Vitamin D Deficiency Aggravates Hepatic Oxidative Stress and Inflammation during Chronic Alcohol-Induced Liver Injury in Mice.

Vitamin D deficiency has been reported in alcoholics. This study is aimed at evaluating the effects of vitamin D deficiency on chronic alcohol-induced liver injury in mice. Mice were fed with modified Lieber-DeCarli liquid diets for 6 weeks to establish an animal model of chronic alcohol-induced liver injury. In the VDD+EtOH group, mice were fed with modified diets, in which vitamin D was depleted. Vitamin D deficiency aggravated alcohol-induced liver injury. Furthermore, vitamin D deficiency aggravated hepatocyte apoptosis during alcohol-induced liver injury. Although it has a little effect on hepatic TG content, vitamin D deficiency promoted alcohol-induced hepatic GSH depletion and lipid peroxidation. Further analysis showed that vitamin D deficiency further increased alcohol-induced upregulation of hepatic inducible nitric oxide synthase (inos), two NADPH oxidase subunits p47phox and gp91phox, and heme oxygenase- (HO-) 1. By contrast, vitamin D deficiency attenuated alcohol-induced upregulation of hepatic antioxidant enzyme genes, such as superoxide dismutase (sod) 1 and gshpx. In addition, vitamin D deficiency significantly elevated alcohol-induced upregulation of hepatic proinflammatory cytokines and chemokines. Taken together, these results suggest that vitamin D deficiency aggravates hepatic oxidative stress and inflammation during chronic alcohol-induced liver injury.

Association among placental 11ß-HSD2, PPAR-γ, and NF-κB p65 in small-for-gestational-age infants: A nested case-control study.

PROBLEM: 11ß-Hydroxysteroid dehydrogenase 2 (11ß-HSD2) catalyzes active glucocorticoids into their inactive products, preventing the passage of glucocorticoids into the fetus from maternal circulation. Peroxisome proliferator-activated receptor (PPAR)γ is a member of the nuclear receptor superfamily that regulates the expression of placental 11ß-HSD2. Nuclear factor-kappa B (NF-κB) is a transcription factor that regulates inflammatory signaling. This study aimed to investigate the association among 11ß-HSD2, PPAR-γ, and NF-κB p65 in small-for-gestational-age (SGA) infants. METHOD OF STUDY: Forty-six SGA and 46 appropriate-for-gestational-age (AGA) infants were enrolled in this study. Both newborns and placentas were weighed. Placental 11ß-HSD2 levels were measured using Western blotting. Placental PPAR-γ and NF-κB p65 were detected by immunohistochemistry. Placental inflammatory cytokines were evaluated by real-time RT-PCR. RESULTS: 11ß-HSD2 levels were lower in SGA placentas than those in AGA placentas. Placental PPAR-γ-positive nuclei were less in SGA than those in AGA. By contrast, placental NF-κB p65-positive nuclei were more in SGA than those in AGA. The levels of CRP, TNF-α, IL-8, and IL-1ß, several inflammatory cytokines, were higher in SGA placentas. Correlation analysis showed that neonatal weight was positively associated with PPAR-γ and 11ß-HSD2 in SGA placentas. By contrast, neonatal weight was inversely correlated with NF-κB p65 in SGA placentas. 11ß-HSD2 was positively correlated with PPAR-γ in SGA placentas. CONCLUSIONS: Inflammation-associated downregulation of placental PPAR-γ and 11ß-HSD2 may be involved in SGA.

Vitamin D Deficiency Attenuates Acute Alcohol-Induced Hepatic Lipid Accumulation in Mice.

Vitamin D deficiency has been frequently reported in chronic liver disease. However, its influence on hepatic lipid accumulation in alcoholic liver disease remains unclear. The present study investigated the effects of vitamin D deficiency on acute alcohol-induced hepatic lipid metabolism in mice. Mice were fed with vitamin D deficient diet, in which vitamin D was depleted for 12 weeks to establish an animal model of vitamin D deficiency. Some mice were administered a single gavage of alcohol (4 g/kg bodyweight) before they were euthanized. Results show that feeding mice with vitamin D deficient diet did not induce hepatic lipid accumulation. In contrast, vitamin D deficiency markedly reduced alcohol-induced triacylglycerol (TAG) content and prevented hepatic lipid accumulation. Moreover, vitamin D deficiency significantly attenuated alcohol-induced sterol-regulated element-binding protein (SREBP)-1c activation, which regulates genes for hepatic fatty acid (FA) and TAG synthesis, and the expression of its target genes fatty acid synthase (Fasn) and acetyl-coenzyme- A carboxylase (Acc). In addition, vitamin D deficiency alleviated alcohol-induced downregulation of hepatic nuclear peroxisome proliferator-activated receptor (PPAR)α, which governs FA transport and ß-oxidation, and the expression of Carnitine palmitoyltransferase (Cpt)-1α, cytochrome P450, family 4, subfamily a, polypeptide (Cyp4a)10, and Cyp4a14, which are key enzymes for hepatic fatty acids ß-oxidation and ω-oxidation. Taken together, these results suggest that vitamin D deficiency is not a direct risk factor for hepatic lipid accumulation. Vitamin D deficiency alleviates acute alcohol-induced hepatic lipid accumulation through inhibiting hepatic de novo fatty acid syntheses and promoting fatty acid ß-oxidation and ω-oxidation.

Lipopolysaccharide Downregulates 11ß-Hydroxysteroid Dehydrogenase 2 Expression through Inhibiting Peroxisome Proliferator-Activated Receptor-γ in Placental Trophoblasts.

It is increasingly recognized that excessive glucocorticoids induce fetal intrauterine growth restriction (IUGR). Placental 11ß-hydroxysteroid dehydrogenase 2 (11ß-HSD2), a glucocorticoid-catalyzing enzyme, prevents active glucocorticoids from maternal circulation into the fetus, thus protecting against IUGR. Previous studies demonstrated gestational LPS exposure caused fetal IUGR. The aim of the current study was to investigate the effects of LPS on 11ß-HSD2 in mice placentas and human placental trophoblasts. Pregnant ICR(CD-1) mice were i.p. injected with LPS (200 µg/kg) on gestational day 16. As expected, gestational LPS exposure downregulated 11ß-HSD2 in mice placentas. In vitro, LPS downregulated 11ß-HSD2 in human placental trophoblasts. Additional experiment showed that LPS, which activated NF-κB, suppressed rosiglitazone-induced activation of peroxisome proliferator-activated receptor-γ (PPARγ) in mice placentas and human placental trophoblasts. Moreover, NF-κB p65 knockdown and specific NF-κB inhibitor attenuated LPS-induced suppression of PPARγ nuclear translocation in human placental trophoblasts. In addition, NF-κB p65 knockdown attenuated LPS-induced downregulation of 11ß-HSD2 in human placental trophoblasts. Mechanically, LPS promoted physical interaction between NF-κB p65 and PPARγ in the cytoplasm and nucleus of placental trophoblasts. Finally, pretreatment with rosiglitazone, a PPARγ agonist, partially alleviated LPS-induced reduction of fetal weight and crown-rump length. Taken together, these results suggest that LPS downregulates 11ß-HSD2 through suppressing PPARγ in placental trophoblasts. Placental 11ß-HSD2 downregulation may contribute partially to LPS-induced fetal IUGR.

Perinatal sulfamonomethoxine exposure influences physiological and behavioral responses and the brain mTOR pathway in mouse offspring.

Sulfamonomethoxine (SMM) is widely used in the veterinary field in China. Although some clinical surveys have revealed that sulfonamide antibiotics cause adverse nervous system symptoms, the related mechanisms of maternal SMM exposure on the neurobehavioral development of offspring remain unclear. Here, we investigated the effects of perinatal SMM exposure on the physiological and behavioral responses of pubertal offspring mice and the underlying mechanisms. We randomly allocated pregnant mice into the groups treated with SMM at different doses and the saline-treated groups. Maternal mice were orally administered SMM daily from gestational day 1 to postpartum day 21. On postnatal day (PND) 22, the parameters of growth, endocrine hormones, and brain amino acid composition were assessed, as well as the brain transcript levels of key genes involved in the mammalian target of rapamycin (mTOR) signaling pathway. From PND 50 to 55, a battery of behavioral tests relevant to anxiety and memory were then administered. Analysis of the results indicated that the pups, particularly the pubertal female offspring, showed anxiety-like behavior. Moreover, the pubertal offspring showed cognitive impairments and fat accumulation. Furthermore, the relative mRNA expression of genes involved in the mTOR signaling pathway in females on PND 22 was elevated, whereas the expression of N-methyl-d-aspartate receptor 2B (NR2B) was reduced. Together, the results showed that perinatal SMM exposure perturbs neuroendocrine functions, and further alters gene expression in the mTOR pathway and NR2B gene expression early in life, which may contribute to brain dysfunction in pubertal life.

Obeticholic Acid Protects against Lipopolysaccharide-Induced Fetal Death and Intrauterine Growth Restriction through Its Anti-Inflammatory Activity.

Farnesoid X receptor (FXR) is expressed in human and rodent placentas. Nevertheless, its function remains obscure. This study investigated the effects of obeticholic acid (OCA), a novel synthetic FXR agonist, on LPS-induced fetal death and intrauterine growth restriction. All pregnant mice except controls were i.p. injected with LPS (100 µg/kg) daily from gestational day (GD) 15 to GD17. Some pregnant mice were orally administered with OCA (5 mg/kg) daily from GD13 to GD17. As expected, placental FXR signaling was activated by OCA. OCA pretreatment protected against LPS-induced fetal death. In addition, OCA pretreatment alleviated LPS-induced reduction of fetal weight and crown-rump length. Additional experiments showed that OCA inhibited LPS-evoked TNF-α in maternal serum and amniotic fluid. Moreover, OCA significantly attenuated LPS-induced upregulation of placental proinflammatory genes including Tnf-α, Il-1ß, IL-6, Il-12, Mip-2, Kc, and Mcp-1 By contrast, OCA elevated anti-inflammatory cytokine IL-10 in maternal serum, amniotic fluid, and placenta. Further analysis showed that OCA blocked nuclear translocation of NF-κB p65 and p50 subunits in trophoblast giant cells of the labyrinth zone. These results provide a mechanistic explanation for placental FXR-mediated anti-inflammatory activity. Overall, this study provides evidence for roles of FXR as an important regulator of placental inflammation.

Maternal cadmium exposure reduces placental zinc transport and induces fetal growth restriction in mice.

Cadmium (Cd) is linked with increased risk of fetal growth restriction (FGR). Nevertheless, the mechanism remains unknown. This study established a mouse model of Cd-induced FGR through two exposure methods. Pregnant mice were either administered with CdCl2 (5, 50 and 250ppm) throughout pregnancy through drinking water or intraperitoneally injected with CdCl2 (4.5mg/kg) on GD9. As expected, fetal weight and crown-rump length were reduced in a gender-independent manner. Interestingly, Mt1 and Mt2, two metallothionein genes, were up-regulated in maternal liver. Correspondingly, Cd accumulated mainly in maternal liver and kidney, and only trace amounts of Cd could pass from dam to placentas and fetuses. Further analysis showed that placental Zn concentration was elevated. Conversely, embryonic Zn concentration was reduced. Moreover, placental Znt1 and Znt2, two zinc transporters, were down-regulated in Cd-exposed mice. These results suggest that maternal Cd exposure during pregnancy reduces placental Zn transport and induces fetal growth restriction.

Rosiglitazone pretreatment protects against lipopolysaccharide-induced fetal demise through inhibiting placental inflammation.

Peroxisome proliferator-activated receptor (PPAR)-γ is highly expressed in human and rodent placentas. Nevertheless, its function remains obscure. The present study investigated the effects of rosiglitazone, a PPAR-γ agonist, on LPS-induced fetal death. All pregnant mice except controls were intraperitoneally injected with LPS (150 µg/kg) daily from gestational day (GD)15 to GD17. As expected, maternal LPS injection caused placental inflammation and resulted in 63.6% fetal death in dams that completed the pregnancy. Interestingly, LPS-induced fetal mortality was reduced to 16.0% when pregnant mice were pretreated with RSG. Additional experiment showed that rosiglitazone pretreatment inhibited LPS-induced expressions of tumor necrosis factor (Tnf)-α, interleukin (Il)-1ß, Il-6, macrophage inflammatory protein (Mip)-2 and keratinocyte-derived chemokine (Kc) in mouse placenta. Although rosiglitazone had little effect on LPS-evoked elevation of IL-10 in amniotic fluid, it alleviated LPS-evoked release of TNF-α and MIP-2 in amniotic fluid. Further analysis showed that pretreatment with rosiglitazone, which activated placental PPAR-γ signaling, simultaneously suppressed LPS-evoked nuclear factor kappa B (NF-κB) activation and blocked nuclear translocation of NF-κB p65 and p50 subunits in trophoblast giant cells of the labyrinth layer. These results provide a mechanistic explanation for PPAR-γ-mediated anti-inflammatory activity in the placentas. Overall, the present study provides additional evidence for roles of PPAR-γ as an important regulator of placental inflammation.

Maternal serum lipid levels during late pregnancy and neonatal body size.

OBJECTIVE: The aim of this study was to determine the predictive value of maternal serum lipid levels during late pregnancy for neonatal body size. METHODS: This study was conducted from January 1, 2011 to July 31, 2012 at a Maternal and Child Health Hospital. Fasting blood glucose, serum triglyceride, total cholesterol, HDL and LDL were estimated in maternal collected before delivery. Detailed anthropometry of the neonates was performed. RESULTS: Women who delivered a large for gestational age baby were older, taller, had a higher pre-pregnancy weight, higher pre-pregnancy BMI, and higher weight gain during pregnancy than women who delivered an appropriate for gestational age infant. After adjusting for maternal age, pre-pregnancy BMI, weight gain during pregnancy, parity, neonatal sex and gestational age at delivery, we found that only maternal HDL level was inverse associated with birth weight, length and head circumference (p<0.05). On logistic regression analysis, the significant metabolic predictors of large for gestational age was HDL (OR 0.57, 95%CI: 0.38-0.84, per 1 mmol/L increase) after adjusting for the confounders. CONCLUSIONS: Maternal serum HDL level determined in maternal blood taken close to delivery was independently associated with neonatal size and was the independent predictor for large for gestational age.

Identification of Variable-Number Tandem-Repeat (VNTR) Sequences in Acinetobacter pittii and Development of an Optimized Multiple-Locus VNTR Analysis Typing Scheme / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To develop a multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) assay for Acinetobacter pittii typing.</p><p><b>METHODS</b>Polymorphic VNTRs were searched by Tandem Repeats Finder. The distribution and polymorphism of each VNTR locus were analyzed in all the A. pittii genomes deposited in the NCBI genome database by BLAST and were evaluated with a collection of 20 well-characterized clinical A. pittii strains and one reference strain. The MLVA assay was compared with pulsed-field gel electrophoresis (PFGE) for discriminating A. pittii isolates.</p><p><b>RESULTS</b>Ten VNTR loci were identified upon bioinformatic screening of A. pittii genomes, but only five of them showed full amplifiability and good polymorphism. Therefore, an MLVA assay composed of five VNTR loci was developed. The typeability, reproducibility, stability, discriminatory power, and epidemiological concordance were excellent. Compared with PFGE, the new optimized MLVA typing scheme provided the same and even greater discrimination.</p><p><b>CONCLUSION</b>Compared with PFGE, MLVA typing is a faster and more standardized alternative for studying the genetic relatedness of A. pittii isolates in disease surveillance and outbreak investigation.</p>

Gender difference in the association of hyperuricemia with hypertension in a middle-aged Chinese population.

In this study, we report the relationship between hyperuricemia and hypertension in a middle-aged Chinese population, emphasizing the difference of gender. The cross-sectional study was conducted among 1776 adults aged 45-60 years, who participated in the Hefei Nutrition and Health Study (2012). Hyperuricemia was defined as serum uric acid (SUA)> 420 µmol/l for men, and > 360 µmol/l for women. Hypertension was defined as systolic blood pressure (SBP) ≥ 140 mmHg or diastolic blood pressure (DBP) ≥ 90 mmHg. Anthropometric measurements and biochemical data were collected using standardized procedures. Multivariate logistic regression analysis was performed to determine the relationship between hyperuricemia and hypertension with adjustment of potential confounding factors. Body mass index (BMI), waist circumference (WC), SBP, DBP, fasting glucose, SUA and the prevalence of hyperuricemia and hypertension were significantly higher in male than in female (p < 0.001). Females had significantly higher levels of triglycerides (TG) and high-density lipoprotein (HDL)-cholesterol (5.23 ± 0.87 vs 5.12 ± 1.01, p < 0.05, 1.50 ± 0.37 vs 1.28 ± 0.41, respectively.) than males. Simple correlation analysis showed that SUA was positively associated with WC and TG. In addition, after adjusting for potential confounders, hyperuricemia was associated with increased risk of hypertension in both males and females, with odds ratios (95% CI) of 1.680 (1.110-2.543) and 1.065 (1.012-1.118), respectively. Conclusions: The association of hyperuricemia with hypertension was stronger in males than in females, and middle-aged men with hyperuricemia had greater association with hypertension. Our findings remain to be confirmed in future prospective studies.

[Influence of different dietary patterns on bone mineral density and body mass index of college freshmen in urban and rural areas of China].

OBJECTIVE: To find out the differences of dietary patterns among freshmen coming from urban and rural areas that might have influenced their bone mineral density and body mass index (BMI). METHODS: With stratified random sampling method, dietary patterns and their bone mineral density, BMI of 1319 freshmen were studied. RESULTS: (1) The ratios of urban freshmen who chose "western food" pattern (χ(2) = 31.548, P = 0.000; χ(2) = 13.068, P = 0.001), "animal food" pattern (χ(2) = 8.279, P = 0.016; χ(2) = 41.137, P = 0.000) or "calcium food" pattern (χ(2) = 37.254, P = 0.000; χ(2) = 15.651, P = 0.000) were higher than that of rural freshmen, and the ratios of rural freshmen who chose "Chinese traditional" pattern (χ(2) = 36.194, P = 0.000; χ(2) = 25.936, P = 0.000) were higher than that of urban freshmen. (2) The average height, weight, BMI, speed of sound (SOS) of male freshmen from rural areas were lower than that from the city and the differences were statistically significant (P < 0.05). Among those female freshmen, only height and weight were significantly different (P < 0.05). (3) In both rural and urban freshmen, the factor scores of "western food" pattern had a positive correlation with BMI, with the correlation coefficients as 0.187, 0.192, 0.551, 0.465 (P < 0.001). The factor scores of "calcium food" pattern were positively related to bone mineral density (SOS values) with correlation coefficients as 0.680, 0.342, 0.841, 0.786, P < 0.001 respectively. The factor scores on "Chinese traditional" pattern were negatively correlated with BMI, with correlation coefficients as -0.223, -0.093 (P < 0.05) which were positively related to bone mineral density (SOS values) in both rural and urban male freshmen, with correlation coefficients as 0.905, 0.711 (P < 0.001). CONCLUSION: Different dietary patterns chosen by urban and rural freshmen had a significant impact on both bone mineral density and BMI.

Proteome analysis of Neisseria meningitidis serogroup strains C associated with outbreaks in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>During 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China. With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed.</p><p><b>METHODS</b>Clinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3-10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry.</p><p><b>RESULTS</b>502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426, with lower express volumes too. No Opa and Opc were detected in 053426.</p><p><b>CONCLUSIONS</b>The different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.</p>

Primary analysis on protein maps of Helicobacter pylori strains associated with gastric carcinoma / 中华流行病学杂志

<p><b>OBJECTIVE</b>To identify specific proteins of Helicobacter pylori (H. pylori) that associated with gastric carcinoma.</p><p><b>METHODS</b>The whole-cell proteins of H. pylori were separated by two-dimensional electrophoresis (2-DE). The protein maps of four H. pylori strains associated with gastric carcinoma and nine strains that isolated from patients with non-gastric carcinoma were then compared by ImageMaster 2D v3.1. MALDI-TOF mass spectrometry was performed to identify the proteins of interest. The proteins were searched by software mascot and identified by peptide fingerprint map.</p><p><b>RESULTS</b>Three proteins seemed to be associated with gastric carcinoma including acylneuraminate cytidylyltransferase with Mowse score 79 with the sequence coverage of 32%. The other two had no unambiguous protein to match.</p><p><b>CONCLUSION</b>Acylneuraminate cytidylyltransferase seemed to be a specific H. pylori protein associated with the presence of gastric carcinoma. Other two were novel proteins that might be associated with gastric carcinoma. However, the mechanism needs to be explored.</p>