Background Human retinal pigment epithelial (RPE) cell transplantation treating retinal degenerative diseases is a researching topic,and the source of human RPE cells is a key problem.Many biological carriers can be used for the preparation of RPE cell layer.However,some advantages,such as cytotoxicity,lack of stability and immunologic reaction etc.are still existed.To study an ideal biological carrier is very important.Objective This experimental was to determine the effects of amniotic membrane on the proliferation and differentiation of human RPE cells and the possibility as a scaffold for RPE cell transplantation.Methods ARPE19 cell line cells were cultured and passaged in DMEM/F12 medium with 10％ fetal bovine serum,and 8-12generation of cells were used.The cells were divided into two groups.One group of cells were incubated on the denuded amniotic membrane,and the other group of cells were cultured in the medium (control group).MTT was performed to detect the A492 value of RPE cells for the evaluation of cell proliferation ability 24,48,72,96 hours after culture.Cell morphology was compared by histopathological examination 3 weeks after culture.The mRNA expression of pigment epithelium-derived factor (PEDF),N-cadherin,β-catenin and cell connection related proteins in the cells of both groups were assayed using reverse transcription polymerase chain reaction (RT-PCR).Ultrastructure of the cells was observed under the transmission and scan electronic microscope 3 weeks after culture.Results The number of ARPE-19 cells cultured on denuded amniotic membrane was decreased significantly in comparison with the normal culture plate(F=41.760,P =0.000).Histopatholy also showed that the cell density on amniotic membrane was lower than of normal cells on plate surface.Moreover,the expression level of claudin 1 mRNA,N-cadherin mRNA and PEDF mRNA were significantly up-regulated in denuded amniotic membrane group in comparison with control group (t=15.828,P=0.000 ;t=6.839,P=0.002 ;t=14.667,P=0.000),but the expression of Connexin 43 mRNA was down-regulated in denuded amniotic membrane group compared with control group(t=3.358,P=0.024).Ultrastructural examination revealed that ARPE-19 cells cultured on amniotic membrane exhibited a polygonal epithelial phenotype with cilium on the apical side,however,the cells cultured on normal culture plate displayed fusiform shape and uneven thickness.Conclusions Amniotic membrane plays a promoting effect on the differentiation of ARPE-19 cells and a inhibitory effect on the proliferation of ARPE-19 cells,suggesting that amniotic membrane might be an useful scaffold for the preparation of functionally mature RPE cells for clinical transplantation.

Breast Neoplasms ; metabolism ; pathology ; Cell Polarity ; Epithelial Cells ; metabolism ; pathology ; Epithelial-Mesenchymal Transition ; Eye Proteins ; metabolism ; Female ; Humans ; Membrane Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Receptors, Proteinase-Activated ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

Carcinoma, Renal Cell ; immunology ; pathology ; surgery ; Humans ; Keratin-19 ; metabolism ; Kidney Neoplasms ; immunology ; pathology ; surgery ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Nephrectomy ; Vena Cava, Inferior ; pathology ; surgery

AIM: To detect the differentiation effects of retinal cells or extracts on bone marrow-derived mesenchymal stem cells (BMSC).METHODS: Human fetal BMSC were previously labeled by carboxyfluorescein succinimidyl ester (CFSE), and co-cultured with retinal pigment epithelial （RPE） cells which were pre-treated with ultraviolet irradiation at a ratio of 1∶1 to induce the differentiation of BMSC for up to 14 days. In some assays, a retinal extract of bovine retinal extract (BRE) was added to detect the potential effects of retinal component on the differentiation of BMSC. In addition, neuron-specific enolase (NSE), Nestin and Glial fibrillary acidic protein (GFAP) immunostaining were performed to determine the characteristics of BMSC.RESULTS: The results indicated that by co-cultured with RPE cells, fetal BMSC were differentiated into neural-like cells expressing special neuronal markers Nestin, GFAP and NSE. And the expression of these markers was obviously increased by BRE.CONCLUSION: Retina derived cells and extracts can induce the differentiation of BMSC into neural-like cells.

Objective To investigate the change of plasma visfatin level and the relationship of plasma visfatin level to body mass index(BMI),waist hip ratio(WHR),blood glucose,plasma insulin levels as well as other factors in the subjects with different glucose tolerances.Methods Fasting and glucose loading 2 h plasma visfatin levels were assayed by ELISA in patients with type 2 diabetes and impaired glucose tolerance(IGT),and controls.The relationship of plasma visfatin level with BMI,HOMA-IR,blood lipids,plasma glucose,plasma insulin,free fatty acids(FFA)levels were also analyzed.Results Fasting and glucose loading 2 h plasma visfatin levels in the type 2 diabetic patients were significantly reduced compared with controls[(11.63?7.48)?g/L vs (16.82?6.06)?g/L and (12.02?6.86)?g/L vs (16.26?7.78)?g/L,both P＜0.05],and the level of plasma visfatin in IGT was lower than that in diabetics and higher than controls.The level of visfatin in obese subjects was modestly higher than that in non-obese subjects,but the difference is not significant.Plasma visfatin level was positively correlated with WHR(r=0.42,P＜0.01),and negatively with 2 h plasma glucose after glucose loading(2hPG)and HbA_(1C)(r=-0.33,P＜0.01 and r=-0.25,both P＜0.05).Multiple regression analysis showed that fasting plasma glucose,WHR,2hPG,high density lipoprotein cholesterol and HbA_(1C) were independent related factors in influencing plasma visfatin levels.Conclusion The change of visfatin level is associated with glucose metabolism,which may contribute,in part,to the pathogenesis of diabetes and obesity.

OBJECTIVETo explore the differences of electroacupuncture (EA) on onset time and symptom improvement for treatment of different types of functional constipation.

METHODSThirty-eight cases of constipation were selected, including 9 cases of constipation-predominant irritable bowel syndrome (IBS-C), 10 cases of slow transit constipation (STC), 10 cases of pelvic floor dyssynergia (PFD) and 9 cases of inadequate defecatory propulsion (IDP). The electroacupuncture was applied at Tianshu (ST 25), Fujie (SP 14), Shenshu (BL 23), Dachangshu (BL 25) and so on in abdominal and lumbosacral area, 5 times per week, 10 times as a treatment course. The onset time, score of clinical symptoms of constipation and improvement of every symptom in each group were compared.

RESULTS(1) The onset time was (1.78 +/- 0.83) days in IBS-C type, (3.11 +/- 1.90) days in IDP type, (4.10 +/- 1.85) days in STC type and (4.30 +/- 2.00) days in PFD type, indicating statistical differences between IBS-C type and STC type, IBS-C type and PFD type (both P < 0.05). (2) Compared before the treatment, the total scores of symptoms on the onset day in each group were all improved (P < 0.05, P < 0.01), and score of IBS-C type was superior to the rest 3 types (P < 0.05, P < 0.01). (3) EA improved desire to defecate or frequency of defecation in each type (P < 0.05, P < 0.01), in which both were improved in STC type and PFD type, and the improvement of defecation frequency was more significant in STC type (P < 0.01). EA relieved unsmooth defecation or pendant-expansion feeling in each type (all P < 0.05), in which both were improved in IBS-C type (both P < 0.05). EA relieved abdominal distension and pain in IBS-C type, STC type and PFD type (all P < 0.05), while its effects were not obvious on defecation difficulty, defecation time and defecation texture (all P > 0.05).

CONCLUSIONThe electroacupuncture for treatment of 4 types of constipation is characterized by rapid onset; the improved symptoms are not identical in the electroacupuncture treatment plan for each type of defecation; the main improvement of symptoms are lied on desire to defecate and frequency of defecation, unsmooth defecation or pendant-expansion feeling and abdominal distension and pain. Meanwhile the improvements of defecation texture, defecation difficulty and defecation time were not signi-ficant.

Adult ; Aged ; Constipation ; etiology ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Irritable Bowel Syndrome ; complications ; Male ; Middle Aged ; Pelvic Floor Disorders ; complications ; Treatment Outcome ; Young Adult

By retrieving the clinical research literature of treatment functional dyspepsia by traditional Chinese medicine (TCM) from January 2004 to December 2014 based on China National Knowledge Internet (CNKI), we would establish a TCM decoction database for treating functional dyspepsia in this study. One hundred and sixty-four literature were included, involving 159 prescriptions, 377 medicines, in a total of 1 990 herbs. These herbs can be divided into 18 categories according to the effectiveness; and qi-regulating herbs, blood circulation herbs, and antipyretic herbs ranked top three ones according to the frequency of usage of the herbs, whose medicine usage frequency accounted for 51.81%. Usage frequency of 16 herbs was over 30, and Atractylodes, Radix, Poriaranked top three according to the usage frequency. Medicinal properties were divided into 9 kinds according to the frequency statistics, and the top three were warm, flat, and cold. Taste frequency statistics were classifiedinto 9 kinds, and the top three were acrid, sweet, and bitter. In frequency statistics of the meridian tropism of herbs, it was classifiedinto 11 kinds, and the top three were spleen, stomach, lung. The analysis can provide a reference for treatment and study of TCM of functional dyspepsia.
China ; Databases, Bibliographic ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Dyspepsia ; drug therapy ; physiopathology ; Humans ; Internet ; Spleen ; physiopathology ; Stomach ; physiopathology

Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.

Objective To study the effects of a rotating magnetic field(RMF)on Li pilocarpine-in- duced seizure activity and the expression of mGluR1 and mGluR5 in the hippocampus.Methods Thirty rats were divided into 5 groups.Each rat in the model(M),short treatment(ST)and long treatment(LT)groups was treated with intra-peritoneal injections of lithium chloride(60 mg/kg),followed by an intra-peritoneal injec- tion of pilocarpine(35 mg/kg)24 h later.The rats in the ST group were exposed to 20 mT RMF for 20 min ev- ery day for 3 d before seizure induction,while the rats in LT group were exposed to the same RMF for 8 d.The latency,severity and duration of seizure,as well as accompanying symptoms and electroencephalogram data, were recorded,and the expression of mGluR1 and mGluR5 was calculated using an electrophoretic imaging anal- ysis system.Results The duration,times and accompanying symptoms of seizure were significantly decreased in the LT group.The mGluR1 mRNA level and mGluR1/mGluR5 ratio in the M group were markedly increased, but the mGluR5 mRNA level was obviously decreased,while the expression of mGluR1 in the ST and LT groups was decreased,and mGluR5 was increased.Conclusions Seizure activity in rats can be inhibited by 20 mT RMF,and the expression of mGluRl and mGluR5 in the hippocampus of rats suffering seizures can be markedly influenced by longer-term RMF.