Retinal degenerative disease is the leading cause of visual loss,the mechanism is not clear and has no effective treatment method.In recent years,the field of stem cell research has made great progress.Stem cells have the potential of differentiating into all the body cells.Embryonic stem cells (ESCs) can be used to differentiate a variety of retinal cells,which has brought a new promise for the treatment of retinal degeneration disease.However,the most important step of this treatment is how to differentiate ESCs into photoreceptor cells and retinal pigment epithelium (RPF) cells.In this paper,we introduced the recent progress in the differentiation methods of ESCs into retinal cells,which contains spontaneous differentiation method,co-culture method,growth factors and small molecules induced method,adherent monoculture method,three dimensional differentiation method.

Objective To find out the differences on the knowledge,attitude and behavior related to Hepatitis B transmission between Hepatitis B patients and control group( healthy people) and to analyzing the risk factors of Hepatitis B transmission. Method Adopted case-control study, 50 people who were Hepatitis B patients and 100 people as control group were selected to conduct retrospective investigation. Results Skin wound, disqualified medical treatment,unhealthy habits and manners are high risk factors to Hepatitis B transmission.The knowledge,attitude and behavior of Hepatitis B are very important to prevent Hepatitis B transmission. Conclusion There was a misunderstanding on the knowledge about Hepatitis B transmission among Hepatitis B patients. They thought daily contact was the key way of HBV transmission. They underestimated the blood transmission and sexual transmission.We need to conduct more propaganda on the knowledge of Hepatitis B prevention to change peoples' unhealthy habits and behavior.

Objective To investigate the possible effects of COL8A1 on the proliferation, invasion and drug sensitivity of murine hepatocarcinoma cell line Hca-F, we used an RNA interference (RNAi) approach to silence COL8A1 expression. Methods The expression levels of COL8A1 in HcaF/siRNA cells were assessed by RT-PCR and Western blot. The inhibitory effect of RNAi on Hca-F cell invasion in vitro was demonstrated by ECM invasion assay. The in vitro proliferative ability and drug sensitivity of COL8A1-deficient cells were determined by MTT assay. Results The expression of COL8A1 was significantly reduced in COL8A1/siRNA cells after 30h transfection, compared with both the RNAi control and the Hca-F cells. The reduced COL8A1 expression also attenuated the proliferative, invasive ability, as well as increased drug sensitivity of Hca-F/siRNA cells. Conclusion Our current results indicate that the expression of COL8A1 functionally mediates tumor cell proliferation, invasion, and drug sensitivity, and is a potential target for therapeutic anti-cancer drugs.

Objective To investigate the effect of ellagic acid extracted from gallnut on multiple myeloma SP2 / 0 cell line and related mechanisms. Methods Multiple myeloma SP2 / 0 cell line was treated for 48 h with different concentrations of ellagic acid in vitro. Cell morphology,proliferation,apoptosis and cell cycle were analyzed with microscope,MTT experiment and flow cytometry,respectively. Tumor cell proliferation and apoptosis-related gene expression of COX-2 were detected by Western blotting. Results Cell cycle was arrested at the G1 phase 48 h after treatment with ellagic acid, the cell in G1 were (55. 21±3. 01)% , (64. 48 ± 0. 43)% , (75. 10 ± 2. 46)% , respectively, with significant difference as compared with control group [(34. 04±1. 74)% ,P<0. 01]. Cell suppression rate (21. 18±5. 92)% ,(44. 58±3. 43)% and (70. 15±2. 90)% ,respectively, in 20,40 and 60 μg·mL-1 ellagic acid treatment groups. Compared with the control group,the differences were significant (P<0. 01). Cell apoptosis rate was (9. 60 ± 0. 56)% , (19. 30 ± 1. 51)% and (35. 10 ± 5. 26)% , respectively, in 20,40 and 60 μg·mL-1 ellagic acid treatment groups,with significant differences as compared with the control group[(3. 23±0. 85)% ,P<0. 01]. With the increase of drug concentration,COX-2 expression was decreased. Conclusion Ellagic acid can inhibit myeloma SP2 / 0 cell proliferation and promote apoptosis.

Objective Production of autotoxin protein in sf9 insect cells with biological activity. Methods Autotaxin cDNA was cloned into pFastBacTMHTA from melanoma cell by extraction of total RNA using TRIzol method and RT-PCR. Bacmid-ATX is isolated from transformed competent bacterial DH10 which carries Bac genomic sequences and transfected into sf9 using lipofectamine 2000. Recombinant ATX virus was amplified in sf9 and further used for infection and expression of ATX protein. Two step purification product using HistrapTMHP and Hiload 16/600 Suerdex 200pg was determined for lysophospholipase D (lysoPLD) activity. Results Correct insertion of PCR fragment is confirmed by BamH I/Xho I digestion and sequencing. ATX virus can infect sf9 and induced enzymatic activity. Column purification and SDS-PAGE resulted 95% in purity and 6mg/liter in yield with significant lysoPLD activity. Conclusion ATX Baculovirus was successfully constructed that can infect sf9 cells and express active lysoPLD. Production of active ATX can be used for crystalography studies and screening for small pharmaceutical inhibitors.

Objective:To develop a two-step method for purification of monoclonal antibody rhTF243 protein from mouse ascites by using hydrophobic charge induction chromatography(HCIC) and affinity chromatography with protein A sepharose CL-4B.Methods:The ascites was first purified by HCIC after centrifugation and filtration.Then the fraction containing the protein of interest was directly purified by affinity chromatography with protein A sepharose CL-4B.Results:The purity of the obtained monoclonal antibody was up to 97% with recovery of 73% and of high activity.Conclusion:The method for purification of monoclonal antibody is developed using HCIC and Protein A affinity chromatography and the obtained antibodies are of high purity and activity.

Objective To ascertain the utility and difference of sonography with echo-tracking (ET) technique and pulse-Doppler to assess vascular stiffness in rats with hypercholesterolemia and atherosclerosis.Methods Sonography associated with ET technique and pulse-Doppler were used to measure stiffness parameter (β),arterial compliance (AC),distensibility coefficient (DC),one-point pulse wave velocity (PWVβ),resistence index(RI),peak systolic velocity(PSV),end-diastolic velocity(EDV) and EDV/PSV of the aorta in cholesterol-fed SD rats (group T1,n =10,for 4 weeks;group T2,n =10,for 12 weeks) and normal control rats(group C1,n =10;group C2,n =10).All parameters and blood biochemical markers[total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-CH) and highdensity lipoprotein cholesterol (HDL-CH)] among groups were analyzed with ANOVE factor analysis.Correlation was analyzed with Pearson analysis.Light microscopic evaluation were used to demonstrate atherosclerotic changes in the aorta.Results The PWVβ value and PSV of the aorta between group T1 and T2 were significantly different (P =0.001,P ＜0.05).The β,PWVβ values of the aorta in group T1 and T2 were significantly higher than those of group C1 and C2 (P ＜0.05).AC and DC values of the aorta in group T1 and T2 were significantly lower than those of group C1 and C2 (P ＜0.05).Correlation analysis showsed that RI was positively correlated with systolic pressure(P ＜0.05).All parameters had correlated with each other among β,PWVβ,AC,DC,TG,TC,systolic pressure and diastolic pressure.DC and AC were negatively correlated with β and PWVβ,also DC was negatively correlated with TG.Light microscopy confirmed morphologic typical changes of aortic atherosclerosis in group T1 and T2.Conclusions Sonography with the ET method compared with pulse-Doppler is much more sensitive and it can be used to evaluate tissue elastic changes in arterial walls associated with atherosclerosis and hypercholesterolemia.PSV can reflect atherosclerosis of rat's abdominal aorta well,but pulse-Doppler is limited in the diagnosis of earlier atherosclerosis period.

The local field potentials (LFPs) underlying specific behavior were recorded and analyzed in this paper from primary motor cortex (M1) with several medium, such as the self-made single channel micro-electrodes, the system of multi-channels physiological signal acquisition and processing and so on. During the experiment, the specific behavior was divided into four periods according to the changes of the recorded LFPs and the changes of the specific behavior recorded simultaneously in rats. The four periods were named prophase of catching period, planning period, catching period and the completion period, respectively. Then several methods were used for the analysis of the LFPs by MATLAB, such as time domain analysis, power spectral distribution analysis and time-frequency analysis. The results suggested that the LFPs which were caused by different behavior from a large number of movement-related neurons of M1 during the specific behavior in the process of catching play an important part in the "code" guiding role in rats. The results demonstrat that the LFPs of M1 may provide a feasibility to discriminate the motor behavior of forelimb.
Animals ; Brain-Computer Interfaces ; Electrodes, Implanted ; Evoked Potentials, Motor ; physiology ; Feeding Behavior ; physiology ; Male ; Microelectrodes ; Motor Cortex ; physiology ; Rats ; Rats, Wistar

Objective To investigate clinical characteristics and emergency managements of postrenal acute renal failure(ARV)induced by melamine in infant.Method Fluid therapy for urine alkalization and hydration,cistoscope drainage and peritoneal dialysis step by step were exerted in those who had both a history of certain milk intake and ARF according to the definition of pediatric ARF which developed by Pediatric Nephrology Assembly of Chinese Pediatric Association in 1994.Results Thirty-four postrenal ARF cases with anuria due to melamine in Nanjing Children's Hospital of Nanjing Medical University were involved in the study.Seventy cases(50%)re-ceived fluid therapy only.Nine cases(26.5%)received fluid thempy and eistoscope drainagemand 4 cases (11.8%)received fluid therapy and cistoscope drainage and peritoneal dialysis.Four cases(11.8%)received ur-gent peritoneal dialysis due to severe hyperkalemia.All cases(100%)survived.The urine pH at the first day.the second day,and after the second day in those who just pass away urine were 6.1±1.0、6.5±0.7.5.3±0.4,respectively(F=4.563,P=0.026).Conclusions Fluid therapy for urine alkalization and hydration and stop sequential thempy are effective in infant with postrenal ARF induced by melamine.

Recording and extracting characteristic brain signals in freely moving animals is the basic and significant requirement in the study of brain-computer interface (BCI). To record animal's behaving and extract characteristic brain signals simultaneously could help understand the complex behavior of neural ensembles. Here, a system was established to record and analyse extracellular discharge in freely moving rats for the study of BCI. It comprised microelectrode and micro-driver assembly, analog front end (AFE), programmer system on chip (PSoC), wireless communication and the LabVIEW used as the platform for the graphic user interface.
Animals ; Behavior, Animal ; physiology ; Brain ; physiology ; Electroencephalography ; methods ; Microelectrodes ; Rats ; Signal Processing, Computer-Assisted ; instrumentation ; Telemetry ; instrumentation ; User-Computer Interface