BACKGROUND:Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Tol-like receptor 1 in which wil provide possible experimental basis for the future potential of celltherapy strategies.
OBJECTIVE:To investigate the role of Tol-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells.
METHODS:Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4+T cells. CD4+T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Tol-like receptor 1 expression levels related genes;number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4+T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Tol-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry.
RESULTS AND CONCLUSION:Tol-like receptor 1 mRNA was expressed in both CD4+T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4+T cells than in CD4+T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4+T cells compared with CD4+T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P<0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4+T cells was significantly higher than that in CD4+T cells cultured alone, (4.53±1.27)%vs. (2.39±0.80)%(P<0.01). Tol-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells.

Objective]To explore the essence of doubts differentiation from‘Neiwaishangbianhuolun’,the monumental work of the medical practitioner Dongyuan Li in Jin and Yuan Dynasties, and relativity between internal damage and exogenous diseases.[Methods] By means of researching the background of‘Neiwaishangbianhuolun’, analyzing the 13 main differentiations, think about the essence of doubts differentiation; meanwhile, by connecting to the explicate of mechanism of ascending yangqi belonging to exterior-releasing medicine(wind-expel ing drugs) in his representative work‘Piweilun’, discuss the contrary and transforming relationships between exogenous wind-cold and internal damage of the stomach and spleen, comparing to academic thoughts of Zhongjing Zhang’s‘Shanghanlun’. [Results]Realize essence of doubts differentiation from ‘Neiwaishangbianhuolun’ is not exterior or interior of disease location,but deficiency and excess of disease character.By analyzing the functional mechanism of wind-expel ing drugs and relationship among exogenous diseases,wind-expel ing drugs and the stomach and spleen, raise the relativity between internal damage and exogenous diseases. [Conclusions] The revealing of the essence of deficiency and excess differentiations from‘Neiwaishangbianhuolun’, together with knowledge of the functional essence of wind-expel ing drugs and the relevance of internal damage and exogenous diseases, may open the mind partly for syndrome differentiation and medications in the clinic.

BACKGROUND:Previous studies have found that embryonic bone marrow mesenchymal stem cel s can promote human Th17 cel proliferation, but the inherent regulatory mechanisms stil need to be elucidated. OBJECTIVE:To investigate the role of Tol-like receptor 3 in the immunoregulation of Th17 cel s by mesenchymal stem cel s.
METHODS:Human CD4+T cel s from healthy donors were isolated by immunomagnetic bead method, and then cultured alone or co-cultured with embryonic bone marrow mesenchymal stem cel s for 4 days. The mRNA expression level of interleukin-17, Tol-like receptor 3 and MyD88 was detected by real-time PCR.
RESULTS AND CONCLUSION:Compared with CD4+T cel cultured alone group, the mRNA level of interleukin-17 was significantly higher in the co-culture group (3.59±0.11 vs. 1.14± 0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 3 mRNA was detected in the co-culture group compared with the CD4+T cel cultured alone group (3.10±1.60 vs. 0.94± 0.01, P<0.05). Furthermore, MyD88 in the co-culture group was significantly higher than that in CD4+T cel cultured alone group (2.29±0.05 vs. 1.85±0.31, P<0.01). Tol-like receptor 3 may be involved in the immunoregulation of Th17 cel s by embryonic bone marrow mesenchymal stem cel s, which provides experimental evidence for potential cel therapeutic strategy.

Objective To study the effect of somatostatin analogue (SMS201-995,SMS) on apoptosis of human cholangiocarcinoma SK-ChA-1 cells and apoptosis-regulated gene bcl-2 and bax. Methods The changes of apoptosis and bcl-2/bax gene expression were detected by DNA gel electrophoresis, flowcytometric analysis and immunohistochemistry, in situ hybridization methods in cholangiocarcinoma SK-ChA-1 cells after treated with SMS(100?ng/ml) for 6, 12 and 24 hours.Results The characteristic ladder was not detected by agarose gel electrophoresis after treated with SMS(100?ng/ml) for 6 hours. When the cells were treated for 12 and 24 hours, the ladder of DNA electrophoresis was observed. While the cells were treated with SMS for 6, 12 and 24 hours, the percentage of annexin V-positive cells were (22?5)% ,(39?7)% and(58?10)%. Besides,SMS inhibited bcl-2 protein expression and increased bax protein and mRNA expression. Conclusion SMS can induce SK-ChA-1 cell apoptosis mainly with regulation of bcl-2 and bax gene expression.

Objective] To explore core thought of Li Dongyuan’s spleen and gastric theory, grasp the basic spirit of Dongyuan theory, and better guide the clinic. [Method]Basing on some relevant theories from NeiJing, the eminent physician in Jin and Yuan Dynasties established the academic thought centering on regulating the spleen and stomach in treatment of the deficiencies. [Result]The thought revealed the cooperative function between invigorating Yang and tonifying middle-energizer and Qi. It also proposed the concept of “ Yin-fire” and the method of relieving high fever with drugs of sweet flavor and warm nature, creating a series of prescriptions of invigorating Yang and tonifying middle-energizer, which broadening the clinical treatment of traditional Chinese medicine. [Conclusion]The academic thought centered on adjusting spleen and stomach by raising Yang and tonifying Qi, broadens the TCM clinical therapy, today still effectively guides clinical practice.

0.05).The iris diaphragm was flat to mild concave which lead the width of anterior chamber angle increased postoperation, no peripheral synechia was found. Conclusion: Phacoemulsification and foldable posterior chamber intraocular lens implantation through clear corneal incision had no significant influence on the long term intraocular pressure in simple senile cataract patients, the width of anterior chamber angle increased postoperation.

With the development of modern vitrectomy,some new ideas have been proposed concerning the treatment of Terson syndrome.Developed neurosurgery has offered more chances of survival to patients with subarachnoid hemorrhage(SAH).Terson syndrome,as the initial presentation,has gained more and more attention from ophthalmologists,though strict guidelines for its treatment have not yet been established.The paper reviews the progress in the clinical researches of Terson syndrome.

Objective:To evaluate the experience of perioperative care of phacofragmentation with vitrectomy.Methods:During 2000 to 2005,A group of 29 eyes in 29 patients underwent phacofragmentation with vitrectomy:Their lens nucleus were sank in vitreous cavity for different kinds of causes.The proficient nursing cooperation and perioperative care were retrospected analyzed.Results:The therapeutic effects after the operations were satisfactory.The post-operative best corrected visual acuities were between 0.1-0.5.No severe complications occurred.Conclusion:Right pre-operative instructions,sufficient preparation,and proficient nursing cooperation can ensure the success of the operation.

Objective:To determine whether laser in situ keratomileusis(LASIK) affects the central 30-degree visual field.Methods:This clinical trial comprised 31 patients(61 eyes)scheduled to have LASIK for myopia or myopic astigmatism.Automated static perimetry was performed before and 1 week,6 months after the surgery using the Octopus 1-2-3 perimetry.The main outcome measures were the meao sensitivity(MS) change in the central 15-degree visual field and the MS change in the 15-to 30-degree visual field.The preoperative and postoperative MS values were calculated,and the difference between them(delta MS) was determined.Statistical analysis were performed by SPSS12.0 using the randomized complete block design ANOVA(two factors analysis of variance).Differences among variables were evaluated by q-test. Results:One week after the operation,all uncorrected visual acuity equal to or better than 4.7.Six months after operation,visual acuity distributed between 4.9 and 5.2. There was no significant change in the central 15-degree visual field MS,neither 1 week nor 1 month after the operation;between 15 and 30 degrees,there was a statistically significant decrease.One week after the operation,the delta MS value was(-0.91?1.42)dB(mean?standard deviation);6 month after the operation,the delta MS value was(-0.87?1.38)dB. Conclusion:Automatic static perimetry can detect decreased sensitivity in the 15-to 30-degree visual field after myopic LASIK.It may be a useful quantitative subjective test for measuring the vision quality after refractive surgery in future.

BACKGROUND: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied.OBJ ECTIVE: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells.DESIGN: Observational-contrast study.SETTING : Department of Labor Hygiene, the Third Military Medical University of Chinese PLA.MATERIALS: Vascular endothelial cell strain; H332PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters(Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden).METHODS: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. ① Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37℃ and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes.Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. ②Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly.MAIN OUTCOME MEASURES: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; ②Results of active analysis and expressional measurement of protein kinase C in radiation group and control group.RESULTS: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation.Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorylation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group,respectively (t = 2.974, 4.209, 4.047, P ＜ 0.05), and then, fallen down to normal value 24 hours later. ② Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group,especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36%, 93% and 47% in radiation group as compared with that in control group, respectively (t =2.801, 3.654, 3.035, P ＜ 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation.CONCLUSION: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C.Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.