Objective Evaluating the effect of situational questions for qualifying the students'abilities in physiolo-gy examination .Methods Comparing the difficulty coefficients and discrimination indexes between situational and traditional choice questions .Results Compared with the traditional choice questions , the difficulty coefficient of situational questions increased , while the discrimination indexes were more reasonable .The discrimination indexes of situational understanding questions were higher than those of the traditional memory and understanding questions . There were no difference between discrimination indexes of the situational application questions and those of tradi -tional application questions .Conclusions Situational questions not only improved the quality of examinations , but also facilitate evaluating students'learning ability .

The applied value of serum hepcidin in differential diagnosis of infection fevers versus tumor fevers was explored.A total of 432 fever patients were selected according to the domestic fever of unknown origin (FUO) diagnostic criteria from our hospital between June 2010 and November 2013.Venous blood samples were taken on the day 1,5,10 after admission.The infection group (98 cases) and the tumor group (50 cases) were set up based on the clinical and laboratory findings.ELISA was used to determine the serum hepcidin and IL-6 levels.SPSS 13.0 was used for statistical analysis.Hepcidin showed obvious descending trend on the 10th day in both the bacterial infection group (66 cases) and the virus infection group (32 cases),and the descending trend was similar to that of inflammatory indexes such as procalcitonin (PCT),hypersensitive C-reactive protein (h-CRP),erythrocyte sedimentation rate (ESR),white blood cell (WBC),and ferritin.Serum hepcidin showed no obvious differences in the tumor group on the day 1,5,10 after admission.In the infection groups,serum hepcidin was positively correlated with IL-6 (r=0.687,P=0.000) and CRP (r=0.487,P=0.026),but had a poor correlation with blood sedimentation,ferritin,PCT and WBC (P＞0.05).Monitoring dynamic changes of hepcidin and related inflammatory factors in patients with fever is expected to be used for clinical identification of infection fever and tumor fever.

Objective To study the separation and purification of total flavonoids from Polygonum hydropiper Linn. by macroporous adsorption resin; To investigate the effects of various factors in the process of static and dynamic adsorption on effects of adsorption and desorption to determine the optimum capability. Methods The total flavonoids from Polygonum hydropiper Linn. was extracted; the contents of flavonoids of ethyl acetate part (FEA) and flavonoids of n-butanol part (FNB) were detected; the adsorption experiment of FEA and FNB was conducted by macroporous adsorption resin. D101, AB-8, DM130, and XDA-8 macroporous resin were used to optimize the separation and purification process. Results The XDA-8 macroporous resin was selected to enrich total flavonoids from Polygonum hydropiper Linn. The optimum process was: adjusting the pH value of FEA flavonoids to 6, the concentration of sample was 750 g/mL, eluting with 75% ethanol to elute 5 bed volumes at a flow rate of 1 bed per hour; adjusting the pH value of FNB flavonoids to 6, the concentration of sample was 1 mg/mL, eluting with 60% ethanol to elute 5 bed volumes at a flow rate of 1 bed per hour. Conclusion XDA-8 macroporous adsorption resin is helpful to separate and purify the total flavonoids of Polygonum hydropiper Linn.by the best process.

Objective To observe the effect of flavonoids ethyl acetate(FEA)from Polygonum hydropiper L.on biochemical indexes and inflammatory cytokines in mice with endotoxemia; To expore the mechanism. Methods Total flavonoids in the whole plant of Polygonum hydropiperum L. were extracted by enzymolysis-ultrasonic coupling method. The FEA part were obtained by extracting and separating, followed with macroporous resin purification and enrichment. The animal model of endotoxemia was established by stimulation of lipopolysaccharide (LPS) in mice. Experimental mice were divided into blank group, model group, FEA high-, medium-, and low-dose groups. Each administration group was given the corresponding concentration of herb liquor. The concentration of malondialdehyde (MDA) and myeloperoxidase (MPO) in intestinal tissue, total antioxidant capacity (T-AOC), superoxide dismutase (T-SOD) activity and glutathione peroxidase (GSH-Px) activity in liver tissue, glutathione (GSH), lysozyme (LZM) and acid phosphatase (ACP) in serum were determined. The contents of tumor necrosis factor-α (TNF-α), interferon (IFN)-α, IFN-γ and interleukin-2 (IL-2) in lung tissue were detected by RT-PCR. Results Compared with blank group,the levels of MDA, MPO in intestinal tissue and serum ACP of model mice were increased, while T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM levels were decreased; TNF-α in serum, intestinal and liver tissues were increased, the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissue were increased. Compared with the model group, the levels of MDA, MPO in intestinal tissue and serum ACP were decreased in all dose of FEA groups;The levels of T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM of FEA medium and high-dose groups were increased. The content of TNF-α in mice serum, intestinal and liver tissues of all dose of FEA groups were significantly reduced, and the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissues were significantly decreased. The pathological morphology of mice lung, ileum and colon tissues of FEA high-dose group were significantly ameliorated than model group. Conclusion FEA can attenuate inflammation injury of endotoxemia mice induced by LPS, which has protective effects for organism.

OBJECTIVETo establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells.

METHODSWe cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31(+) cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay.

RESULTSThis approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel.

CONCLUSIONSThe improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.

Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Line ; Collagen Type I ; Culture Media ; Embryonic Stem Cells ; cytology ; Endothelial Cells ; cytology ; Humans

OBJECTIVETo establish a high efficient human coagulation factor VIII (FVIII) eukaryotic stable expression system using lentiviral vector, and determine its biosafety.

METHODSLentiviral transfer plasmid carrying human B-domain-deleted FVIII(BDDhFVIII)-IRES-GFP(BDDhFVIII/pXZ9)or IRES-GFP(pXZ9) was constructed. Lentivirus particles were produced by transiently co-transfected 3-plasmids into 293FT cells and further concentrated via ultracentrifugation. CHO cells were infected, 72h later, the FVIII antigen (FVIII:Ag) concentration in the medium was examined by ELISA, the activity was detected via one stage coagulation,and the transcription of FVIII in the infected CHO cells was determined by RT-PCR.Virus infection ability in the medium and the gag gene in CHO cells were determined to evaluate the model's biosafety.

RESULTSLentiviral transfer plasmid BDDhFVIII-IRES-GFP(BDDhFVIII/pXZ9)carrying human B-domain-deleted FVIII or IRES-GFP (pXZ9) was successfully constructed, and high titer lentiviruses has been prepared. The lentivirus could infect CHO cells efficiently, after an additional 72 h, the FVIII:Ag concentration had up to (1724.9±283.7) mU/ml, the FVIII:C level increased to (10.58±1.55)%, and transcripts of BDDhFVIII mRNA could be measured by RT-PCR. Neither the gag gene nor the virus in the supernatant was detected.

CONCLUSIONLentivirus-mediated human coagulation factor VIII could be expressed efficiently in CHO cells. The system couldn't produce offspring virus, proving a good biosafety.

Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Factor VIII ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection

BACKGROUNDTumor intrinsic chemoradiotherapy resistance is the primary factor in concomitant chemoradiotherapy failure in advanced uterine cervical squamous cell carcinoma. This study aims to identify a set of genes and molecular pathways related to this condition.

METHODSForty patients with uterine cervical squamous cell carcinoma in International Federation of Gynecology and Obstetrics stage IIb or IIIb, treated with platinum-based concomitant chemoradiotherapy between May 2007 and December 2012, were enrolled in this trial. Patients included chemoradiotherapy resistant (n = 20) and sensitive (n = 20) groups. Total RNA was extracted from fresh tumor tissues obtained by biopsy before treatment and microarray analysis was performed to identify genes differentially expressed between the two groups.

RESULTSMicroarray analysis identified 108 genes differentially expressed between concomitant chemoradiotherapy resistant and sensitive patients. Functional pathway cluster analysis of these genes revealed that DNA damage repair, apoptosis, cell cycle, Map kinase signal transduction, anaerobic glycolysis and glutathione metabolism were the most relevant pathways. Platelet-derived growth factor receptor alpha (PDGFRA) and protein kinase A type 1A (PRKAR1A) were significantly upregulated in the chemoradiosensitive group, while lactate dehydrogenase A (LDHA), bcl2 antagonist/killer 1 (BAK1), bcl2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), single-strand-selective monofunctional uracil-DNA glycosylase 1 (SMUG1), and cyclin-dependent kinase 7 (CDK7) were upregulated in the chemoradiotherapy resistant group.

CONCLUSIONWe have identified seven genes that are differentially expressed in concomitant chemoradiotherapy resistant and sensitive uterine cervical squamous cell carcinomas, which may represent primary predictors for this condition.

Aged ; Carcinoma, Squamous Cell ; drug therapy ; genetics ; radiotherapy ; Chemoradiotherapy ; Female ; Humans ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; drug therapy ; genetics ; radiotherapy

OBJECTIVETo explore the effects of di-butyl phthalate (DBP) on the reproductive system of adolescent male rats.

METHODSSprague-Dawley (SD) rats aged 5 weeks were assigned to receive corn oil (vehicle control) or DBP orally at 10, 100 and 500 mg/(kg x d) for 30 days. After the exposure, the testis, epididymis, liver and pituitary of the rats were weighted and their ratios to the body weight obtained. Histopathological changes of the testis and epididymis were examined by Hematoxylin-eosin staining, the levels of testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the serum were measured by radioimmunoassay, and the relative mRNA expressions of the steroidogenesis acute regulatory protein (StAR), proliferating cell nuclear antigen (PCNA), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc) and scavenger receptor (SR) were detected by real-time quantitative RT-PCR.

RESULTSDBP induced significant histopathological changes in the testicular tissue at 100 and 500 mg/(kg x d), and decreased the testicular and epididymal weights, inhibited the mRNA expressions of StAR and PCNA, reduced the levels of T and LH, and elevated the level of FSH at 500 mg/(kg x d). At the dose of 10 mg/(kg x d), DBP increased serum LH and FSH and the mRNA expression of P450scc. While the SR mRNA expression showed no significant changes in all the groups.

CONCLUSIONHigh level of DBP has apparent toxic effect on reproductive system of male rats. Low - dose DBP can increase the level of serum gonadotropin LH and affect the mRNA expression of P450scc in the testis.

Animals ; Cholesterol Side-Chain Cleavage Enzyme ; metabolism ; Dibutyl Phthalate ; administration & dosage ; toxicity ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Phosphoproteins ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Scavenger ; metabolism ; Testis ; drug effects ; metabolism

OBJECTIVETo analyze the global gene expression profile of primary cultured nasopharyngeal carcinoma (NPC) cells using cDNA microarray techniques to screen new candidate genes related to the occurrence and progression of NPC.

METHODSA NPC cell line C666 and primary cultured NPC cells from biopsy specimens in 5 cases were analyzed with microarray techniques in comparison with 3 normal nasopharyngeal epithelial (NPE) biopsy specimens. Several differentially expressed genes identified from the microarray results were verified by fluorescence real-time PCR (FQ-PCR) and immunohistochemistry (IHC).

RESULTSPrimary cultured cells of both NPC and NPE were verified by cytokeratin IHC, EBER1 in situ hybridization and EBV-DNA real-time PCR. Compared with NPE cells, a total of 493 genes in at least 4/6 of the samples were identified to be differentially expressed in the primary cultured NPC cells, including 264 up-regulated and 229 down-regulated ones. Several differentially expressed genes according to the microarray results were confirmed by real-time PCR and IHC.

CONCLUSIONcDNA microarray technique provides an effective and accurate means for global gene expression profiling of primary cultured NPC cells to screen the differentially expressed genes, which may serve as an important basis for studying the mechanism, classification and diagnosis of NPC at the molecular level.

Animals ; Cells, Cultured ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; RNA ; isolation & purification

OBJECTIVETo study the related risk factors of anemia of rural elderly women in Huangling county, Shanxi, northwest of China.

METHODSElderly women aged 50-75 years in Huangling (northwest of China) were selected as study objects. Finger hemoglobin (Hb) was measured and basic health survey was face-to-face questioned. Two-hundred anemia elderly women were entered into the case group; and by age-matching, 200 with normal Hb concentration were entered into the control group. Dietary survey, health and lifestyle questionnaire were undertaken, and related blood indexes were tested.

RESULTSIn case and control group, annual income was (446.1 +/- 107.9) vs (903.8 +/- 179.1) yuan (t = 3.06, P < 0.01), daily average physical active time was (9.6 +/- 3.2) vs (10.3 +/- 3.1) hours (t = 1.94, P < 0.05), proportion of experiencing food scarce period was 31.8% vs 22.6% (chi2 = 4.14, P < 0.05), waist circumference was (76.2 +/- 7.3) vs (79.5 +/- 8.9) cm (t = 4.08, P < 0.01), respectively; the total protein was (78.0 +/- 5.8) vs (81.9 +/- 6.0) g/L(t = 5.94, P < 0.01), serum iron was (13.9 +/- 5.7) vs (16.1 +/- 5.0) micromol/L (t = 4.19, P < 0.01), serum ferritin was (94.9 +/- 76.4) vs (116.6 +/- 85.2) microg/L (t = 2.58, P < 0.01), saturation of transferrin was 22.9% +/- 10.0% vs 25.6% +/- 8.7% (t = 3.16, P < 0.01), respectively. Multifactor conditioned logistic regression analysis showed that the odd ratio (OR) for anemia with annual income, whether experiencing food scarce period, daily average physical active time, staple food, soybean products, energy was 0.57, 4.74, 0.06, 0.59, 0.55, 0.65, respectively; their confidence interval (CI) was 0.45 - 0.71, 0.73 - 30.56, 0.01 - 0.52, 0.38 - 0.91, 0.34 - 0.87, 0.44 - 0.98, respectively.

CONCLUSIONThe quality of diet, health status and related blood indexes in anemia elderly women were lower than those in control group; lower income, less active time, less staple food, soybean products and energy intake should be risk factors of anemia.

Aged ; Anemia ; epidemiology ; Case-Control Studies ; China ; epidemiology ; Diet Surveys ; Female ; Hemoglobins ; analysis ; Humans ; Middle Aged ; Nutritional Status ; Risk Factors ; Rural Population ; Surveys and Questionnaires