AlM: To investigate expressions of matrix metalloproteinase-2 ( MMP-2 ) and extracellular matrix metalloproteinase inducer ( EMMPRlN) in retinoblastoma (Rb) and the relationships between MMP-2, EMMPRlN and tumor development.METHODS:lmmunohistochemical technique was used to detect expressions of MMP-2 and EMMPRlN in 39 cases of paraffin embedded Rb samples. Quantitative analysis of expressions of MMP-2 and EMMPRlN were assessed by measuring the mean gray scale of Rb tissue with LElCA lM50 Color Pathologic Analysis System. The differences of expressions of MMP-2 and EMMPRlN in each clinical and pathological stage were statistically analyzed, and the same step was also undertaken to study the relationship between Rb with MMP-2 positive expression and that with EMMPRlN positive expression.RESULTS:The positive expression rate of MMP-2 was 90% (Gray value: 109. 64 ± 14. 52; 35/39), and that of EMMPRlN was 85% (Gray value:108. 01±13. 60;33/39). The expressions of MMP - 2 and EMMPRlN were significantly higher in tumors of glaucomatous stage (Gray value:108. 21±11. 47 and 107. 56±14. 32) than those in intraocular stage ( Gray value: 121. 13 ± 11. 32 and 119. 34 ± 12. 66; P<0. 01 and P<0. 05). And the same conclusion can be concluded between those in extraocular stage (Gray value: 91. 03 ± 11. 71 and 92. 26 ± 12. 93) with those in glaucomatous stage (P<0. 01 and P<0. 05). The expressions of MMP-2 and EMMPRlN were significantly higher in tumors with optic nerve invasion (Gray value:103. 89±13. 39 and 105. 23±14. 00) than those without optic nerve invasion ( Gray value: 118. 39 ± 15. 11 and 117. 53±16. 13) (P<0. 01 and P<0. 05).CONCLUSlON:The positive expression levels of MMP-2 and EMMPRlN may correlate with tumor infiltration and metastasis.

Objective To study the effect of Cinepazide Maleate Injection on blood rheology of acute cerebral infarction patients. Methods 152 patients with cerebral infarction which attacked in 72 h after onset were separated into experimental group (n=88) and control group (n=64). 320 mg Cinepazide Maleate Injection was given to the experimental group and 20 mL Danshen Injection was given to control group, both by intravenous drip, once a day, for 14 d. The clinical efficacy and the changes of blood rheology were observed and compared in the 2 groups. Results The results of Chinese Stroke Scale (CSS) in the experimental group were significantly better than those in the control group (P<0.05), and the decrease in the parameters of blood rheology was much more obvious in the experimental group than in the control group (P<0.05) 2 weeks later. The excellent rate was 62.5% in the experimental group, which was higher than 35.9% in the control group significantly (P<0.05). Conclusion The Cinepazide Maleate Injection is effective in the treatment of acute cerebral infarction and can reduce the blood viscosity.

Objective To compare the diagnostic value of high frequency ultrasound,mammography and breast blood-oxygen function imaging system in diagnosis of breast masses.Methods The images of 89 breast masses by high frequency ultrasound,mammography and breast blood-oxygen function imaging system were reviewed.The study compared the sensibilities and specificities of the three methods in the masses and analyzed their cause of missed diagnosis.Results The sensitivity of the diagnosis of breast cancer by high frequency ultrasound,mammography and breast blood-oxygen function imaging system were 80.00％,70.00％ and 50.00％,respectively.The specificity of the diagnosis of breast cancer by them were 94.94％,98.73％ and 97.47％,respectively.There were sigmfiacant differences in diagnosis of breast benign lesion among the high frequency ultrasound,mammography and breast blood-oxygen function imaging system.The sensitivity of high frequency ultrasound in detecting fibroadenoma of breast was 98.11％ higher than the other methods.Mammography was insensitive to the benign breast lesions,but its specificity was high.Conclusions High frequency ultrasound,mammography and breast blood-oxygen function imaging system are the highly effective diagnostic tool for breast cancer,but the diagnostic value of mammography and breast blood-oxygen in breast fibroadenoma and in cystic disease of breast aren' t high.Breast blood-oxygen function imaging system can enhance the accuracy of the diagnosis of mammary inflammation.

Objective To re-evaluate the immunogenicity of meningococcal serogroups A and C polysaccharide vaccine among a healthy population of age 5 to 59.Methods Pre and post-vaccination ser-um samples were collected from the subjects involved in a randomized , controlled clinical trial conducted in 2005 at Hechi, Guangxi, China.Enzyme-linked immunosorbent assay (ELISA) was performed for the quan-titative detection of specific anti-capsule IgG antibody against meningococcal serogroups A and C in serum samples.Serum bactericidal assay ( SBA) was used to measure the bactericidal antibody activity against se-rougroups A and C bacterial strains .Results Geometric mean concentrations (GMCs) of specific anti-cap-sule IgG antibody were 23.66 μg/ml and 78.83 μg/ml for serogroup A (P<0.05), and 1.23 μg/ml and 51.25 μg/ml for serogroup C (P<0.05) in serum samples of pre and post-vaccination, respectively.Be-fore immunization , 99% of serum samples ( 97/98 ) showed serogroup A-specific IgG concentration ≥2μg/ml, which reached up to 100%(98/98) after vaccination (P>0.05).The percentage of serum sam-ples with serogroup C-specific IgG concentration ≥2 μg/ml rose from 20%to 99% after vaccination ( P<0.05).The ratio of positive serum samples with at least four times of increases in concentration of specific IgG against serogroup A and serogroup C after vaccine immunization were 34% and 100%, respectively . The rSBA geometric mean titers ( GMTs) for serogroups A and C in serum sample of post-vaccination were respectively elevated from 1164 to 5530 (P<0.05), and from 4 to 6225 (P<0.05).The percentage of se-rum samples with rSBA GMTs≥128 increased from 91% to 99% for serogroup A (P>0.05), and from 14%to 96%for serogroup C (P<0.05) after vaccination.The rSBA GMTs with at least four times of in-creases after vaccination were detected respectively in 53% and 100% of serogroups A and C vaccinated subjects.Pearson correlation coefficient between IgG concentration and rSBA GMTs was r=0.15 (pre) and r=0.23 (post) for serogroup A, and r=-0.14 (pre) and r=0.58 (post) for serogroup C (P<0.05). Conclusion This study has demonstrated an efficient and sustained immunogenicity with meningococcal sero -groups A and C polysaccharide vaccine as evidenced by the data from standardized assays of ELISA and SBA .

OBJECTIVETo study the biologic process of energy metabolism in brain during acupuncture using positron emission tomography (PET) with 18F-2-desoxyglucose (18FDG) for further elucidation of the relationship between acupunctural signal and nerve system.

METHODSElectroacupuncture (EA) was applied on right lateral of a healthy volunteer and paralytic limbs of 4 patients with cerebral infarction at acupoints L14, LI11, ST36 and SP6 using Hans acupoint-nerve stimulator. PET imaging was conducted on the healthy subject or patients with the same posture before and during EA with GE Advance II PET system.

RESULTSPET showed that in the healthy subject, before EA, the glucose metabolism (GM) in bilateral cerebral cortex, bilateral thalamus, basal nuclei and cerebellum was almost symmetrical, but during EA, the GM in contralateral thalamus, contralateral frontal lobe and parietal lobe (motor and sensory area) increased obviously. While in the patients before EA, the GM in the infarcted area was significantly lower than that in the non-infarcted area, as compared with that observed with CT and MRI, it showed a similar figure but with bigger abnormal area. During EA, GM in the infarcted area increased with apparent reduction of size. Increased GM of focal area, widened cerebral cortex and decreased edematous area were shown in patients with larger infarction area. Quantitative analysis revealed evident change in local/total ratio of glucose and increase of GM change rate.

CONCLUSION(1) EA on limb acupoints of healthy subject could induce obvious increase of regional GM in brain and contralateral thalamus, contralateral frontal lobe and parietal lobe (motor and sensory area). (2) EA on acupoints of paralytic limbs could cause increase of GM in contralateral thalamus, contralateral frontal lobe and parietal lobe. Besides, GM also increased in the area with lowered GM before EA, accompanied with shrinkage or disappearance of lesion. (3) Acupuncture could evoke the function of brain cells and raise the GM in them.

Aged ; Brain ; diagnostic imaging ; metabolism ; Cerebral Infarction ; diagnostic imaging ; metabolism ; Electroacupuncture ; Energy Metabolism ; Female ; Fluorodeoxyglucose F18 ; Glucose ; metabolism ; Humans ; Male ; Middle Aged ; Radiopharmaceuticals ; Random Allocation ; Tomography, Emission-Computed

OBJECTIVETo screen the stage-specific expression proteins during rats spermatogenesis, and to investigate the beta-actin expression and localization in the tissues of rat testicular.

METHODSHighly enriched type A spermatogonia, pachytene spermatocytes and round spermatids were isolated by STAPUT method (sedimentation velocity at unit gravity, with 2% - 4% BSA gradient in DMEM/F12 medium) respectively to get the total proteins. The difference of protein expression between the three kinds of cells was analyzed by two-dimensional electrophoresis. Then the distribution of beta-actin in rat testicular tissues was investigated using specific anti-beta-actin antibodies by immunohistochemical method.

RESULTSbeta-actin was identified as a stage-specific expression protein by two-dimensional electrophoresis. beta-actin protein was more strongly expressed in type A spermatogonia and pachytene spermatocytes, but not in round spermatids. The immunohistochemical results showed that beta-actin was mainly located in the cytoplasm of type A spermatogonia and pachytene spermatocytes and in the nuclei of nearly mature spermatids.

CONCLUSIONbeta-actin protein is a stage-specific expressed protein and may play an important role in spermatogenesis.

Actins ; biosynthesis ; Animals ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; physiology ; Testis ; cytology ; metabolism

BACKGROUNDGlioblastoma is the most common and lethal cancer of the central nervous system. Global genomic hypomethylation and some CpG island hypermethylation are common hallmarks of these malignancies, but the effects of these methylation abnormalities on glioblastomas are still largely unclear. Methylation of the O6-methylguanine-DNA methyltransferase promoter is currently an only confirmed molecular predictor of better outcome in temozolomide treatment. To better understand the relationship between CpG island methylation status and patient outcome, this study launched DNA methylation profiles for thirty-three primary glioblastomas (pGBMs) and nine secondary glioblastomas (sGBMs) with the expectation to identify valuable prognostic and therapeutic targets.

METHODSWe evaluated the methylation status of testis derived transcript (TES) gene promoter by microarray analysis of glioblastomas and the prognostic value for TES methylation in the clinical outcome of pGBM patients. Significance analysis of microarrays was used for genes significantly differently methylated between 33 pGBM and nine sGBM. Survival curves were calculated according to the Kaplan-Meier method, and differences between curves were assessed using the log-rank test. Then, we treated glioblastoma cell lines (U87 and U251) with 5-aza-2-deoxycytidines (5-aza-dC) and detected cell biological behaviors.

RESULTSMicroarray data analysis identified TES promoter was hypermethylated in pGBMs compared with sGBMs (P < 0.05). Survival curves from the Kaplan-Meier method analysis revealed that the patients with TES hypermethylation had a short overall survival (P < 0.05). This abnormality is also confirmed in glioblastoma cell lines (U87 and U251). Treating these cells with 5-aza-dC released TES protein expression resulted in significant inhibition of cell growth (P = 0.013).

CONCLUSIONSHypermethylation of TES gene promoter highly correlated with worse outcome in pGBM patients. TES might represent a valuable prognostic marker for glioblastoma.

Azacitidine ; analogs & derivatives ; pharmacology ; Brain Neoplasms ; drug therapy ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytoskeletal Proteins ; genetics ; DNA Methylation ; Glioblastoma ; drug therapy ; genetics ; pathology ; Humans ; LIM Domain Proteins ; genetics ; Promoter Regions, Genetic ; Treatment Outcome

To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells (BMSCs) at defined stages, subtractive cDNA library was established by means of suppression subtractive hybridization. The BMSCs cultured for 12 and 21 d were used as driver and tester, respectively. A subtract library was successfully constructed and five positive clones were selected from the library. Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank. It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1. RT-PCR was performed to confirm the differentially expressed genes. The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day, while the expression of decorin was only detected in the cells of 21-day.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cell Differentiation ; genetics ; Cells, Cultured ; Decorin ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Library ; Humans ; Membrane Proteins ; genetics ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology

An about 700 bp DNA fragment was amplified from genome DNA of S. aureus TSTw by PCR. This fragment was cloned into pGEM-7Zf(+) and the recombinant plasmid was transformed into E. coli DH5 alpha. The sequencing result of the recombinant plasmid demonstrated that it contains seb gene with 717 bp (without signal encoding region of 81 bp) which has the same nucleotide sequence as described in literature. The seb gene was cloned into expression vector 7ZTS and was transformed into E. coli JM109 (DE3). The expression level of SEB was as high as 33.3% of the cell total proteins.
Cloning, Molecular ; Enterotoxins ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Engineering ; Recombinant Proteins ; biosynthesis

The rare codons of a fragment in staphylococcal enterotoxin A gene were turned into the most high usage frequency codons in E. coli by overlap PCR technique. Genes of sea and seam were cloned into 7ZTS expression vector and transformed into JM109(DE3), respectively. The result shows that expression level of sea gene was very low, but the expression level of seam was as high as 15% of total cell proteins. The expression product shows activity of antitumor in vivo.
Animals ; Base Sequence ; Codon ; genetics ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; Gene Expression Regulation, Bacterial ; Male ; Mice ; Molecular Sequence Data ; Neoplasms, Experimental ; drug therapy ; pathology ; Point Mutation ; Recombinant Proteins ; genetics ; metabolism ; pharmacology