BACKGROUND:A series of studies have demonstrated that ventral mesencephalic dopamine precursors may differentiate into considerable dopaminergic neurons (DNs) available for cell transplantation for Parkinson's disease.However,the detailed molecular characteristics of these cells remain undear.OBJECTIVE:To observe the expression of main marker genes of DNs cultured in vitro.DESIGN,TIME AND SETTING:This observation experiment was performed at the Brain Tumor Laboratory in the Second Affiliated Hospital of Soochow University and Shanghai Biochip Company from December 2005 to December 2008.MATERIALS:Inbred pregnant Wistar rats (F+36) weighing 300 g approximately were used for in vitro culture of mesencephalic cells.METHODS:Ventral mesencephalic cells from inbred pregnant Wistar rats were cultured with basic fibroblast growth factor,and were induced to differentiate in vitro with L-Ascorbic acid-2-phosphate sesquimagnesium salt.Total RNA were extracted before and 4,7 days after the induction.Affymetrix microarrays were utilized to detect the expression changes of main marker genes of DNs.MAIN OUTCOME MEASURES:Morphologic changes of differentiated ventral mesencephalic precursors;Expression of DN-ralated common marker genes before and after the induced differentiation.RESULTS:Rat ventral mesencephalic precursors differentiated into numerous tyrosine hydroxylase-positive cells,which expressed multiple marker genes of DNs,such as AHD2,calcium binding protein 1 (Cab1),DDC,DAT,Drd2,Girk2,VMAT-2,with variously degrees and with different tendencies.Among those genes,only Cab1 was significantly up-regulated and only Girk2 was significantiy down-regulated.CONCLUSION:Rat ventral mesencephalic precursors can be induced to differentiate into plenty of tyrosine hydroxylase-positive DNs,mainly originating in A10 regions.

This article describes the development,components and function of the laboratory automation system (LAS) and laboratory information system(LIS). And it also discusses the construction and application laboratory automation,and the operation of the automated laboratory information system.

AIM: To evaluate the relationship between the medial rectus cells counts in concomitant exotropia and surgical results. METHODS: A total of 32 pieces of medial rectus muscle were collected for HE staining in this study, of which 18 pieces were from patients with concomitant exotropia and 14 pieces were from healthy individuals. A method of strabismus score was used to assess the operative effect.RESULTS: The difference of strabismus score before and after the operation in the intermittent exotropia group was significantly higher than that in constant exotropic group (P<0.01). Under light microscope, the loosen muscle fibers and the increased stromal components in the cross sectional area of medial rectus were observed in strabismic group. The muscle cells counts was obviously lower in strabismic group than in control group (P<0.01), which was related to the difference of strabismus score before and after the operation (P<0.05).CONCLUSION: The decreased medial rectus cells counts induce concomitant exotropia directly. It is the crucial causes of the bad surgical results.

Aim To construct and express the fused gene of anti-glioma single chain Fv(39ScFv)/ human tumor necrosis factor α (TNF-α ). Methods The mature peptide cDNA of TNF-α was fused to 3′ -terminus of ScFv gene. The 39ScFv-TNFα fusion gene was cloned into expression vector pET20b(＋ ) and induced to express in E.coli. Results The fusion protein expressed in E.coli account for 20％ of the total bacterial protein. It displayed a single band of Mr 44 000 by reducing SDS-PAGE and Western blot, and retained both of its anti-glioma immunoreactivity and biological activity of TNF. Conclusion The fusion gene was constructed and expressed successfully in E.coli. The fusion protein possessed bifunctional activities, it may prove useful in targeting immunotherapy against glioma.

To investigate the protective effects of Tangxinping Capsule, a compound traditional Chinese herbal medicine, on diabetic cardiomyopathy in rats.

Objective To discuss the clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope and its surgical techniques. Methods Clinical data of 161 patients with chronic subdural hematoma from August 2009 to December 2015 was analyzed retrospectively. 74 of them experienced surgeries assisted by rigid neuroendoscope (endoscope group) and other 87 cases were operated without neuroendoscope (routine group) during the same period. Results Although there were significant difference in operative duration between the two groups, complications, ratio of total removal of hematoma after surgery, postoperative inpatient duration and recurrent rate of hematoma were more advantageous in endoscope group. The operative duration of endoscope group with (112.68 ± 34.86) min was longer than that of routine group with (74.11 ± 28.23) min (t = 7.75, P = 0.000), while the postoperative inpatient duration of endoscope group with (8.23 ± 2.01) d was shorter than that of another group with (10.79 ± 5.02) d (t = -4.12, P = 0.000). There were no surgical associated complications in endoscope group, but 1 patient in routine group experienced intracerebral hematoma of frontal lobe and associated aphemia. Total removal of hematoma was confirmed in endoscope group with 98.65% (73/74), which was higher than that in routine group with 86.21% (75/78) (χ2 = 8.34, P = 0.004). Hematoma recurrence was found in 16 cases of routine group (18.39%), but more superiority in endoscope group with 1.35% (χ2 = 12.29, P = 0.000). Outpatient follow-up was carried out in all patients from 6 to 38 months with an average duration of 30.06 months. In 17 cases with recurrent hematoma during follow-up, 15 of them were cured by a second surgery, and another 2 patients were cured by atorvastatin. Conclusion As a simple, safe and effective technique, the application of rigid neuroendoscope during surgery for chronic subdural hematoma is more advantage than routine surgery. A self-made suction with adjustable soft curved tip is suitable for such procedure.

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

Objective To investigate whether the malignant transformation of macrophages ( Mφ) in glioma mesenchyme was induced by the fusion of glioma cells ( SU3 ) and Mφ.Methods SU3 cells transfected with red fluorescent protein genes were co-cultured in vitro with Mφexpressing enhanced green fluorescent protein.The cell lineages with RFP+/GFP+dual-color fluorescence were established by using monoclonal selection method.A series of tests for analyzing cancer-related phenotypes, tumorigenicity and specific markers for murine macrophage were performed.Results (1) A few of dual-color fluorescent cells were observed in the co-culture.Three monoclonal cell lineages (C3, C4 and C12) were obtained success-fully.(2) Three types of cells including RFP+, EGFP+and RFP+/EGFP+cells were formed during the cul-ture of monoclonal C12 cell lineage.The percentage of EGFP+cells was increased along the extended culture time and increased passages.Then, EGFP+cells gradually became the predominant cell population.Nota-bly, the percentage of RFP+/EGFP+cells were decreased and maintained at a low level, but the RFP+cells almost disappeared.(3) EGFP+cells from monoclonal C12 cell lineage showed the malignant characteristics such as loss of contact inhibition, rapid proliferation andchromosome aneuploidy, as well as high tumorigenic rate in nude mice (5/5).They also expressed macrophage specific marker CD68 and showed a large number of telocentric chromosomes.Conclusion The results of this study suggested that the malignant transforma-tion of host macrophages as previously observed in solid tumor might be induced by cell fusion occurred be-tween human glioma cells and macrophages.Along with the previous evidences showing the isolation of the malignantly transformed macrophages ( ihCTC) from solid tumor tissue of tumor-bearing mice, the results confirmed an objective existence of malignant transformation of host macrophages in tumor microenvironment. The malignant transformation of host cells induced by fusion with tumor cells revealed not only a new under-standing for the progression of tumor and cancer heterogeneity, but also new targets for cancer therapy.

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics