By means of cell hybridization in situ with digoxigenin labeled probe, we detected the expression of C-myc protooncogene in peripheral blood mononuclear cells from 31 SLE patients and 10 healthy persons. The results showed that the level of C-myc mRNA in SLE patients was significantly higher than that in normal controls, and the increased level correlated with disease activity. There was also a positive correlation between C-myc mRNA and serum levels of ANA, anti-dsDNA, IgG, C3. Our study indicates that C-myc gene is important for the pathogenesis of SLE.

Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Methyl Ethers ; analysis ; Workplace

Objective To discuss the clinical significance of the diffuse calcified distribution in diagnosis of benign and malignant breast lesions .Methods 379 patients with different benign and malignant breast lesions confirmed by surgery underwent digital X‐ray mammography .The morphology ,distribution ,number ,diameter ,concentration and density of calcification in lesions ,the maximum range of the calcified area and other accompanied manifestations in benign and malignant breast lesions were analyzed .Results As for the morphology of calcification ,tiny polymorphic calcification was found in 58 .5% of malignant lesions ,meanwhile ,dot‐like one was found in 49 .3% of benign lesions .Fine linear calcification or branched linear one occurred in malignant lesions ,however ,round one occurred in benign lesions .As for the calcification distribution ,regional distribution was found in 43 .9% of malignant lesions , meanwhile the clustered distribution was found in 58 .4% of benign lesions .And all lobar or segmental distribution was seen in malig‐nant lesions .As for the calcification diameter ,the calcification with the diameter less than 0 .5 mm occurred in 71 .6% of malignant lesions ,however ,that with diameter more than 1 .0 cm occurred in 69 .3% of benign lesions .As for the concentration of calcification , lesions with calcification more than 25 were 71 .8% of malignant ones ,whereas those with calcification of 15-25 were 58 .4% of be‐nign ones .As for calcified density ,uneven calcification occurred in 94 .2% of malignant lesions ;however the even one occurred in 63 .4% of benign lesions .The maximum diameter of calcification ranged from 40 mm to 80 mm was found in 59 .0% of malignant lesions , whereas that ranged from 0 mm to 40 mm was in 77 .2% of benign lesions .Conclusion Statistical differences have been found in the morphology ,distribution ,diameter ,concentration ,density and maximum diameter of calcification between the breast benign and ma‐lignant lesions .

AIM: To study the effect of double gene transduction mediated by lentiviral vectors on the characteristics of human embryonic stem cells (hESCs). METHODS: Using the backbone from inducibal dual and excisable transgene vector (iDuet101) , lentiviral vectors overexpressing cytotoxic T lymphocyte - associated molecule - 4 immuno-globulin (iDuet101 - CTLA4Ig) , indoleamine 2, 3 dioxygenase (iDuet101 - IDO) , and ubiquitin - C promoter - lueifer-ase - ires - puromycin ( ULIP) were constructed and packaged according to the standard recombinant techniques. Human embryonic stem cells (hESCs) were first transduced with iDuet101, iDuet101 - CTLA4Ig or iDuet101 - IDO, then, after the selection, were transduced again with ULIP. The expression and function of the exogenous genes were detected. Immu-nohistochemistry, RT - PCR and flow cytometry were applied for detection of embryoid bodies ( EB) formation in vitro and in vivo teratoma formation. RESULTS: Double - transduced hESCs showed typical shape of cell clones and positive staining of tumor rejection antigen -1 - 60 ( Tra -1 - 60 ) and octomer transcription factor - 4 ( OCT - 4 ). The formation of EB was observed, in which a - fetoprotein (AFP), paired box gene 6 ( Pax6) and Musashi 1 ( MSI1) were positively expressed. The cells formed teratomas, and the luciferase signals existed until 28 days after xeno - transplantation. CONCLUSION : Double transduction of non - transcriptional factors mediated by lentiviral vectors does not affect the cell growth rate and their differentiation ability.

Objective To explore the method and clinical effect of laparoscopic anatomical right liver resections. Method The candidates for laparoscopic right hepatic lobectomys were 4 cases including 3 cases of liver hemangioma and 1 case of hepatorrhexis. Results The laparoscopic right hepatic lobectomy we performed saccess bully in all the 4 patients, operation time was (470±42.7)min. The blood loss in operation was ( 1950± 881.3) ml. The postoperative hospital stay was ( 15 ± 2.9) days.There was not complcation. Conclusions Laparoscopic right hepatic libectomy is feasible and safe.For the patients with benign liver disease, it is an operation with less operation wound.

Objective To upgrade the current X-ray vehicle.Methods Modern digital X-ray radiography was analyzed and 3 kinds of X-ray detectors were compared.Canon CXDI digital X-ray detector was used to update field X-ray vehicle into a field digital X-ray radiography vehicle.Results Real-time X-ray signal synchronization was realized.Digital images could be edited and labeled with detection information.Conclusion The working efficiency is enhanced and the support ability is improved.

Objective To analyze the characteristics of erectile dysfunction(ED)in aged men.Methods Erectile dysfunction(ED)was diagnosed according to the International Index of Erectile Function(IIEF).78 aged men(average 65.9 yrs)as the study group,and 82 young patients(average 36.0 yrs)as the control group,all with ED,Were compared in complicating diseases,self-rating SDS scores,penile-brachial indexes(PBI),the time of achieving erection,and ejaculatory latency time.Results The main complicating diseases in study group were cardiovascular diseases(42 cases,53.9%),lower urinary tract symptoms(LUTS)(31 cases,39.7%)and diabetes(26 cases,33.3%).In the control group,the main complicating diseases were chronic prostatitis(52 cages,63.4%),premature ejaculation(PE)(32 cases,39.0%)and depression(12 csses,14.6%).SDS scores of study group and control group were(29.13±5.63)and(39.59±13.31),PBI were(0.78±0.12)and(0.91±0.06),the time of achieving erection were(13.85±5.75)min and(3.61±4.29)min,ejaculatory latency time were(7.03±5.35)min and(3.81±5.53)min.All with significant difference(P＜0.01).Conclusion Most of the ED old men were complicated with organic diseases,such as the time of achieving erection,PBI low scores and longer ejaculatory latency time.

Objective To investigate whether baicalein inhibits the proliferation, cell cycle of and pseudopod formation in A431, a skin squamous cell carcinoma cell line, by suppressing the expression of ezrin protein. Methods A431 cells were grouped to be transfected with ezrin-targeting siRNA (siRNA group), treated with baicalein of 5, 10, 20, 40 μmol/L, respectively (baicalein group), or remain untreated (control group). After additional culture, wound healing assay and Transwell assay were performed to observe the migration and invasion of A431 cells, RT-PCR to detect the mRNA expression of ezrin in A431 cells, Western blot and immunoflu-orescence to measure the expression of ezrin protein and its phosphorylation. The pseudopod formation in A431 cells was observed by using scanning electron microscopy. Results After 24-hour culture, wound healing assay displayed that the percent wound closure was 13.3 ± 1.7, 7.6 ±1.6 and 5.9 ± 1.3, respectively, in A431 cells treated with baicalein of 5, 10, 20μmol/L, significantly lower than that in untreated A431 cells (16.3 ± 2.3, all P < 0.01), and the inhibition of baicalein on the migration of A431 cells was concentration-dependent. In the Transwell assay, a significant decrease was observed in the number of A431 cells per high power field permeating through the artificial basement membrane in the groups treated with baicalein of 5, 10, 20 μmol/L for 48 hours compared with the control group (46.5 ± 3.8, 34.3 ± 3.4, 25.3 ± 2.3 vs 56.3 ± 3.8, all P < 0.01), whereas no significant difference was noted between these baicalein-treated groups and siRNA-transfected group (28.3 ± 2.1, all P > 0.05). RT-PCR analysis showed that the mRNA expression of ezrin in baicalein-treated A431 cells significantly decreased compared with that in untreated cells (all P< 0.01), but showed no difference from that in siRNA group (P > 0.05). A statistical difference was also observed in the expression of ezrin and phosphorylated ezrin protein between baicalein-treated A431 cells and untreated cells (all P< 0.05), but not between 40 μmol/L baicalein-treated A431 cells and siRNA-transfected cells (P> 0.05). Furthermore, the suppression of baicalein on ezrin protein and mRNA expression was concentration dependent. The number of pseudopod per cell was significantly lower in 20 μmol/L baicalein-treated A431 cells and siRNA-transfected cells than that in untreated A431 cells (5.3 ± 1.9, 4.5 ± 2.8 vs 22.6 ± 2.8, both P < 0.01), while no significant difference was observed between the former two groups of cells (P > 0.05); the length of pseudopodia also reduced in baicalein-treated cells. Conclusions Baicalein may inhibit the proliferation and invasion of A431 cells by directly or indirectly suppressing the expression of ezrin and phosphorylated ezrin, which in turn contributes to the effect of baicalein against tumor proliferation and metastasis.

AIM:To investigate NF-?B p65 activation and I?B-? expression in keloid fibroblasts(KFB)and normal skin fibroblasts(NSF)stimulated with TNF-? and to explore the underlying molecular pathogenesis of keloid formation.METHODS:Primary KFB was cultured.The location of NF-?B p65 and I?B-? in KFB and NSF at quiescent condition and the nuclear translocation of NF-?B p65 after TNF-? stimulation were observed by immunofluorescence technique.NF-?B p65 DNA binding activity was detected with TransAMTM NF-?B p65 kit.The I?B-? protein level was determined by means of Western blotting technique.RESULTS:After stimulated with TNF-?,NF-?B p65 translocated into the nucleus.NF-?B p65 DNA binding activity increased to its maximum at 1 h and was dropped to normal at 4 h.TNF-? induced most degradation of I?B-? at 15 min and became detectable in cytoplasm after 4 h.KFB showed more sensitive ability to TNF-? stimulation than NSF.CONCLUSION:NF-?B may play a role in keloid pathogenesis.

AIM:To explore a method of isolation,purification,culture and identification of human microglia.METHODS:The brain tissue from abortive fetus was sterilely obtained,then chopped and triturated gently.The suspension was filtered and centrifuged to separate and isolate mixed glia cells.The cell density was adjusted to 2?1010 cells/L with DMEM/F12 complete medium and cultured in 75 cm2 flask at 37 ℃ in CO2 incubator(marked as first 0 d).After 7-10 days culture,floating cells including microglia and oligodendrocytes appeared in the flask.The first time purification was carried out to obtain highly purified microglia,for which the flasks were shaken gently.The floating cells were collected and transferred into a new flask(marked as second 0 d).4-5 days later,when microglia and oligodendrocytes grew in confluent state,the second purification was performed,for which the mixed cultured cells were digested with trypsin-EDTA and cell suspension was centrifuged,then the cell pellet was suspended by DMEM/F12 complete medium and cultured in flask(marked as 0 h).After 2 h,non-microglial cells including oligodendrocytes and cell debris were removed,and fresh medium was added into the adherent cells.In this way,the whole procedure of microglia purification was completed and the highly purified microglial cells were obtained.After purification,the expression rate of CD45,CD11b and CD68 on/in microglia were detected by laser-confocal microscopy and flow cytometry on the basis of immuno-fluorescence staining to identify the purity of microglial cells.Microglial phagocytotic function was evaluated by phagocytosis of fluorescent microspheres.The level of activation was identified by the phenomena of membrane ruffling in combination with fluorescent phalloidin staining.RESULTS:More than 98% of the microglias that we isolated and purified expressed CD45,CD11b and CD68.Almost all of the microglias could phagocytize fluorescent microbeads CONCLUSION:We succeed in isolating and purifying human microglias,which express CD45,CD11b and CD68.Almost all of the cells exhibit phagocytic function.So these cells will be useful for further functional and proteomic studies.