Cell therapy has a very promising potential for end-stage liver diseases (ESLD).Fetal liver stem/progenitor cells (FLSPCs) have advantages of safety,high survival and proliferation rates,and a small volume,all which make them ideal for liver disease stem cell therapy.During the early phase of our study,we applied a three-step separation method to enrich FLSPCs and obtained a separation efficiency similar to that of the flow-cell sorting method.Additionally,using a fulminant hepatic failure model in rats,we have demonstrated that FLSPCs can contribute to morphological and functional recovery of the liver.This manuscript will discuss how FLSPCs can be induced to accurately differentiate into hepatocytes and cholangiocytes and how FLSPCs maintain self-renewal.The Notch signaling plays a critical role in regulating the differentiation and self-renewal of many types of stem cells.Our previous findings have shown that the Notch signaling plays an important role in FLSPCs differentiation into hepatocytes.Therefore,the Notch signaling might be involved in the differentiation and self-renewal of FLSPCs.We conducted a study on the regulatory effects and relative molecular mechanisms of the Notch signaling on FLSPCs and found the corresponding interfering target,which might become an index for the clinical application of FLSPCs.

The proliferating cell nuclear antigen (PCNA) gene expression was blocked and retinal pigment epithelium (RPE) proliferation was inhibited by using antisense oligonucleotides (AS-ODN) mediated by liposome, to find a new genetic therapy of proliferative vitreoretinopathy (PVR). RPE cells cultured in vitro were transfected with synthetic fluorescence labled AS-ODN mediated by liposome-Lipofectamine, and the intracellular distribution and persistence time of AS-ODN were dynamically observed. AS-ODN (0.07, 0.28 and 1.12 micro mol/L and sense oligonucleotides (S-ODN with the same concentrations as AS-ODN) mediated by liposome were delivered to the RPE cells cultured in vitro, and CPM values were measured by 3H-TdR incorporation assay and analyzed statistically by variance by comparison with blank control group. Expression of PCNA mRNA in RPE cells was detected by in situ hybridization after the treatment of different concentrations of PCNA AS-ODN and S-ODN, and the average optic density (AOD) was measured by image analysis system and was subjected to q-test and correlation analysis with CPM. Our results showed that AS-ODN mediated by liposome could quickly aggregate in cellular plasma and nuclei in 30 min and 6 h, and stayed for as long as 6 days. AS-ODN (0.28 and 1.12 micro mol/L) markedly suppressed proliferation of RPE cells in a dose-dependent manner with the difference being statistically significant (P < 0.05 and P < 0.01, repectively) as compared with blank control group. AOD was well correlated with CPM (r = 0.975). It is concluded that liposome could increase transfection efficiency of AS-ODN in RPE cells, and AS-ODN could sequence-specifically suppress PCNA mRNA expression and proliferation of human RPE cells.

Objective:To compare the results of two different measures assessing prospective memory (PM) in elderly schizophrenia,and explore their diagnostic validity.Methods:Fifty patients who were diagnosed as schiz-ophrenia according to the criteria of Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-Ⅳ)and 50 normal elderly people were enrolled in the study.Laboratory paradigm prospective memory task and the Chinese version of the Cambridge prospective memory Test (C-CAMPROMPT)were administered to measure thelevel of EBPM and TBPM.The Wechsler Adult Intelligence Scale-Fourth Edition,Continuous Performance Test-I-dentical Pairs were used to evaluate IQ and attention.Results:Whether in the laboratory paradigm or C-CAM-PROMPT,schizophrenia group performed worse than normal controls on PM total scores[7(0,16)vs.12(0,16),14 (4,34)vs.25(11,36)],EBPM[3(0,8)vs.6(0,8),7(2,16)vs.14(4,18)]and TBPM [3(0,8)vs.6(0,8),6(2,18) vs.12(4,18)],and patients had comparable performances in PM subtypes[-1.0(-2.2,1.0)vs. -1.0(-2.1, 0.8),-1.9(-3.4,0.8)vs. -1.8(-2.9,1.6)].In the C-CAMPROMPT,the scores of EBPM and TBPM were negatively related to age(r=-0.36 --0.40,P<0.001 ),but there was no significant relationship between PM performance and age in the laboratory paradigm task.The area under ROC curve of the laboratory paradigm and C-CAMPROMPT were 0.73 and 0.85.While 8 and 19 as the cut off value,the sensitivity were 0.60 and 0.74,and the specificity were 0.76 and 0.90,respectively.Conclusion:The two measures have confirmed that the elderly schizophrenia have comparable performances in EBPM and TBPM.Both the C-CAMPROMPT and the laboratory paradigm have moderate level of diagnostic validity,but the former is slightly higher and more susceptible to aging.

Background:Visceral hypersensitivity is considered to be one of the major pathophysiological mechanisms of irritable bowel syndrome. Aims:To investigate the expression of TRPV1 and electrophysiological characteristics of colon-specific dorsal root ganglion( DRG)neurons in rat model of visceral hypersensitivity. Methods:Twenty 10-day-old rats were randomly divided into two groups. In model group,visceral hypersensitivity was induced by colorectal administration of acetic acid;while in control group the same amount of saline was administered. Colon-specific DRG neurons were labeled retrogradely by injection of DiI,a fluorochrome,into the colon wall. Expression of TRPV1 in DRG neurons was detected by immunofluorescence and the electrophysiological characteristics of DRG neurons was detected by using patch-clamp technique. Results:In model group,the expression rate of TRPV1 in colon-specific DRG neurons was significantly higher than that in controls(46. 1% vs. 36. 6% ,P <0. 01),the average rheobase was significantly decreased[(57. 80 ±1. 32)pA vs.(73. 45 ± 4. 51)pA,P < 0. 05],while the frequency of action potentials(APs)in response to doubling rheobase stimulation was significantly increased[(8. 20 ± 1. 10)Hz vs. (4. 54 ± 0. 66)Hz,P < 0. 05]. Score of abdominal withdrawal reflex(AWR)under a 60 mm Hg colorectal distention was positively correlated with the expression rate of TRPV1 and the frequency of APs in response to doubling rheobase stimulation(r = 0. 87 and r = 0. 73,P < 0. 01),but was negatively correlated with the rheobase(r = - 0. 81,P < 0. 01)in model group. Conclusions:Increased expression of TRPV1 and excitability in colon-specific DRG neurons might be a crucial step in formation of visceral hypersensitivity.

Objective:To preliminarily study the characteristics of ventricular synchronicity in patients of idiopathic pulmonary arterial hypertension (IPAH) with different heart function.
Methods:Our research included in 2 groups:IPAH group, n=40 patients intermittently collected in our hospital from 2010-10 to 2014-12 and Control group, n=23 normal subjects from the same period of time. Tissue Doppler echocardiography was conducted to compare interventricular systolic and diastolic time to peak (Ts and Te), their differences (Te-Ts) and (Te-Ts) of left ventricle (LV), right ventricle (RV), interventricular septum (IVS) between 2 groups.
Results:Ts and Te of RV were longer than Ts and Te of LV and IVS in both groups, P<0.05. Compared with Control group, by heart function reducing, IPAH group showed different Ts and Te of LV and IVS, P<0.05 and Ts was gradually prolonged, Te was gradually shorter;while Te of RV became longer firs tand then became shorter thereafter. In IPAH group, the patients with heart function grade IV had the longer Ts of LV and IVS than those with heart functionII, III;the patients with heart function IV had the shorter Te of LV, RV and IVS than those with heart function II, III, all P<0.05. In IPAH group with different heart function, Te between RV-IVS and RV-LV were different from Control group, all P<0.05;Te between LV-IVS in heart function IV patients had the largest difference from Control group, P<0.05. Compared with Control group, IPAH group had gradually decreased Te-Ts of LV and IVS by heart function reducing asgrade II>III>IV, all differences had statistic meaning.
Conclusion:IPAH patients with different right heart function may have interventricular systolic and diastolic de-synchronicity;by reduced heart function, interventricular Te-Ts could be specifically shortened.

Objective:To compare structure and function of mite Der f 3,Der p 3 and Eur m 3.Methods: Obtained mite allergens amino acid sequences from the International Union of Immunological Societies nomenclature database .Then physiochemical characterization,sequence alignment,secondary structure,three dimensional (3D) structure and epitopes of three proteins were analyzed by bioinformatics methods.Results:According to results of alignment ,Der f 3,Der p 3 and Eur m 3 displayed 88.51%sequence iden-tity.Der f 3,Der p 3 and Eur m 3 all contained three active sites and two trypsin functional domains ,which showed high identity of amino acid residues.Active sites of three proteins ,which closing to each other in three dimensional (3D) structure,constituting the active center of the enzyme.Secondary and 3D structure of three proteins all contains α-helices,β-sheets and random coils.Epitopes analysis revealed that Der f 3,Der p 3 and Eur m 3 all have 5 main potential epitopes located in random coils.Epitope sequences of Der f 3,Der p 3 and Eur m 3 overlapping in three domains (peptides of 79-81aa,129-135aa and 172-174aa),but the residues in these three domains were not identical.Conclusion:These studies lay the foundation for biochemical and genetic analysis of these 3 allergens,and may contribute to vaccine development for allergen-specific immunotherapy.

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Objective To investigate the clinical effects of biofeedback therapy on constipation-predominant irritable bowel syndrome(IBS-C).Methods 30 patients with IBS-C were given to biofeedback therapy,5 times every treatment course.and then their clinical symptoms,mental state and quality of life before the treatment and at the end of treatment with biofeedback therapy were respectively evaluated by symptom scores,self-rating anxiety scale (SAS),self-rating depression scale (SDS),and short form 36 health survey questionnaire (SF-36).Results Post-treatment with biofeedback therapy,there were very significant decrease in total and subscales scores of bowel symptom including abdominal pain/discomfort,abdominal distension,abnormal appearance of defecation,abnormal process of defecation((12.31±2.01) vs (19.16±2.17),(2.95±0.57) vs (5.04±1.04),(2.64±0.92) vs (4.25±1.09),(3.66±1.09) vs (5.10±0.57),(3.06±1.08) vs (4.77±0.95) respectively,P＜0.01).The integration of SAS and SDS were obviously decreased after treatment((39.53±6.39) vs (44.43±7.89),P＜0.05;(40.70±8.38) vs (46.46±8.74),P＜0.05),the SF-36 scores were also improved in five dimensions including rolephysical,social-functioning,vitality,role-emotional and mental health((74.16±21.25) vs (57.0±39.40),(86.21±13.54) vs (75.54±20.96),(75.16±13.42) vs (64.66±20.54),(78.87±28.36) vs (57.76±46.26),(81.60±16.08) vs (71.20±22.04) respectively,P＜0.05).Conclusion Biofeedback therapy can improve the clinical symptoms,alleviate the moods of anxiety and depression,and improve the quality of life in patients with IBS-C.

Objective To investigate the roles of glutamate signaling pathway in melanin transfer.MethodsEpidermal melanocytes and keratinocytes were isolated from human foreskin tissue followed by purification and primary culture.Immunofluorescence microscopy was conducted to observe the intracellular distribution of N-methy-D-aspartate receptor 1 (NMDAR1) and NMDAR2A in melanocytes.Some melanocytes were classified into 4 groups to be pretreated with MK801 (the NMDAR antagonist dizocilpine maleate) at 100μmol/L for 5 minutes followed by treatment with NMDA(an NMDAR agonist) at 100 μmol/L (MK801-pretreated group 1),pretreated with MK801 at 100 μmol/L for 1 hour followed by treatment with NMDA at 100μmol/L (MK801-pretreated group 2),treated with MK801 at 100 μmol/L for 5 minutes (MK801 group),treated with NMDA at 100 μmol/L for 5 minutes (NMDA group),respectively,then,confocal microscopy was performed to measure the intracellular calcium (Ca2+) concentration of the melanocytes.The distribution of β-tubulin was visualized by confocal microscopy in melanocytes treated with MK801 at 100 μmol/L for 24 hours.Some melanocytes and keratinocytes were cocultured with or without MK801 at 100 μmol/L for 24 or 48 hours,then,scaning microscopy was carried out to observe the junction structure between melanocytes and keratinocytes,and alkali method coupled with spectrophotometric analysis to determine melanin content in keratinocytes.Results The intracellular calcium concentration of melanocytes was decreased by MK-801,but increased by NMDA at 100 μmol/L,and the increase was blocked by the pretreatment with MK-801 for 5 minutes or 1 hour.After incubation with MK-801 at 100 μmol/L for 24 hours,a more intense staining for β-tubulin was observed around the nuclei of melanocytes.There was a significant reduction in the number of filopodia on the surface of and between melanocytes and keratinocytes after treatment with MK-801 at 100 μmol/L for 48 hours.Also,the content of melanin(represented as the absorbance value at 375 nm) transferred from melanocytes into keratinocytes was statistically reduced in coculture system treated with MK-801 at 100 μmol/L compared with that without treatment (0.158 ± 0.003 vs.2.203 ± 0.006,t =6.323,P ＜ 0.01 ).Conclusions The glutamate signaling pathway exerts a regulatory effect on intracellular calcium concentration of distribution of β-tubulin in,filopodia formation of melanocytes and melanin transfer between melanocytes and keratinocytes.

Objectives To compare prospective memory (PM) deficits with retrospective memory (RM) deficits and to explore the correlation between PM and RM in chronic schizophrenia. Methods Fifty chronic schizophrenia pa?tients and fifty healthy controls were recruited. The PM performance [event-based PM (EBPM) and time-based PM (TB?PM)] were evaluated by the Chinese version of the Cambridge Prospective Memory Test (C-CAMPROMPT); working memory (WM) was evaluated by the digital span subtest (DS);immediate auditory logical memory (IALM), delayed audito?ry logical memory (DALM), immediate visual reproduction memory (IVRM) and delayed visual reproduction memory (DVRM) were evaluated by the logical memory and visual reproduction subtest. The score of each test was transformed to comparable standard score. Results Patients performed significantly worse on EBPM [(7.9 ± 3.4) vs. (13.7 ± 2.9)], TBPM [(6.9±3.6) vs. (13.0±3.2)], DS [sequence:(5.8±2.0) vs. (7.5±2.2);backward:(6.5±1.9) vs. (8.2±2.8)], IALM [(8.3±3.1) vs. (11.9 ± 2.5)], DALM [(7.4 ± 3.7) vs. (11.8 ± 2.6)], IVRM [(8.0 ± 2.7) vs. (11.2 ± 3.8)], and DVRM [(7.7 ± 3.5) vs. (10.8 ± 2.7)] scores than controls (P<0.05). The extent of deficits of EBPM and TBPM were greater than those of DS (sequence and backward), IALM, DALM, IVRM and DVRM (P<0.05), but not DALM (P>0.05). The performance of PM in chronic schizophrenia was significantly related to DS (sequence and backward), IALM, DALM and DVRM (P<0.05), but not IVRM (P=0.155). Conclusion:There are greater prospective memory deficits than retrospective memory deficits in chron?ic schizophrenia and the prospective memory deficits are correlated with the retrospective memory deficits in chronic schizophrenia.