Objective: To compare the volatile compounds extracted from Rhodiola renulata respectively by HS-SPME and SD. Methods:The volatile constituents from Rhodiola crenulata were extracted respectively by HS-SPME and SD, and then the contents and the names were confirmed by GC-MS. Results:Totally 39 compounds were identified from Rhodiola crenulata by HS-SPME while 16 ones were identified by SD. Among them, 4 common compounds were detected. Conclusion: There are some differences between the two methods. Compared with SD, HS-SPME is obviously better because more volatile constituents can be extracted from the herb, furthermore, HS-SPME has notable advantages of higher retrieval matching and sensitivity.

OBJECTIVETo develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.

METHODSA pair of common primers of 18 HPV subtypes for PCR, was designed in HPV conservative L1 region. Genotyping probes for detecting 15 high-risk HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, together with 3 low-risk HPV 6, 11 and 42 were selected respectively from Genbank and fixed on membrane to make DNA chip. PCR amplification and DNA chip technology were optimized. 100 clinical samples were used to investigate the effect of HPV genotyping DNA chip. Veracity of the genotyping results was verified by sequencing.

RESULTSFrom the 100 clinical samples, 30 were found to be HPV positive, including high-risk HPV subtypes 16, 18, 33, 45, 51, 58, and 66, and low-risk HPV 6, 11 and 42. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.

CONCLUSIONThe HPV genotyping system developed here with DNA chip showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.

DNA Probes, HPV ; genetics ; DNA, Viral ; genetics ; Female ; Genotype ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; virology

Objective To analyze the urine trace albumin(mALb)/creatinine(Cr) ratio and blood uric acid(UA),and other various metabolic index level in patients with diabetic nephropathy(DN),combined with clinical data such as patients' age,body mass index(BMI),course of diseases,to explore the related mechanism of occurrence and development of DN.Methods 76 DN patients were selected.The microalbuminuria group(urinary mALb/Cr<300μg/mg) had 46 cases,the clinical albuminuria group(urinary mALb/Cr≥300μg/mg) included 30 cases,another 49 diabetic patients without kidney damage were seleted as control group.The urinary mALb/Cr,blood UA,fasting blood glucose(FBG),triacylglycerol(TG),total cholesterol(TC),high-density lipoprotein(HDL),low density lipoprotein(LDL),glycosylated hemoglobin(HbA1c) levels were determined.The BMI and the length of the course of the disease calculate.Results The patients' age,course of the disease,urinary mALb/Cr,blood UA,FBG,TC,TG,LDL,HbA1c and BMI level in the clinical albuminuria group and microalbuminuria group were significantly higher than those in the control group,the differences were statistically significant (F=6.18,12.48,141.43,12.48,8.49,4.98,6.18,3.89,3.17,3.89,all P<0.05).The high uric acid hematic disease rates of the clinical albuminuria group and microalbuminuria group were 26.09% and 26.09%,which were significantly higher than 10.20% of the control group,the differences were statistically significant(x2=4.074,24.833,all P<0.05).Urinary mALb/Cr was positively correlated with age,duration,BMI,UA,TG,TC,LDL,FBG,HbA1c(r=0.120,0.299,0.148,0.340,0.157,0.149,0.103,0.487,0.103).Multiple linear stepwise regression analysis suggested that duration,blood UA,FBG were independent risk factors of urinary mALb/Cr;TG,BMI,urinary mALb/Cr were independent risk factors for blood UA.Conclusion Urinary mALb/Cr and blood UA are the independent risk factors,high uric acid hematic disease may participate in the development process of DN,and diabetes duration,UA,BMI,TG,TC,LDL,FBG,HbA1C associated with increased urinary mALb/Cr excretory DN patients,the effective monitoring can improve the symptoms of DN and quality of life.

OBJECTIVEIn order to explore the roles of Huoxueruanjian compound on liver fibrogenesis and its molecular mechanism, this paper has investigated the Influence of blood serum with such traditional Chinese medicine compound on the expression of Smad3, Smad7 and procollagen alpha2(I) gene in hepatic stellate cell (HSC).

METHODSHSC-T6 was deal with different Concentration of blood serum medicine with Heluoshugan which was made by routine way. Then expression change of Smad3, Smad7 and procollagen alpha2(I) mRNA among each groups were observed by RT-PCR. Furthermore, the expression change of Smad3 protein were examined by Western blot.

RESULTSExpression of Smad3 and procollagen alpha2(I) mRNA as well as Smad3 protein had been downregulated after treating with blood serum medicine of Heluoshugan (P<0.01, P<0.05, respectively). The expression of procollagen alpha2(I) mRNA changed at the same tendency as those of Smad3. The role of blood serum medicine was significant difference between different concentration, P<0.05. And the expression of procollagen alpha2(I) mRNA changed in concentration-dependent manner. Blood serum medicine has no effects on the Smad7 mRNA.

CONCLUSIONThe anti-fibrosis roles of HuoXueruanjian Compound maybe influence the function of TGF-beta and Smad by nonspecific action, thereby inhibit the transcription of procollegan alpha2(I) mRNA and decrease the production of ECM. As regards Smad3, it may be facilitating the development of liver fibrosis when its expression increases. Otherwise, it manifest with anti-fibrosis role.

Animals ; Collagen Type I ; genetics ; DNA-Binding Proteins ; genetics ; Liver Cirrhosis ; drug therapy ; etiology ; pathology ; Medicine, Chinese Traditional ; RNA, Messenger ; analysis ; Rats ; Smad3 Protein ; Smad7 Protein ; Trans-Activators ; genetics

BACKGROUNDMono-(2-ethylhexyl) phthalate (MEHP), the metabolite of di-(2-ethylhexyl) phthalate (DEHP), was suspected to be toxic to human embryos. This study contributes to investigating its toxic effects by an embryonic stem cell test (EST) based on two human embryonic stem cell (hESCs) lines.

METHODSCH1 established in our own lab and H1, a federally registered cell line were two human embryonic stem cell lines used in this test. Four endpoint measurements were performed consisting of cell viability, proliferation ability, apoptosis as well as changes of gene expression patterns after spontaneous differentiation were determined. For measuring effects on the first three endpoints, the cells were treated with various concentrations of MEHP dissolved in dimethyl sulfoxide (DMSO) and only with DMSO which served as control and harvested after 5 days. For measuring effects during spontaneous differentiation, the RNA of embryoid bodies (EBs) formed after 8 days' MEHP exposure was collected and changes in differentiation specific gene expression patterns were analyzed by quantitative real time RT-PCR.

RESULTSAs a result the viability and proliferation ability of both cell lines decreased significantly at 1000 µmol/L MEHP, while there was no effect on apoptosis or cell morphology. In addition MEHP also changed the gene expression pattern in the EBs of both cell lines.

CONCLUSIONMEHP in a high dose was cytotoxic and affected the development of hESCs, which indicates its embryo toxicity in human embryos.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Diethylhexyl Phthalate ; analogs & derivatives ; toxicity ; Dose-Response Relationship, Drug ; Embryonic Stem Cells ; drug effects ; pathology ; Humans

OBJECTIVETo observe the effects of Nrf2 gene expression induced by RU486 at different doses on A549 cell damage induced by paraquat (PQ).

METHODSAfter A549 cells transfected with Ad-RUNrf2 were treated by RU486 at the doses of 10(-10), 10(-9), 10(-8) and 10(-7) mol/L for 6 h, A549 cell cultures were exposed to 10(-3) mol/L of PQ for 48 h. Then qRT-PCR and EMSA assays were used to detect the expression of Nrf2 gene, and qRT-PCR and ELISA assays were utilized to measure the effects of Nrf2 gene on the expression of the inflammatory cytokines IL-6, IL-10 and TNF-α, apoptotic factors Caspase-3, Caspase-9 and Cytochrome C. The oxidation factors (CAT and MDA protein contents) were observed by Chemical Colorimetric Analysis.

RESULTSNrf2 gene relative expression and protein contents increased with RU486 concentrations, and the above expression was the highest when the concentration of RU486 was 10(-7) mol/L, which was significantly higher than those in control and PQ exposure groups (P < 0.01 or P < 0.05). The relative gene expression and protein expression of IL-6 and TNF-α enhanced with the reduced concentrations of RU486, which were the lowest when RU486 concentration was 10(-7) mol/L, as compared with control and PQ exposure groups (P < 0.01 or P < 0.05), while the change of IL-10 content was the opposite. The relative expression of Caspase3, Caspase9 and Cytochrome C genes also increased with the reduced concentrations of RU486, which were the lowest when RU486 concentration was 10(-7) mol/L, as compared with control and PQ exposure groups (P < 0.01 or P < 0.05). The content of CAT enhanced with RU486 concentration, which was the highest when RU486 concentration was 10(-7) mol/L, as compared with control and PQ exposure groups (P < 0.05). But the change of MDA content was the contrary.

CONCLUSIONNrf2 expression induced by RU486 can promote the balance of oxidation-antioxidation system in A549 cells and inhibit the inflammation and apoptosis factors, which has a protective effect on A549 cell injury induced by PQ.

Cell Line ; Gene Expression ; drug effects ; Humans ; Interleukin-10 ; metabolism ; Interleukin-6 ; metabolism ; Mifepristone ; administration & dosage ; pharmacology ; NF-E2-Related Factor 2 ; genetics ; Paraquat ; toxicity ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo demonstrate the effect of bromoxynil on membrane potential and respiratory control rate (RCR) in isolate mitochondria from mice liver tissue in vitro and the intervention of NAC.

METHODSThe mitochondrial was randomized to control group, bromoxynil-poisoned group and NAC-protected group. S3, S4 and RCR of the mitochondria in each sample was detected by the method of oxygen electrode. Each sample was stained by JC-1 and the changes of membrane potential of mitochondria were observed under fluorescence microscope.

RESULTSThe S3 [(0.031 +/- 0.008) nano atoms oxygen x mg(-1) x min(-1)], RCR (1.820 +/- 0.181) of bromoxynil-poisoned group and RCR (4.253 +/- 0.210) of NAC-protected group were significantly lower than those of control group (P<0.01); the S4 [(0.017 +/- 0.004) nano atoms oxygen x mg(-1) x min(-1)] of NAC-protected group was significantly higher than control group (P<0.01). The S3 [(0.046 +/- 0.005) nano atoms oxygen x mg(-1) x min(-1)] and RCR of NAC-protected group were significantly higher than group B (P<0.01), S4 [(0.011 +/- 0.001) nano atoms oxygen x mg(-1) x min(-1)] of NAC-protected group was significantly lower than bromoxynil-poisoned group (P< 0.01). Observation under fluorescence microscope: the red fluorescence of mitochondria was dim or disappeared in bromoxynil-poisoned group while brightened in NAC-protected group but still dimmer than control group.

CONCLUSIONIn vitro, the mitochondrial RCR and the mitochondrial membrane potential are decreased after the mitochondria is incubated with bromoxynil, and NAC could improve it.

Acetylcysteine ; pharmacology ; Animals ; Electron Transport ; drug effects ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mice, Inbred ICR ; Mitochondria, Liver ; drug effects ; metabolism ; Nitriles ; toxicity

BACKGROUNDMechanical stress plays an important role in the maintenance of bone homeostasis. Current hypotheses suggest that interstitial fluid flow is an important component of the system by which tissue level strains are amplified in bone. This study aimed to test the hypothesis that the short-term and appropriate fluid shear stress (FSS) is expected to promote the terminal differentiation of pre-osteoblasts and detect the expression profile of microRNAs in the FSS-induced osteogenic differentiation in MC3T3-E1 cells.

METHODSMC3T3-E1 cells were subjected to 1 hour of FSS at 12 dyn/cm(2) using a parallel plate flow system. After FSS treatment, cytoskeleton immunohistochemical staining and microRNAs (miRNAs) were detected immediately. Osteogenic gene expression and immunohistochemical staining for collagen type I were tested at the 24th hour after treatment, alkaline phosphatase (ALP) activity assay was performed at 24th, 48th, and 72 th hours after FSS treatment, and Alizarin Red Staining was checked at day 12.

RESULTSOne hour of FSS at 12 dyn/cm(2) induced actin stress fiber formation and rearrangement, up-regulated osteogenic gene expression, increased ALP activity, promoted synthesis and secretion of type I collagen, enhanced nodule formation, and promoted terminal differentiation in MC3T3-E1 cells. During osteogenic differentiation, expression levels of miR-20a, -21, -19b, -34a, -34c, -140, and -200b in FSS-induced cells were significantly down-regulated.

CONCLUSIONThe short-term and appropriate FSS is sufficient to promote terminal differentiation of pre-osteoblasts and a group of miRNAs may be involved in FSS-induced pre-osteoblast differentiation.

Actins ; chemistry ; Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Core Binding Factor Alpha 1 Subunit ; genetics ; Cyclooxygenase 2 ; genetics ; Gene Expression Profiling ; Mice ; MicroRNAs ; physiology ; Osteoblasts ; cytology ; Osteogenesis ; Stress, Mechanical ; Stress, Physiological

Objective To investigate the innovation ability status of undergraduate students in Third Military Medical University,and put forward some effective suggestions.Methods A questionnaire was applied to survey the innovation ability status of undergraduate students in Third Military Medical University.A total of 210 valid questionnaires were collected.The questionnaires covered five aspects including undergraduate students' basic information,innovative consciousness,innovation thinking,innovation skills and basic knowledge.The results were assessed by using SPSS 19.0 statistical software for t-test or ANOVA of students from different grades,majors and academic years.Inspection level was α=0.05.Results The total score of innovation ability in undergraduates was (70.5 ± 8.2) point,and no significant difference was observed in the total score of undergraduates' innovation ability within different grades (P=0.435).However,the innovation thinking ability of undergraduates in Grade Four was significantly higher than that in Grade Two [(77.0 ± 10.7) vs.(72.6 ± 10.9),P=0.030)],and the score of basic knowledge of undergraduates in Grade One was significantly higher than that in Grade Four [(76.2 ± 6.0) vs.(69.3 ± 8.7),P=0.014)].The total score of innovation ability of undergraduates from clinical medicine was significantly higher that of undergraduates from preventive medicine and other majors [(72.5 ± 8.8) vs.(69.9 ± 7.5),P=0.035;(72.5 ± 8.8) vs.(66.7 ± 7.9),P=0.004].There were no significant differences in total score of innovation ability or score of any first level index of undergraduates between eight-and five-year system of preventive medicine (P＞0.05).Conclusion The overall innovation ability of undergraduates in military medical university was relatively high,and undergraduates from different grades,majors and academic years have their own special advantages in innovative consciousness,innovation thinking,innovation skills and basic knowledge,and it is necessary to carry out more researches focusing on educational and training mechanism of innovation ability according to the personality of undergraduates in military medical universities.

OBJECTIVE: To investigate the efficiency of spectral computed tomography (CT) optimal monochromatic images in improving imaging quality of liver vessels. MATERIALS AND METHODS: The imaging data of 35 patients with abdominal CT angiography were retrospectively analyzed. Hepatic arteries, portal veins, and hepatic veins were reconstructed with mixed energy (quality check, QC), 70 keV and optimal monochromatic mode. Comparative parameters were analyzed including CT value, image noise (IN), contrast-to-noise ratio (CNR), signal-to-noise ratio (SNR), and subjective qualitative analysis. RESULTS: The optimal monochromatic value for assessment of the common hepatic artery, portal vein, and hepatic vein ranged between 49 keV and 53 keV, with a mean of 51 keV. There were statistically significant differences (p < 0.001) among the optimal monochromatic, 70 keV and QC images with regards to the hepatic vascular CT value, IN, CNR, SNR, and subjective qualitative score. CNR of the common hepatic artery in the optimal monochromatic, 70 keV and QC groups was 24.6 ± 10.9, 18.1 ± 8.3, and 11.6 ± 4.6, respectively (p < 0.001) with subjective scores of 4.7 ± 0.2, 4.0 ± 0.3, and 3.6 ± 0.4, respectively (p < 0.001). CNR of the hepatic portal vein was 6.9 ± 2.7, 4.3 ± 1.9, and 3.0 ± 2.1, respectively (p < 0.001) with subjective scores of 4.5 ± 0.3, 3.9 ± 0.4, and 3.3 ± 0.3, respectively (p < 0.001). CNR of the hepatic vein was 5.7 ± 2.3, 4.2 ± 1.9, and 2.7 ± 1.4, respectively with subjective scores of 4.3 ± 0.3, 3.8 ± 0.4, and 3.2 ± 0.3, respectively (p < 0.001). CONCLUSION: Optimal monochromatic images can lead to improvement in the imaging parameters and optimization of the image quality of the common hepatic artery, hepatic portal vein and hepatic vein compared with conventional mixed kV and with 70 keV datasets.
Angiography ; Dataset ; Hepatic Artery ; Hepatic Veins ; Humans ; Liver ; Noise ; Portal Vein ; Retrospective Studies ; Signal-To-Noise Ratio ; Tomography, X-Ray Computed