopathological characteristics of both PBC and AIH.Accurate andprompt diagnosis of overlap syndrome patients should be based on the clinical presentation,biochemicaland immune indexes,and hepalic pathological changes.
7.Experimental Establishment of Life Cycle of Clonorchis sinensis
Chi LIANG ; Xuchu HU ; Zhiyue LV ; Zhongdao WU ; Xinbing YU ; Jin XU ; Huanqin ZHENG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To establish and maintain the life cycle of Clonorchis sinensis in laboratory.Methods Adult worms and eggs of Clonorchis sinensis were collected from naturally infected cats.Eggs were ingested by freshwater snails in aquarium.When the cercariae were released from infected snails, they invaded into freshwater fishes.From the 30th day on after the release of cercariae, the infection rate and metacercariae density in freshwater fishes were determined.Results After 95 days the infected snails began shedding cercariae in a temperature range of 24.3-37.2 ℃, and no cercariae were found under 20 ℃.The infection rate in the snails Parafossarulus striatulus and Alocinma longicornis was 12.5% and 18.0%, respectively.Metacercariae were found in fish at 30 days after cercariae infection, and matured metacercariae were detected in 45 days.The number of metacercariae per gram of fish meat in Pseudorasbora parva, Ctenopharyngodon idellus, Rhodeus sinensis, Hypophthalmichthys nobilis, Cirrhinus molitorella, Carassius auratus, Cyprinus carpio and Oreochromis niloticus was 1 792, 16, 8, 6, 5, 4, 4, and 2, respectively.Rats and cats were fed with metacercariae from fish to receive adult worms.Conclusion Life cycle of Clonorchis sinensis has been established and maintained in the laboratory.
8.Molecular mechanism of familial hypertriglyceridemia caused by lipoprotein lipase gene mutation (C310R/E396V)
Yu LUN ; Xiaofang SUN ; Ping WANG ; Jingwei CHI ; Xu HOU ; Yangang WANG
Chinese Journal of Endocrinology and Metabolism 2017;33(8):656-661
Objective The purpose of this study is to investigate the molecular mechanisms of p.C310R(c.T928C) and p.E396V(c.A1187T) lipoprotein lipase(LPL) gene mutations in vitro, which may help to construct the spectrum of LPL gene mutations and phenotype. It also can provide accurate early diagnosis for high-risk population of familial hypertriglyceridemia and provide the basis for the development of gene targeted therapy. Methods Genomic DNA was extracted from proband′s family members′ peripheral blood cells and screened by whole-exome sequencing to verify candidate gene variations. PCR products were afterwards directly sequenced again to confirm corresponding LPL variants. At the cellular level, lentiviruses containing LPL mutations were constructed and then transfected into COS-1 cells. Functional significance of the mutants was corroborated by analyzing LPL activity and mass in the cell medium and lysates via ELISA and enzyme-fluorescent method. mRNA was assayed by RT-PCR to confirm the effect on gene transcription. Results DNA sequence analysis revealed that the proband was a heterozygote for a novel c.T928C mutation in exon 6 of LPL gene, while his nephew was a compound heterozygote for the c.T928C mutation in exon 6 and a novel c.A1187T mutation in exon 8. In vitro studies, these two mutations can cause decreased activity and mass of extracellular LPL(P<0.05). Moreover, further investigation indicated that LPL C310R mutation tremendously affected post-transcriptional modification of LPL gene, whereas LPL E396V mutation dampened intracellular LPL trafficking. Conclusion Both the mutations are pathogenic by reducing the activity and mass of LPL in the plasma, which affected normal metabolism of triglycerides.
9.Analysis of diagnosis and treatment of adrenal lymphangioma
Dianbin SONG ; Jingjing ZHANG ; Man YU ; Qiang CHI ; Hui XU ; Yan MAO ; Zhiyong WANG
Chinese Journal of Urology 2021;42(2):144-146
By summarizing and analyzing the clinical diagnosis and treatment experience of 17 cases of adrenal lymphangioma, the imaging characteristics and pathological types of the disease were discussed. The results showed that the imaging of adrenal lymphangioma was non-specific, and the appearance was similar to that of general cysts.Howerer, the density of the cyst was slightly higher than that of simple cysts. Some cases showed calcification on the cyst wall, and a few showed adenoma-like appearance. The diagnosis mainly depends on pathological examination. For those patients with tumors ≥4.0 cm, endocrine function, suspected malignancy, or obvious clinical symptoms, surgery is recommended.
10.Intravascular ultrasound in stent implantation for coronary artery disease
Zhuhua NI ; Xinchun YANG ; Lefeng WANG ; Yonggui GE ; Hongshi WANG ; Kun XIA ; Weiming LI ; Li XU ; Yonghui CHI ; Yu LIU
Chinese Journal of Tissue Engineering Research 2008;12(30):5979-5984
BACKGROUND: Stent under-expansion and procedurally related abnormal lesion morphologies (e.g. dissection, thrombus) are associated with stent restenosis and acute, subacute and chronic thrombosis.OBJECTIVE: To explore whether larger post-procedural final minimum stent area can be acquired and more procedurally related complications can be identified in stent implantation guided by intravascular ultrasound.DESIGN, TIME AND SETTING: Retrospective analysis was performed at the Heart Center of Beijing Chaoyang Hospital, Capital Medical University between January 2004 and February 2005.PARTICIPANTS: Fifty patients with coronary artery disease with 52 lesions were enrolled in the study and underwent stenting guided by intravascular ultrasound. The patients were characterized as non-diffused lesion with vessel diameter ≥ 2.5 mm. Patients with severe left main lesion were excluded.METHODS: Qualitative and quantitative analyses were carried out in 50 patients with 52 lesions before and after stent implantation. The stent diameter and the end-point of therapy were determined by intravascular ultrasound standard.MAIN OUTCOME MEASURES: The differences of end point for stent implantation and the enlargement of lumen area gained by stent implantation were compared between cardioangiography and by intravascular ultrasound.RESULTS: The average stent diameter guided by intravascular ultrasound was larger than by cardioangiography (P=0.011); the peak balloon pressure was higher in intravascular ultrasound group than cardioangiography group (P < 0.001), and area stenosis percentage measured by quantitative coronary angiogram was smaller in intravascular ultrasound group than cardioangiography group (P=0.044). ②Cardioangiography showed success rate was 96.2% and intravascular ultrasound showed the success rate was only 37.7% after first balloon high-pressure dilation. Intravascular ultrasound subgroup analysis showed higher peak balloon pressure (P < 0.001), larger lumen diameter (P < 0.001), larger lumen area (P < 0.001), and smaller area stenosis percentage (P < 0.001). No obvious stenosis was found at the proximal and distal segments of the stent observed by cardioangiography, while atherosclerotic lesions at proximal segment were found in 39 cases (75.0%) and at distal segment were in 23 cases (44.2%) observed by intravascular ultrasound. The lumen area was larger in non-fatty plaque than in fatty plaque after stent implantation (P < 0.001). Compared with non-fatty plaque, the enlargement of vessel area was 1.30 mm2 smaller, while plaque compression was 0.48 mm2 larger. CONCLUSION: Stent implantation guided by intravascular ultrasound can acquire larger final lumen area and identify more procedurally related complications.