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

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 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

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.

Objective To investigate the efficacy and safety of different acoustic power of high-intensity focused ultrasound (HIFU) in treating human pancreatic xenograft models. Methods Human pancreatic cancer cells (YY-1) were implanted subcutaneously in nude mice to establish animal models. The tumor bearing mice were divided into low-power HIFU treatment group (200 W,n=10), high-power HIFU treatment group (300 W,n=10) and blank control group (n=10). The change of tumor volume, the tumor growth rate and side effects were recorded. The apoptosis rate of tumor cells of each group was determined by TUNEL method. Results The tumor volume and the tumor growth rate of the low-power group and the high-power group were significantly lower than those of the control group (P<0.05), while no statistically significant differences in the tumor volume and the tumor growth rate existed between the low-power group and the high-power group (P>0.05). Compared with the low-power group, the incidence of side effects in the high-power group was significantly higher (P<0.05), including mainly skin burn (60%) and acoustic channel injury (20%). At the 7th and 14th day after the treatment, the apoptosis rates of tumor cells in both the low-power group and the high-power group were significantly higher than that of the control group (P<0.05), but the difference in the apoptosis rates of tumor cells was not statistically significant between the low-power group and the high-power group (P>0.05). Conclusion For the treatment of human pancreatic xenograft tumor in nude mice models, HIFU with low power is effective and safer.

Objective To investigate the management and prevention of the complications of sigmoid rectal pouch for urinary division after radical cystectomy. Methods The clinical data of 34 patients who underwent a sigmoid rectal pouch procedure were analyzed retrospectively, and the clinical experience was summarized in the management and prevention of the complications of sigmoid rectal pouch for diversion. Results Twenty-six patients were followed up for 2 months to 11 years, and 10 patients lost in follow-up. The early follow-up results were as follows:3 patients had postoperative high fever with unilateral the kidney water, 1 patient had retropubic bleeding and need to stop bleeding, 3 patients suffered from wound split open and were performed relaxation suture, and 1 patient had sigmoid colon rectum bladder fistula 10d after operation. The late follow-up results were as follows:1 patient had urethral neoplasms recurrence, 5 patients developed distance metastases, and 5 patients developed nocturnal incontinence and worn safety pad. There were no hyperchloremic acidosis requiring clinical treatment, hydronephrosis as well as retrograde pelvis infection. Conclusions The operation of sigmoid rectal pouch for urinary division is fairly simple, with no serious complication. It is a better alternative diversion procedure, and should be accepted gradually by patients and surgeons.

BACKGROUND:The decel ularized porcine smal intestinal submucosa is a kind of bioactive extracel ular matrix, which is mainly composed of col agen, glycoprotein, proteoglycan and rich in col agen, glycosaminoglycan and various growth factors, and these components play an important role in promoting the differentiation and proliferation of tissue cel s. OBJECTIVE:To prepare the injectable smal intestinal submucosa and to investigate its co-culture with rat adipose-derived mesenchymal stem cel s in vitro. METHODS:The injectable smal intestinal submucosa and rat adipose-derived stem cel s were prepared. Cel counting kit-8 test for cel proliferation:Passage 3 adipose-derived stem cel s were seeded onto the injectable smal intestinal submucosa (experimental group) and cel s cultured under normal condition as control group. The cel proliferation was observed at 1, 3, 5 and 7 days of incubation. Live/dead staining test for the survival of cel s:Passage 3 adipose-derived stem cel s were respectively cultured in the injectable smal intestinal submucosa extracts (experimental group) and complete culture medium (control group). Cel survival was determined at 1, 3, 5 and 7 days of culture. RESULTS AND CONCLUSION:Scanning electron microscope oval and strip adipose-derived stem cel s adhered onto the material. The absorbance values in the experimental group were higher than those in the control group at 1 and 5 days of incubation (P<0.05). Cel survival:The number of cel s appeared to be in a rising trend with time in both two groups;after 1-day co-culture, al cel s in the two groups survived. Then dead cel s appeared in both two groups, showing no significant difference. These results show that the injectable smal intestinal submucosa exhibits a good cytocompatibility.

Objective To investigate the changes of quantitative parameters of dynamic enhanced CT in non-small cell lung cancer before and after targeted therapy,and compare them with the traditional evaluation criteria,in order to find the parameters which can be exploited for timely,objective evaluation of the effect of targeted therapy.Methods The study included 21 patients with targeted therapy who had received dynamic enhanced CT before and after treatment.Enhancement time-density curves were obtained based on the CT values of the lesion at individual time points,and the functional indices:peak height (PH),the time to peak height (Tp),the ratio of PH of the mass to aorta (M/A) and perfusion value were calculated.The effects of the treatment on these indices were evaluated and compared with the effect of the treatment on lesion diameter. Results Twenty-one patients had 33 rechecking results. There was a statistically significant agreement between lesion diameter-based treatment evaluation and perfusion-based treatment evaluation ( U =8.761,P ＜ 0.01 ). The perfusion value decreased in patients with disease regression[before treatment:(0.28 ±0.11 ) ml · min-1 · ml-1,after targeted therapy(0.18 ±0.09) ml ·min-1 · ml-1,t =- 3.2722,P =0.0042],but increased in patients with disease progression[before treatment(0.21 ±0.08) ml · min-1 · ml-1,after targeted therapy:(0.34 ±0.11 ) ml · min-1 · ml-1,t =2.6064,P =0.0403].Conclusions On dynamic enhanced CT in non-small cell lung cancer patients after targeted therapy,perfusion value changed in the same trend as the diameter of tumor.The effectiveness of targeted therapy may be evaluated by perfusion value changes.

Colitis, Ulcerative ; complications ; drug therapy ; Humans ; Male ; Middle Aged ; Psoriasis ; complications ; drug therapy ; Pyoderma Gangrenosum ; drug therapy ; etiology