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.

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 evaluate the effect of selenomethionine (Se-Met) against ultraviolet B (UVB)-induced oxidative damage to human HaCaT keratinocytes,and to explore its possible mechanisms.Methods Cultured HaCaT cells were divided into several groups:normal control group receiving no treatment,Se-Met groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours respectively,UVB groups irradiated with UVB of 30,60 and 90 mJ/cm2 respectively,Se-Met + UVB groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours firstly,then irradiated with UVB of 30,60 and 90 mJ/cm2 respectively.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to estimate cellular proliferative activity,flow cytometry to detect cell apoptosis,colorimetry to evaluate superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and to determine malondialdehyde (MDA) levels.Statistical analysis was carried out by using factorial design analysis of variance (ANOVA),one-way ANOVA and least significant difference (LSD) test.Results Factorial design ANOVA showed that UVB radiation had an inhibitory effect on the proliferative activity of HaCaT cells (F =128.04,P ＜ 0.05),which significantly decreased along with the increase of UVB doses,with significant differences between the three UVB groups (P ＜ 0.05).Se-Met pretreatment also affected cellular proliferative activity (F =5.95,P ＜ 0.05),which was significantly increased in Se-Met (10 nmol/L-1 μmol/L) + UVB groups compared with the UVB groups at corresponding doses (all P ＜ 0.05).There was no significant interaction effect on cellular proliferative activity between UVB radiation and Se-Met pretreatment (F =1.65,P ＞ 0.05).The apoptosis rate of HaCaT cells in the 30-mJ/cm2 UVB group was 31.9％ ± 2.67％,significantly higher than that in the normal control group (4.1％ ± 0.67％,P＜ 0.05) and in the 10-,50-,100-,200-nmol/L and 1-μmol/L Se-Met + 30-mJ/cm2 UVB groups (21.9％ ± 3.72％,17.2％ ± 1.67％,4.6％ ±-0.85％,7.5％ ± 1.86％ and 13.5％ ± 1.95％ respectively,all P ＜ 0.05).Similarly,SOD and GSH-Px activities were significantly weaker (both P ＜ 0.05),while MDA levels were higher (all P ＜ 0.05) in the 30-mJ/cm2 UVB group than in the normal control group;however,there was a significant increase in SOD and GSH-Px activities but a decrease in MDA levels in the Se-Met (10 nmol/L-1 μmol/L) + 30-mJ/cm2 UVB groups compared with the 30-mJ/cm2 UVB group (all P ＜ 0.05).Conclusions Se-Met can reduce UVB-induced oxidative damage to HaCaT cells,likely by enhancing antioxidase activity and decreasing oxygen radicals.

Objective To evaluate the efficacy,adverse events of gemcitabine vs.5-FU with radiotherapy for locally advanced pancreatic carcinomas.Methods Between January 2007 and January 2011,a total of 56 patients with locally advanced pancreatic carcinomas was included and clinical data were retrospectively analyzed.All patients received 3-DCRT radiotherapy with individual dose of 1.8 ～ 2 Gy,5 times per week,and total dose of 45 ～ 50.4 Gy for 25 ～ 28 fractions,and received concurrent chemotherapy (5-FU or gemcitabine).The patients (n =30) in gemcitabine group were treated with gemcitabine (500 rng/m2 at the 1st,8th,15th,22nd day,at 10 mg · (m2)-1 · min-1,through micro-pump) during radiotherapy; 3 weeks after radiotherapy the patients received gemcitabine infusion at a dose of 800 mg · (m2)-1 · d-1,one time per week,for 3 or 4 cycles.The patients (n =26) in 5-FU group were treated with 5-FU (500 mg/m2 at the 1 ～ 5th day per week,IV),the cycle was repeated every 2 weeks during radiotherapy; 3 weeks later the patients received 5-FU infusion at a dose of 800 mg · (m2)-1 · d-1,the 1st ～5th day per week,the cycle was repeated every 2 weeks ; with a total of 3 or 4 cycles.The efficacy and adverse events were observed,and the patients were followed until June 2013,and the median survival,1 year and 2 year survival was calculated.Results Of the 56 patients,the overall response (CR + PR) rate was 73.2％,and it was 65.3％ in radiotherapy with 5-FU group,80.0％ in radiotherapy with gemcitabine group (P ＜ 0.05).The overall one and two year survival rate was 48.2％ and 14.3％,while median survival was 15.2 months,and the corresponding values were 42.3％,11.5％,13.3 months in radiotherapy with 5-FU group,and 53.3％,16.7％,16.6 months in radiotherapy with gemcitabine group,and the survival difference between the two groups was not statistically significant (P =0.071).At the end of treatment,the pain-relief rate (VAS score ＜4) of the 56 patients was 83.3％,it was 75.0％ in 5-FU group and 90.0％ in gemcitabine group.In radiotherapy with gemcitabine group,the incidence of 3～ 4 grade myelosuppression was significantly higher than that in radiotherapy with 5-FU group,and the difference between the two groups was statistically significant (20.0％ vs 7.6％,P ＜ 0.05).Conclusions For the locally advanced pancreatic carcinomas,radiotherapy with gemcitabine can improve pain-relief and prolong survival compared with radiotherapy with 5-FU,but the incidence of adverse event of myelosuppression is higher.

Objective To study the role of human MutS homolog 2 (hMSH2), a newly identified protein ligand that was recognized by Vγ9δ2 T cells , in innate anti-gastric carcinoma immunity .Methods Flow cytometry and laser confocal microscopy were used to identify hMSH 2 that ectopically expressed on gas-tric carcinoma cell line 803.An anti-hMSH2 antibody was used to block hMSH2 to evaluate its effects on the cytotoxicity of Vγ9δ2 T cells and their cytokines secretion .Subcellular distribution of hMSH 2 in gastric car-cinoma tissues was examined by tissue microarray immunohistochemistry analysis .Results Ectopic mem-brane expression of hMSH 2 was observed on 803 cells at a relatively high level .Vγ9δ2 T cells blocked with specific anti-hMSH2 antibody showed a decreased cytotoxicity and a reduced IFN-γbut an increased TNF-αsecretion.Ectopic expression of hMSH2 was found in various types of gastric carcinoma tissues at different stages.Enhanced expression of hMSH2 was detected in specimens collected from patients with chronic super-ficial gastritis.Conclusion Ectopically expressed hMSH2 served as a stress-induced endogenous ligand which could promote the cytotoxicity of Vγ9δ2 T cells against gastric carcinoma cells and enhance their IFN-γsecretion.hMSH2 played an essential role in innate anti-gastric carcinoma immunity .

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 .