AIM: To evaluate the efficacy of combined therapy for amblyopia in children by making use of pattern visual evoked potential ( P-VEP) game.
METHODS: This was a prospective case control study. These asthenopic children were divided into two groups. The control group ( 66 eyes of 49 patients ): occlusive therapy with glasses, cover, precision work, red light treatment and so on, later the stereo vision training was added. The experimental group (72 eyes of 52 patients):conventional methods mentioned above with P - VEP games.
RESULTS: The total effective rate and cure rate of experimental group in 6mo were higher than those of control group. The overall effective rate was 94. 4% in the experimental group and 83. 3% in the control group. There was a statistically significant difference between them (P<0. 05).
CONCLUSION: The comprehensive therapy by making use of P-VEP game is an individualized effective new way in treating amblyopia.

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

OBJECTIVETo investigate the effect of aluminum trichloride on the abnormal phosphorylation of tau protein in SH-SY5Y cells.

METHODSSH-SY5Y cells were assigned to control group and aluminum trichloride exposure groups (200, 400, and 800 µmol/L Al(3+)). The cell morphology was observed after 48 hours of exposure; the cell viability was measured by CCK-8 assay; total protein was extracted from the cells, and the expression of phospho-tau (p-tau) 181, 231, 262, and 396 and tau 5 was measured by Western blot.

RESULTSAs the Al(3+) concentration rose, the number of living SH-SY5Y cells decreased, and the synapses of the cells retracted. The viability of cells exposed to 800 µmol/L Al(3+) was significantly lower than that of the control group (P < 0.05). The 200, 400, and 800 µmol/L Al(3+) exposure groups showed significantly higher expression of p-tau 181, 231, and 396 and tau5 than the control group (P < 0.05), and the 800 µmol/L Al(3+) exposure group showed significantly higher expression of p-tau 262 than the control group (P < 0.05).

CONCLUSIONUnder the present experimental conditions, aluminum trichloride has toxic effect on SH-SY5Y cells and can lead to abnormal expression of p-tau 181, 231, and 396 and tau 5 at low Al(3+) concentration.

Aluminum Compounds ; toxicity ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chlorides ; toxicity ; Humans ; Phosphorylation ; tau Proteins ; metabolism

The aim of this article is to provide methods for the isolation and identification of pancreatic stem cells and cell source for research and therapy of diabetes. ICCs were isolated by collagenase IV digesting and then cultured; epithelial cells were purified from monolayer cultured ICCs. The growth curve of the epithelial cells was measured by MTT. The expression of molecular markers in the cells was identified by immunohistochemical staining. The surface markers in the epithelial cells were analyzed by FACS. Epithelial cells were purified from isolated human fetal ICCs and passaged 40 times, and 10(6) - 10(8) cells were cryopreservated per passage. The growth curve demonstrated that the epithelial cells proliferated rapidly. The epithelial cells expressed PDX-1, PCNA, CK-7, CK-19, Nestin, Glut2, and Vimentin, but Insulin was undetected. The cells expressed CD29, CD44, and CD166, but did not express CD11a, CD14, CD34, CD45, CD90, CD105, and CD117. Taken together, these results indicate that self-renewable epithelial cells can be isolated and purified from human fetal pancreas. These also show that the epithelial cells originate from ducts and have the characteristics of pancreatic stem cells.
Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; Cell Survival ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; Fetus ; Flow Cytometry ; Homeodomain Proteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Integrin beta1 ; analysis ; Islets of Langerhans ; Proliferating Cell Nuclear Antigen ; analysis ; Stem Cells ; cytology ; metabolism ; Trans-Activators ; analysis

ABSTRACT:OBJECTIVE To observe the neuroprotective effects of ilex pubescens total flavonoids(IPTF) on the rats with cerebral ischemic preconditioning(CIP) and the expressions of BDNF,GDNF and VEGF in the cortex and hippocampus CA1 area,to explore the mechanisms of IPTF on improving cerebral ischemic tolerance.METHODS CIP was performed by bilater-al common carotid artery occlusion for 10 min in rats.72 h later,cerebral ischemia reperfusion(I∕R)was established by middle cerebral artery occlusion(MCAO)for 2 h.Wistar rats were randomly devided into five groups,Sham group,I∕R group,CIP+MCAO group,IPTF high dose group(200 mg∕kg) and IPTF low dose group(100 mg∕kg).The behavioral injury was detected by neurologic deficit scores(NDS),and the infarct areas were detected by TTC staining.The expressions of BDNF,GDNF and VEGF in cortex and hippocampus CA1 area were measured by immunohistochemical staining.RESULTS IPTF decreased the neurologic deficit scores and the infarct areas.It also increased the positive area and integral optical density of BDNF,VEGF in cortex and CA1 area of rats with CIP.CONCLUSION The mechanisms of IPTF on improving cerebral ischemic tolerance may be related with the up-regulation of the endogenous protein BDNF and VEGF.

Objective To evaluate the homogeneity of the proficiency test samples to verify whether it meets the requirements of the comparison in international reference laboratory.Methods According to the Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing (CNAS-GL03),14 biochemical indexes including ALT,AST,ALP,GGT,CK,LDH,TP,T-Bil,Urea,Cr,UA,Glu,TG and TC in the past three years (from 2014 to 2016)were tested by the Roche detection system Modular P800 Biochemical analyzer.The mean ((x)),standard deviation (s)and coefficient of variation (CV)of the samples were calculated.One-way analysis of variance (ANOVA)was performed and the guideline of Ss ≤0.3σ was used to evaluate the between-bottle differences.Results The results showed that the CVs of AST in RELA 2014A and B were higher than 2.0％.The CVs of CK were over 2％ in all tests except for RELA 2016B.The results of ANOVA for RELA samples demonstrated that the F value of CK was over the critical value 4.39,which was statistically significant (P ＜ 0.05).The F values of the ALT and T-Bil in 2015B and the Cr in 2014A were also over 4.39 (P ＜ 0.05) respectively,while the F values of other measurements were less than the critical value of F,indicating there was no statistical significance (P ＞ 0.05).The CK measurement data Ss ＞ 0.3σ in all the samples by the guideline of Ss ≤ 0.3σ,suggesting that there was a between-bottle difference in CK.The other indexes were Ss ≤ 0.3σ,showing no between-bottle difference in those items.Conclusion There were significant differences between the bottles of the CK item in the past three years,and the homogeneity of all the other items in the samples could meet the requirements of Proficiency Testing for the international reference laboratory.

Objective To understand the characteristics of sexual network of 321 HIV positive MSM and HIV infection status among sexual partners in Zhejiang Province. Methods A cross-sectional study selected HIV positive MSM diagnosed between September, 2015 and September, 2016 were conducted in Hangzhou, Ningbo, Wenzhou in Zhejiang Province using self-administrated questionnaire. Information related to socio-demographic characteristics, risky behavior, size of sexual network were collected. Results A total of 321 HIV positive MSM were recruited in this study. 71.76％ aged 20 to 40 years old and 51.71％ were native. The rate of self-reported sexual behavior with stable homosexual partners, casual homosexual partners, spouse and other heterosexual partners at investigation were 37.78％, 45.00％, 36.56％ and 66.67％. The rate of never using condom with stable homosexual partners, casual homosexual partners, spouse were 27.91％, 16.36％ and 44.00％. HIV positive rate among stable homosexual partners, casual homosexual partners, spouse and other heterosexual partners were 28.26％ , 3.77％, 11.43％ and 0.00％. Conclusion The size of sexual network was large, especially in casual sexual network. Unsafe sexual behavior, like consistent condom use was low and multiple sexual partners . HIV infection was high among stable homosexual partners and spouse.

Objective:To investigate the value of autoantibodies and serum levels of IgG4 and CA19-9 in the diagnosis of IgG4 associated cholangitis (IgG4-SC).Methods:Detect the serum IgG4 and CA19-9 of 41 clinical cases of IgG4-SC patients,162 clinical cases of non IgG4-SC patients and 40 healthy human serum samples by immunoassay and direct chemiluminescence methods, also detect the antinuclear antibodies (ANA),anti neutrophil antibody (ANCA),anti smooth muscle antibody (SMA) and anti mitochondrial antibody (AMA) of the above serum samples by indirect immunofluorescence and analyze the detection results.Results:①The positive rates of ANA,ANCA,SMA and AMA in patients with IgG4-SC were 41.46%,7.32%,0 and 2.44%.Among them,the positive rate of ANA was significantly different from that of the normal control group(P<0.01),and the positive rate of SMA and AMA was significantly different from that of non IgG4-SC group(P<0.01),and so as the positive rate of ANCA do with that of PSC group.②The number of serum IgG4 and CA19-9 increased samples were significantly compared with the normal control group (P<0.01);the area under the ROC curve (AUC) was 0.979 and 0.646,respectively,and P<0.05.Conclusion:The high level of serum IgG4 and CA19-9 and autoantibody detection are of great accuracy and important clinical value in the differential diagnosis of IgG4-SC.

OBJECTIVETo study the relationship between the percentage of polypronuclear zygotes and clinical pregnancy following IVF.

METHODSWe collected the data of 954 IVF cycles, and according the percentage of polypronuclear zygotes in the IVF cycles, allocated them to Groups A (without polypronuclear zygotes) , B (with < 30% polypronuclear zygotes) and C (with > or = 30% polypronuclear zygotes). Then we analyzed the relationship between the percentage of polypronuclear zygotes and the rate of clinical pregnancy.

RESULTSCompared with Group A, Group C showed a significantly lower rate of clinical pregnancy (43.2% vs 28. 1%, P < 0.05), while Group B exhibited a markedly higher rate (43.2% vs 52.36%, P < 0.05) and obviously decreased polypronuclear zygote formation with the increase of age (35.6% vs 24.1%, P < 0.05).

CONCLUSIONThe percentage of polypronuclear zygotes in IVF cycles may serve as a prognostic indicator of the clinical outcome.

Adult ; Age Factors ; Female ; Fertilization in Vitro ; methods ; Humans ; Ovulation Induction ; Pregnancy ; Pregnancy Rate ; Zygote

OBJECTIVETo evaluate the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from HLA-matched sibling donor (MSD allo-HSCT) for severe aplastic anemia (SAA).

METHODSThe clinical data of 41 SAA patients received MSD allo-HSCT from May. 2003 to Aug. 2011 were analyzed retrospectively. 24 patients were male, 17 were female. Median age was 23 (5 - 43) years old. 28 patients had SAA-I, 9 had SAA-II, and 4 had post-hepatitis aplastic anemia. 17 patients received allogeneic bone marrow (BM) transplantation (allo-BMT), and 24 received allogeneic peripheral blood stem cell (PBSC) transplantation (allo-PBSCT). The conditioning regimens: 20 patients received cyclophosphamide (CY) + anti-thymocyte globulin (ATG) + fludarabine (Flu), 21 received CY + ATG + Flu+ cytarabine (Ara-C) ± busulfan (Bu)/melphalan (Mel). Prophylaxis for graft-versus-host disease (GVHD): 25 patients received cyclosporine (CSA) plus short-term methotrexate (MTX), 16 received tacrolimus (FK506) plus short-term MTX. The median number of infused CD34(+) cells were 3.48 (2.39 - 4.80)×10(6)/kg in allo-BMT and 2.95 (1.27 - 5.98)×10(6)/kg in allo-PBSCT, respectively.

RESULTSHematopoietic reconstitution was observed in all 41 patients (100%). The median time of neutrophils (ANC) reached to 0.5×10(9)/L and platelets (PLT) reached to 20×10(9)/L were 14 (10 - 23) days and 19 (8 - 38) days, respectively. 12 patients developed acute GVHD (aGVHD), out of which 11 developed grade I-II aGVHD, and one developed grade IV. 2 patients occurred chronic GVHD (cGVHD), out of which one with local cGVHD and the other with extensive. 4 patients occurred graft rejection (GR), all of them recovered haemopoiesis and survived after donor PBSC infusion. 5 patients (12.2%) died, out of which one died of extensive cGVHD, and 4 died of invasive fungal infections (IFI). Median follow-up time was 23 (3 - 79) months. 36 patients survived. 5-year estimated overall survival (OS), disease free survival (DFS), and transplant-related mortality (TRM) was (81.1 ± 9.0)%, (68.4 ± 11.0)%, and (18.9 ± 9.0)%, respectively. Univariate analysis showed that lover OS had significant correlation with receiving PBSCT, occurrence of aGVHD, the number of infused CD34(+) cells no more than 2.5×10(6)/kg, the number of red blood cell (RBC) transfusion before transplant more than 30 U and occurrence of IFI after transplantation (P = 0.034, 0.001, 0.006, 0.000, 0.001, respectively). Occurrence of aGVHD had significant correlation with the disparity between donor and recipient ABO blood groups, the number of PLT transfusion more than 100 U, and the number of RBC transfusion more than 30 U before transplantation, the number of infused CD34(+) cells no more than 2.5× 10(6)/kg (P = 0.019, 0.038, 0.005, 0.005, respectively). The occurrence of GR had significant correlation with the number of PLT transfusion more than 100 U before transplantation (P = 0.038).

CONCLUSIONMSD allo-HSCT is an effective therapy for patients with SAA. Lower number of blood transfusion before transplantation, use of BMT, more number of infused CD34(+) cells can effectively prevent and treat aGVHD and IFI after transplantation, which may improve the efficacy of MSD allo-HSCT for SAA.

Adolescent ; Adult ; Anemia, Aplastic ; therapy ; Child ; Child, Preschool ; Female ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Retrospective Studies ; Siblings ; Tissue Donors ; Treatment Outcome ; Young Adult