Background Marrow mesenchymal stem cells (MSCs) has been successful induced to differentiate into corneal cells,retinal ganglion cells (RGCs) and retinal neuron-like cells in recent years,but there are few studies about MSCs induced into photoreceptor cells and their microenvironment.Objective The aim of this study was to explore the induce and differentiation of bone marrow mesenchymal stem cells (BMSCs) into photoreceptor-like cells in vitro and microenvironment.Methods The second generation of human BMSCs strain and RPE cells strain were cultured and passaged,respectively,and the fourth generation of BMSCs and the third generation of RPE cells were used in the experiment.BMSCs were cocultured using the mesenchymal stem cells medium (MSCM) contained 20 μg/L basic fibroblast growth factor (bFGF),20 μg/L epithelial growth factor (EGF)and 20 μg/L brain derived neurotrophic factor (BDNF) with RPE cells to induce the differentiation of BMSCs in the induced group,and BMSCs were cultured in MSCM only in the control group.The morphology of induced and differentiated cells were observed under the inverted microscope.Inmmunocytochemistry was used in induced for 3-,5-,7-day cells to detect the expression rate of rhodopsin protein for the identification of phenotype of the differentiated cells.RT-PCR was used in induced for 5-,7-day cells to detect the expressions of rhodopsin mRNA and recoverin mRNA.Results Cultured BMSCs grew well with the spindle shape,and passaged RPE cells showed the uniform size and polygon shape with the abundant pigment in the cells.Some induced cells appeared to be the neuron-like cells with round shape and long prominence and the secondary reticular branches.The expression rates of rhodopsinin the cells were (5.83±0.29)％,(20.36±0.32)％ and (29.80±2.30)％ in the third,fifth and seventh day after induce,which were significantly higher than (0.71 ±0.35) ％,(2.56±0.24) ％ and (2.32±0.42) ％ of control cells (t3 d =41.510,t5d =107.290,t7 d =30.036,P＜0.01).The grey scales of rhodopsin mRNA and recoverin mRNA were significantly elevated in the induced and differentiated cells compared with control cells in the fifth and seventh day (rhodopsin mRNA:t5 d =103.506,t7 d =122.584,P＜0.01 ; recoverin mRNA:t5 d =106.674,t7 d =189.992,P＜0.01).Conclusions BMSCs can be successfully induced to differentiate into photoreceptor cells after cocultured by conditioned medium with RPE cells.

Acupuncture Therapy ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Enuresis ; drug therapy ; etiology ; Female ; Humans ; Male ; Phytotherapy

Objective To explore the effects of Tianqijiangtang capsule combined with Metformin on the levels of IL‐6 ,TNF‐α and C‐RP protein in patients with T2DM. Methods 80 T2DM were randomized into control group (treatment with Metformin ,n= 40) and treatment group (treatment with Tianqijiangtang in combination with Metformin ,n=40). The changes of levels of IL‐6 ,TNF‐α and C‐RP were observed before and after the treatment. Results Compared to the pretreatment in the same group and after treatment in control group ,IL‐6 ,TNF‐αand C‐RP of treatment group were reduced significantly [(6.79 ± 0.60) ,(7.15 ± 0.62) vs (5.08 ± 0.43)pg/ml;(8.94 ± 0.55) ,(8.61 ± 0.66) vs (5.18 ± 0.45) pg/ml;(6.79 ± 0.60) ,(6.58 ± 0.59) vs (5.56 ± 0.30) mg/L ,P<0.01]. Conclusion Tianqijiangtang capsule combined with Metformin can decrease the levels of IL‐6 ,TNF‐αand C‐RP in T2DM patients.

Objective:To investigate the intracellular localization and association of chicken major histocompatibility complex class Ⅰsubunits with invariant chain.Methods:At first,the sequences of MHCⅠ ? and ?2m subunits were amplified with RT-PCR.Secondly,we constructed the eukaryotic expression plasmids of MHCⅠ ? and MHCⅠ?2m that was fused with red or enhanced green fluorescent protein,respectively.The recombinant plasmids of MHC Ⅰsubunits and pEGFP-C1-Ii that could express enhanced green fluorescent protein were transiently transfected into COS-7 cells with Lipofectamine 2000.Immunofluorescence microscopy was carried out to detect the expression and intracellular localization of Ii and MHC Ⅰsubunits,and immunoprecipitation was used for analyzing their association.Results:The colocalization was found in endocytic compartments when GFP-Ii,MHCⅠ?-RFP and MHCⅠ?2m-RFP were co-expressed in COS-7 cells.Immunoprecipitation of Ii showed that GFP-Ii co-isolated with MHCⅠ?-GFP and MHCⅠ?2m-GFP subunits when COS-7 cells were transiently transfected with pEGFP-C1-Ii,pEGFP-N1-MHCⅠ? and pEGFP-N1-MHCⅠ?2m.Conclusion:Chicken Ii can not effectively associate with single ? or ?2m subunits from chicken MHC Ⅰmolecules,but Ii and the integrated MHC Ⅰmolecule can form the complexes that are colocalized in endomembrane system of COS-7 cells.

Objective To dynamic monitoring the changes of serum cortisol,adrenocorticotropic hormone (ACTH) in children with sepsis and servere sepsis,and to assess it's relationship with clinical features.Methods Thirty children with sepsis were enrolled in this study from Sep 2010 to Jan 2011 in our hospital,and were divided into severe sepsis group (n =10) and sepsis group (n =20).The serum cortisol and ACTH were analysed in acute and recovery period respectively in both two groups,and compared with 10 non-sepsis children as control group.Results In the acute period,the level of serum cortisol in severe sepsis group [ (33.97 ±6.83) μg/ml ] was significantly higher than those in sepsis group [ (26.30 ± 10.43) μg/ml]and control group [ ( 19.06 ± 6.91 ) μg/ml ] ( P ＜ 0.01 ) ; the level of serum ACTH in severe sepsis group [ (25.47 ± 9.41 ) pg/ml ] was significantly higher than those in sepsis group [ ( 15.52 ± 5.85 ) pg/ml ] and control group [ (9.16 ± 3.02 ) pg/ml ] ( P ＜ 0.01 ).Conclusion Stress response exists in the early period of sepsis,and the levels of serum cortisol and ACTH are increased significantly.The levels of serum cortisol and ACTH are positive correlation with the severity of diseases,and they could be used as serious index and a marker for evaluating prognosis in children with sepsis.

Chinese medicinal formulae( CMF) were often used in the clinics of traditional Chinese medicine (TCM) which were critical for modernization of Chinese medicine to shed light on the interaction between CMF and biological organisms. In current studies, correlation between system and part, macroscopic actions and microcosmic mechanism, ADME process and pharmacologic actions were often neglected. Thus, we put forward integrative pharmacology, which could integrate the correlation between CMF and biological organisms from multi-levels and multi-dimensional views. Integrative pharmacology would reveal the molecular mechanism of CMF for ailments treatment and screen out effective material systematically, which would be the new paradigm of TCM research.
Absorption ; Computational Biology ; Medicine, Chinese Traditional ; methods ; trends ; Pharmacology ; methods ; trends ; Tissue Distribution

OBJECTIVETo observe the intestinal movement of transient receptor potential vanilloid 1 or vanilloid receptor 1 (TRPV1) knockout mice after stimulated by electroacupuncture (EA), and to primarily explore the roles of TRPV1 receptor in the jejunal motility regulated by acupuncture.

METHODSNormal wild-type CL57BL/6 and TRPV1 gene knockout mice were recruited in two groups, the B6 group and the TRPV1 group, 15 in each group. The thermal threshold and the mechanical pain threshold were respectively detected using JL-F digital photo thermal analyzer instrument and ALMEMO2450 machine. The difference between the two thresholds were compared. Meanwhile, a self-made pressure head was placed in the jejunum. The internal pressure was monitored. When the pressure was stable, 2 mA 2/15 Hz EA at Quchi (LI11), Tianshu (ST25), Shangjuxu (ST37), and Dachangshu (BL25) to observe the changes of intestinal pressure value between before and after EA. The curve of internal pressures was recorded.

RESULTS(1) The stimulation of light/heat and mechanical stimulation were obviously slowed in the TRPV1 group than in the B6 group (P < 0.01). (2) In the intestinal pressure observation experiment, the bowel movement was not obviously seen in the two groups when acupunctured at Quchi (LI11) and Shangjuxu (ST37) (P > 0.05). (3) Acupuncture at Tianshu (ST25) and Dachangshu (BL25) of TRPV1 knockout mice could lead to intestinal movement (P < 0.05), mainly inhibited bowel movement. The changing degree was equivalent to that of B6 mice.

CONCLUSIONSUnder the physiological condition, TRPV1 might be mediated by thermal and mechanical stimulation. But TRPV1 mediated acupuncture effect was quite complex, indicating TRPV1 mice might be one of intestinal movement mediating factors.

Animals ; Electroacupuncture ; Gastrointestinal Motility ; Jejunum ; physiology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; TRPV Cation Channels ; physiology

Objective To characterize mycosis fungoides (MF)-associated serum proteins by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) technique. Methods Serum samples were collected following informed consent from 15 patients with MF and 15 patients with chronic eczema or neurodermatitis. Serum protein profiles were detected by CM10 protein chip combined with SELDI-TOF-MS. The proteins differentially expressed between MF and chronic eczema or neuroder matitis were assessed by the Zhejiang University Cancer Institute - ProteinChip Data Analysis System (ZUCIPDAS). Results A total of 329 protein peaks were detected from these patients, significant difference was observed in only 30 protein peaks. The optimal diagnostic model was developed by support vector machine algorithm with two protein peaks at Mass/Charge (m/z) 3939 and 5909 respectively. The expression of protein peak at m/z 3939 was higher in chronic eczema and neurodermatitis than in MF, while that at m/z 5909 appeared to express in an opposite way. In simulation training, this model proved to be able to distinguish MF from chronic eczema and neurodermatitis with both the specifity and sensitivity being 100 percent. The leave-one-out cross-validation also revealed a specificity and sensitivity of 100 percent for this model in the comparison of MF with chronic eczema and neurodermatitis. Conclusion These results suggest that SELDITOF-MS technique combined with bioinformatics is highly specific and sensitive in the diagnosis of MF.

Apoptosis ; Gene Expression Profiling ; Hepatic Stellate Cells ; pathology ; Humans ; Liver ; metabolism ; Liver Cirrhosis ; genetics ; metabolism ; pathology ; Liver Diseases ; genetics ; metabolism ; pathology ; MicroRNAs ; metabolism ; physiology ; Transforming Growth Factor beta1 ; metabolism

Objective:To explore the antibacterial effect of Belamcanda Chinensis DC and Por tulaca oleracece L on P.aeruginosa (PA) in vitro.Methods:Fourty six strains PA were tested for minimum inhibition concentrati on ( MIC) by water decoct agents of the two drugs.MIC50 and MIC90 were st atistically studied.Results:For Belamcanda Chinensis DC,MIC was 31.25～3.90 g/L;MIC50 was 7 .81 g/L,and MIC90 was 15.62 g/L;whereas， for portulaca oleracece L,MIC was 31 .25～7.81 g/L, MIC50 was 15.62 g/L,and MIC90 was 31.25 g/L.Conclusion:Both of the two drugs have stronger antibacterial effects on P.ae ruginsosa in vitro.