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.

This study was conducted to investigate the physicochemical properties of Polyporus umbellatus sclerotial exudate. Morphological characteristics of the sclerotia and its exudate were observed during different stages of sclerotial formation. The pH of the exudate was detected at different time during cultivation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of P. umbellatus sclerotial exudate during cultivating time. Additionally, the protein content was measured by means of BCA protein assay. Furthermore, CAT content was detected using ultraviolet absorption method. That the protein content of the exudate and CAT specific activity rose gradually during the passage of the cultivating time indicated a high level of oxidative stress during P. umbellatus sclerotial exudate formation. The results showed that the pH of the exudate increased gradually and then dropped down during sclerotial formation. That the pH of the exudate maintained the acidity state during the cultivation indirectly indicated that acidic environment would help sclerotial formation. The exudate produced gradually and was absorbed by the sclerotia itself.
Culture Media ; chemistry ; metabolism ; Fungal Proteins ; chemistry ; metabolism ; Fungi ; chemistry ; metabolism ; Hydrogen-Ion Concentration ; Medicine, Chinese Traditional ; methods ; Oxidative Stress ; Polyporus ; chemistry ; metabolism ; Polysaccharides ; chemistry ; metabolism

Objective The significance of serum cytokine profile in coronary heart disease patients have been evaluated by observing the variation cytokines Methods The serum IL-1 a,IL-1 ?,IL-2,IL-4, IL-6 ,IL-8,IL-10,VEGF,IFN-?,,TNF-?,MCP-1 and EGF in 76 patients and 26 matched healthy volunteers have been observed.Serum cytokines had been measured by protein chips methods in EVIDENCE 180 automatic biochips analyzer and cytokine profile were got by bioinformatics.Results The serum level of IL-1?,IL-6,IL-8,and TNF-? were (4.32?8.14) ng/L ,(13.16?16.81) rig/L,(375.53?292.14) ng/L,(15.46?15.38) ng/L respectively,which increased obviously in patients (P

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

Objective To explore the incidence and clinical treatment of related complications caused by implantable venous access port(IVAP) in patients with breast cancer during chemotherapy.Methods The data of 755 patients with breast cancer recieved chemotherapy by which caused some related complications in our hospital from January 2014 to March 2016 were retrospectively analyzed.Results 753 patients IVAPs were implanted succussfully.The total placement time of implantable venous access port was from 110 days to 940 days,with median placement 147.33 days.The related complications of IVAP were catheter malposition(0.79％,6/755),catheter-related thrombosis(27.81％,210/755),catheter fracture(0.13％,1/755),port exposure(0.93％,7/755) and IVAP-related bloodstream infection(0.13％,1/755).The IVAP-related complications and thrombosis rate were significant higher when IVAPs implanted in the left internal jugular veincompared with that in right internal jugular vein(34.88％ vs.25.74％,33.10％ vs.24.68％).Conclusion Application of IVAP in patients with breast cancer during chemotherapy is a safe and effective operation.The most common complication is asymptomatic mural thrombus formation around the catheter,which should be paid attention to.

Ergosterol is the important component of the fungal membrane, and having stable structure. This makes it a suitable indicator for growth of fungi. In the paper, isolation and determination techniques of ergosterol as the indicator of the fungal biomass were reviewed. The methods of extracting ergosterol include traditional saponification and refluxing, rapid physical disruption, rapid ultrasonication, supercritical fluid extraction and so on. The ergosterol determination methods are high performance liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and thin-layer chromatography, et al. The application of these techniques was also introduced. Finally, the paper prospected the feasibility of applying the ergosterol as the indicator of fungal biomass in composting.

BACKGROUND: Nucleotide-binding oligomerization domain (NOD)-like receptor plays an important role against inflammatory responses caused by pathogens and non-pathogens, as well as in the initial stage of autoimmune response. Meanwhile, NOD2 and NOD-like receptor protein 3 (NLRP3) are the representative proteins of NOD-like receptor family. OBJECTIVE: To detect the expression of NOD2 and NLRP3 in a rabbit model of osteoarthritis. METHODS: Thirty New Zealand rabbits were randomly divided into six groups (n=5 per group), including five experimental groups (2, 4, 6, 8 and 10 weeks) and one control group. The model of osteoarthritis was established by fixing the left knee joints using plaster cast, and were sacrificed at postoperative 2, 4, 6, 8 and 10 weeks. The controls received no intervention, and were killed at 10 weeks postoperatively. The left distal femur articular cartilage was taken for safranin-fast green staining. The pathological changes were evaluated by Mankin's scores, and the expression levels of NOD2 and NLRP3 were detected by immunohistochemistry. RESULTS AND CONCLUSION: The Mankin's scores in the experimental groups were significantly higher than those in the control group (P <0.01). Moreover, the scores in the experimental groups were significantly increased with time (P < 0.01). The expression levels of NOD2 and NLRP3 in the chondrocytes were also increased with time (P < 0.01). These results indicate that the expresison of NOD2 and NLRP3 in the cartilage cells is positively correlated with the pathological changes of osteoarthritis, which may be through promoting apoptosis in cartilage cells, thus accelerating the development of osteoarthritis.

ObjectiveTo investigate the clinical effect and treatment experience of complicated acetabular fractures treated with open reduction with combined ilio-inguinal and Kocher-Langenbeck (K-L) approaches.MethodsSixty-six patients with complicated acetabular fractures were treated surgically with combined ilio-inguinal and K-L approaches from February 2004 to December 2009.Among them,45 cases were followed up for 8-45 months,with the average 26 months.ResultsOperation time was 1.4-5.7 h,with the average of 2.8 h.Intraoperative blood loss was 530-2300 ml,with the average of 1250 ml.According to Matta's score system,the excellent and good rate was 86.7％.According to modified Merle d' Aubigne and Postel score system,the excellent and good rate of function of hip was 86.7％.According to Brooker heterotopic ossification standard assessment system:Ⅰ degrees in 6,Ⅱ degrees in 3.According to Letournel and Judet clinical classification system,postoperative traumatic arthritis were 6 patients.Among them,3 patients with Ⅲ- Ⅴ period with symptoms of serious hip pain and joint activities obstacles were treated with total hip replacement secondly.Six cases of sciatic nerve injury and 9 cases of lateral femoral cutaneous nerve damage were recovered gradually after six months to a year.All incisions healed primarily.All got bony union,the healing time was 8 to 16 weeks,with the average of 10.4 weeks.None was complicated with femoral head necrosis.ConclusionFor the patients with complicated acetabular fractures,open reduction with combined ilio-inguinal and K-L approaches can get clinical advantages of revealed fully,fracture restoration convenient,fixed tightly,fewer complications and satisfactory result.

AIM: To analyze the effect factors of trabeculectomy combined with intraoperative application of mitomycin C in the treatment of neovascular glaucoma.
METHODS: Fifty patients (50 eyes) with neovascular glaucoma collected from January 2013 to August 2015 in our hospital were treated by trabeculectomy combined with intraoperative application of mitomycin C. Single factor and multi factor variables analysis were used for effect factors of trabeculectomy combined with intraoperative application of mitomycin C in the treatment of neovascular glaucoma.
RESULTS: By results of single factor variable analysis,< 50 years old, preoperative intraocular pressure ( IOP) was ≥45mmHg and postoperative occurrence of anterior chamber hemorrhage were risk factors for treatment failure ( P < 0. 05 ), and gender, proliferative diabetic retinopathy and previous cataract surgery and prior photocoagulation were not the risk factors for failure (P>0. 05 ). By multivariate analysis, < 50 years old and postoperative occurrence of anterior chamber hemorrhage were risk factors for treatment failure ( P < 0. 05 ), and preoperative IOP≥45mmHg was not a risk factor (P>0.05). CONCLUSION: For patients < 50 years old with neovascular glaucoma, should be careful on the selection of surgical treatment. For high- risk patients, we should strengthen the monitoring and give timely intervention, which are helpful to improve the prognosis.

Objective To study the effects of nitric oxide synthase(NOS) and nitric oxide(NO) in post-asphyxial renal injury in neonatal rats.Methods Forty-eight Wistar neonatal rats were randomly divided into 4 groups:controls,2 h,24 h and 48 h post-asphyxia groups (12 in each group).The rats were decapitated in different times(2 h,24 h and 48 h) after asphyxia for 30 minutes.The renals were dissected to determined the concentrations of NO and NOS.And the scores of renal tubules were measured under light microscope.Results Compared with control group,the levels of NO and NOS significantly increased at 2 h and 24 h after asphyxia.The scores of renal tubules were significant difference at 24 h and 48 h after asphyxia compared to controls.Conclusion These findings suggest NOS and NO may play an important role in the development of post-asphyxia renal injury.