Objective To investigate the clinical application of Fibroscan to noninvasive detection of liver fibrosis in asymp‐tomatic carriers(ASCs)with chronic hepatitis B virus(HBV).Methods Clinical data of 195 chronic HBV ASCs were retrospec‐tively analyzed ,including liver function ,HBV DNA quantitation ,liver stiffness(LS) ,color Doppler ultrasound results of the liver and spleen and histopathological findings of the liver.LS was evaluated by transient elastography(Fibroscan).Liver biopsy was performed in 80 carriers for histological examination.Results The LS value was(6.22 ± 2.50)kPa in 195 cases.There was no significant difference in the LS value between HBV DNA <500 copies/mL and HBV DNA≥500 copies/mL groups.The LS value in patients with HBV carrying duration>15 years was significantly higher than that in patients with HBV carrying dura‐tion≤5 years(P<0.01).It was (6.72 ± 2.33) kPa in patients(n=49)with liver tissue inflammation and necrosis

Objective To investigate the effect of PBL combined with EBM applied in the teaching of fever of unknown origin. Methods PBL combined with EBM teaching was applied in fever of unknown origin course for 30 clinical medicine specialty(eight years) students of Tongji class of grade 2009(experiment group), while PBL teaching was applied in fever of unknown origin course for 30 clinical medicine specialty (eight years) students of Tongji class of grade 2008 (control group). After teaching, the theory examination for both basic knowledge and case analysis was organized for all students of both groups. At the same time the questionnaire survey was conducted to 30 students of grade 2009 to evaluate the teaching effect. The results were assessed by using SPSS 18.0 statistical software for the T-test of the experimental group and the control group.Inspection level was α=0.05. Results The theory test score of students in the experimental group was (93.5±3.2) point, signifi-cantly higher than that of the students in the control group(84.7±2.8). There was statistically signifi-cant difference between the scores of the two groups of students (P=0.00). Survey results showed 19 students ( 63 . 33%) thought that the development of PBL teaching combined with evidence-based medicine teaching had its necessity, and 16 students(53.33%) thought that the teaching method im-proved their clinical thinking ability of logical reasoning. Conclusion The concept of PBL combined with EBM has achieved significant resultsinthe teaching offever of unknown origin, and it is necessary to carry out this teaching mode in medical colleges with certain teaching strength.

Objective To explore the difference of gene expression profiling between normal basilar arteries and basilar arteries of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) in rabbits. Methods cDNA chip of normal basilar arteries and basilar arteries of CVS after SAH in rabbits were downloaded from GEO database. The chip was analyzed and screened by Bioconductor software, and function enrichment and pathway analysis of the differentially expressed genes were analyzed by Cytoscape software. Then 6 adult male Japanese rabbits were used, and randomly divided into normal control group (n=3) and SAH model group (n=3). Rabbit SAH models were established by cisterna secondary-blood-injection method. RNA data of normal basilar artery specimens on the 0 day and basilar artery specimens after SAH on the 5-day were used to validate the parts of differentially expressed genes by qRT-PCR. Results A total of 4356 differentially expressed genes were found in normal basilar arteries and basilar arteries of CVS after SAH in rabbits. Among them, 920 genes were considered to be significant with P-value<0.05, such as GRIK1, MYH13, ZNF45, SAA3, RLN1, MSR1 and others. Function enrichment analysis indicated that the differentially expressed genes were involved in regulation of Ca2+transmembrane transporter activity, negative regulation of ion transmembrane transport, regulation of potassium ion transport, positive regulation of JAK-STAT signaling cascades and other biological processes. Pathway analysis showed that calcium signaling pathway, cGMP-PKG signaling pathway, HIF-1 signaling pathway, PI3K-Akt signaling pathway and other signaling pathways maybe related with the differentially expressed genes. qRT-PCR verification showed that the expression of MSR1 in SAH model group was consistent with that of the chip result. Conclusion The gene expressions of basilar arteries of CVS after SAH in rabbits are significantly different, and MSR1 gene can be used as a potential target for studying the pathological mechanism of CVS.

The noninvasive measurement of liver stiffness (LS) was evaluated by transient elastography (FibroScan) and the possible influencing factors from the patients' clinical situations including age, gender, liver inflammation represented by alanine transaminase (ALT) and total billirubin (TBIL) level, HBV replication (HBV DNA loads), portal vein pressure (portal vessel diameter, PVD), splenic thickness (SPT) and body mass index (BMI) were analyzed in patients with chronic hepatitis B (CHB). A total of 466 patients including 31 patients with acute-on-chronic liver failure (ACLF), and 435 patients with chronic hepatitis B (CHB) among which 82 patients were diagnosed with liver cirrhosis (LC) by clinical manifestations and liver B-type ultrasonic inspection were enrolled at Tongji Hospital from April to December 2009. LS was measured by a FibroScan device (EchoSens, France). Simultaneously, ALT and TBIL levels, HBV DNA loads, PVD, SPT and BMI in all patients were also tested. Forty-one healthy volunteers served as controls. The values of LS were correlated positively with ages of CHB patients and significantly higher in males than in females. In patients with BMI>28 kg/m(2) (obesity) and abnormal levels of ALT and TBIL, LS values were significantly increased as compared with those having normal levels of ALT and TBIL. The patients with ACLF had the highest LS value. Furthermore, LS values in the patients with LC were significantly higher than those in patients without LC. It is concluded that noninvasive measurement of liver fibrosis by FibroScan provides an alternative method to evaluate liver fibrosis of patients with CHB. In order to properly illustrate the stiffness value taken by transient elastography, patients' gender should be taken into consideration and it is also suggested to avoid possible influencing factors including liver inflammation (high levels of ALT and TBIL) and obesity (high BMI).

Objective To observe the changes of islet cell apoptosis and oxidation-antioxidation before the transplantation, and to explore the pathways of islet protection. Methods Fifteen human pancreases were perfused with the Hanks solution containing collagenase, then digested and isolated. During the procedure, islet cell apoptosis was detected by TUNEL, SOD and MDA in the pancreas were measured by colorimetric method, and the morphologic changes were observed by H-E staining and dithizone staining. Results In the procedure of human islet isolation, especially in the stage of digestion, the apoptosis of human islet cells occurred. In the stages of perfusion and digestion, the MDA contents reached the high levels (6. 18 ± 2. 38 and 9. 21 ± 2. 75 umol/mg protein respectively),and the structures of the islets and tissues around the islets were damaged. Conclusion In the stages of perfusion and digestion, apoptosis of islet cells can be caused by oxidation. It suggests that antioxidation is a pathway for protection of islets before transplantation.

OBJECTIVETo explore the distribution, combination and evolution of various syndromic etiologies of erectile dysfunction (ED) based on the syndrome etiology theory.

METHODSUsing the ED Syndromic Etiology Scale, we collected the clinical data on the Chinese medicine diagnoses of 297 cases of ED, extracted the core syndromic etiologies by analysis of principal components and factors, and analyzed the patterns of distribution, combination, and evolution of ED syndromic etiologies according to the general information of the patients.

RESULTSThrough analysis of principal components and factors, 9 core syndromic etiologies were extracted, i. e. , liver constraint with qi stagnation, kidney yin deficiency, damp-heat, liver constraint transforming into liver-fire, blood stasis, kidney yang deficiency, heart-spleen paired deficiency, qi-yin paired deficiency, and phlegm-damp. Each of these syndrome etiologies exhibited its own specific distribution patterns. Of the total number of cases studied, 51.52% had 2 or 3 core syndromic etiologies and 36.03% had only one.

CONCLUSIONIn the early stage of ED, its syndromic etiologies are usually liver constraint with qi stagnation, kidney yin deficiency, damp-heat, liver constraint transforming into liver-fire, and blood stasis. With the natural progres- sion of the disease, its syndromic etiologies gradually evolve into kidney yang deficiency, heart-spleen paired deficiency, qi-yin paired deficiency, phlegm-damp, and blood stasis, and finally into yin-yang deficiency of the heart, spleen and kidneys, combined with phlegm-damp and blood stasis.

Adult ; Erectile Dysfunction ; diagnosis ; drug therapy ; etiology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged

OBJECTIVETo investigate the regulatory effect of ligustrazine and Radix Astragalus (Chinese drugs for supplementing qi and activating blood circulation) on uterine mucosa stromal cyto-chemotactic factor RANTES and receptor CCR5 expression.

METHODSThe eutopic and ectopic endometrial stromal cells (EMSC) isolated and purified from 10 patients with ovarian endometrial cyst were selected as the experimental group, while those from 10 patients with benign lesion were selected as the control group. After they were intervened by different factors, including astragalus injection (AI), ligustrazine injection (LI), AI + LI, and Danazol, the expression levels of RANTES and CCR5 in the cells were detected by ELISA and RT-PCR.

RESULTSRANTES expressions in eutopic and ectopic EMSC were different insignificantly (P > 0.05). The ectopic EMSC RANTES expression was 13.602 +/- 3.358 ng/L and eutopic EMSC RANTES was 12.850 +/- 7.997 ng/L in the positive control group, which was significantly higher than that in the negative control group (both being 0.027 +/- 0.016 ng/L, P <0.05), and the ectopic EMSC expressions after intervened with Chinese drugs were significantly lower than that in the positive control group (P <0.05). CCR5 expression in ectopic EMSC was 0.759 +/- 0.039 ng/L, which was higher than that in eutopic EMSC (0.249 +/- 0.026 ng/L, P <0.05). Ectopic EMSC CCR5 expression was higher in the positive control group than that in the negative control group (0.759 +/- 0.039 ng/L vs 0.478 +/- 0.094 ng/L, P <0.05). Similar situation also was shown between the positive and negative control groups in terms of eutopic EMSC CCR5 expression (0.249 +/- 0.026 ng/L vs 0.131 +/- 0.021 ng/L, P < 0.01), and the expression was significantly lower in all the Chinese drugs treated EMSC groups as compared with that in the positive control group (P < 0.01).

CONCLUSIONCCR5 expression was higher in ectopic EMSC than that in eutopic EMSC. Ligustrazine and Radix Astragalus could down-regulate the auto-secretion of RANTES and CCR5 in patients with endometriosis.

Adult ; Astragalus Plant ; chemistry ; Cells, Cultured ; Chemokine CCL5 ; genetics ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Endometriosis ; drug therapy ; genetics ; metabolism ; pathology ; Endometrium ; cytology ; metabolism ; pathology ; Female ; Humans ; Pyrazines ; pharmacology ; Receptors, CCR5 ; genetics ; metabolism ; Stromal Cells ; drug effects ; metabolism

AIM:To observe the expression of long noncoding RNA TTTY 15 in osteosarcoma tissues and cell lines and to explore its effect on the viability and invasion ability of osteosarcoma cell lines.METHODS:qPCR was used to detect the expression of TTTY15 in 11 cases of osteosarcoma and its adjacent tissues.The mRNA levels of TTTY15 in os-teosarcoma cell lines(143B,Saos2,MG-63,U2OS and HOS)and human osteoblast cell line hFOB1.19 were also tested. TTTY15 was down-regulated after transfected with small interfering RNA in MG-63 cells,the cell line with the highest level of TTTY15.The effect of TTTY15 knockdown on the viability of MG-63 cells was measured by CCK-8 assay.The cell cycle distribution was analyzed by flow cytometry.The effect of TTTY15 knockdown on the cell invasion ability was detected by Transwell assay.The levels of miR-216b-5p and FOXM1 mRNA were detected by qPCR, and the changes of the related proteins were determined by Western blot.RESULTS:Compared with the adjacent tissues,the expression of TTTY15 in-creased in the osteosarcoma tissues(P<0.01).Compared with the human osteoblast cell line,the expression of TTTY15 increased in the osteosarcoma cell lines(P<0.05), and the level of TTTY15 in the MG-63 cells was the highest(P<0.01).After knockdown of TTTY15 expression in the MG-63 cells,the cell viability was decreased(P<0.05),cell cycle progression was inhibited(P<0.01), and the cell invasion ability was decreased(P<0.01).The expression of miR-216b-5p was increased(P<0.01)and the expression of FOXM1 mRNA was decreased(P<0.01).The protein expres-sion of FOXM1,CDK4,cyclin D1,MMP-2 and N-cadherin was decreased,while the protein expression of E-cadherin was increased(P <0.05).CONCLUSION: The expression of TTTY15 is increased in the osteosarcoma tissues and cell lines.The low expression of TTTY15 inhibits the cell viability and invasion ability of osteosarcoma cells.The possible mechanism is that the knockdown of TTTY 15 expression results in the increase in miR-216b-5p expression and the down-regulation of FOXM1 expression.

Objective: To demonstrate the effect of structured and autonomous sports activities on children s basic motor skills in a preset outdoor field for a certain time and frequency, and to provide a reference for improving children s development of basic motor skills. Methods: Selected 141 typical developing children from 4 public kindergartens in Nanning, and randomly divided them into experimental group(71) and control group(70). The experimental group participated in physical activities for 10 weeks (3 times a week, 1 hour each time) in the preset outdoor field (each physical activity was composed of 30 minutes of autonomous physical activity and 30 minutes of structured physical activity), while the control group had no intervention of site preset and specially designed activities. Before and after the experiment, 9 basic motor skills (including 3 fine motor skills and 6 gross motor skills) were tested. Results: There were significant differences in double hand throwing, single leg balance (left), single leg balance (right), swing bridge balance and beam walking before and after intervention in experimental group(t=7.82,4.25,9.69,-10.01,-28.28,P<0.05), but there was no statistical significance in the improvement of heel toe coordinated walking(t=-0.60,P>0.05). There was no significant improvement in coin throwing (right hand), coin throwing (left hand) and square stacking(t=0.91,1.03,0.42,P>0.05), while in the control group, except for the balance of swinging bridge(t=4.27,P<0.05) and heel and toe walking(t=3.96,P<0.05), the improvement of other basic motor skills was not statistically significant(t=1.21, 0.74, 0.31, 0.97,P>0.05). Conclusion The quality of sports environment has a potential impact on the basic motor skills of preschool children. The basic motor skills of the pre-school children can be greatly improved by conducting structural and autonomous physical activities with a certain time and frequency.

One pair of degenerate primer was designed according to conserved motifs of the psaB (A2 subunit of photosystem I) of Chlamydomonas reinhardtii, Chlamydomonas moewusii, Chlorella vulgaris and Mesostigma viride, and a total RNA of Dunaliella salina (D. salina) was extracted with TRIzol reagent. A cDNA fragment, about 1.8kb in length, from green algal D. salina was obtained through RT-PCR method. The resulting PCR product was cloned into T-vector and screened to determine its sequence. Homologous analysis of the deduced amino acid sequence was performed by BLAST and subsequeqtly compared with GenBank data. The obtained cDNA sequence was 1815 bp long, which encodes 605 amino acids (GenBank accession number: AY820754). The sequence shared high homologue with the following psaB: Chlamydomonas reinhardtii 92%, Chlamydomonas moewusii 91%, Chlorella vulgaris 86%, Mesostigma viride 85%, Physcomitrella patens subsp. Patens 85% and Nephroselmis olivacea 84%. It can be concluded that the cloned sequence is psaB cDNA fragment from D. salina.
Algal Proteins ; genetics ; Amino Acid Sequence ; Animals ; Chlamydomonas reinhardtii ; genetics ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Molecular Sequence Data ; Photosystem I Protein Complex ; genetics ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid