BACKGROUND:The morphological structure of nanofiber scaffold which fabricated by electrospinning technique is similar to the natural extracelular matrix, which provides a good microenvironment for cel growth and proliferation, and can also enhance cel adhesion, migration, proliferation and differentiation. OBJECTIVE: To observe the biocompatibility of double-layer poly(L-lactic acid) electrospun nanofiber scaffold and human periodontal ligament cels. METHODS: Electrospinning technique was used to prepare double layers poly(L-lactic acid) electrospun nanofiber scaffold. The toxicity of different concentrations of (100%, 75%, 50%, 25%) double-layer poly(L-lactic acid) electrospun nanofiber scaffold extracts to human periodontal ligament cels was evaluated by MTT assay. The double-layer poly(L-lactic acid) electrospun nanofiber scaffold was co-cultured with human periodontal ligament cels. The cel adhesive capacity in early stage was determined by MTT assay. The growth of cels on the scaffold was observed by scanning electron microscopy. RESULTS AND CONCLUSION: Different concentrations of double-layer poly(L-lactic acid) electrospun nanofiber scaffold extracts did not create any toxicity to human periodontal ligament cels. After co-culture for 2, 6, 24 hours, human periodontal ligament cels were poorly adherent onto the double-layer poly(L-lactic acid) electrospun nanofiber scaffold. After 7 days of co-culture, human periodontal ligament cels adhered wel on the loose surface of scaffold, maintained the original shape, stretched wel, and interconnected processes were observed; on the dense surface of the scaffold, multi-layer cels were observed. The cels showed fusiform or polygonal appearance and were connected together. These results demonstrate that the double-layer poly(L-lactic acid) electrospun nanofiber scaffold has good biocompatibility with human periodontal ligament cels.

Objective:To investigate the expressions of Smad2 and Smad4 in breast carcinoma tissue,and to analyze their relationships with oncogenesis and development of breast carcinoma and significances.Methods:Fifty-three samples of breast ductal carcinoma tissue and 50 samples of surrounding normal tissue were selected.The expression levels of Smad2 and Smad4 in cancer tissue and surrounding normal tissue were detected with immunohistochemical S-P method,and the relationships between the expression levels of Smad2 and Smad4 and the clinicopathologic parameters of breast carcinoma were evaluated.Results:The expression level of Smad2 protein in the breast carcinoma tissue was significantly higher than that in the surrounding normal tissue (z = - 2.08,P <0.05);the expression level of Smad4 protein in breast carcinoma tissue was lower than that in the surrounding normal tissue (z= - 5.01,P < 0.01).In breast carcinoma tissue,the Smad2 and Smad4 expressions were not significantly correlated with age (r=0.035,P >0.05;r=-0.077,P >0.05),tumor size (r= 0.128,P >0.05;r=0.133,P >0.05),lymph node invasion (r =0.163,P >0.05;r =0.006 P >0.05),distant metastasis (r =0.113,P >0.05;r = 0.126,P > 0.05),ER expression (r = 0.056,P > 0.05;r = 0.047,P > 0.05) and PR expression (r=0.129,P >0.05;r=0.107,P >0.05).However,the expression levels of Smad2 and Smad4 were negatively correlated with the expression of HER2 (r = - 0.388,P < 0.01;r = - 0.360,P < 0.01 ) and pathological grade (r = - 0.331,P < 0.05;r = - 0.388,P < 0.01 ).The expression of Smad2 was positively correlated to the expression of Smad4 in breast carcinoma (r=-0.83,P <0.01).Conclusion:The expressions of Smad2 and Smad4 may play an important role in the development of breast carcinoma,and they may be used as the potential biological markers for evaluating the degree of malignancy and prognosis of breast carcinoma.

OBJECTIVE: To establish a method for the content determination of domiphen by potassium chromate indicator method.METHODS: The contents of domiphen were determined based on the theory that bromide ion in domiphon could react with AgNO3 to produce silver bromide precipitation.The method was compared with the sodium tetraphenylborate method issued in China Pharmacopeica(2005 edition).RESULTS: The RSD of contents was 0.18%,and the average recovery was 100.2% in the potassium chromate indicator method.There was no significant difference between the results of two determination methods by t-test.CONCLUSION: The potassium chromate indicator method is simple,fast and accurate,which can be used for the content determination of domiphen.

This study is to explore the active ingredients of traditional Chinese medicine rhubarb with antiproliferative activity on hypertrophic scar fibroblasts (HSF). Rhubarb was extracted with Soxhlet extraction method by different polar solvents. MTS method was used to screen rhubarb solvent extracts (25 microg x mL(-1)) with anti-proliferative activity on HSF, and flow cytometry was used to detect their influences on cell cycle. Then, the active ingredients were analyzed by HPLC. The components with high activity were identified by UPLC-Q/TOF and verified by HE staining. The results showed that the ethyl acetate extract of rhubarb had higher anti-proliferative activity (P < 0.01), increased significantly the proportion of cells in G0/G1 phase (P < 0.01), and reduced the proliferation index (PI) (P < 0.01). The main active ingredients were anthraquinones. The results of confirming experiment showed that emodin, rhein and gallic acid could inhibit cell proliferation in a dose-dependent manner. In conclusion, the ethyl acetate extract of rhubarb showed anti-proliferative activity on HSF, and the anti-proliferative ingredients might be anthraquinones.

Objective:To explore the formula and preparation technology parameters of Shengmai dispersible tablets. Methods:With the granulation status, disintegration time, friability, taste and and so on as the testing indices, the formula and preparation tech-nology of Shengmai dispersion tablets were optimized by uniform design. Results:The optimized formula of Shengmai dispersible tab-lets was as follows:25% extract powder, 58% MCC, 8% CCMC-Na, 4% CMS-Na, 2% L-HPC, 2% magnesium stearate and 1%sweetener. L-HPC and magnesium stearate were added after the granulation, and the tablet hardness was controlled at 25N. The opti-mized dispersible tablets could disintegrate uniformly within 3 min. Conclusion: The optimization of the prescription and preparation process parameters of Shengmai dispersing tablets is stable and reliable, and has good repeatability, and the process is feasible.

OBJECTIVE:To optimize enzymatic extraction technology of polysaccharide from Schisandra chinensis. METH-ODS:Using pH value of enzymatic extraction solution,the amount of enzyme,extraction temperature as response factor,S. chi-nensis polysaccharide as response value,on the basis of single-factor experiments,3-factor,5-level central composite experimental design was adopted for the experiment. Validation test was also conducted. RESULTS:The optimal extraction technology was as pH value of 5.7,enzyme dosage of 1.3%,extraction temperature of 53 ℃. In validation test,the extraction rate of S. chinensis polysaccharide was 14.30%(RSD=1.84%,n=6). CONCLUSIONS:The optimized extraction technology is simple,reasonable and stable,and can be used for the extraction of polysaccharide from S. chinensis.

Objective This study aimed to explore the clinical value of human papilloma virus ( HPV) E6/E7 mRNA tests in identifying precancerous lesions of the uterine cervix-cervical intraepithelial neoplasia 2 or more CIN2+( CINⅡand CINⅢ).Methods This study is a cross-sectional survey design , between December 2011 to December 2013.The specimens were collected from the First People′s Hospital of Huizhou and the Third People′s Hospital of Huizhou in Department of Obstetrics and Gynecology outpatient and inpatient of cervical disease suspected patients , with thin-prep cytologic test ( TCT ) and histopathological results as reference , detected 345 patients of exfoliated cervical epithelial cells by using the branched DNA (b-DNA) technology to evaluate the application value of high risk HPV E 6 /E7mRNA in the clinical diagnosis of CIN.Using spss 19.0 software for data analysis.Results (1)Compared with TCT, the positive rate of E6/E7 mRNA in 325 samples were grading by cytology as follows: no intraepithelial lesion cells (NILM) 21.1%(40/190), atypical squamous cells (ASC) 38.5%(15/39 ), low-grade squamous intraepithelial lesions ( LSIL ) 76.9% ( 30/39 ) , atypical squamous cells can not exclude high-grade intraepithelial lesions (ASC-H) (8/10), high-grade squamous intraepithelial lesions (HSIL) 72.3%(34/47), TCT grades and HPV E6/E7 mRNA positive rate showed linear association (χ2 =67.654,P<0.01;r=0.497, P<0.01 ); and with HPV E6/E7 mRNA copy number was also relevant ( r =0.511, P <0.01).(2) Compared with pathological results , the positive rate of E6/E7 mRNA in 164 women samples were grading by pathology as follows:with NILM was 27.8%(10/36), with CIN Ⅰwas 65.9%(29/44), with CINⅡwas 80.6%(54/67), and with CINⅢwas 82.4%(14/17), pathological grades and HPV E6/E7 mRNA positive rate showed a linear correlation (χ2 =26.426, P<0.01; r=0.438, P<0.01); and the number of copies correlated with the increase of pathological grades too (r=0.543, P<0.01).(3) Screening effectiveness analysis results showed , the sensitivity of HPV E6 /E7mRNA was 84.6% while TCT was 47.7%.The sensitivity and specificity were 40.0% and 91.1% respectively when HPV E6/E7 mRNA and TCT processed as sequential detection test.The CIN2 +( CINⅡand CIN Ⅲ) best diagnostic critical point of 890.26 copies/ml,was established using receiver operating characteristic ( ROC) curve.The sensitivity and specificity were 58.5% and 93.7%, respectively.Conclusions The sensitivity of HPV E6/E7 mRNA test is better than TCT, the specificity is high in HPV E6/E7 mRNA and TCT processed as sequential detection test.Using the optimal cut-off value of ROC curve to detect CIN 2+has high sensitivity and specificity, so the detection of HPV E6/E7 mRNA may have some clinical value in screening and risk assessment of precancerous lesions of the uterine cervix.

Objective To explore the sleep characteristics of submariners during a long-term voyage, so as to provide scientific evidence for ensuring submariners with good sleep during long-term voyages. Methods The sleep status of submariners who participated in a long-term voyage was tested by Self-Rating Scale of Sleep (SRSS) before the voyage, and before and after each voyage section during the voyage. The sleep status variation of submariners who performed different types of tasks, from the beginning to the end of each voyage section and of each resting-on-the-sea section was analyzed respectively. Comparison of sleep scores was performed between submariners and surface ship crew in the second voyage section. Numbers of submariners with sleep problem were compared in each voyage section. Results Generally speaking, submariners' sleep status at the end of voyage section was significantly worse than that at the beginning of voyage section and that before the whole voyage (P<0.001, P<0.01), and the sleep status at the beginning of the third voyage section was significantly worse than that before the whole voyage (P<0.05). Submariners had a steady sleep status when taking a resting-on-the-sea before starting their first voyage section, which was no significant difference from that before the whole voyage (P>0.05). After finishing a voyage section and taking a resting-on-the-sea, submariners' sleep status returned to the level of pre-voyage (P>0.05), and was significantly better than that before the resting-on-the-sea (P<0.05, P<0.01). After finishing two voyage sections and then taking a resting-on-the-sea, the submariners' sleep status showed no obvious variation (P>0.05). Compared with that of surface ship crew who accomplished the same voyage section, submariners had an obviously better sleep status after taking a resting-on-the-sea (P<0.05). Meanwhile, submariners who finished a voyage section showed a significantly worse sleep status than those resting on the sea (P<0.01) and surface ship crew who finished a same voyage section (P<0.05). In each voyage section, submariners with sleep problems who finished resting-on-the-sea were significantly less than those who finished navigation (P<0.001, P<0.05). There was no significant difference in the number of submariners with sleep problems between those who taking non-resting and taking resting-at-dock after finishing the first voyage section (P>0.05), but the latter was significantly more than the former when the second voyage section was finished (P<0.05). During the resting-on-the-sea period, the numbers of submariners with sleep problems in both the second and the third voyage section were significantly more than those in the first voyage section (P<0.05, P<0.01). The numbers of submariners with sleep problems who implemented the third voyage section were significantly more than those who implemented the first and the second voyage section (P<0.01). Conclusions Generally, the sleep quality of submariners is significantly worse after accomplished a voyage section task, and the degree of sleep problems may be accumulated to worse and worse along with the increase of long-term voyage time. Whereas, submariners may have a significantly better sleep status after taking a resting-on-the-sea, implying that resting-on-the-sea is an effective way to ensure submariners a good sleep during a long-term voyage.

AIM To observe the effects of Huangdi Anxiao Capsules (Puerariae lobatae Radix,Eriobotryae Folium,Notoginseng Radix et Rhizoma,etc.) on blood glucose and insulin resistance in GK rats with type 2 diabetes mellitus (T2DM) and its possible mechanism of action.METHODS GK rats with T2DM included in the experiment were randomly divided into model group,rosiglitazone (1.44 mg/kg) group,Shenqi Jiangtang Granules (1.08 g/kg) group,Huangdi Anxiao Capsules high,medium and low (12,6,3 g/kg) group and normal control group of Wistar rats.After six weeks of consecutive administration,fasting blood glucose (FBG),serum insulin (FINS),fasting plasma glucose (FPG),and insulin resistance index (HOMA-IR) were measured in all groups.Serum biochemical indexes of (TG,TC,HDL-C,LDL-C),glycosylated hemoglobin (HbA1c),glucagon-like peptide (GLP-1) and insulin-like growth factor (IGF-1) were measured.The pancreatic tissue was obtained by routine paraffin embedding and HE staining to observe the pathological changes.RESULTS Compared with the normal group,FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1c of the model group were significantly higher (P ＜0.01),and GLP-1 and IGF-1 expressions were markedly decreased (P ＜ 0.01).Compared with the model group,the levels of FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1 c in Huangdi Anxiao Capsules high-,medium-and low-dose group were significantly decreased (P ＜ 0.05,P ＜0.01);GLP-1 and IGF-1 expressions were markedly increased (P ＜0.05,P ＜0.01),compared with the model group,the structure changes of pancreatic tissue in Huangdi Anxiao Capsules high-,medium-and low-dose groups largely reduced.CONCLUSION Huangdi Anxiao Capsules can reduce GK rats fasting blood glucose,insulin resistance,and its mechanism may be related to promoting the emergence and proliferation of pancreatic islet cells,improving the function of islet beta cells,and increasing insulin secretion.

Objective To investigate the boosting efficiency of a subunit vaccine consisting of the fusion protein Ag85B-Mpt64190-198-Mth8.4 (AMM) , dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaceharide nucleic acid (BCG-PSN) on the primed inoculation with BCG. Methods The AMM subunit vaccine was composed of fusion protein AMM, adjuvant DDA and BCG-PSN. The first mouse experi-mental group was immunized with BCG first, then boosted with the AMM subanit vaccine in the 10th week. The second experimental group was boosted with the AMM subunit vaccine in the 8th week and the 10th week respectively with a two weeks interval after the primed with BCG. Two control groups were treated re-spectively with physiological saline alone and BCG alone. After the primed inoculation, ELISPOT and ELISA were used for the detection of the cell-mediated and humoral immune response in week 14 and week 22 re-spectively. Furthermore, the immunized mice were challenged with live BCG to mimic tuberculosis infection in the 22nd week after the primed inoculation. Subsequently the T cell typing and humoral response were de-tected by flow cytometry and ELISA, respectively. Results ( 1 ) The level of secreting IFN-γ: 14 weeks af-ter the primed inoculation,with the stimulation of the specific antigen-Ag85B, the number of cells secreting IFN-γ in the second experimental group (135±14) was more than BCG alone immunized group (19±16), t = 10. 98, P < 0.01. In the 22nd week, the number of cells secreting IFN-γ in the second experimental group (208±11) was still more than BCG alone group (57±18), t =6.43, P <0.01. (2) The level of humoral immune response: the IgG1 antibody titer in the second experimental group was obviously higher than that in the first experimental group. However, the ratio of IgG2a to IgG1, as the index reflecting the Thl-type immune response, in the experimental group 2 was lower than that in the experimental group 1. (3) The contents of CD4+ CD25+ T cells after challenged with live BCG strain: the first and the second ex-perimental groups were both higher than the BCG alone group (t1 = 3.08, t2 = 3.16, P < 0.05 ). Conclu-sion Boosting the BCG-pfimed mice with tuberculosis AMM subunit vaccine twice can induce higher level of cell-mediated and humoral immune response than BCG alone, which could activate the regulative immune response at the same time.