Adolescent ; Adult ; Air Pollutants, Occupational ; adverse effects ; analysis ; Cadmium Compounds ; adverse effects ; analysis ; Eye ; drug effects ; Humans ; Oxides ; adverse effects ; analysis ; Young Adult

Objective To investigate the effects of substrate stiffness on the proliferation of human umbilical vein endothelial cells ( HUVEC) during dengue virus infection.Methods Polyacrylamide gels were prepared for cell culture [(0±4) kPa].The proliferation of HUVEC cultured on substrates with differ-ent stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-etrazolium,inner salt ( MTS) assay.The cycle and apoptosis of HUVEC were determined by flow cytometry analysis.Dengue virus serotype 2 (DENV-2) strains were propagated and identified by con-ventional assays.The HUVEC were infected with DENV-2 strains at a MOI of 10 and cultured on traditional plastic and hydrogel substrates, respectively.The levels of nitric oxide (NO) and endothelin-1 (ET-1) were detected by nitric acid reductase assay and double antibody sandwich ELISA.Results Young′s modulus E value of the hydrogels was (3030 ±0.44) Pa.The proliferation of HUVEC and the expression of NO and ET-1 were enhanced along the increased substrate stiffness.However, no significant differences with the cell cycle and apoptosis were observed between cells cultured on different substrates.Conclusion The stiffness of substrates affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection.The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury.The establishment of hydrogel substrates as the model of vascu-lar basement membranes might provide a new way for the in vitro investigation of the pathogenesis of DENV infection.

Objective To analyze the effects of dengue virus 2 ( DENV-2 ) infection on the ex-pression of IL-29 in primary human umbilical vein endothelial cells ( HUVECs) cultured on hydrogel sub-strates .Methods Primary HUVECs were isolated and cultured on hydrogel substrates .DENV-2 stains were used to infect the primary HUVECs at a multiplicity of infection( MOI) of 10.Flow cytometry analysis was performed to detect the apoptosis and infection rate of HUVECs after 48 hours of culturing .The gene chip profiling was performed to analyze mRNA expression .The expression of IL-29 at mRNA and protein levels were measured by real-time fluorescent quantitative PCR analysis and double antibody sandwich ELISA as -say, respectively.Results Compared with 96.36%of baby hamster kidney (BHK) cells that were infected with DENV-2 stains, only 4.71%primary HUVECs cultured on hydrogel substrates were infected .The pri-mary HUVECs cultured on hydrogel substrates with or without DENV-2 infection showed no significant differ-ences with the rates of cell apoptosis and infection (P>0.05).A significant difference was observed with the expression of IL-29 at mRNA level between primary HUVECs cultured on hydrogel substrates and the cells cultured in plastic bottles (P<0.05).The results of the real-time quantitative PCR analysis and ELISA as-say showed that IL-29 was highly expressed in DENV-2 infected primary HUVECs cultured on hydrogel sub-strates as compared with those in control groups (P<0.05).Conclusion The expression of IL-29 was de-tected in DENV-2 infected primary HUVECs cultured on hydrogel substrates , which was significantly differ-ent from that in DENV-2 infected primary HUVECs cultured in plastic bottles .The successful establishment of hydrogel substrates as the model of vascular basement membranes might provide a new way for the investi -gation of the pathogenesis of DENV infection .

This study is to study is to investigate the coumarins from Fruit of Cnidium monnieri and their cytotoxic activities. The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for their cytoxic activities by MTT method. Eleven compounds were isolated and identified as osthole (1), bergaptan (2), xanthotoxol (3), xanthotoxin (4), imperatorin (5), isopimpinellin (6), osthenol (7), psoralen (8), 5,7-dimethoxycoumarin (9), oxypeucedaninhydrate (10), and swietenocoumarin F (11). Compounds 7, 9-11 were isolated from the Cnidium genus for the first time. Compounds 1,5,10 and 11 showed significant cytotoxic activities against L1210 cell lines at a concentration of 1 x 10(-5) mol x L(-1) with inhibitory rates of were 70.13, 63.10, 55.77, and 75.08% respectively.
Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cnidium ; chemistry ; toxicity ; Coumarins ; chemistry ; isolation & purification ; toxicity ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Fruit ; chemistry ; toxicity ; Mice ; Molecular Structure

Objectives To provide prenatal diagnosis and genetic counseling for four athigh-risk pregnant women with a suspected family or personal history of fragile X syndrome (FXS) by genetic screening of fragile X mental retardation (FMR1) gene.Methods This study was conducted on four pregnant women (No.l to 4) who received outpatient treatment in Peking University First Hospital from August 2014 to June 2016.Genomic DNA was extracted from peripheral blood samples of the pregnant women and six of their family members,four of which were suspected or confirmed FXS and the other two were FMR1 gene carriers.Amplide X kits were used to detect CGG repeat size in FMR1 gene.Two amniocytes and one chorionic villi samples were collected from three pregnant women to extract DNAs for FMR1 gene and karyotyping analyses.Results There were patients diagnosed with FXS in all the families by detecting CGG repeat numbers in FMR1 gene.The pregnant woman No.1 was a permutation carrier;No.2 carried normal FMR1 alleles while her brother had a mutation with over 20 CGG repeats in FMRI gene at chromosome X.No.3 and 4 were full mutation carriers with over 200 CGG repeats in FMR1 gene.After genetic counseling,No.3 decided to terminate the pregnancy due to abnormal fetal karyotype (47,XY,+21) and full mutation of FMR1 alleles.No.1 and 4 continued to pregnancy as their fetuses were normal in FMR1 alleles and karyotype.No.2 continued to pregnancy as her fetus was free of FXS risk.Conclusions Prenatal diagnosis and genetic counseling should be conducted on women at highrisk for FXS to avoid birth defects.People with a family history of FXS should be tested for FMR1 gene carrier status.

This research adopted silt as the sample,and the five highest hydrogen production performing strains contained in the sample were isolated. The strain whose hydrogen production was the highest was identified as Enterobacter cloacae by the analysis of 16S rDNA sequencing and comparison. It is showed by Plackett-Burman Experimental Design that only glucose,citric buffer and reducing agent had significant effects on hydrogen production by Enterobacter cloacae FML-C1. The path of steepest ascent was undertaken to approach the optimal response region of those three factors. Central Composite Design(CCD) and Response Surface Methodology(RSM) were employed to investigate the interaction of the variables and to ascertain the optimal values of the factors,which finally led to the maximum hydrogen production(VH2) . The theoretical optimal medium conditions were:glucose 21.5 g/L,citric buffer 13.6 mL/L,reducing agent10.0 mL/L. The five tentative tests matched this model well. The final VH2 was up to 2347.4 mL/L,which was 127.42% enhanced in comparison to the original. The result shows that PB experiment design and RSM analytical method work well in selecting factors which have significant influences on the hydrogen production and,moreover,achieve the ideal optimal result.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

Objective To evaluate the protective effect on lung by using continuous pulmonary artery perfusion with oxygenated blood and L-arginine during cardiopulmonary bypass(CPB).Methods Forty five cases received mitral valve replacement were randomly divided into 3 groups and each group involved 15 cases. Group I(control group), patients received routine procedure of CPB. Proup Ⅱ, patients received rcontinuous pulmonary artery perfusion with oxygenated blood. Group Ⅲ,continuous pulmonary artery perfusion with oxygenated blood containing L-arginine (200 mg/kg) (n=15). All cases received routine procedure of CPB and continuously infused from the root of pulmonary artery until releasing aortaoaic clamp. Blood samples were collected from the radial artery respectively at the following time points:after the induction of anaesthesia, 1 hour after opening of aorta, 0, 6, 12, 24 hours after patients being taken back to ICU. ELISA test was used to detected the expression of tmmor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10). Lung tissue samples (1.0 cm ×1.0 cm×1.0 cm) were obtained from right lower lobe. The pathologicl changes of lung tissues were observed under light mi-croscope by using HE staining. Results at each time points, the expression of TNF-α, IL-6 in group Ⅱ and group Ⅲ weresignificantly lower than that in group Ⅰ (P<0.05). The level of TNF-α, IL-6 in group Ⅲ were lower than in group Ⅱ(P<0.05). However, the expression of IL-10 in group Ⅱ and group Ⅲ were higher than in group Ⅰ, and the level of IL-10 in group Ⅲ were higher than that in group Ⅱ(P<0.05). In the group Ⅰ: HE staining showed marked pulmonary interstitial edema, intra-alveolus neutrophilic granulocyte exudation with karyorrhexis. In the group Ⅱ, light capillary vessel hyperaemia and pulmonary interstitial lymphocyte exudation were detected. Nearly normal lung tissue were observed in group Ⅲ. Conclusion Continuous pulmonary artery perfusion with oxygenated blood and L-arginine could inhibit the synthesis of inflammatory factors significantly and increase the releasing of anti-inflammatory factors during CPB. Therefore, it may reduces pulmonary inflammatory reaction and have protective effects on lung tissue.

ObjectiveTo study the effect of sub-chronic exposure to deltamethrin(DM) on neural behavior and expression of NMDA receptor in hippocampus of mice.Methods 60 Female SPF Kunming mice were divided into 4 groups and given DM 60 days by gavage.Hot-plate,rotarod,grip strength,hing limb landing foot splay were used to examine the sensory and motor change of mice.Autonomic activity test was used for detecting the functional status of the central nervous system in mice.Passive avoidance test for detection of the behavior changes of learning and memory,and RQ-PCR was employed to measure the expression of NMDA receptor in hippocampus of mice.ResultsThe behavior of sensory and motor of mice sub-chronic exposure to deltamethrin did not have changes significantly(P ＞ 0.05 ).In the test of autonomic activity test,the average of autonomic activity times were (93 ± 18) times,(107 ± 13) times,(105 ±22) times.Compared with the control group,the average of autonomic activity times in middle-and high-dose groups were increased significantly (P ＜ 0.05 ).The latent periods in poisoning groups were (175.4 ±38.4) s,(146.4 ±51.2)s,(132.3 ±45.0) s,and the error times were (0.7 ±0.3)times,( 1.4 ± 0.5 ) times,( 1.8 ± 0.9) times.Compared with the control group,latent periods of high-dose group were decreased and the error times of middle-and high-dose groups were increased significantly (P ＜ 0.05 ).Compared with the control group,the relative expression levels of NR1 and NR2A mRNA in hippocampus of middle and high-dose groups were increased significantly (P＜ 0.05 ),and the relative expression level of NR2B mRNA in highdose group was decreased significantly(P ＜ 0.05).ConclusionSub-chronic exposure to DM could increase the excitability of mice,damage the function of learning and memory,and influence the expression of NMDA receptor in hippocampus of mice.

Objective To explore the clinical factors on the functional recovery of spinal cord cavernous hemangioma operation. Methods Fifty cases patients with complete spinal cord cavernous hemangioma were retrospectively analyzed their clinical data and its influence on prognosis of 2 years. Single factor and multiple factors analysis were performed. Results Compared with the preoperative stage,the grade of Aminoff-Logue score of 28 cases(50. 9%)had improved at least one grade,14 cases(25. 5%)patients with no significant changes and 13 cases(23. 6%)with decreased one grade at least. Logistic regression model showed that patients with clinical types(OR=3. 27,95%CI 2. 34-15. 83,P=0. 006),the time of the attacks to the surgery(OR=2. 93,95%CI 1. 41-10. 61,P=0. 012),and the attacks to the operation time( OR=4. 31,95%CI 2. 46 -12. 32,P = 0. 019 ) were related to neurological function recovery of spinal cord cavernous hemangiomas. Conclusion For patients with SCH dysfunction,regardless of what kind of clinical types of neurological dysfunction extent,should receive surgical treatment to improve neurological function recovery rate as soon as possible.