Objective To study the effects of tonifying kidney-yin on activities of embryo stem cell. Methods Embryonic stem cell line CRL-1825 was choosed as model cell in present study, and tonifying kidney-yin herb serum was added to CRL-1825 cell. The influence of tonifying kidney-yin on stem cell differentiation, cell cycle, apotosis and express of key genes were observed. Results Tonifying kidney-yin can inhibit stem cell differentiation and apotosis, promote stem cell proliferation and progression of cell cycle. In addition, tonifying kidney-yin can up-regulate Wnt, Oct4 gene expression, and down-regulation P16INK4a expression. Conclusions Tonifying kidney-yin can promote stem cell proliferation and maintain the state of stem cell.

Objective　To investigate p16 gene expression in salivary adenoid cystic carcinoma (ACC) and the relation between p16 gene expression and lung metastasis. Methods　p16 gene protein was detected in 36 cases of ACC by ABC immunohistochemical method. Results　58.3 % (21/36) cases showed positive staining. Higher incidence for positive staining was found in ACC without lung metastasis 76 % (19/25) than those with lung metastasis 36.6 % (4/11) with remarkable statistical difference. Expression of p16 was correlated with the P-TNM stageing (P＜0.05), but was not with pathologic pattern (P＞0.05). Conclusion　The p16 gene may play an important role in suppressing the expression of metastatic potential in human salivary ACC.

Objective To investigate the treatment of symptomatic patent ductus arteriosus (PDA) in very low birth weight preterm infants. Methods From January 1, 2008 to December 31, 2010, 78 very low birth weight preterm infants (birth weight＜1500 g) were diagnosed as symptomatic PDA. Among which, 42 cases administered orally with indomethacin (0.2 mg/kg, every 12 hrs for three times) were taken as treatment group, while five cases in this group who failed to indomethacin treatment were interrupted with video-assisted thoracoscopic surgery. And 36 cases who did not receive treatment for ductus arteriosus were taken as control group. The clinical outcomes, complications and prognosis of these patients were observed. Results There were no significant differences between the gentle percentage, gestational age, diameter of ductus arteriosus, rate of complicated with heart failure, sepsis, neonatal respiratory distress syndrome and intraventricular hemorrhage of two groups (P>0.05, respectively). The ductus arteriosus closed in 33 patients of treatment group (78.6%) and in nine patients of control group (25.0%)(χ2=22.39,P=0.000). There were no significant differences in serum creatinine level and platelet count between before and after the treatment in treatment group(P>0.05). Compared with control group, the treatment group had lower incidence of intraventricular hemorrhage (z=1.167, P=0.030), shorter duration of oxygen therapy [(8.0±5.5) d vs (13.3±9.3) d, t=2.225, P=0.032] and shorter hospital stay [(39.0±7.7) d vs (43.6±10.6) d, t=2.229, P=0.029]; while the incidence of bronchopulmonary dysplasia and necrotizing enterocolitis were similar (P>0.05). The five cases of PDA who received video-assisted thoracoscopic surgery were successfully interrupted with no residual shunt left, while three of them had lung infections and one had pleural effusion, but no pneumothorax and infant death associated with surgery occurred. Conclusions Symptomatic PDA of very low birth weight preterm infants should be treated actively. Oral indomethacin was an effective and safe method to cure the PDA in these infants. Surgical ligation under video-assisted thoracoscopic surgery after failure of indomethacin treatment might be a good option.

Objective To evaluate the effects of 131I treatment for Graves hyperthyroidism patients with leucopenia and the alteration of WBC levels after treatment.Methods A total of 257 Graves hyperthyroidism cases were retrospectively studied after 131I treatment.Based on baseline WBC count,119 cases with WBC count ＜4.0 × 109/L before 131I treatment were diagnosed with leucopenia and 138 cases had normal WBC.There were no significant differences in age,weight of thyroid,131I uptake rate in 24 h,dose of 131I and course of the disease between the two groups (all t ＜ 0.972,all P ＞ 0.05).WBC,lymphocyte,neutrophil and platelet counts were recorded before and 1,3,6 and 12 months after 131I therapy.The therapeutic effects were graded as clinical cure,improvement,invalidation and hypothyroidism.Statistical analyses,including independent samples t test,x2 test and one-way analysis of variance,were performed using SPSS 13.0.Results The WBC levels in the leucopenia group were (3.49 ± 0.43) × 109/L,(4.06 ±0.98) × 109/L,(4.20 ±1.04) × 109/L,(4.37 ±0.93) × 109/L and (4.88 ± 1.20) × 109/L before 131I treatment and 1,3,6,12 months after 131I treatment,respectively; while,those in the normal WBC group were (5.70 ± 1.08) × 109/L,(5.50 ± 1.14) × 109/L,(5.74 ±0.99) × 109/L,(5.95 ± 1.14) × 109/L and (6.07 ± 1.17) × 109/L,respectively.There was no statistically significant difference of WBC levels before and 1 month after 131 I treatment (t =1.662,P ＞ 0.05) in the normal WBC group,but WBC levels at those timepoints were significantly different in the leucopenia group (t =3.816,P ＜ 0.05).In the leucopenia group,there was no significant change of lymphocytes before and after 131I treatment,while the average neutrophil count showed an increasing trend during the 1,3,6 and 12 months after 131I treatment (F =40.583,t:1.468-11.264,all P ＜ 0.05).The average platelet counts at 6 and 12 months after 131I treatment were (187.80 ± 36.03) × 109/L and (206.88 ± 26.04) × 109/L respectively,which were higher than that before 131I treatment (F =9.735,t =2.604,4.892,all P ＜0.05).In the normal WBC group,there were no statistical differences of WBC changes before and after 131I treatment except for a lower lymphocyte count at 1 month after 131I treatment than that at baseline ((1.79 ± 0.37) × 109/L vs (1.99 ±0.63) × 109/L;F =12.868,t =3.284,both P ＜ 0.05).The treatment effects of the two groups were not significantly different (x2 =0.739,P ＞ 0.05).Conclusions 131I treatment presents similar therapeutic effects on Graves hyperthyroidism patients with leucopenia and those with normal WBC levels.WBC levels in patients with leucopenia may recover to baseline during the post-treatment follow-up.Thus 131I treatment is a safe and effective method for Graves hyperthyroidism patients with leucopenia.

Objective: To observe the value of texture analysis based on gray level co-occurrence matrix in differential diagnosis of glioblastoma multiform (GBM) and primary central nervous system lymphoma (PCNSL). Methods: Image data of 46 cases of GBM (GBM group) and 36 cases of PCNSL (PCNSL group) confirmed by pathology were retrospectively analyzed. MaZda software was used to manually draw ROI on the maximum level of tumor on enhanced-T1WI and ADC images, and then texture parameters including angular second moment energy (AngScmom), Entropy, Contrast, correlation (Correlat) and inverse difference moment (InvDfMom) were extracted respectively. Multivariate Logistic regression model was constructed for texture feature parameters with statistically significant differences between 2 groups, and ROC curve was used to analyze differential diagnostic efficiency of GBM and PCNSL based on texture parameters and Logistic regression model. Results: There were significant differences of AngScMom, Contrast, Correlat and Entropy on enhanced-T1WI images, also of AngScMom, Correlat and Entropy on ADC images between GBM group and PCNSL group (all P<0.01). Parameters with statistical significances between 2 groups were brought into the binary Logistic regression analysis, and the Logistic regression model was obtained. ROC curve showed that the efficiency of Entropy for identifying GBM and PCNSL was the highest both on enhanced-T1WI and ADC images, AUC was 0.81 and 0.72, the sensitivity was 78.26% and 56.52%, and specificity was 77.78% and 80.56%, respectively. AUC of Logistic regression model for identifying GBM and PCNSL was 0.92, the sensitivity and specificity was 91.30% and 83.33%, respectively. Conclusion: Texture feature based on gray level co-occurrence matrix was helpful for differential diagnosis of GBM and PCNSL.

Objective To elucidate the function of phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway underlying the regulation of Na+-I-symporter (NIS) and the influence of different levels of iodine on PI3K-AKT signaling pathway in lactating breast cells.Methods The primary cultured mammary gland cells were divided into three groups:①control group [0 μmol/L LY294002 + 0 μg/L insulin-like growth factor Ⅰ (IGF-Ⅰ)];②stimulation group (50 μg/L IGF-Ⅰ);③inhibition group (40 μmo]/L LY294002 + 50 μg/L IGF-Ⅰ).In addition,the cells were treated with different iodine contents (0,5,50,1 000,3 000 μg/L) for low iodine groups 1 and 2,iodine group,high iodine groups 1 and 2,and IGF-Ⅰ (50 μg/L) was used to stimulate PI3K-AKT signaling pathway.The expressions of AKT and NIS mRNA and protein were determined by real-time quantitative PCR and Western blotting,respectively.Results The expression of AKT mRNA (1.497 ± 0.550) in stimulation group was higher than that in inhibition group (0.777 ± 0.108,P ＜ 0.05),while the expression of NIS mRNA and protein in stimulation group (0.783 ± 0.187,0.618 ± 0.103) was lower than those in inhibition group (2.430 ± 1.423,1.417 ± 0.250,all P ＜ 0.05).With the iodine concentration increasing,except high iodine group 1 (1.090 ± 0.356),the expression of AKT mRNA in low iodine groups 1 and 2,iodine group,high iodine group 2 (1.758 ± 0.893,1.320 ± 0.538,1.003 ± 0.006,0.745 ± 0.307) tended to decline;total AKT protein (0.640 ± 0.106,0.601 ± 0.081,0.583 ± 0.089,0.555 ± 0.097,0.532 ± 0.023) and NIS mRNA (2.259 ± 0.682,1.823 ± 0.332,1.409 ± 0.366,1.321 ± 0.405,1.150 ± 0.454) tended to decline in low iodine groups 1 and 2,iodine group,high iodine groups 1 and 2;except low iodine group 2 (0.484 ± 0.179),NIS protein expression tended to decline (0.556 ± 0.199,0.502 ± 0.179,0.455 ± 0.126,0.435 ± 0.138);however,except low iodine group 2 (0.076 ± 0.045),the p-AKT protein expressions (0.078 ± 0.049,0.079 ± 0.040,0.085 ± 0.055,0.095 ± 0.051) were on the rise.Conclusion PI3K-AKT signaling pathway may play an inhibition role in the expression of NIS in lactating breast cells.

This article introduces the basic theories about atomic force microscope (AFM) and electron microscope (EM), respectively. New applications of each microscopic technology in regenerative medicine, covering both material science and life science, are discussed. The advantages or disadvantages of the kinds of microscopes in working conditions, sample preparation, resolution and the like, are discussed and compared systematically to make clear each scope of applications. This could be a useful guide for selecting the appropriate microscopic analysis in research work about regenerative medicine.
Humans ; Microscopy, Atomic Force ; methods ; trends ; Microscopy, Electron ; methods ; trends ; Regenerative Medicine ; methods ; trends

Objective To investigate the role of PI_3 K/Akt signal pathway in Ephrin-Al gene mediated invasion,metastasis of Huh-7 cells.Methods Western blot was used to test the protein expression of phosphatidylinositol 3-kinase(PI_3 K)and mitogen-activated protein kinase(MAPK)after Huh-7 cells were treated with Ephrin-A1/Fc fusion protein.According to the protein expression,LY294002 was used to block PI_3 K/Akt pathway specifically,then p-Akt protein expression,mobility and invasive ability of Huh-7 cells were examined.Results In Huh-7 cells actived by Ephrin-Al/Fc fusion protein,p-Akt expression was higher than that in control group(t=4.564,P＜0.05),but there was no difference of p-p38MAPK expression between Ephrin-Al/Fc fusion protein group and IgG/Fc fusion protein group(P＞0.05).PI_3 K/Akt pathway was specifically blocked by LY294002,the p-Akt protein expression decreased in Huh-7 cells,and the mobility and invasive ability mediated by Ephrin-Al in Huh-7 cells decreased(P＜0.05).Conclusions PI_3 K/Akt pathway effects an important role in mobility and invasive ability of Huh-7 cells mediated by Ephrin-A1.

Objective To explore the effect of wild type or mutant parkin gene expression on the growth of human hepatocellular carcinoma cell line Huh-7. Methods The parkin (wild type or mutant) expression vector and empty vector were transferred into Huh-7 cell lines with LipofectAMINE 2000 reagents. The positive clones that expressed parkin gene stably were chosen by G418 and checked by reverse transcription-polymerase chain reaction (RT-PCR) to check the DNA sequences. The cytobiological behaviors of those positive clones were analyzed by cell proliferation assay and tumorigenesis in nude mice. Results Huh-7 cell lines that expressed wild type or mutant parkin gene stably were successfully established. The growth of wild type parkin-expressed cells was obviously inhibited compared with the control cells transfected with empty vectors(t= 3. 875, P= 0. 031).The volume of tumor formed by wild type parkin-expressing cells in nude mice was also significantly reduced (t=8. 228,P=-0. 003). Mutant parkin gene expression had a slight effect on the growth of Huh-7 cells in vitro and in vivo (P＞0.05). Conclusion The re-expression of wild type parkin gene can favor the malignant phenotype revision of Huh-7 cells. Therefore, it might be a good candidate for tumor suppressor gene associated with HCC.

AIM:To investigate the specific anti-tumor effects of mature dendritic cells ( DCs) transfected with amplified mucin 1 ( MUC1) mRNA in vitro.METHODS:DCs separated and purified from the peripheral blood mononu-clear cells were induced in vitro and then identified by flow cytometry .pcDNA3.1(+)-MUC1 plasmid was constructed and was able to transcribe MUC1 mRNA in vitro.The MUC1 mRNA was transfected into DCs by electroporation .MUC1-trans-fected DCs were used to induce T cells to be cytotoxic T-lymphocytes .Quantitative real-time PCR was performed to assess MUC1 mRNA expression in transfected DCs .The proliferation of T cells was examined by MTT assay .The proportion of CD8 +cells in the T cells was determined by flow cytometry and the specific cytotoxicity was measured by LDH assay .The secretion of IFN-γwas detected by ELISA .RESULTS: The marker gene expression in the DCs transfected with MUC 1 mRNA was significantly increased compared with control group , peaking at 24 h.The transfection group showed the higher capacity to stimulate the proliferation of T cells compared with control group when the ratio of DCs to T cells was 1∶10.The proportion of CD8 +cells in transfection group was higher than that in control group .The lethal effect of special cytotoxic T-lymphocytes on target cells in transfection group was stronger than that in control group .The level of IFN-γin the cell su-pernatant of transfection group was higher than that in control group .CONCLUSION:DCs plus MUC1 mRNA by electri-cal transfection induces specific anti-tumor effects , which provides an experiment evidence of using MUC 1 as a target for immunotherapeutic strategy against non-small cell lung cancer .