AlM: To explore the roles of neuronal axon-guidance molecules Slit3 and Robo4 receptor in corneal neovascularization ( CNV ) by study their expression in neovascularized cornea of rats.
METHODS: CNV models were established by implantation pellets containing basic fibroblast growth factor ( bFGF ) into corneal stroma. CNV models were measured by biomicroscopy photography. lmmunohistochemical staining and imaging analysis system were used to detect the expression of Slit3 and Robo4 in the models after 1, 4, 7, 10 and 14d.
RESULTS:The area of CNV and the expression of Slit3, Robo4 were increased in CNV models compared to that in normal cornea and reached highest level on 7d. And the expression level of Slit3 and Robo4 were significantly correlated with the size of CNV on every time point except 1d (r=0. 84-0. 91, all P<0. 05).
CONCLUSlON: The expression of Slit3 and Robo4 may be related to the CNV development. They are potential therapeutical target for CNV.

Objective Acquiring genetic information of Y-SNPs and Y-STRs genetic makers from samples with the surname of Kong, which is useful for exploring the correlation between surname and Y chromosome in forensic applications studies.Methods Two multiplex genotyping assays and SNaPshot assay were used to analyze 255 unrelated male blood samples who share the same surname Kong and 330 unrelated male blood samples obtained randomly. 17 Y-STRs were typed for the surname Kong population samples. The software Arlequin 3.5.1.2 and the program Network 4.6.1.1 were used for data statistical analysis.Results 13 haplogroups were observed. The highest haplogroup frequency in the two populations were O3a2c1a-M117 (21.57%, 14.85%). 196 haplotypes in Kong population deifned by 17 Y-STRs locus were obtained and the haplotype diversity was 0.9939. 14-12-25-28-19-15-12-19-12-11-12-22-12-11-14-10-19 is the typical haplotype. Median Joining algorithm and Mismatch Distribution were adopted to analyze the Y-STR haplotye under haplogroup O3-M122, and the result shows that there are two “central star” distribution. Conclusion Combined with Y-SNP and Y-STR analysis showed that the Kong population had experienced complicated exchanges and expansion or continued growth, which has more than one surname origin. Hence, its population genetic structure and historical differences have potential applications in forensic science.

Adenocarcinoma ; secondary ; Aged, 80 and over ; Cryptorchidism ; pathology ; Humans ; Male ; Prostatic Neoplasms ; pathology ; Testicular Neoplasms ; secondary

Objective Study on the absorption,distribution,transformation,excretion and toxicity of four forms of arsenic in rats.Methods Four thousand and fifty data were obtained from 28 days animal metabolism experiments of 81 rats exposed to the four As species.Distribution,metabolism,excretion,and subacute toxicity of 4 As species were compared by analyzing the arsenate [As(V)],arsenite [As(Ⅲ)],methyl arsenate (MMAV) and dimethyl arsenate (DMAV) content of rat feces (excrement and urine),blood,seven organs,and liver and kidney pathology.Results After oral administration of each As species,82.9％ of As(Ⅲ),85.1％ of As(V),95.0％ of MMAV and 96.2％ of DMAV were accumulatively secreted via feces and urine,while 16.2％ of As(Ⅲ),14.1％ of As(V),4.65％ of MMAV and 0.120％ of DMAV were detected in blood.The DMAV accumulated in blood and urine after dosing As(Ⅲ),As(V) and MMAV,and the content of DMAV in the blood was 8 times greater than urine.Four kinds of arsenic had effects on rat liver and kidney function,the influence on liver was acute phase effect while that on renal was cumulative effects.The effect of As(Ⅲ) was the most significant,followed by MMAV.Four kinds of arsenic could cause tissue deformation and inflammatory cells infiltration,but showed no difference between groups.Conclusion The distribution,metabolism,excretion,and toxicity of the four kinds of arsenic in rats were different in varying degrees.The research of health risk for arsenic should consider its different forms.

Objective To observe the levels of alanine aminotransferase(ALT), total bilirubin(TBIL), hyaluronic acid (HA), procollagen type Ⅲ aminoterminal peptide (P Ⅲ NP) and larninin (LN) in the sera of rats infected with Clonorchis sinensis (C. sinensis) after treatment of albendazole combined with milkvetch root. Methods Thirty-two healthy adult Wistar rats were randomly divided into four groups with 8 in each based on body mass: control group, non-treatment group, Albendazole group(ALB group) and albendazole combined with milkvetch root group(ALB+MR group). The rats in non-treatmen, ALB and ALB+MR groups were infected orally with metacercariae of C. sinensis 50 per rat. The rats in control group were mock-infected with saline. The rats in ALB group were treated to each rat with 50 mg/kg alhendazole for 5 days, and ALB+MR groups were given to same treat with albendazole, meanwhile each rat injected with 800 mg/kg milkvetch root intraperitoneally for 30 days. All rats were killed after infestation 14 weeks and their sera samples were collected to detect ALT, TBIL, HA, PⅢNP, LN. Results There were statistically significant differences in the levels of ALT and LN in the sera of rats between groups(F=31.40,11.82, P<0.01). Compared with control[ (47.88±4.88)U/L, (51.20±4.12)μg/L], the levels of ALT and LN in rats in non-treatment group [(85.50±9.65)U/L, (64.20±4.18) μg/L] and ALB group [(65.29± 7.78) U/L, (58.23±2.55) μg/L] were significantly increased (P<0.05). Compared with non-treatment group, the levels of ALT and LN in rats in ALB group and ALB+MR groups[(50.25±9.29)U/L, (53.68±5.63)μg/L] were significantly decreased(P<0.05), and they decreased more obviously in ALB+MR group (P<0.05). There were statistically significant differences in the levels of TBIL, PⅢNP and HA in the sera of rats between groups (χ2=15.309,21.418,19.759, P<0.01). Compared with control[(0.700±0.350)μmol/L, (26.085±4.075)μg/L, (81.935±42.550)μg/L], the levels of TBIL, PⅢNP and HA in rats in non-treatment group(2.400 μmol/L, 46.220 μg/L,310.885 μg/L) and ALT group(1.200 μmol/L,36.540 μg/L, 178.010 μg/L) were significantly increased(P<0.05). Compared with non-treatment group, the level of TBIL in ALT+MR group(0.750 μmol/L), the levels of pⅢNP and HA in ALT and ALT+MR group(30.470,100.240 μg/L) were significantly decreased(P< 0.05). The levels of TBIL, PⅢNP and HA decreased more obviously in ALB+MR group(P<0.05). Conclusions The liver function in rats infected with C. sinensis is abnormal. The liver function and fibrosis are improved after treatment with albendazole or albendazole combined with milkvetch root. The treatment of albendazole combined with milkvetch root is more effective.

Apoptosis Regulatory Proteins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; virology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; virology ; Newcastle disease virus ; physiology ; Oncolytic Viruses ; physiology ; Tumor Cells, Cultured

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

OBJECTIVE To study the effect of polygonatum polysaccharide on zebrafish with Alzheimer disease. METHODS Zebrafish were trained in T maze for 7 d. The 40 zebrafish successfully trained were divided into 4 groups:blank group, model group, positive group and polygonatum polysaccharide group. Model group, positive group and polygonatum polysaccharide group were put in AlCl3100μg·L-1 for 6 d. The positive group was exposed to Huperzine A solution 4μg·L-1, and the polygonatum polysaccharide group was exposed to polygonatum polysaccharide solution 6 g·L-1 for 6 d. The model group was not treated, and the blank group was not treated. Each stage of zebrafish was recorded by video, and the time of each group in the EC region was analyzed. After administration, the brain tissue was taken out and the expression of N-cadherin, P38 and p-P38 protein factors was determined by Western blotting. RESULTS In behavior, the analysis of the time spent in the EC area, the blank group, the positive group and the polygonatum polysac?charide group were compared with the model group, respectively, there were statistically significant differences (P<0.05). At the protein level, compared with the model group, the P38 and p-P38 proteins in the positive group and the polygonatum polysaccharide group were down-regulated, while the N-cadherin protein was up-regulated, with statistical difference (P<0.05). CONCLUSION Polygonatum polysaccharide can improve the learning and memory ability of zebrafish with Alzheimer disease by up regulating the protein level of N-cadherin and hindering P38 phosphorylation.

Objective To report the phototoxicity effects of a novel photosensitizer ZnPcS4-BSA on photodynamic therapy (PDT) towards human U251 glioma cells in vitro. Methods The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra to determine the optimal incubation time. Human U251 glioma cells were incubated with ZnPcS4-BSA of various concentrations and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay.Flow cytometer was used to detect apoptosis.Gene expressions of vascular endothelial growth factor (VEGF) were detected by Real-Time PCR in the U251 cells after PDT and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results The uptake of ZnPcS4-BSA by U251 glioma cells reached the maximum after incubation for 4 hours.ZnPcS4-BSA of different concentrations without laser irradiation had no significant effects on cell survival rates (P＞0.05).Without ZnPcS4-BSA incubation,compared with 0,25,50,100,200 J/cm2 groups, the cell survival rate of the 400 J/cm2 group was significantly lower (P＜0.05), whereas no significant difference was found between any other two groups. When the U251 glioma cells incubated with 30 μ mol/L ZnPcS4-BSA for 4 hours underwent laser irradiations of 25,50,100,200 J/cm2,the cellular survival rates significantly decreased with the increased energy densities (P＜0.05). When the U251 glioma cells incubated with ZnPcS4-BSA of 20,40,60,80,100 μ mol/L for 4 hours underwent laser irradiation of 200 J/cm2, the cellular inhibition rates significantly increased with the increased concentrations (P ＜0.05). Compared with controls, the cellular apoptosis and VEGF expression significantly increased in the U251 glioma cells incubated with ZnPcS4-BSA of 20 μmol/L after laser irradiation of 100 J/cm2 (P＜0.05). Conclusion The novel ZnPcS4-BSA is a good photosensitizer for PDT towards U251 glioma cells,because the ZnPcS4-BSA-mediated PDT can induce effective apoptosis of the targeted cells.

OBJECTIVESTo evaluate the three different methods in monitoring the lamivudine-resistant HBV mutants in lamivudine-treated patients with chronic hepatitis B.

METHODSThe sensitivity and specialty of melting curve assay and polymerase chain reaction microplate nucleotide hybridization-enzyme linked immunosorbent assay (PCRmnh-ELISA) were compared with those of mismatch polymerase chain reaction-restriction fragment length polymorphism (mPCR-RFLP) and sequence analysis, through detection of HBV YMDD mutants in 44 serums from chronic hepatitis B patients receiving lamivudine monotherapy at the time of viral breakthrough.

RESULTSmPCR-RFLP assay was more sensitive (10(4) copies/ml) than both PCRmnh-ELISA (10(5) copies/ml) and melting curve assay (10(6) copies/ml). 26 YMDD mutants and 18 wild-types were determined by the means of mPCR-RFLP. Among the 26 mutants, only 16 and 18 mutants were found by melting curve assay and PCRmnh-ELISA, respectively. Whereas, out of the 18 wild-types, 2 and 13 mutants were detected by melting curve assay and PCRmnh-ELISA, respectively. To confirm the different results determined by the three methods in 16 samples, sequence analysis was conducted and showed that the rate of consistency with sequencing was 93.8% by mPCR-RFLP, 43.8% by melting curve, and 18.8% by PCRmnh-ELISA, respectively (chi2=18.7, P<0.01).

CONCLUSIONSThe mPCR-RFLP assay is reliable to monitor HBV YMDD mutations. Melting curve assay and PCRmnh-ELISA should be further improved to increase their sensitivity and specialty.

Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Drug Resistance, Viral ; Female ; Gene Products, pol ; genetics ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sensitivity and Specificity