BACKGROUND: Electrostatic layer-by-layer (LbL) self-assembly is a simple, effective and novel approach to carrying macromolecules into microcapsule carriers.OBJECTIVE: To prepare chitosan (CS) microcapsules to control the releasing of heparin (Hep) by LbL.METHODS: CS microspheres were fabricated by a sodium sulfate-based precipitation process and then used as positively charged templates for polyelectrolyte multilayer coatings to carry heparin by the LbL self-assembly technique. The polycation was CS and the polyanion was Hep. The microcapsule was formed via coating {CS/Hep}3 on CS hydrogel template. The microcapsules composed of {CS/ Hep}3 on CS hydrogel templates were confirmed by fluorescence inverted microscopy, confocal laser scanning microscopy and laser particle size analysis. The assembly of polyelectrolyte layers on CS hydrogel microspheres was monitored by Zeta Potential Analyzer.RESULTS AND CONCLUSION: The resulting microcapsules were about 1 μm in average diameter, and allowed spontaneous loading of heparin through electrostatic interaction, with the encapsulation efficiency and carrying capacity of 83.8% and 3.05%, respectively.

Objective To study the expression of APE/Ref-1 in gastric cancer. Methods Detection of APE/Ref-1 protein was performed with gastric cancer tissue microarray (TMA) by IHC in gastric cancer, non-neoplastic mucosa and lymph node with metastasis. Results The positive rates of APE/Ref-1 in cytoblast in non-neoplastic mucosa, tumor tissue, and metastatic lymph nodes were 96.6%, 97.1% and 98.9%, respectively, while the positive rates in cytoplasm were 71.8%, 21.4% and 10.0%, respectively and in both cytoblast and cytoplasm were 71.4%, 21.4% and 10.0%, respectively. Compared to non-neoplastic mucosa, tumor lesion had lower APE/Ref-1 expression in cytoblast and cytoplasm. In 35.9% of patients it was decreased slightly and in 11.2% of them it was decreased markedly in cytoblast (P

Objective To verify the authors' previous cDNA micro-array results and to further investigate the moleculer mechanisms of gastric cancer. Methods Quantitative real-time RT-PCR was employed to detect the expressions of three S100A calcium-binding genes in 22 fresh surgical samples of gastric tumor tissue and non-cancerous mucosa from the same patients. Results The transcription level of S100A2 in primary cancer lesion was elevated in 80% of samples when compared with matching non-neoplastic mucosa (P=0.018) and the average up-regulation level was 10.78 fold. 55% of cancer lesions showed higher transcription level of S100A4 than their adjacent non-neoplastic mucosa, the average up-regulation level was 2.31 fold. S100A6 transcription level was higher in 74% (P=0.01) of primary cancer lesion with an 2.25 fold up-regulation than the adjacent non-neoplastic mucosa. After rectified by ?_2-microglobulin, the relative expression levels of S100A2, S100A4 and S100A6 were 2.83?10~ -4 , 6.44?10~ -2 and 0.41, respectively. According to the Spearman correlation coefficient analysis there were significant positive correlations between S100A2 and S100A4, and S100A2 and S1006 (P value were 0.00 and 0.017, respectively). Conclusion The changes in S100A2 and S100A6 genes may be an early event in a majority of gastric cancer patients, while S100A4 may be associated with the infiltration of gastric cancer. Further study on the three genes might be helpful for understanding the nature of gastric carcinoma.

Objective To study the expression of S100A4 in gastric cancer and normal gastric tissue, and analyze its correlation with the clinico-pathological features and prognosis of gastric cancer. Methods Real-time quantitative reverse transcription PCR (real time qRT-PCR) was used to detect the S100A4 expression in 20 fresh surgical samples of gastric cancer and normal gastric tissue as controls. The microarray of gastric cancer tissue was established for the analysis of the S100A4 expression immunohistochemically in 208 gastric cancer tissue and isogeneic normal gastric mucosa and lymph node with metastasis. Results The S100A4 expression was increased in 55% (11/20) of gastric cancer samples with an average of 2.31 fold up-regulation of that of the normal mucosa. Patients with lymph node metastasis showed a higher percentage of elevated S100A4 transcription than those without metastasis (P=0.024). As displayed by immunohistochemistry, the positive rate of S100A4 in non-neoplastic mucosa, primary tumor and lymph node with metastasis was 9.4%, 28.1% and 32.2%, respectively (P≤0.01). A higher percentage of elevated S100A4 expression was shown in patients in advanced stage than in patients in early stage (P=0.004). In primary tumor lesions, the S100A4 expression correlated significantly with the depth of invasion (P=0.003) and poorer prognosis (P=0.034). S100A4 expression in lymph node with metastasis was also associated with poor outcome (P=0.002). Multifactorial Cox's regression analysis showed that TNM stage (P=0.029) and the expression levels of S100A4 (P=0.024) in lymph node were independent influence factors for prognosis. Conclusions Expression of S100A4 may be a late event which is associated with the progression and prognosis of gastric cancer. The analysis of S100A4 expression in lymph-node metastasis is helpful in judging the prognosis of gastric cancer.

Background Dry eye is the most common ocular surface disease.Anterior segment pattern of optical coherent tomography (OCT) is able to be used to assess the tear meniscus directly and accurately.Objective This study was to evaluate the correlation between measuring outcomes of tear meniscus by anterior segment module of OCT with signs and symptoms of dry eye.Methods A retrospective serial cases-observational study was designed.Thirty-six patients who completed Ocular Surface Disease Index (OSDI) and got a score more than 20 were enrolled in the First Hospital of Peking University from September to October,2010 under the informed consent.The upper and lower tear meniscus height (U/LTMH,μm),depth (U/LTMD,μm) and cross sectional area (U/LMA,mm2) were measured with the anterior segment module of OCT (RTVue.pitch).A series of clinical tests were performed,including tear film interferometry,break-up time (BUT),fluorescein corneal staining,lissamine green conjunctival staining and Schirmer Ⅰ test (S Ⅰ t).The correlations of measuring outcomes of tear meniscus by anterior segment module of OCT with dry eye-related examinations were analyzed with Spearman correlation coefficients.Results The values of UTMH,UTMD,UMA were (232±56) μm,(156±53) μm,(0.021 ±0.008) mm2,respectively,and the values of LTMH,LTMD,LMA were (222 ± 72) μm,(163 ± 65) μm,(0.020 ± 0.006) mm2,respectively.No significant differences were found in the measuring values between the upper and the lower tear meniscus (all at P＞0.05).The quantitative values of tear meniscus were not significantly correlated with the OSDI scores or tear film interferometry grade.Significant correlations were seen between UTMH,UTMD,UMA and BUT (r=0.46,P=0.01 ;r=0.40,P=0.02;r=0.43,P=0.01).LTMH,LTMD,LMA were positively correlated with BUT (r=0.61,P=0.00;r=0.64,P=0.00;r=0.64,P=0.00) and S Ⅰ t (r=0.54,P=0.02 ;r=0.46,P=0.01;r=0.55,P=0.01) as well as negatively correlated with fluorescein cornealstaining (r=-0.37,P=0.04;r=-0.37,P=0.04;r=-0.41,P =0.05) and lissamine green conjunctival staining (r =-0.56,P =0.01 ; r =-0.37,P =0.04 ; r =-0.47,P =0.00).Conclusions Anterior segment module of OCT is an effective approach to the measurement of tear meniscus.The measuring outcomes of anterior segment of OCT are associated with dry eye-related signs.

Objective To observe the morphologic protection effect of intra-articular injection of osteoprotegerin (OPG)on articular cartilage in a rabbit model of osteoarthritis (OA).Methods Sixty male New Zealand rabbits were randomly divided into 3 groups:OPG group (n=20),sham-operated group (n=20) and PBS group (n=20).In OPG group and PBS group,each rabbit underwent anterior cruciate ligament transection (ACLT) in left knee joint,then 0.1 ml OPG solution or PBS were injected into the left knee for 8 weeks (5 times a week) in OPG group and PBS group,respectively.In sham-operated group,the anterior cruciate ligament was just exposed without transection,and then the incision was closed.All rabbits were sacrificed 12 weeks after operation,and the left knee joints were obtained.The Pelletier score and Mankin score were used to evaluate the macroscopic and microscopic cartilage morphology.Results The score of femoral condyle cartilage and tibial plateau cartilage was 1.80±0.89 and 1.80±0.77 in OPG group,respectively,and 3.10±0.97 and 3.20±0.77 in PBS group.However,there was no statistical difference in Pelletier score between the sham-operated group and the OPG group.Consistent with the macroscopic data,the OPG group has a significantly lower Mankin score involving cartilage structure (2.65±0.88),chondrocyte (1.35±0.71),Safranin O staining (1.83±0.83),tidemark integrity (0.30±0.47) and total score (6.13±1.97) compared with the PBS group (4.52±1.09,1.85±0.63,2.80±0.75,0.65±0.49,and 9.83±1.98,respectively).The OPG group has a significantly higher total Mankin score compared with the sham-operated group (4.80±1.25),however,there were no statistical differences in four subitem scores between the two groups.Moreover,the cartilage thickness in OPG group was 371.84±38.94 μm,255.09±74.82 μm in PBS group,and 404.68±15.97 μm in the sham-operated group,and the differences were statistically significant between three groups.Conclusion OPG has a protective effect on articular cartilage and can slow the progression of OA by reducing the grade of synovitis,decreasing the volume of osteophytes,and suppressing the loss of cartilage thickness.

Objective To study the effect of intra-articular injection of dehydroepiandrosterone (DHEA) on the balance of ADAMTS/TIMP-3 system in a rabbit osteoarthritis models.Methods Sixty rabbits were underwent bilateral anterior cruciate ligament transection (ACLT).Rabbits were randomizedn to the following treatment:one knee of each rabbit was treated with 100 μmol/L DHEA dissolved in dimethylsulphoxide (the experimental group) and the other knee was treated under the same schedule using dimethylsulphoxide (the control group) 4 weeks after transection,once a week for eight weeks.Twelve weeks after ACLT,all rabbits were killed after X-ray assessment and the knee joints were evaluated by gross morphology and histology.The concentration of hydroxyproline and glycosaminoglycan in the cartilage were analyzed.The mRNA expression of ADAMTS-4,ADAMTS-5,tissue inhibitor of metalloproteinases-3 (TIMP-3),transforming growth factor (TGF)-β1.Aggrecan and Collagen Ⅱ in the cartilage were analyzed using reverse transcription polymerase chain reaction (RT-PCR).The protein expression of aggrecan ARGxx and Collagen Ⅱ in the cartilage were analyzed by Western blot.Results By Mann-Whitney test,Gross morphologic scores on femoral condyle and tibial plateau in the control group were significantly higher than the experi-mental group (Z=-3.517,P＜0.01 ; Z=-2.518,P＜0.05).By unpaired Student's t test,histological evaluation showed that the grade of cartilage damage in the experimental group [(5.3±1.2) μg/ml] were less severe than that in the control group (10.1 ± 1.3,P＜0.01).The concentration of hydroxyproline [(5.7±0.3,23.6± 1.7) μg/ml] and glycosaminoglycan (30±4) in the experimental group increased significantly when compared with the control group [(4.6±0.5),(18.5±1.4),(24±4) μg/ml,P＜0.01].The mRNA expression of ADAMTS-4 (0.15±0.03)and ADAMTS-5 (0.10±0.04) in the experimental group decreased significantly compared with the control group (0.29±0.08,0.15±0.05; all P＜0.05).The mRNA expression of TIMP-3 (0.85±0.10),TGF-β1(1.2±0.4),Aggrecan (0.87±0.31) and Collagen Ⅱ (2.74±0.59) in the experimental group increased significantly when compared with the control group (0.70±0.13,0.8±0.4,0.49±0.16,2.2±0.5; all P＜0.05).The protein expression of Aggrecan ARGxx (0.53±0.10) in the experimental group decreased significantly when compared with the control group (0.81±0.12,P＜0.01).The protein expression of Collagen Ⅱ (2.3±0.7) in the experimental group increased significantly when compared with the control group (1.7±0.5,P＜0.05).Conclusion DHEA protects against cartilage degradation and inhibits the progression of OA,TGF-β1,Aggrecan and Collagen Ⅱ in cartilage may be the mechanism of the protective effect of DHEA on OA.

Objective To investigate the proliferation inhibition and the apoptosis induction effect of brucine on human chronic myeloid leukemia cell line HL-60 cells.Methods HL-60 cells were exposed to various dosages of brucine 24,48,72 h respectively,MTT method was used to assay the growth inhibition effect of brucine on HL-60 cells and the IC50 of brucine was evaluated at the same time.The morphology was observed by AO/EB stains.The cell apoptosis and cell cycle was tested by flow cytometry with Annexin V-FITC/PI double staining and PI labeling respectively.Results The results indicated that the brucine displayed significant anti-proliferative effect on HL-60 cells in a dose-and time-dependent manner,with IC50 value of 211.8 μg/ml(24 h),107 μg/ml(48 h),83 μg/ml(72 h)respectively.The most significant inhibition was observed at 320 μg/ml for 48 h.In this condition,apoptosis morphology was induced by brucine with nuclear chromatin condensation,most of the nuclears were orange stained and condensation-like or bead-like,which was consistent with the Annexin V-FITC/PI results.The cell apoptosis rates were(2.1±1.1)％,(21.3±1.2)％,(38.6±1.3)％,(58.5±4.1)％,(75.3±0.87)％and(66.2±0.75)％in different dose of brucine,respectively.At the same time,the cell cycle analysis results showed that the cell ratio in G0/G1 phase was decreased while in G2/M and sub-G0 phases was increased,comparing with blank control group.Conclusion Brucine can inhibit cell growth dramatically,which may be related to the cell apoptosis and the cell cycle arrest.

Objective Learn about prevalence of depressive symptoms and to determine the relationship of depressive symptoms with social capital and social networks among migrant female adolescents.Methods A cross-sectional survey was conducted among 329 single females selected by convenience sampling who were aged from 16 to 28 years old,having worked for from one month to ten years.Center for Epidemiologic Studies Depression Scale(CES-D) was used to evaluate the status of depressive symptoms that were sentenced by total score ≥20.Self-compiled items were applied to assess social capital with six aspects involving organizational characteristics,exclusion,etc.and social networks included size and density with score ranging from 1 to 15 and from 0 to 1 respectively.Results The prevalence of depressive symptom was 52.9% in participants.Working in restaurants(OR=0.554),being younger(OR=0.506),high scores of solidarity,trust and cooperation of neighborhood(OR=0.964,0.758) were associated with low risk of depressive symptoms.Conclusions Female migrant adolescents from rural to urban should be paid more attention to in depression prevention.Improving solidarity,trust and cooperation in neighborhood in city could probably decrease risk of their depressive symptoms.

Background and purpose:One of the reasons why cancer cells are resistant to chemotherapy is the existence of cancer stem cells. The purpose of this study was to investigate the chemoresistance of CD44+/CD24+ Siha cells to cisplatin and its mechanisms.Methods:Siha cells were cultivatedin vitro. The CD44+/CD24+ Siha cells were sorted out by fluorescence activated cell sorter (FACS) andin vitro proliferation was detected by MTT assay after treatment with the different concentrations of cisplatin. The cell apoptosis rate was detected by flow cytometry after 10 μg/mL cisplatin acted on CD44+/CD24+ Siha cells for 24, 48 and 72 h. The relative mRNA and protein expressions of Bcl-2, Oct-4 and ABCG2 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results:The survival rates of CD44+/CD24+ Siha cells treated with different concentrations of cisplatin (0.1, 1, 5, 10, 15 and 20 μg/mL) were higher than those of their parental Siha cells [(88.42±1.51)%vs (92.87±1.5)%, (79.94±1.05)%vs (84.72±1.09)%, (69.78±0.81)%vs (75.13±2.86)%, (58.97±0.70)%vs (65.79±2.71)%, (49.60±0.88)%vs (52.10±0.52)%, (45.13±0.69)%vs (48.84±1.02)%,P<0.05]. Compared with their parental Siha cells, the apoptosis rates of CD44+/CD24+ Siha cells were lower after 10 μg/mL of cisplatin acting on them for 24, 48 and 72 h, respectively [(3.05±0.16)%vs (5.17±0.27)%, (17.94±2.02)%vs (32.60±4.28)% and (40.14±3.01)%vs (56.62±5.32)%,P<0.05]. The results from both qRT-PCR and Western blot indicated that Oct-4, ABCG2 and Bcl-2 were highly expressed on CD44+/CD24+ Siha cells. A significant difference was found in Oct-4, ABCG2 and Bcl-2 expression between CD44+/CD24+Siha cells and their parental cells (P=0.015<0.05).Conclusion:CD44+/CD24+ Siha cells could be resistant to apoptosis induced by cisplatin and expressed high levels of cancer stem cell markers such as Oct-4 and ABCG2. This study lays the basis for useful isolation and further targeted therapy of cervical cancer stem cells.