Construction of SNP-STR Multiplex Amplification System with Genetic Markers and Its Forensic Application / 法医学杂志

Objective To select and develop a SNP-STR multiplex amplification system with genetic markers compatible with current STR databases. To understand its genetic polymorphisms in Sichuan Han population and its application value in DNA mixture analysis. Methods Based on the STR genetic markers in commercial kits, SNPs adjacent to these STR markers were selected to be SNP-STR genetic markers. A SNP-STR multiplex amplification system with genetic markers based on allele-specific amplification was constructed using allele-specific amplification primers. The genetic polymorphism of the system in the Sichuan Han population was investigated and the efficiency of systems with different numbers of loci to detect the two individual DNA mixture samples was evaluated. Results An allele-specific multiplex amplification system constituted of 13 SNP-STR genetic markers was selected and constructed. In Sichuan Han population, the heterozygosity of each locus ranged from 0.76 to 0.88, and the combined discrimination power reached 0.999 999 999 999 999 968. In the analysis of the two individual DNA mixture samples： for single-locus amplification, the genotype of the minor components can still be detected when the mixture ratio reaches 1 000∶1; for multiple loci multiplex amplification, the maximum mixture ratio can reach 500∶1. As the number of loci in the system increased, the detection efficiency of the minor components in the DNA mixture decreased. Conclusion SNP-STR genetic markers have a higher polymorphism than STR. The multiplex amplification system made of SNP-STR genetic markers has a better analysis efficiency for mixed samples than traditional STR multiplex amplification system.

Exposure status of multiple environmental pollutants during pregnancy: based on biomonitoring data from Laizhou Wan Birth Cohort in Shandong / 上海交通大学学报（医学版）

Objective · To evaluate the levels of environmental pollutants including lead, mercury, organophosphorus pesticides (OPs), perfluoroalkyl and polyfluoroalkyl substances (PFASs) and triclosan (TCS) and further analyze the correlation between these pollutants in pregnant women.Methods · Pregnant women were recruited from the Laizhou Wan Birth Cohort (LWBC) in Shandong from September 2010 to December 2013. A total of 149 pregnant women were finally enrolled who completed questionnaires and provided sufficient biological samples for pollutants measurement including blood lead, blood mercury, urinary metabolites of OPs[dimethylphosphate (DMP), dimethylthiophosphate (DMTP), diethylphosphate (DEP), diethylthiophosphate (DETP), etc.], serum perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), as well as urinary TCS. Spearman correlation analysis and cosine cluster analysis were used to explore the correlation between pollutants. Results · The detection rates of lead, DMP, PFOA and PFOS were all 100.0％. And the detection rates of mercury, DMTP, DEP, DETP and TCS were 89.3％, 81.2％, 97.3％, 96.6％ and 59.1％, respectively. The median and range of concentrations for lead, mercury, PFOA, PFOS and TCS were 28.40 (11.30–65.70) μg/L, 0.85 (＜LOD–10.98) μg/L, 39.54 (1.16–273.68) μg/L, 4.56 (0.55–15.38) μg/L, 0.58 (＜LOD–58.01) μg/g, respectively. The median and range of concentrations for DMP, DMTP, DEP and DETP were 36.33 (0.55–1 331.04) μg/g, 2.65 (＜LOD–128.84) μg/g, 14.70 (＜LOD–585.05) μg/g, 1.84 (＜LOD–86.21) μg/g, respectively. The concentrations of DMP and DEP were generally higher than those in developed countries. The concentration of PFOA was much higher than those in foreign studies, while the concentrations of PFOS and TCS were relatively lower. Correlation analysis and cosine cluster analysis revealed that mercury was positively correlated with PFOA (r=0.36, P=0.000) and PFOS (r=0.42, P=0.000). Conclusion · The population in LWBC is widely exposed to multiple pollutants and there are certain correlations between mercury and PFASs, suggesting that attention should be paid to emerging pollutants besides traditional ones.

Polarity of ameloblasts and odontoblasts and their related regulators / 华西口腔医学杂志

The polarity of ameloblasts and odontoblasts is crucial for their differentiation and function. Polarity-related molecules play an important role in this process. This review summarizes the process of polarity formation of ameloblasts and odontoblasts and their related regulators.

Association between exposure to organophosphate pesticides and levels of oxidative stress in pregnant women with different paraoxonase 1 genotypes / 上海交通大学学报（医学版）

Objective·To investigate whether paraoxonase 1 (PON1) genotypes were effect modifiers in the relationship between exposure to organophosphate pesticides (Ops) and oxidative stress level in pregnant women.Methods · A total of 204 pregnant women recruited from a hospital in Shandong Province were included in the study.Four nonspecific dialkyl phosphate (DAP) metabolites of Ops were measured in each urine sample.Levels of two oxidative stress biomarkers [total free sulfhydryl (-SH) and malondialdehyde (MDA)] were measured in serum samples.Blood samples were also analyzed for detecting PON1 genotypes (PONI-108,PON1192 and PON155).Separate linear regression models were conducted to explore the relationship between DAP metabolite levels and oxidative stress levels in all 204 pregnant women or women within each PON1 genotype.Results· There was no significant association between DAP metabolite levels and oxidative stress levels in all 204 women.Levels of dimethyl phosphates [β (95％ CI):-104.10 (-191.31,-16.88)] and dialkyl phosphates [f (95％ CI):-111.78 (-221.84,-1.72)] were negatively associated with-SH level among pregnant women with PON1192RR genotype,but this association was not found among women with other genotypes.Conclusion· OP exposure may be associated with a higher oxidative stress level among pregnant women with PONI192RR genotype.

Effects of fenvalerate exposure during puberty on oxidative stress in male rat testis / 上海交通大学学报（医学版）

Objective· To study the effects of fenvalerate exposure during puberty on oxidative stress in rat testis.Methods· Fifty male Sprague-Dawley (SD) rats were randomly divided into the control group (corn oil),low dose group (0.02 mg/kg fenvalerate),moderate dose group (1 mg/kg fenvalerate),high dose group (50 mg/kg fenvalerate) and intervention group (50 mg/kg fenvalerate+100 mg/kg N-acetyl-L-cysteine),ten rats for each group,for two months by gavage at four weeks of age.Malondialdehyde (MDA) content,activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in testis and testicular tissue morphology were detected.Results· Compared with the control group,the rat body weight and activities of GSH-Px and SOD in testis were significantly decreased in high dose group while MDA content was increased (all P＜0.05).Compared with the high dose group,MDA content was decreased and GSH-Px activity was increased in the intervention group (both P＜0.05).The results of testicular histology showed that with the increasing exposure dose,the spermatogenic cells were arranged loosely,the number of layers was decreased and the inner diameter of seminiferous tubules was increased.Conclusion· Exposure to fenvalerate during puberty may induce oxidative damage in testis tissue of male rats.

The role of bone morphogenetic protein signaling pathway in tooth root development / 华西口腔医学杂志

The bone morphogenetic protein (BMP) family is an important factor in the regulation of cell ular life activities and in the development of almost all tissues. BMP-mediated signaling plays an important role in tooth root development, which is a part of tooth development. Epithelial and mesenchymal interactions are involved in tooth root development, but the BMP signaling pathway has a different effect on tooth root development in epithelial and mesenchymal. This review summarizes the advances of BMP signaling in tooth root development.

Amelioration of Tripterygium wilfordii Polycoride on TNBS/Ethanol-induced Ulcerative Colitis via Inhibiting Lipid Peroxidation and Its Downstream Inflammatory Meditors / 中草药·英文版

Objective To study the protective effect of Tripterygium wilfordii polycoride (TWP) against TNBS/ethanol-induced ulcerative colitis (UC) rat model. Methods TNBS and ethanol enema were adopted to build TNBS/ethanol-induced UC rat model. Ninety male Wistar rats were divided into six groups: normal, model, low-, medium-, high-dose TWP and azathioprine (AZA) groups, each for 15 rats. All rats were administered by corresponding medicine for 14 d. After 14 d, corresponding colon tissues underwent general and microscopic evaluation. Blood samples were taken from heart and serum was separated by centrifugation. MDA, SOD, GSH, IL-1β and TNF-α levels in serum were tested by ELISA. Colonic samples underwent RT-PCR and Western blotting analysis. Results DAI, general and microscopic evaluation all showed that TWP could promote colonic mucosa healing and such effect was equal to AZA. ELISA results about lipid peroxidation indicated that TWP could decrease MDA level and increase SOD and GSH levels in a dose-dependent manner. TWP with high dose could strongly decrease the MDA level and increase the SOD and GSH levels (P 0.05), whereas slightly stronger towards terminal inflammatory cytokines (P > 0.05). Conclusion TWP could significantly lower the infiltration of inflammatory cells under microscope, eventually led to mucosa healing, the mechanism of which was to inhibit lipid peroxidation, then further inhibit NF-κB activation, eventually lower the expression of inflammatory meditors locally and systemically.

Regulatory effect of Tripterygium wilfordii polycoride (TWP) towards TLR4/MyD88 independent pathway in TNBS/ethanol ulcerative colitis (UC) rat model / 中国中药杂志

In order to study the regulatory effect of Tripterygium wilfordii polycoride (TWP) towards TLR4/MyD88 independent pathway in TNBS/ethanol ulcerative colitis (UC) rat model, TNBS/ethanol enema was adopted to build TNBS/ethanol UC rat model. After the successful modeling procedure, 90 male Wistar rats are were divided into 6 groups, including namely normal group, model group, TWP low, middle, high dose groups (3, 6, 12 mg•kg⁻¹)and azathioprine (AZA) group (6 g•kg⁻¹), with 15 rats in each group. All rats in each group were administrated with corresponding medicines for 14 days. After 14 days of administration, corresponding colon tissues were taken for general and microscopic evaluation. Western blotting analysis and RT-PCR were adopted to test the mRNA and protein expressions of TLR4/MyD88 independent pathway-related molecules, namely TLR4, TRAM, TRIF, NF-κB and IFN-γ. The results showed that DAI, general and microscopic evaluations all indicated that TNBS/ethanol UC rat model was successful. TWP can improve UC-related clinical manifestation and heal colonic mucosa, which was equal to AZA. RT-PCR and WB results showed that the expression of TLR4/MyD88 independent pathway-related molecules in model group were significantly superior to that in normal group at either mRNA or protein level (P<0.01). Compared with model group, TWP can inhibit the expression of each node in TLR4/MyD88 independent pathway in a dose-dependent manner. The inhibitory effect of TWP with high dose towards the above molecules was inferior to that in model group at either mRNA or protein level (P<0.05). The inhibitory effect of TWP with high dose towards upstream molecules of TLR4/MyD88 independent pathway (TLR4, TRAM, TRIF, NF-κB) was slightly superior to AZA group at either mRNA or protein level. However, such inhibitory effect towards terminal inflammatory cytokines (IFN-γ) was inferior to AZA group at either mRNA or protein level. All the above differences had no statistical significance. Therefore, in TNBS/ethanol UC rat model, TLR4/MyD88 independent pathway took part in regulating inflammation. TWP exerted its anti-inflammation effect by inhibiting the expression of TLR4/MyD88 independent pathway in a dose-dependent manner.

Study on substance basis of Yinhuang Qingfei Capsule in vitro by HPLC-Q-TOF-MS/MS / 中草药

Objective: To identify and analyze the chemical constituents in Yinhuang Qingfei Capsule by HPLC-Q-TOF-MS/MS method. Methods: The HPLC method was used with the conditions that the column was Inertsil ODS-2 C18 (250 mm × 4.6 mm, 5 μm). Columu and electrospray ion (ESI) source was employed for the qualitative analysis under positive ion mode. These components were further analyzed by MS spectra, and by comparing with the corresponding reference substances and literature data. Results: According to the MS principle and literature data, 54 compounds were identified from the sample of Yinhuang Qingfei Capsule. Conclusion: An efficient HPLC-Q-TOF-MS/MS approach has been established for studying the chemical constituents in Yinhuang Qingfei Capsule, which paves a way for the quality control and further substance basis studies of the preparation.

Study on effects of Tripterygium wilfordii polycoride in resisting macrophage inflammation and regulating inflammation via TLR4/NF-κB / 中国中药杂志

To investigate the effect of Tripterygium wilfordii polycoride (TWP) on LPS-induced macrophage inflammatory response, particularly the inhibitory effect on inflammatory factors TNF-α and IL-1β and the regulatory effect on inflammation via TLR4/NF-κB. The MTT method was adopted to test the effects of tested drugs, TWP, dexamethasone (DXM) and azathioprine (AZA) on cell growth to define the appropriate concentration. LPS was used to induce the inflammatory reaction in mouse RAW264. 7 cell lines. The Elisa kit was adopted to test the release level of TNF-α and IL-1β. The Western blotting was applied to test the protein expressions of TNF-α and IL-1β. The RT-PCR was adopted to test the expressions of TLR4 and NF-κB. According to the results, TWP could inhibit the release of macrophage inflammatory factors TNF-α and IL-1β in a dose dependent manner. All of TWP groups showed a weaker efficacy than that of the DXM group. But the TWP high dose group revealed a better effect on TNF-α and equal effect on IL-1β compared with the AZA group. TWP show an equal or better effect in down-regulating TLR4 and NF-κB p65 expressions in a dose dependent manner than DXM and AZA. In conclusion, TWP could inhibit TLR4 and NF-κB p65, which may be related to the down-regulation of TLR4 and NF-κB p65 receptor expressions.

Role of sclerostin in the bone loss of postmenopausal chinese women with type 2 diabetes / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.</p><p><b>METHODS</b>The postmenopausal patients suffering from type 2 diabetes mellitus and age, body mass index, and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion. The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine (L1-L4), femoral neck, and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry, respectively. Meanwhile, the clinical and laboratory indexes of bone mineral metabolism were analyzed. Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.</p><p><b>RESULTS</b>Finally, 265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group, respectively. Serum sclerostin level of the diabetic group was significantly higher than that of the control group (48.2±19.4 vs. 37.2±18.6 pmol/L, P<0.001) and was increased with age in both groups (diabetic group, r=0.374, P<0.001; control group, r=0.312, P<0.001). In type 2 diabetes patients, serum sclerostin concentration was positively correlated with hemoglobin A1c level (r=0.237; P=0.021). Biochemical bone turnover markers, intact parathyroid hormone and bone-specific alkaline phosphatase, were negatively associated with serum sclerostin level (r=-0.138, P=0.078 and r=-0.265, P<0.001). Conversely, the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients (r=0.354, P<0.001). Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine, femoral neck, and total hip (r=0.324, 0.367, and 0.416, respectively; all P<0.001).</p><p><b>CONCLUSIONS</b>Sclerostin might participate in the pathogenesis of bone loss of type 2 diabetes. The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.</p>

Saturated fatty acid induces insulin resistance partially through nucleotide-binding oligomerization domain 1 signaling pathway in adipocytes / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the potential role of nucleotide-binding oligomerization domain 1 (NOD1), a component of the innate immune system, in mediating lipid-induced insulin resistance in adipocytes.</p><p><b>METHODS</b>Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments. The effect of oleate/palmitate mixture on nuclear factor-κB (NF-κB) activation was analyzed by reporter plasmid assay. The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[3H] glucose uptake assay. Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA (siRNA) targeting NOD1 upon fatty acids treatment were analyzed.</p><p><b>RESULTS</b>Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6, tumor necrosis factor-α, and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4, and these effects were blocked by siRNA targeting NOD1. Furthermore, saturated fatty acids decreased the ability of insulin-stimulated glucose uptake. Importantly, siRNA targeting NOD1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake.</p><p><b>CONCLUSION</b>NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes, suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.</p>

Effect of maternal BDE-209 exposure on sexual development in male offspring rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effect of maternal decabromodiphenyl ether (BDE-209) exposure on the sexual development in male offspring rats.</p><p><b>METHODS</b>Twenty-four pregnant Sprague-Dawley rats were randomly divided into three BDE-209 exposure groups and one control group. The three BDE-209 exposure groups were given BDE-209 (100, 300, and 900 mg/kg) by gavage on gestational days 12∼18, and the control group was given corn oil. The body weight and body length of each newborn male rat was measured at postnatal days 4, 10, 16, and 21. Twelve newborn male rats were randomly selected from each group; anogenital distance was measured at postnatal day 21, serum testosterone was measured, and the organ coefficient of testis was calculated.</p><p><b>RESULTS</b>The newborn male rats in all exposure groups showed declining trends in body weight and body length compared with those in the control group, and the 900 mg/kg BDE-209 exposure group had significantly lower body weight and body length than the control group at postnatal days 4, 10,16, and 21 (P < 0.01). At postnatal day 21, the 100, 300, and 900 mg/kg BDE-209 exposure groups had anogenital distances of 17.82±2.35 mm, 16.32±1.66 mm, and 15.80±1.34 mm, respectively, demonstrating a significant decrease with increased exposure dose (P < 0.05), but no significant difference was found when comparing these values with that of the control group (16.64±2.38 mm) (P > 0.05). There were no significant differences in serum testosterone and organ coefficients of testis and epididymis between the control group and BDE-209 exposure groups (P > 0.05).</p><p><b>CONCLUSION</b>Maternal exposure to BDE-209 has adverse effect on the growth of male offspring rats, but it leads to no significant changes in sexual development.</p>

Effect of carbon disulfide on mitochondrial apoptosis-related proteins in cytoplasm of testicular tissue among male rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the role of mitochondrial pathway in the apoptosis of spermatogenic cells induced by inhalation of carbon disulfide in male rats.</p><p><b>METHODS</b>Twenty-four male Sprague-Dawley rats (clean grade) were divided into four groups according to their body weights: three CS(2) exposure groups (CS(2) concentrations: 50, 250, and 1250 mg/m(3)) and a control group. The rats in CS(2) exposure groups were exposed to CS(2) by static inhalation for 10 weeks (2 h/d, 5 d/w), while the rats in control group were exposed to air. Then, all rats were sacrificed by decapitation; testicular tissues were collected, and cytoplasmic proteins were extracted; the levels of apoptosis-inducing factor (AIF), cytochrome c (cyto c), Bcl-2, Bax, procaspase-9, and procaspase-3 were measured by Western blot, and the activities of caspase-9 and caspase-3 were measured using a test kit.</p><p><b>RESULTS</b>Compared with the control group, all CS(2) exposure groups had significantly increased levels of cyto c in the cytoplasm of testicular tissue (P<0.05); in the 250 mg/m(3) CS(2) exposure group, the Bax/Bcl-2 ratio and activities of caspase-9 and caspase-3 increased significantly (P<0.05), and the content of procaspase-9 and procaspase-3 decreased significantly (P<0.05); in the 1250 mg/m(3) CS(2) exposure group, the relative expression levels of Bax and AIF in cytoplasm increased significantly (P<0.05), and the expression level of Bcl-2 decreased significantly (P<0.05).</p><p><b>CONCLUSION</b>Mitochondrial pathway plays an important role in the CS(2)-induced apoptosis of spermatogenic cells in testicular tissue among male rats.</p>

Linoleic acid activates GPR40/FFA1 and phospholipase C to increase Ca2+i release and insulin secretion in islet beta-cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To elucidate GPR40/FFA1 and its downstream signaling pathways in regulating insulin secretion.</p><p><b>METHODS</b>GPR40/FFA1 expression was detected by immunofluorescence imaging. We employed linoleic acid (LA), a free fatty acid that has a high affinity to the rat GPR40, and examined its effect on cytosolic free calcium concentration ([Ca2+]i) in primary rat beta-cells by Fluo-3 intensity under confocal microscopy recording. Downregulation of GPR40/FFA1 expression by antisense oligonucleotides was performed in pancreatic beta-cells, and insulin secretion was assessed by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>LA acutely stimulated insulin secretion from primary cultured rat pancreatic islets. LA induced significant increase of [Ca2+]i in the presence of 5.6 mmol/L and 11.1 mmol/L glucose, which was reflected by increased Fluo-3 intensity under confocal microscopy recording. LA-stimulated increase in [Ca2+]i and insulin secretion were blocked by inhibition of GPR40/FFA1 expression in beta-cells after GPR40/FFA1-specific antisense treatment. In addition, the inhibition of phospholipase C (PLC) activity by U73122, PLC inhibitor, also markedly inhibited the LA-induced [Ca2+]i increase.</p><p><b>CONCLUSION</b>LA activates GPR40/FFA1 and PLC to stimulate Ca2+ release, resulting in an increase in [Ca2+]i and insulin secretion in rat islet beta-cells.</p>

Genome-wide analysis of DNA methylation changes in the rat ovary after prenatal exposure to di-(2-ethylhexyl)-phthalate / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of prenatal exposure to Di-(2-ethylhexyl)-phthalate (DEHP) on genome-wide epigenetic alterations in ovary of adult offspring rat.</p><p><b>METHODS</b>Pregnant Wistar rats were randomly treated with DEHP (1000 mg/kg) or con oil at 12 - 17 days upon pregnance. DNA methylation changes in the ovary for the adult offsprings which were 70 days old were detected by Rat DNA methylation promoter plus CpG island arrays CpG island chip. Gene ontology (GO) method was performed to analyze the function of genes which were significantly different between exposed group and control group. Gene Igfbp1 (insulin-like growth factor binding protein 1) and Itga3 (integrin alpha 3) were randomly selected and the methylation status were verified by bisulfite genomic sequencing (BSP).</p><p><b>RESULTS</b>The methylation status were significantly different between exposed and control group in 406 genes (71 genes as hypermethylation and 335 genes as hypomethylation) (P < 0.05). GO analysis revealed that molecular transducer activity, cell part, cell, cellular process, multicellular organismal process, response to stimulus, biological regulation, regulation of biological process, reproduction, reproductive process, and rhythmic process were involved. The sequencing results were consistent with the data obtained by chips.</p><p><b>CONCLUSION</b>This study provides evidence that prenatal exposure of DEHP may be associated with methylation changes on the genes in the rat ovary. Genes related to reproductive process have highly significant methylation changes, which may shed new light on mechanisms of reproductive and developmental toxicity after prenatal exposure to DEHP.</p>

Effects of in utero exposure to di(2-ethylhexyl) phthalate on sexual development in female offspring / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate the ability of di(2-ethylhexyl) phthalate (DEHP) with inducing damage in sexual development of female offspring rats after maternal exposure.</p><p><b>METHODS</b>On gestational day (GD) 12, pregnant Wistar rats were weighed, encoded and randomly assigned to 5 groups (10 dams per group). From GD 12 through GD 17 each dam was dosed daily by gavage with either corn oil (vehicle control, 1 mgxkg(-1)xd(-1)) or DEHP (1, 250, 750 and 1000 mgxkg(-1)xd(-1)). Then female offspring were monitored for eye opening on postnatal day (PND) 14-17, organ coefficient on PND 22 and the time to vaginal opening on PND 30 - 38 (if vagina did not open during the period, observation time should extent to adult), as well as body weight, time to first estrus.</p><p><b>RESULTS</b>No significant changes were observed on eye opening at any dose, which were (15.8 +/- 0.4) d, (16.3 +/- 0.6) d, (16.0 +/- 0.6) d, (15.9 +/- 0.6) d, (15.8 +/- 0.4) d respectively in control, 1, 250, 750 and 1000 mgxkg(-1)xd(-1) (F = 1.363, P = 0.262). However, 62.50% (15/24), 81.25% (26/32) female offspring were permanently absence of vaginal orifice in 750 and 1000 mgxkg(-1)xd(-1) groups respectively, while control, 1 and 250 mgxkg(-1)xd(-1) groups developed normally with vaginal orifices (chi(2) values were 84.92, 132.79, respectively, P < 0.01). The ages of vaginal opening were (32.7 +/- 1.3) d, (33.3 +/- 1.5) d, (32.2 +/- 1.5) d, (33.1 +/- 1.3) d, (33.3 +/- 1.2) d and the body weight were (91.56 +/- 6.65) g, (93.79 +/- 6.28) g, (92.98 +/- 8.48) g, (100.57 +/- 6.47) g, (103.83 +/- 8.24) g in control, 1, 250, 750 and 1000 mgxkg(-1)xd(-1). After covariance adjustment for body weight, which can statistically influenced the age of vaginal opening (F = 40.857, P < 0.05), difference were found at the age of vaginal opening (F = 3.075, P < 0.05), and 250 mgxkg(-1)xd(-1) group was advanced than control (t = -2.056, P < 0.05).</p><p><b>CONCLUSION</b>Exposure to DEHP in utero from GD 12 - 17 can result in abnormalities of sexual development such as the time to vaginal opening and vaginal atresia.</p>

Phosphoinositide 3-kinase/Akt and nuclear factor-kappaB are involved in Staphylococcus aureus-induced apoptosis in U937 cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To explore the mechanisms involved in Staphylococcus aureus (S. aureus) invading human monocytic U937 cells.</p><p><b>METHODS</b>S. aureus were added to U937 cells at multiplicity of infections (MOI) of 20:1 for 0, 15, 30, 60, and 90 minutes, respectively. Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry analysis. Akt and nuclear factor-kappaB (NF-kappaB) activities were detected by Western blotting.</p><p><b>RESULTS</b>Infection of U937 cells with S. aureus induced rapid cell death in a time-dependent manner, and the cells displayed characteristic features of apoptosis. S. aureus-induced apoptosis was associated with a prominent downregulation of activated (phosphorylated) Akt and NF-kappaB. The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-kappaB in a dose-dependent manner. Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells.</p><p><b>CONCLUSIONS</b>S. aureus can stimulate the apoptosis of U937 cells. S. aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-kappaB.</p>

Exploration and Practice on the Reformation for the Microbiology Experiment Teaching Methods / 微生物学通报

In order to meet the requirements of cultivating the practical abilities and creativities of students who receive higher education, we initiated the reformation of education in the microbiology experiment teaching methods, implementing a system for module-based education, carefully monitoring every link in teaching, combining the encouragement and strict requirements together, adopting a proper way of assessment. It is proven that the implementation of the educational reformation mobilizes the interests of students and enhances the comprehensive qualities of students, which accomplishes the purposes of teaching.

Involvement of p38 mitogen-activated protein kinase in E. coli-induced U937 apoptosis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate whether the effect of E. coli on U937 cell lines apoptosis is mediated via p38 mitogen-activated protein kinase (MAPK) activation.</p><p><b>METHODS</b>The U937 cell lines were treated with E. coli at different time or together with SB203580, an inhibitor for p38. Cell apoptosis was analyzed by flow cytometry. p38 activities were detected by Western blotting.</p><p><b>RESULTS</b>E. coli induced apoptosis in cultured U937 cell lines in a time-dependent manner. The phosphorylation of p38 was induced after 10 minutes infection, reached the peak after 20 minutes, and started to decline after 30 minutes. In contrast, the level of total p38 protein was not changed in whole experimental period. Inhibition of p38 with SB203580 significantly inhibited E. coli induced apoptosis in U937 cells.</p><p><b>CONCLUSION</b>The activation of the p38 MAPK in U937 cell lines by E. coli is a major pathway to mediate the apoptosis.</p>