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 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 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 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.

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.

OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

To explore the distribution of leukemia mortality and its relationship with age of residents who live in city Urumqi, data of leukemia mortality of Urumqi residents from 1985 years to 2000 years was analysed. On the basis of multiple cross-sectional analysis,a cohort study was applied to further analysis. The results showed that the total mortality and male's mortality of leukemia in Urumqi residents were almost the same as that of national level, while the female's mortality of leukemia was lower than that of national level. The cohort study showed that mortality of adolescent with leukemia descended with the increase of age, but mortality of gerontic leukemia ascended with the increase of age. In conclusion, there is a significant relationship between the morbific age of leukemia and prognosis of patients with leukemia.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Female ; Humans ; Infant ; Infant, Newborn ; Leukemia ; mortality ; Male ; Middle Aged ; Time Factors