Objective: This work aimed to construct stable MCF-7 cell sublines from which staphylococcal nuclease domain con-taining 1 (SND1) expression was interfered to analyze the effect of SND1 silencing on the proliferation and metastasis of MCF-7 cells. Methods: The lentivirus that could mediate SND1 silencing was prepared. MCF-7 cells were infected with the lentiviruses to construct stable sub-cell lines. Quantitative real-time polymerase chain reaction and Western blot analysis were employed to determine SND1 ex-pression level. MTS, wound healing, and transwell assays were applied to analyze the effect of SND1 silencing on the proliferation, mi-gration, and invasion of MCF-7 cells. Results: A lentivirus expression vector that contains sequences encoding shRNAs targeting SND1 and an shRNA negative control were successfully established. The lentiviruses (LV-SH1, LV-SH2, LV-SH3, and 和 LV-NC) were then collected and packaged. Stabilized MCF-7 sublines were prepared through infection with lentiviruses. The most efficient MCF-7 stable cell subline, MCF-SH3, was selected for SND1 silencing. Compared with the control cell, the proliferation, migration, and inva-sion potential of MCF-SH3 were significantly decreased. Conclusion: SND1 could promote the proliferation, migration, and invasion of breast cancer cells. Thus, silencing SND1 expression will inhibit such proliferation, migration, and invasion. These results indicated that the unusual expression of SND1 is associated with breast cancer and may participate in cancer progression by affecting prolifera-tion, migration, and invasion.

Objective To establish a high performance liquid chromatography (HPLC) method for the determination of baicalin and naringin inQinbei mixture.Methods The HPLC system consisted of the Fortis-C18(4.6 mm × 250 mm, 5μm) column, and the mobile phase consisted of MeOH:0.4% H3PO4 (42:58), and the flow rate was 1.0 ml/min, and the UV detector was set at 280 nm, and the column temperature was 30℃.Results The linear response range of baicalin was 0.062-0.930μg. The linear response range of naringin was 0.033-0.492μg. The average recovery of baicalin was 98.11% (RSD=1.62%). The average recovery of naringin was 96.78% (RSD=1.74%).Conclusions The method is simple, rapid, accurate and repeatable. It can be applied in determination of baicalin and naringin inQinbei mixture.

Objective To investigate the effects of ursolic acid (UA) on cisplatin (CDDP)-induced expres_sion of transient receptor potential vanilloid 1 (TRPV1) in mouse cochlea .Methods Sixty BALB/c mice were ran_domly divided into 4 groups (15 mice in each group) and received introperitoneal injection once daily for 5 days:Control group (normol saline) ,UA group (80 mg/kg/day) ,CDDP group (4 .5 mg/kg/day) ,and CDPP (4 .5 mg/kg/day) plus UA group (80 mg/kg/day) .The expression of TRPV1 in mouse cochlea was determined by immuno_histochemistry ,microscope image analysis and western blot ,and auditory thresholds were evaluated by auditory brainstem response (ABR) measurement .ResuIts The expression of cochlea TRPV1 and ABR threshold shift was significantly increased in the mice treated with CDDP (P< 0 .05) ,as compared with control mice .These effects were prevented by UA treatment (all P<0 .05) .Furthermore ,a linear relationship analysis revealed that the ex_pression of cochlea TRPV1 was significantly correlated with ABR threshold shift(|r|>0 .7 , P<0 .05) .ConcIusion UA effectively attenuated CDDP -induced ototoxicity and improved auditory function through inhibition of TR_PV1 .

Objective:To evaluate the bone alteration subject to remodeling and analyze the esthetic result following immediate implant placement of incisors.Methods:In this study,20 patients (1 3 women,7 men)were involved,who needed implants for incisors of maxilla.The patients received 23 im-mediate implants totally.On the day of surgery and 6 months after the implants were placed,Cone beam CT (CBCT)was taken.The thickness of the alveolar ridge and the vertical change of marginal bone levels onthe mesial and distal aspects of theimplants were measured using the computer software (Planme-caRomexis Viewer 3.6.0.R).The evaluation of esthetic result by labial convexity score (LCS)and pa-pilla index score (PIS)were analyzed pre-operation and one year after the final crown was delivered.The statistics with paired-t test for the measurement data and Willcoxon test for rating data were done by SPSS 20.0.Results:The survival rate in the two-year follow-up was 1 00%.The measuring point 1 (MP1 ), MP2,MP3 and MP4 (0,2,4,6 mm apical to the implant platform,respectively)got significant altera-tions after 6 months of the follow-up.These differences were statistically significant (P<0.05 ).The major alteration happened at MP1 and MP4,which got (-0.89 ±2.06)mm and (-0.75 ±1 .28)mm reduction of the alveolar,respectively.The marginal alveolar ridge resorption was (-0.42 ±1 .24)mm and(-0.91 ±1 .96)mm for Ankylos System and Replace System,respectively,and the difference was not statistical significant .The esthetic results were quite acceptable.Before treatment,1 8 incisors rated 3 for LCS,and 2 incisors rated 4 for LCS;after final restoration,only 5 incisors rated 3 for LCS,and 1 4 incisors rated 2 for LCS.Before treatment,1 5 incisors rated 3 for PIS;after final restoration,1 3 incisors rated 3 for PIS.There was no statistically significant difference for the PIS pre-operation and 1 year after final restoration,while there was statistically significant negative change for LCS.Conclusion:Even fol-lowing the proper surgical technique,the alveolar ridge wall still can’t be maintained after immediate im-plant placed in fresh socket of incisors.The inter-dental papilla could be well maintained,while due to the remodeling of labial bone,labial convexity will inevitably collapse.Therefore immediate implant still has esthetic risk.

OBJECTIVEThis study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.

METHODThe HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.

RESULTThe results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.

CONCLUSIONThis method is able to be a scientific basis of quality assessment according to its convenient and reliable.

To investigate the effect and possible mechanism of echinacoside-containing serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Rat bone marrow mesenchymal stem cells were cultivated by the whole bone marrow adherence method. The 3rd generation of cells were divided into 3 groups: the blank control group, the classic osteogenic-induced group and the 10% echinacoside-containing serum group. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. The ex- pression of ZHX, protein was detected by Western blot technique. RT-PCR technique was used to detect the expression of ZHX₃mRNA. According to the result, the expressions of the alkaline phosphatase and osteocalcin in the classic osteogenic-induced group and the 10% echinacoside-containing serum group were significantly higher than that of the blank control group (P <0. 01). And expressions of the alkaline phosphatase activity and osteocalcin in the 10% echinacoside-containing serum group were significantly higher than that in the classic osteogenic-induced group (P < 0.01). Meanwhile, the classic osteogenic-induced group and the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the black control group, with significant differences (P < 0.01); the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the classic osteogenic-induced group, with a significant difference (P < 0.01). In conclusion, 10% echinacoside-containing serum can promote the differentiation of the bone marrow mesenchymal stem cells cultured in vitro. Its mechanism may be correlated with the increase in the ZHX₃expression.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Glycosides ; blood ; pharmacology ; Homeodomain Proteins ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry ; Transcription Factors ; genetics ; metabolism

Bel1, a transactivator of prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have shown that Bel1 bears a nuclear localization signal (NLS), but its amino acid sequence remains unclear and the corresponding importins have not been identified. In this report, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment could direct nuclear localization, which accords with the consensus sequence K(K/R)X(K/R) of monopartite NLS. Point mutation experiments revealed that K218, R219, and R221 are essential for the nuclear localization of Bel1. The results of the GST-pulldown showed that the Bel1 fragment with residues 215-223, which bears the NLS, interacts with KPNA1, KPNA6, and KPNA7. This result suggests that KPNA1, KPNA6, and KPNA7 maybe involved in Bel1 nuclear translocation.
Cell Line ; Cell Nucleus ; genetics ; metabolism ; virology ; Humans ; Nuclear Localization Signals ; genetics ; metabolism ; Protein Binding ; Protein Transport ; Retroviridae Infections ; genetics ; metabolism ; virology ; Retroviridae Proteins ; chemistry ; genetics ; metabolism ; Spumavirus ; chemistry ; genetics ; physiology ; Trans-Activators ; chemistry ; genetics ; metabolism ; alpha Karyopherins ; genetics ; metabolism

Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; immunology ; Dengue ; virology ; Dengue Virus ; chemistry ; genetics ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Viral Envelope Proteins ; chemistry ; genetics ; immunology

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.

0.05).2.There were significant differences in the levels of TNF-? after treated for 4 weeks(P0.05).4.There were no significances between the 2 groups before and after treated 6 weeks in the change of liver.Conclusions Matrine injection can inhibit HCMV DNA replication.It can also control the expression of TNF-? and regulate the function of the immune system.There fore matrine injection has an antivirus efficacy in HCMV infection.