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.

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.

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.

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 ; HIV ; metabolism ; physiology ; Host-Pathogen Interactions ; Humans ; Viral Proteins ; metabolism

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.

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.

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.

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.

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.

Objective To investigate the ultrasonic characteristics of angiographical normal left main (LM) branch of coronary artery observing with intravaseular ultrasound(IVUS). Methods Seventy-six patients whose coronary angiogram showed the lesions restricted only in left anterior descending (LAD) branch or left cireumflex(LCX) branch and no lesion was found in LM branch were enrolled and IVUS was performed. The plaque burden was measured and the quality of atherosclerosis was identified in lesion site of LAD or LCX by IVUS. Meanwhile,the absence or existence of lesions in LM was identified,and the quality of lesions was analyzed if it showing those existed lesions. The diameter and area of lumen in left main were measured and diameter and area of vessel were also measured. The plaque burden were measured for those who atheroselerosis existed in LM. Results IVUS showed 28 cases completely normal, 12 cases with intimal membrance hyperplesia,36 cases with plaque and 2 cases with intimal membrance flap in patients which LM was angiographically normal. Among those there were 30 eccentric plaques and 6 concentric plaques. For 36 patients whose lesions existed in LM observed by IVUS,there were 25 cases (69.4%) with soft plaque,4 eases (11.1%) with fibrous plaque,2 cases (5.6%) with calcific plaque,5 cases (13.9%)with mixed plaque. IVUS showed lumen diameter was (5.32±0.68)mm and lumen area was (23.34±5.27)mm2 for female patients; and lumen diameter was (5.90±0.50)mm and lumen area was (27.75±4.47)mm2 for male patients. The difference had significane when comparing lumen diameter and lumen area between male and female patients (P=0.042 and P=0.048, respectively). Vessel diameter was (5.90±0.47)mm and vessel area was (27.58±4.21)mm2 in patients with intimal membrance hyperplesia; lumen diameter was (4.39±0.54)mm and lumen area was (17.45±5.23)mm2,vessel diameter was (5.99±0.67)mm and vessel area was(26.61±6.27)mm2 n patients with atherosclerotic plaque.Diameter stenosis percentage was(26.17±7.87)%and plaque burden was(34.79±9.37)%in LM.Conclusions IVUS can find those lesions in LM which CAG cannot detect and identify the quality and severity of lesion precisely.