Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy / 国际脑血管病杂志

Cerebral small vessel disease is a group of heterogeneous diseases with stroke and cognitive impairment as the main clinical features.It can be divided into sporadic type and hereditary type.Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) belongs to hereditary cerebral small vessel disease.It has been reported in China,Japan,Spain,Greece,and other countries.The diagnosis mainly depends on characteristic clinical symptoms,imaging features,and genetic testing.CARASIL manifests as diffuse white matter abnormal signal and subcortical multiple infarcts on MRI,and it caused by HTRA1 gene mutation.

Corin-serine protease and clinical applications in cardiovascular disease / 中华检验医学杂志

Corin is a newly identified membrance protease which responsible for converting pro-B-type natriuretic peptide to biologically active B-type natriuretic peptide.B-type natriuretic peptide play an important role in regulating blood pressure and cardiac function.It is a biological marker of heart failure.So Corin plays a role in reducing blood volume, blood pressure, regulating body fluid balance, and improving cardiac function.Corin may be used as a biomarker for heart failure or other cardiovascular disease.Recent studies have found that Corin is associated with the occurrence and development of cardiovascular diseases, such as heart failure and acute coronary syndrome.The article mainly expound the biological characteristics of Corin, testing methods and its clinical application in cardiovascular disease status.

Expression of PACE4 in non-small cell lung cancer and analysis of its clinicopathologic characteristics / 中国医师杂志

Objective To investigate the expression of paired amino acid cleaving enzyme 4 (PACE4) protein in non-small-cell lung cancer (NSCLC), analyze the correlation of its clinicopathologic characteristics, and investigate its significance in the process of occurrence, development, and metastasis of NSCLC.Methods Between January 2006 and May 2009, the First Affiliated Hospital of Guangzhou Medical College, 172 patients with NSCLC and 15 patients with non-neoplastic tissues were chosen.Immunohistochemistry was used to detect the expression of PACE4, and clinicopathologic characteristics of NSCLC were analyzed.Results (1)The positive expression rate of PACE4 protein in NSCLC was significantly higher than that in non-tumor tissues (P ＜ 0.05).PACE4 expression was observed in 111 of 172 (64.5％) NSCLC.PACE4 had cytoplasmic expression.(2)Clinicopathologically, PACE4 expression was significantly associated with lymph node metastasis (N stage) (P =0.007), and clinical stage (P =0.024).Conclusions High expression of PACE4 indicates that PACE4 may be involved in the processes of occurrence,development, and metastasis of NSCLC.

The relationship between PCSK9 gene I474V polymorphism and ischemic stroke of Uygur and Han ethnic groups in Xinjiang Uygur Autonomous Region / 中华神经科杂志

Objective To investigate the frequency distribution of proprotein convertase subtilisin/kexin 9 (PCSK9) gene I474V polymorphisms and their relationship with patients with ischemic stroke (IS)of Uygur and Han ethnic groups in Xinjiang Uygur Autonomous Region.Methods The I474V polymorphism was identified by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) in 407 patients with IS(including 219 Hans and 188 Uygurs)and 425 health controls (including 255 Hans and 170 Uygurs),and some specimens were sequenced.Results (1) Between IS group and control group,the genotypes Ⅱ and Ⅳ had no statistically significant differences in the levels of triglycerides (TG),high-density lipoprotein cholesterol (HDL-C) ; Total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C) levels had statistically significant differences; LDL-C levels had also statistically significant differences.Between IS and control groups,TC,LDL,HDL-C levels of genotype Ⅱ showed statistically significant difference.In the IS group,TC,LDL-C levels of Ⅳ genotype were significantly higher than the control group,the difference being statistically significant.(2) There was statistically significant difference in the genotype distribution between IS and control groups (9.5％ (77/814) vs 4.5％ (38/850),x2 =16.09,P =0.000).And the distribution of allele frequency was statistically different (18.9％ (77/407) vs 8.9％ (38/425),x2 =17.38,P =0.000).(3) The differences of I474V loci Ⅳ genotype frequency distribution in Xinjiang Uygurs and Hans were statistically significant (27.7％ (52/188) vs 11.4％ (25/219),x2 =17.40,P =0.000; 12.9％ (22/170) vs 6.3％ (16/255),x2 =5.57,P =0.018) ; So did the Ⅴ allele frenquency distribution (13.8％ (52/376) vs 5.7％ (25/438),x2 =15.58,P =0.000; 6.5％ (22/340) vs 3.1％ (16/510),x2 =10.44,P =0.001).(4) There was statistically significant difference in the genotype distribution and allele frenquency distribution between IS group and control group in the Xinjiang Uygurs (27.7％ (52/188) vs 12.9％ (22/170),x2 =11.79,P =0.001 ; 13.8％ (52/376) vs 6.5％ (22/340),x2 =10.44,P =0.001) ; But there was no statistically significant difference in the Hans.Conclusions Ⅱ and Ⅳ genotypes are dominant in the I474V polymorphism loci of PCSK9 gene.The genotype of PCSK9 gene I474V polymorphism is correlated with increasing serum levels of TC and LDL-C.I474V polymorphism is associated with cerebral IS course in Xinjiang region.There is statistically significant difference in the genotype I474V distribution between Uygur and Han groups.I474V polymorphism has a relationship with the occurrence of IS in Xinjiang Uygurs.Ⅳ may be a susceptible genotype and Ⅴ may be a genetic susceptible allele of the Xinjiang Uygurs.

Comparative study of two proteases for mass spectrometric method for the determination HbA1c / 中华检验医学杂志

Objective To explore the difference of two kinds of proteolytic enzymes (GLU-C and Lys-C trypsin),hemoglobin A1c were measured in mass spectrometry.Methods Based on the IFCC recommended hemoglobin A1c reference measurement method by mass spectrometry,the blood samples were preparied (the number of these blood samples:201201,201202,201203).Hemoglobin in the samples were enzymed respectively by two kinds of proteolytic enzymes (Glu-C and Lys-C),before the mass spectra which be used solid phase extraction method,under the optimal experimental conditions,hemoglobin A1c in the sample were measured by the liquid chromatography tandem mass spectrometry and screened of variant hemoglobin with sequest software.All results by mass spectrometry are required to use the Sequest software to search RAW file in the human IPI Database,retrieve four common hemoglobin variant published results and compare hemoglobin variant sites,using statistical software SPSS13.0 and Excel 2003 on the results in the correlation analysis,compare between groups by the independent sample t test.Results In the detection of hemoglobin A1c,respectively Glu-C and Lys-C blood which the samples were enzymed,results of hemoglobin A1c by Lys-C were (34.70 ± 2.80),(51.76 ± 1.60),(73.39 ± 1.11) mmol/mol,results of hemoglobin A1c by Glu-C were(33.12 ± 1.48),(54.54 ± 2.50),(75.40 ± 3.60) mmol/mol,the difference between groups was not statistically significant,the results of two methods of drawing curve,all R2 ＞ 0.995.Searching the human database with sequest software,four common hemoglobin variant were not be finded.Conclusions In the detection of hemoglobin A1c by the mass spectrometry,the application of the above two kinds of proteolytic enzyme in the blood samples,that can be more consistent results,test results of two methods have a good correlation.

Expression and significance of matriptase in ovarian cancer cells with diverse metastatic potential / 中华妇产科杂志

Objective To study the expression and significance of matriptase in different metastatic potential of human ovarian cancer cells.Methods High-metastatic human ovarian cancer cell HO8910PM and ovarian cancer cell HO8910 were collected.The ability of metastatic of the former was stronger than that of the latter.Compared the ability of invasion and migration in HO8910PM and HO8910 by scratch assay and by millicell chamber artificial reconstituted basement membrane invasion assay.Detected the matriptase mRNA and protein expression levels in HO8910PM and HO8910 through reverse transcription(RT)-PCR and immunocytochemistry methods.Results The 24 hours' migration distance(347 ± 8) μm of HO8910PM cells were significantly higher than that in HO8910 group (154 ± 10) μm (P ＜ 0.01) ;The number of HO8910PM cells that penetrated the matrigel after 24 hours' incubation were significantly higher than that in HO8910 group (90.7 ±2.1 vs 63.3 ± 1.5,P ＜0.01).The expression of matriptase mRNA in HO8910PM cells was higher than that in HO8910 group (0.72 ± 0.03 vs 0.38 ± 0.04,P ＜ 0.01).The migration was positively correlated with the matriptase mRNA expression levels (r =0.992,P ＜ 0.01); and the invasiveness was also positively correlated with the matriptase mRNA expression levels (r =0.973,P ＜0.01).As far protein level,the expression of matriptase protein in HO8910PM cells was higher than that in HO8910 group (15.6 ±0.8 vs 7.6 ± 1.3,P ＜0.01).The migration was positively correlated with matriptase protein expression levels (r =0.971,P ＜ 0.01) ;And the invasiveness was also positively correlated with the matriptase protein expression levels (r =0.958,P ＜ 0.01).Conclusions The relationship between the expression levels of matriptase and the metastatic of ovarian cancer cells may be correlative.The function of matriptase in ovarian cancer cells metastatic machanism still need to be confirmed.

Cerebral autosomal recessive arteriopathy with subcortical infarets and leukoencephalopathy / 国际脑血管病杂志

Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is an autosomal recessive hereditary cerebral artery disease in the adolescence,and its main clinical manifestations are dementia,stroke,low back pain and alopecia.At present,most cases are from Japan.3-6 exon point mutations in the HTRA1 gene are associated with the onset of CARASIL.Brain histopathological examination showed a small arterial intimal thickening medial smooth muscle cell loss and hyalinization.Brain MRI showed a diffuse white matter abnormal signal and multiple subcortical infarcts.The diagnosis mainly depends on the characteristic clinical symptoms,imaging characteristics and genetic testing.It should be differentiated from cerebral autosomal dominant arteriopathy with sulcortical infarcts and leucoencephalopathy.

Novel mutation of HTRA1 gene causes cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy: one case / 中华神经科杂志

ObjectiveTo report a novel HTRA1 gene mutation caused cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) with migraine,urinary retention and constipation.MethodsThe patient was a 34-year-old woman who suffered from myalgia with cramp for 16 years,lumbago for 5 years,migraine and mild alopecia for 3 years,right hemiparesis for 5 months,and urinary retention and constipation for 1 month.Vertebral MRI showed degeneration of vertebral bodies with disc herniations.Brain MRI revealed diffuse white matter lesions and lacunar infarcts.Sural nerve and skin biopsies were performed in the patient. HTRA1 gene analysis was performed in patient,her parents and 2 brothers,and Notch3 gene analysis in the patient.ResultsThe sural biopsy demonstrated discontinuous of elastica internal with thickness of intima of arterioles.The capillary basement membranes were thickness with mini granular changes.A homozygous T to A transition at position 524( c.524T ＞ A) was found in HTRA1 gene.The heterozygous c.524T ＞ A mutation appeared in the parents and 2 brothers,but not in the controls.Notch3 mutations were not found in the patient. Conclusion CARASIL caused by novel homozygous c.524T ＞ A mutation of HTRA1 gene can present with migraine,urinary retention and constipation.

Expression and significance of matriptase and HAI-1 in prostate cancer / 中国医师杂志

Objective To investigate the expression and significance of Matriptase and HAI-1 protein in prostate cancer (CaP). Methods Specimens of 46 prostate cancers,20 benign prostate hyperplasias (BPH),10 high-grade intraepithelial neoplasias (PIN),and 10 normal prostates (NP) were used. Expressions of Matriptase and HAI-1 proteins in specimens were detected by SP of immunohistochemistry. The results were analyzed in relation to the clinicopathological data. Results The protein levels of Matriptase in CaP tissues were significantly higher than PIN tissues(Z＝-2.150,P＝0.032),and the expression of matriptase in CaP and PIN was higher than that in BPH and NP (Z＝-3.270,P＝0.001;Z＝-2.817,P＝0.005). No statistically significant difference was observed between BPH and NP group (Z＝-0.895,P＝0.325). A progressive increase in the protein levels of Matriptase was observed with increasing tumor grade (rs＝0.583,P＜0.01) and clinical stages（rs＝0.611,P＜0.01）in CaP specimens. The protein levels of HAI-1 in BPH and NP tissues were significantly higher than CaP and PIN tissues（Z＝-3.277,-3.315,P＜0.01）,the levels of HAI-1 in PIN were higher than CaP (Z＝-2.310,P＝0.020). No statistically significant difference was found between BPH and NP (Z＝-0.872,P＝0.330). A progressive decrease in the protein levels of HAI-1 was observed with increasing tumor grades（rs＝-0.634,P＜0.01） and clinical stages（rs＝-0.521,P＜0.01）. The expressions of Matriptase and HAI-1 in CaP tissues showed negative correlations（rs＝-0.712,-0.560,-0.465,respectively,P＜0.01）. Conclusions The abnormal expressions of Matriptase and HAI-1 proteins may be important events during the progression of CaP in humans. Matriptase and HAI-1 Protein may be used as parameters for assessing the malignancy and prognosis of CaP.

Effects of PR3 and protease-activated receptor-2-activators on the maturation and functions of dendritic cell-like monocytes / 中华风湿病学杂志

Objective To assess the effects of protease 3(PR3)and protease-activated receptor (PAR)-2-activator on the maturation and functions of peripheral blood dendritic cell(DC)-like monocytes.Methods Density gradient centrifugation was used to isolate peripheral blood mononuclear cells(PBMC)from Wegener's granulomatosis(WG)patients and healthy controls(HC).PBMC were stimulated by LPS,human PR3,trypsin,PAR-2-agonist peptide (PAR-2-AP),LPS+PR3 or LPS+trypsin for 24 h.Flow cytometry was used to analyze the expression of PAR-2,CD80,CD83,HLA-DR on stimulated DC-like monocytes-CD14+CD16high monocytes.ELISA kit was used to test the concentration of IL-6 in the culture supernants.Mann-Whitney non-parameteric test was used fur statistical analysis.Resuits No effect of PR3,trypsin and PAR-2-AP on the expression of PAR-2,CD80,CD83,HLA-DR of DC-like monoeytes was found.LPS could significantly induce PAR-2 expression in HC[from(5.8±1.5)%to(24.5±4.5)%,P=0.002]and the expression of CD80,CD83,HLA-DR in HC and WG;PR3,trypsin,PAR-2-AP and LPS could all stimulate the secretion of IL-6.Conclusion PR3 and PAR-2 pathway-activators can not promote PAR-2expression and maturation of DC-tike monocytes,but they can induce the secretion of IL-6.

The relationship between TMPRSS2-Ets fusion gene and prostate cancer / 国际肿瘤学杂志

A unique fusion between the TMPRSS2 gene and the Ets genes ERG,ETV1、ETV4 or ETV5 has been described in prostate cancer, TMPRSS2-ERG is the most high frequency of all. Until now more than 20 kinds of TMPRSS2- ERG hybrid transcripts are found, and they encode 9 kinds of proteins. Deletion is the main mechanism of fusion. Fusion gene can be used to make a diagnosis of prostate cancer , direct clinical medication , then fusion type, fusion number and transcriptive type are also associated with prognosis of dis-ease. So, besides of serum PSA, Gleason score, TMPRSS2- ERG is expected to be another important predicator of prognosis.

The value of the ratio of transmembrane protease serine 2 and ETS-related gene fusion transcripts in the diagnosis of prostate cancer / 中华检验医学杂志

n be used as a potential marker in the diagnosis of PCa. However, it can not be used as an index to monitor tumor progression or prognosis in PCa patients.

Expression and value of Matriptase and HAI-1 in bladder transitional cell carcinoma / 国际肿瘤学杂志

Objective To investigate the expression and clinical significance of Matriptase and HAI-1 in bladder transitional cell carcinoma( BTCC), and to explore their relationship with the clinicopathologieal pa-rameters of BTCC. Methods Expressions of Matriptase and Hal-1 protein in specimens of BTCC and normal bladder tissues were detected by SP method of immunohistochemistry. The results were analyzed in relation to the clinicopathological data. Results The expression of Matriptase and HA1-1 were observed in both normal and tumor tissues of bladder;The protein levels of Matriptase and HAI-1 were significantly higher in normal bladder tissues as compared with BTCC tissues ( P < 0. 05 ) ; a progressive decrease in the protein levels of Hal-1 was observed with increasing of tumor grades (r = -0. 634,P < 0. 001) and clinical stages (r=-0. 521,P < 0. 001 ) ; the protein level of Matriptase was decreased with tumor grade increasing, and was significantly higher in high-grade tissues as compared with moderate and low-grade tissues( P < 0. 05 ) ; Matriptase protein levels showed no significantly difference between different tumor clinical stages ( P > O. 05 ). The correlation co- efficient of Matriptase and HAI-1 in normal tissues was 0. 772(P <0. 001 )and 0. 546 in tumor tissues of blad- der( P < 0. 001 ). Conclusion The abnormal and decreased expressions of Matriptase and HAI- 1 may be im- portant events during the genesis and progression of BTCC in humans; Hal-1 may be used as a new parameterfor assessing the malignancy and prognosis of BTCC; Matriptase protein in BTCC is related with differentiationpromotion, and may be an indicator of differentiation. The correlation between Matriptase and Hal- 1 decreasesin BTCC. The imbalance between HAI-1 and Matriptase may be contributed to the progression of BTCC.

The homotrimeric structure of HtrA2 is indispensable for executing its serine protease activity

Serine protease activity of high temperature requrement 2 (HtrA2) is essential for promoting cell death, as well as for protecting against cellular stresses. An X-ray crystallographic study described the formation of a pyramid shaped homotrimer that is a proteolytically competent form of HtrA2; however, little is known about effects of the trimeric structure of HtrA2 on the natural substrates. In this study, we generated the HtrA2 protein that has a single point mutation at the homotrimerization motif to assess relationship between structure and the proteolytic activity of HtrA2 on its substrates. Using gel filtration, a native gel electrophoresis system, and a co-precipitation assay, we confirm that phenylalanine 149 in HtrA2 is a crucial determinant for the formation of the HtrA2 homotrimeric structure. Moreover, we described that the HtrA2 monomeric form abolished not only autoproteolytic activity, but also the proteolytic activity against XIAP (X-linked inhibitor of apoptosis protein) known as the HtrA2 substrate. Taken together, the results indicate that the homotrimeric structure of HtrA2 is required for executing its serine protease activity.

Cloning of Mouse Enamel Matrix Serine Proteinase Encoding Mature Protein / 吉林大学学报(医学版)

Objective: To clone cDNA of enamel matrix serine proteinase (EMSP1) encoding mature protein from mouse dental germs. Methods: Total RNA was isolated from developing incisors and molars of 7 days mouse pups and reverse-transcribed into cDNA. Two pairs of specific primers was designed to obtain the desired gene by Touchdown PCR and Nested PCR. The segment was inserted into Vector pMD-18T, and recombined vectors was transformed into E.coli JM109.The positive clone was chose and analysed by restriction endonuclease mapping and DNA sequencing. Results:700 bp of cDNA of mouse EMSP1 was sueccessfully cloned from mouse tooth germs tissue. The sequence was consistent with that displayed in PubMed. Conclusion:The mouse EMSP1 cDNA encoding mature protein is obtained for further study.

Effect of Astilbin on expressions of perforin and granzyme B in activated T cells of mouse heart transplantation model with acute rejection / 中国医师杂志

Objective To investigate the effect of astilbin on expressions of perforin and granzyme B in activated T cells of mouse heart transplantation model with acute rejection.Methods Cardiomyocytes of BALB/C mouse and spleen cells of C57BL/6 mouse were harvested and made into single cell suspensions.The cardiomyocytes(2?10~5 ml~(-1))as stimulators and spleen cells(1?10~6 ml~(-1)) as responsers were mixed and cultured.The model of mouse heart transplantation with acute rejection in vitro was therefore established.There were two groups in the experiment.Control group is the mixed culture of the cardiomyocytes and spleen cells;Astilbin group is the mixed culture of the cardiomyocytes and spleen cells with astilbin(15?g/ml).Apoptosis of T cells were analyzed by TUNEL assay.The expressions of perforin and granzyme B were measured by RT-PCR.Results Apoptosis of activated T cells in Astilbin group was significantly increased than that of the control group(P

Effect of perforin and granzyme B in peripheral blood lymphocytes during acute rejection in renal transplantation / 中国医师杂志

Objective To investigate the effect of perforin and granzyme B in peripheral blood lymphocytes during acute rejection in renal transplantation.Methods Sixty-seven recipients of renal allograft were involved in the study.The recipients were divided into four groups: group 1 with 7 cases of acute rejection,group 2 with 8 cases of delayed graft function,group 3 with 27 cases of stable function and group 4 wih 25 cases of long-term survival.The expressions of perforin and granzyme B of peripheral blood lymphocytes were studied by quantitative reverse transcription polymerase chain reaction(RT-PCR).Results The expressions of perforin and granzyme B in acute rejection group were significantly higher than those of the other three groups(P

Construction of serine protease gene mutant and study on pathogenicity of serine protease gene of Entero-coccus faecalis / 中华传染病杂志

Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.

Expression of the novel serine protease SNC19 in different kinds of cells lines and its effects on cellular biological behaviors / 中国病理生理杂志

AIM: To investigate the physiological function of the novel serine protease SNC19 protein and its possible role in cancer invasion and metastasis. METHODS: Monoclonal antibodies directly against SNC19 extracellular domain was prepared. The protein and SNC19 mRNA expression were determined in different kinds of cell lines respectively by Western blot and Northern blot analysis. Cellular migration and adhesion abilities were assayed by monoclonal antibody blocking method. RESULTS: Western blot analysis showed there were two bands of SNC19 protein in BCAP37, COLO205, SW480 cells at about 120 kD and 60 kD while only one band in SW620 cells at 60 kD; Northern blots showed a approximate 3.4-kilobase fragment appearing in most epithelial-derived cell lines with this only form and high levels but no detection was obtained in OV, TCA8113, KB and SGC7901 cells. In antibody blocking experiments, the migration of SW480 cells was significantly inhibited compared with the control and the abilities of (SW480/SW480), SW480/NIH3T3 adhesion increased at the beginning of the experiments, but the difference reduced (along) with the time passed.CONCLUSION: SNC19 protein is closely related with cellular homogeneous and heterogeneous adhesion as well as cellular motility. As a novel serine protease, it may participate both in physiological and pathological processes, such as cell migration, tissue remodeling and cancer invasion and metastasis.

Expression of two primary target antigens (PR3 and MPO) of serum antinutrophil cytoplasmic antibodies in severe preeclampsia women / 中华围产医学杂志

0. 05). No significant difference was found in maternal and neonatal complications between the ANCA( + ) and ANCA(－) subgroups in S-PE subjects. But those 4 cases who developed renal function insufficiency patients were ANCA( + ). Conclusions ANCA might be associated with renal diseases in preeclampsia women, and further studies is required to determine whether ANCA is involved with the pathogenesis of preeclampsia.