[Mechanism of Qingluo Tongbi Formula for regulating immune-bone erosion in rheumatoid arthritis].

OBJECTIVE: To explore the mechanism of Qingluo Tongbi formula for regulating "immune-bone erosion" in rheumatoid arthritis (RA). METHODS: Sixty-four RA patients were randomized into two groups to receive treatment with oral methotrexate or Qingluo Tongbi Formula for 12 weeks. Flow cytometry was used to analyze the changes in the percentages of CD3-CD19+, CD19+CD27 and CD19+BAFFR+B cell subpopulations in peripheral blood of the patients, and serum levels of B cell activating factor (BAFF), RANKL, RANK and osteoprotegerin (OPG) levels were detected using ELISA. Before and after the treatment, serum levels of ß-CTX, TRACP-5b, BGP, BALP, and PINP were measured with ELISA, and bone mineral density was determined with DXEA dual-energy X-ray absorptiometry. In the cell experiment, RAW264.7 cells were induced to differentiated into osteoclasts and treated with Qingluo Tongbi Formula at low-, moderate and high doses (125, 250 and 500 µg/mL, respectively) or with methotrexate (2 µg/mL) for 48 h, and the changes in the expression levels of RANKL, RANK, OPG and c-Fos were detected using Western blotting. RESULTS: The B cell subgroups in RA patients were correlated with the RANKL/RANK/OPG system. Treatment with Qingluo Tongbi Formula obviously down-regulated the percentages of the B cell subgroups, lowered serum levels of BAFF, ß-CTX and TRACP-5b, increased the levels of BGP, BALP and PINP, and improved lumbar bone density of RA patients (P<0.05); All these changes were significantly correlated with the regulation of B cell expressions (P<0.05). In RAW264.7 cells-derived osteoclasts, Qingluo Tongbi Formula significantly decreased the expressions of RANKL, RANK and c-Fos and increased the expression of OPG (P<0.05). CONCLUSION: Qingluo Tongbi Formula inhibits bone erosion in RA possibly by regulating B cell subset percentages and BAFF expression and inhibiting osteoclast differentiation via the RANKL/RANK/OPG pathway.

Effects of dietary niacinamide and CP concentrations on the nitrogen excretion, growth performance, and meat quality of pigs.

Reducing the dietary CP concentration in the formulation of low-protein diets without adverse effects on animal growth performance and meat quality remains challenging. In this study, we investigated the effects of nicotinamide (NAM) on the nitrogen excretion, growth performance, and meat quality of growing-finishing pigs fed low-protein diets. To measure the nitrogen balance, we conducted two trials: in nitrogen balance trial 1, four crossbred (Duroc × Landrace × Large White) barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design with four diets and periods. The diets consisted of a basal diet + 30 mg/kg NAM (a control dose), basal diet + 90 mg/kg NAM, basal diet + 210 mg/kg NAM, and basal diet + 360 mg/kg NAM. In nitrogen balance trial 2, another four barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design. The diets consisted of a basal diet + including 30 mg/kg NAM (control), basal diet + 360 mg/kg NAM, low-protein diet + 30 mg/kg NAM, and low-protein diet + 360 mg/kg NAM. To measure growth performance, two trials were conducted. In growth performance trial 1, 40 barrows (37.0 ± 1.0 kg) were randomly allocated to one of four dietary treatments (n = 10 per group), whereas in growth performance trial 2, 300 barrows (41.4 ± 2.0 kg) were randomly allocated to one of four dietary treatments, with each dietary treatment conducted in five repetitions with 15 pigs each. The four diets in the two growth performance trials were similar to those in nitrogen balance trial 2. Supplementing the diet with 210 or 360 mg/kg NAM reduced urinary nitrogen excretion and total nitrogen excretion and increased nitrogen retention comparted with the control diet (P < 0.05). Compared with the control diet, the low-protein diet with 360 mg/kg NAM reduced faecal, urinary, and total nitrogen excretion (P < 0.05) without affecting nitrogen retention and average daily gain (P > 0.05). Pigs fed the low-protein diet with 360 mg/kg NAM showed a decreased intramuscular fat content in the longissimus thoracis muscle when compared with pigs fed the control diet (P > 0.05). Our results suggest NAM as a suitable dietary additive to reduce dietary CP concentration, maximise nitrogen retention and growth performance, and decrease fat deposition in pigs.

Molecular mechanisms and promising role of dihydromyricetin in cardiovascular diseases.

Vine tea, a Chinese herbal medicine, is widely used in traditional Asian medicine to treat common health problems. Dihydromyricetin (DMY) is the main functional flavonoid compound extracted from vine tea. In recent years, preclinical studies have focused on the potential beneficial effects of dihydromyricetin, including glucose metabolism regulation, lipid metabolism regulation, neuroprotection, and anti-tumor effects. In addition, DMY may play a role in cardiovascular disease by resisting oxidative stress and participating in the regulation of inflammation. This review is the first review that summaries the applications of dihydromyricetin in cardiovascular diseases, including atherosclerosis, myocardial infarction, myocardial hypertrophy, and diabetic cardiomyopathy. We also clarified the underlying mechanisms and signaling pathways involved in the above process. The aim of this review is to provide a better understanding and quick overview for future researches of dihydromyricetin in the field of cardiovascular diseases, and more detailed and robust researches are needed for evaluation and reference.

The effects of dietary sodium butyrate supplementation on the growth performance, carcass traits and intestinal microbiota of growing-finishing pigs.

AIM: This study was carried out to investigate the effects of dietary sodium butyrate supplementation on growth performance, carcass traits and intestinal of growing-finishing pigs. METHODS AND RESULTS: Thirty pigs (27·4 ± 0·4 kg) were randomly assigned to receive one of three diets: basal diet (negative control group), basal diet + 40 ppm zinc bacitracin (positive control group) and basal diet + 0·2% sodium butyrate (sodium butyrate group), respectively. The experiment lasted for 69 days, including 3 days for diet and housing condition adaptation. On day 70, five piglets from each diet group were slaughtered for collecting blood and tissue samples. When compared to the control group, final body weight, daily body weight gain and daily feed intake of pigs in the sodium butyrate group were increased (P < 0·05) and feed intake/body weight gain ratio was decreased (P < 0·05). Carcass weight of pigs in the sodium butyrate group was higher than that of pigs in the negative and positive groups (P < 0·05); backfat thickness of pigs in the positive group was higher than that of pigs in the negative group and sodium butyrate group (P < 0·001). When compared to the negative and positive groups, pigs fed diet supplemented with sodium butyrate showed a increased relative abundance of bacteroidetes in the caecum and a decreased relative abundance of fiemicutes and proteobacteria in the caecum (P < 0·05). CONCLUSION: The results indicated that dietary sodium butyrate supplementation increased growth performance of growing-finishing pigs and improved the carcass traits and intestinal health. SIGNIFICANCE AND IMPACT OF THE STUDY: Antibiotic-free feed has become an inevitable worldwide trend. This study showed that dietary sodium butyrate supplementation improved the growth performance and intestinal health of growing-finishing pigs. Thus, sodium butyrate can be applied in growing-finishing pig feed as an alternative of antibiotics.

Effects of Rhodotorula mucilaginosa fermentation product on the laying performance, egg quality, jejunal mucosal morphology and intestinal microbiota of hens.

AIM: The aim of this study was to investigate the effects of dietary supplementation of Rhodotorula mucilaginosa solid-state fermentation product (RSFP) on the laying performance, egg quality and intestinal microbial flora of hens. METHODS AND RESULTS: In this study, 40-week-old Roman laying hens (n = 216) were randomly assigned to one of the four groups: the control (CON) group, fed 87.5% basal diet +12.5% fermentation substrate; the 0.5% RSFP group, fed 87.5% basal diet +12.0% fermentation substrate +0.5% RSFP; the 2.5% RSFP group, fed 87.5% basal diet +10.0% fermentation substrate +2.5% RSFP; and the 12.5% RSFP group, fed 87.5% basal diet +12.5% RSFP. The effect of each treatment was analysed in six replicates of nine hens. The experimental period was 31 days, which included a 3-day adaptation period. After 31 days of feeding, one hen from each replicate was randomly selected and killed, and the jejunum and digesta in the cecum were collected for the determination of the intestinal morphology and microbial flora respectively. Daily egg mass in the 2.5 and 12.5% RSFP groups and egg production and feed conversion ratio in the 12.5% RSFP group were higher than those in control group (P < 0.05). The yolk colour was improved in hens fed RSFP-supplemented diets (P < 0.05). Hens fed RSFP-supplemented diet showed a decrease in the relative abundances of Proteobacteria, Bacteroides, Helicobacteraceae, Helicobacter and Lachnospiraceae UCG-002, but an increase in the relative abundance of Prevotellaceae UCG-001 in the cecum (P < 0.05). CONCLUSION: Dietary RSFP supplementation improved the laying performance, egg quality and intestinal microflora of hens. SIGNIFICANCE AND IMPACT OF THE STUDY: Dietary supplementing diet with Rhodotorula mucilaginosa solid-state fermentation product, which is rich in carotenoids, improved the yolk colour and increased the carotenoid content, thereby improving the intestinal health of hens.

Effects of dietary calcium pyruvate on gastrointestinal tract development, intestinal health and growth performance of newly weaned piglets fed low-protein diets.

AIM: This study was carried out to investigate the effects of dietary calcium pyruvate supplementation on growth performance and intestinal health of weaned piglets fed low-protein diets. METHODS AND RESULTS: After a 7-day adaptation period, 60 individually housed piglets (Duroc × Yorkshire-Landrace) weaned at 28 days of age were randomly assigned to receive one of three treatments (20 pigs/treatment) for 28 days: control diet (20·0% crude protein [CP]), low-protein diet (15·5% CP), and experimental (15·5% CP + 1·8% calcium pyruvate). At the end of the experiment, six piglets from each diet group were slaughtered and blood and tissue samples were collected. Compared with the control group, feeding piglets with 15·5% CP decreased the daily body weight gain; lengths of the duodenum, jejunum and ileum; and weights of the stomach, duodenum, jejunum and ileum (P < 0·05), while 15·5% CP + 1·8% calcium pyruvate supplementation removed those differences (P > 0·05). Compared with the control group, the diarrhoea incidence and relative richness of Firmicutes in the colon contents of piglets in both the 15·5% CP and 15·5% CP + 1·8% calcium pyruvate groups was decreased. The relative richness of Bacteriodetes in the colon contents of piglets was higher in the 15·5% CP + 1·8% calcium pyruvate group than in the control and 15·5% CP groups (P < 0·05). CONCLUSION: Calcium pyruvate supplementation for four weeks removed the negative effects of a low-protein diet on the gastrointestinal tract development and daily body weight gain of weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed that supplementing a low-protein diet with calcium pyruvate, an effective alternative metabolic fuel to amino acids, was beneficial in improving the intestinal health and maximizing the growth of newly weaned piglets.

Neurobiological mechanisms of TENS-induced analgesia.

Pain inhibition by additional somatosensory input is the rationale for the widespread use of Transcutaneous Electrical Nerve Stimulation (TENS) to relieve pain. Two main types of TENS produce analgesia in animal models: high-frequency (∼50-100 Hz) and low-intensity 'conventional' TENS, and low-frequency (∼2-4 Hz) and high-intensity 'acupuncture-like' TENS. However, TENS efficacy in human participants is debated, raising the question of whether the analgesic mechanisms identified in animal models are valid in humans. Here, we used a sham-controlled experimental design to clarify the efficacy and the neurobiological effects of 'conventional' and 'acupuncture-like' TENS in 80 human volunteers. To test the analgesic effect of TENS we recorded the perceptual and brain responses elicited by radiant heat laser pulses that activate selectively Aδ and C cutaneous nociceptors. To test whether TENS has a long-lasting effect on brain state we recorded spontaneous electrocortical oscillations. The analgesic effect of 'conventional' TENS was maximal when nociceptive stimuli were delivered homotopically, to the same hand that received the TENS. In contrast, 'acupuncture-like' TENS produced a spatially-diffuse analgesic effect, coupled with long-lasting changes both in the state of the primary sensorimotor cortex (S1/M1) and in the functional connectivity between S1/M1 and the medial prefrontal cortex, a core region in the descending pain inhibitory system. These results demonstrate that 'conventional' and 'acupuncture-like' TENS have different analgesic effects, which are mediated by different neurobiological mechanisms.

[Expression of osteoprotegerin and receptor activator of nuclear factor kappa-B ligand in mandibular ramus osteotomy healing with administration of different doses of parathyroid hormone].

Objective: To investigate the effect of parathyroid hormone (PTH) on the bone healing of mandibular ramus osteotomy. Methods: The mandibular ramus osteotomy model was established in sixty rabbits and these rabbits were randomly divided into experimental group A, experimental group B and control group. In the experimental group A and experimental group B, the rabbits were given PTH (20 and 40 µg/kg respectively) every other day after operation. In the control group, 1 ml saline was given. The animals were sacrificed at 1 week, 2 weeks, 3 weeks and 4 weeks postoperatively. The new bone formation was observed by histology and cone bone CT. The expression of osteoprotegerin and receptor activator of nuclear factor kappa-B (RANKL) in the new bone was detected by real-time quantitative PCR. Results: The experimental groups has better osteogenesis and the bone mineral density than the control group in osteotomy area. The experimental group B showed the best osteogenesis.Osteoprotegerin mRNA expression in experimental group A (1.127±0.035, 1.742±0.049, 1.049±0.062, 1.063±0.036) was significantly higher than that in the control group in each period (0.965±0.082, 1.254±0.071, 0.793±0.061, 0.684±0.055) (P=0.010, P=0.000, P=0.001, P=0.020), while group B (1.416±0.205, 2.648±0.168, 1.652±0.091, 1.712±0.070) was significantly higher than group A (P=0.000, P=0.010, P=0.023, P=0.003). RANKL mRNA expression in control group (1.666±0.086, 1.058±0.105, 0.885±0.124, 0.972±0.136) was significantly higher than that of the group A (0.788±0.036, 0.585±0.017, 0.692±0.017, 0.527±0.051) (P=0.001, P=0.006, P=0.003, P=0.028) in each period, while group A was significantly higher than group B(0.247±0.022, 0.240±0.034, 0.134±0.011, 0.103±0.050) (P=0.000, P=0.001, P=0.002, P=0.012). Conclusions: PTH can upregulate the expression of osteoprotegerin and reduce expression of RANKL, thus promoting new bone formation. Intermittent administration of high dose of parathyroid hormone can further promote the healing process after mandibular ramus osteotomy.

Human tissue kallikrein 1 ameliorates erectile function via modulation of macroautophagy in aged transgenic rats.

Previously, we have demonstrated that human tissue kallikrein 1 (hKLK1) improves age-related erectile dysfunction (ED). Autophagy has been implicated in age-related diseases, including ED. However, the molecular mechanisms underlying hKLK1-mediated amelioration of age-related ED via regulation of autophagy remains unknown. To explore the potential mechanism, male wild-type Sprague-Dawley rats (WTR) and transgenic rats harboring human KLK1 (TGR) were bred till 4 or 18 months of age and divided into three groups: young WTR (yWTR) as the control group, aged WTR (aWTR) group, and aged TGR (aTGR) group. The erectile function of each rat was evaluated using cavernous nerve electrostimulation. The ratio of intracavernous pressure/mean arterial pressure (ICP/MAP) and total ICP were also measured. Western blotting, immunohistochemistry, and transmission electron microscopy were performed to detect the levels of autophagy. The expression levels of related signaling pathways were determined by western blotting and immunohistochemistry. We found that hKLK1 improved the impaired erectile function of aged rats. Compared to the yWTR and aTGR groups, the aWTR group showed reduced smooth muscle/collagen ratio, fewer autophagosomes, and lower expression of Beclin 1 and LC3-II, which indicate impaired smooth muscle function and low level of autophagy in the smooth muscle cells. Moreover, the PI3K/Akt/mTOR signaling pathway, which is considered to be a negative regulator of autophagy, was upregulated in the aWTR group. hKLK1 may partially restore erectile function in aged transgenic rats by upregulating protective autophagy via the PI3K/Akt/mTOR pathway. These observations indicate that hKLK1 is a potential gene therapy candidate for age-related ED.

Icariin improves the sexual function of male mice through the PI3K/AKT/eNOS/NO signalling pathway.

We aimed to investigate the effect and mechanism of icariin on male sexual function. Forty-eight Crl:CD1(ICR) male mice were randomly divided into control, low-, medium- and high-dose icariin group (intragastric administration of 50, 100 and 200 mg/kg/d for 21 days). Mating experiment was then performed at a ratio of 1: 3 (male: female). The mating behaviours of male mice were recorded. The genital indexes and serum testosterone, nitric oxide (NO), hypothalamic dopamine (DA) and 5- hydroxy tryptophan (5-HT) concentrations were measured. The expression of endothelial nitric oxide (eNOS), phosphatidylinositol tallow alcohol 3-kinase (PI3K) and phosphorylated protein kinase (p-AKT) in penile tissue was detected by Western blot. All icariin groups exhibited shorter capture latency and ejaculation latency, increased number of capture and ejaculation, higher capture and ejaculation rate, and higher testicular and prostate indexes compared with controls (p < .001). These groups had higher serum testosterone and NO concentrations (p < .001), hypothalamic DA and 5-HT levels, and eNOS, PI3K and phosphorylated AKT expressions in penile tissue (p < .05). The effect of icariin was dose-dependently increased. Our study suggests that icariin improves the sexual function of male mice, which might be associated with the hypothalamic-pituitary-gonadal axis and the PI3K/AKT/eNOS/NO signalling pathway.

Thiamine supplementation facilitates thiamine transporter expression in the rumen epithelium and attenuates high-grain-induced inflammation in low-yielding dairy cows.

An experiment was conducted to uncover the effects of increasing dietary grain levels on expression of thiamine transporters in ruminal epithelium, and to assess the protective effects of thiamine against high-grain-induced inflammation in dairy cows. Six rumen-fistulated, lactating Holstein dairy cows (627 ± 16.9 kg of body weight, 180 ± 6 d in milk; mean ± standard deviation) were randomly assigned to a replicated 3 × 3 Latin square design trial. Three treatments were control (20% dietary starch, dry matter basis), high-grain diet (HG, 33.2% dietary starch, DM basis), and HG diet supplemented with 180 mg of thiamine/kg of dry matter intake. On d 19 and 20 of each period, milk performance was measured. On d 21, ruminal pH, endotoxic lipopolysaccharide (LPS), and thiamine contents in rumen and blood, and plasma inflammatory cytokines were detected; a rumen papillae biopsy was taken on d 21 to determine the gene and protein expression of toll-like receptor 4 (TLR4) signaling pathways. The HG diet decreased ruminal pH (5.93 vs. 6.49), increased milk yield from 17.9 to 20.2 kg/d, and lowered milk fat and protein from 4.28 to 3.83%, and from 3.38 to 3.11%, respectively. The HG feeding reduced thiamine content in rumen (2.89 vs. 8.97 µg/L) and blood (11.66 vs. 17.63 µg/L), and the relative expression value of thiamine transporter-2 (0.37-fold) and mitochondrial thiamine pyrophosphate transporter (0.33-fold) was downregulated by HG feeding. The HG-fed cows exhibited higher endotoxin LPS in rumen fluid (134,380 vs. 11,815 endotoxin units/mL), and higher plasma concentrations of lipopolysaccharide binding protein and pro-inflammatory cytokines when compared with the control group. The gene and protein expression of tumor necrosis factor α (TNFα), IL1B, and IL6 in rumen epithelium increased when cows were fed the HG diet, indicating that local inflammation occurred. The depressions in ruminal pH, milk fat, and protein of HG-fed cows were reversed by thiamine supplementation. Thiamine supplementation increased thiamine contents in rumen and blood, and also upregulated the relative expression of thiamine transporters compared with the HG group. Thiamine supplementation decreased ruminal LPS (49,361 vs. 134,380 endotoxin units/mL) and attenuated the HG-induced inflammation response as indicated by a reduction in plasma IL6, and decreasing gene and protein expression of pro-inflammatory cytokines in rumen epithelium. Western bottling analysis showed that thiamine suppressed the protein expression of TLR4 and the phosphorylation of nuclear factor kappa B (NFκB) unit p65. In conclusion, HG feeding inhibits thiamine transporter expression in ruminal epithelium. Thiamine could attenuate the epithelial inflammation during high-grain feeding, and the protective effects may be due to its ability to suppress TLR4-mediated NFκB signaling pathways.

[The progress in application of parathyroid hormone in craniomaxillofacial bone regeneration study].

Parathyroid hormone(PTH)is synthesized and secreted by chief cell of Gley's glands which possesses dual functions of catabolism and anabolism. It regulates the proliferation and differentiation of multiple cell lines including osteoblast, osteoclast and skeletal lining cells. Furthermore, PTH activates various signaling pathways which control calcium, phosphorous' metabolism and bone conversion, accelerating the bone regeneration and reconstruction. However, the study of PTH in craniomaxillofacial bone regeneration is relatively less and whether the role of parathyroid glands and the mechanism of ossification are consistent with the long bone or not needs further investigation. This review focuses on the progress of PTH in craniomaxillofacial bone regeneration in recent years.

Physcion 8-O-ß-glucopyranoside prevents hypoxia-induced epithelial-mesenchymal transition in colorectal cancer HCT116 cells by modulating EMMPRIN.

Epithelial-mesenchymal transition (EMT) is considered as the most important mechanism that underlies the initiation of cancer metastasis. Here we report that Physicon 8-O-ß-glucopyranoside (PG), a major active ingredient from a traditional Chinese herbal medicine Rumex japonicus Houtt, is capable of preventing human colorectal cancer cells from hypoxia-induced EMT. The treatment of the cells with PG reversed the EMT-related phenotype that has the morphological changes, down-regulation of E-cadherin, and hypoxia-induced cell migration and invasion. The effect was mediated at least in part by inhibiting the mRNA and protein expressions of EMMPRIN via modulation of PTEN/Akt/HIF-1α pathway. In addition, we found that PG-mediated prevention of EMT involved blockade of the hypoxia-induced up-regulation of Snail, Slug and Twist. In summary, this study showed that PG can prevent EMT induced by hypoxia, the environment that commonly exists in the center of a solid tumor. Given the low toxicity of PG to the healthy tissues, our study suggests that PG can serve as a safe therapeutic agent for suppressing cancer metastasis.

In vivo effects of high dietary copper levels on hepatocellular mitochondrial respiration and electron transport chain enzymes in broilers.

The diet of broiler chickens supplemented with increasing concentrations of copper (Cu) was assessed for the effect of Cu on liver mitochondrial function. A total of 160, 1-d-old Cobb 500 broilers (Gallus domesticus) were randomly assigned in equal numbers into 4 groups, which differed in the concentration of copper supplements in the diet; 11 (control), 110, 220 and 330 mg of Cu/kg dry matter. Liver mitochondrial function was recorded at 12, 24, 36, 48 and 60 d of age. Supplementation with 110 mg Cu/kg dry matter enhanced mitochondrial function and activities of complexes I-V, and this was significant at 36 d of age compared with the other diets (P < 0.05). Supplementation with 220 mg Cu/kg dry matter and 330 mg Cu/kg dry matter enhanced mitochondrial function and activities of complexes I-V at 12, 24 and 36 d of age, but displayed reduced function (P < 0.05) at 48 and 60 d of age except in complex IV (P > 0.05). Mitochondrial hydrogen peroxide (H2O2) production was also increased (P < 0.05) with an increase of copper supplementation in the diet. The results indicate that appropriate dietary copper supplements are sufficient for improving mitochondrial function and activities of the respiratory complexes. Higher concentrations of copper, on the other hand, lead to copper toxicity by affecting certain respiratory complexes.

Folic acid supplementation and dietary folate intake, and risk of preeclampsia.

BACKGROUND/OBJECTIVES: Folic acid supplementation has been suggested to reduce the risk of preeclampsia. However, results from few epidemiologic studies have been inconclusive. We investigated the hypothesis that folic acid supplementation and dietary folate intake before conception and during pregnancy reduce the risk of preeclampsia. SUBJECTS/METHODS: A birth cohort study was conducted in 2010-2012 at the Gansu Provincial Maternity & Child Care Hospital in Lanzhou, China. A total of 10,041 pregnant women without chronic hypertension or gestational hypertension were enrolled. RESULTS: Compared with nonusers, folic acid supplement users had a reduced risk of preeclampsia (OR=0.61, 95% CI: 0.43-0.87). A significant dose-response of duration of use was observed among women who used folic acid supplemention during pregnancy only (P-trend=0.007). The reduced risk associated with folic acid supplement was similar for mild or severe preeclampsia and for early- or late-onset preeclampsia, although the statistical significant associations were only observed for mild (OR=0.50, 95% CI: 0.30-0.81) and late-onset (OR=0.60, 95% CI: 0.42-0.86) preeclampsia. The reduced risk associated with dietary folate intake during pregnancy was only seen for severe preeclampsia (OR=0.52, 95% CI: 0.31-0.87, for the highest quartile of dietary folate intake compared with the lowest). CONCLUSIONS: Our study results suggest that folic acid supplementation and higher dietary folate intake during pregnancy reduce the risk of preeclampsia. Future studies are needed to confirm the associations.

Effect of zinc-bearing zeolite clinoptilolite on growth performance, zinc accumulation, and gene expression of zinc transporters in broilers.

This study investigated Zn-bearing zeolite clinoptilolite (Zn-ZCP) as a Zn supplement on performance, organ or tissue development, Zn accumulation in tissues, and gene expression of Zn transporters in the duodenum of broilers. A total of 300 1-d-old Arbor Acres chickens were randomly allocated to 5 dietary treatments with 6 replicate cages of 10 birds per treatment for a 21-d feeding period. The treatments comprised a basal corn­soybean meal diet without supplemental Zn (the control) or similar diets supplemented with 80 mg/kg Zn from ZnSO4 or 20, 40, or 80 mg/kg Zn from Zn- ZCP, respectively. Supplementation of Zn-ZCP had a positive effect on G:F (linear, P < 0.05) and had tendency difference on ADG (linear, P < 0.1) of broilers during 1 to 21 d. Incremental Zn-ZCP inclusion in the diet increased the fresh weight of the pancreas (linear, P < 0.05) and tibia (linear, P < 0.05) in broilers on d 14. The same trend to the fresh weight of the tibia (linear, P < 0.05) and pancreas (linear, P < 0.05) was observed on d 21. Increasing Zn-ZCP level showed a linear response on Zn concentration in the liver (linear, P < 0.001), pancreas (linear, P < 0.05), and tibia (linear, P < 0.05) on d 14. The same trend of Zn concentrations in pancreas (linear, P < 0.05) and tibia (linear, P < 0.001) was observed on d 21. The mRNA levels of Zn transporter 2 (ZnT-2) and Zn transporter 5 (ZnT-5) in the duodenum of chicks fed the diet with 80 mg/kg Zn from Zn-ZCP did not differ from those of chicks fed the control diet, but both were lower (P < 0.05) than those of chicks fed ZnSO4 diet. Metallothionein (MT) mRNA levels of broilers fed the diet supplemented with 80 mg/kg Zn from Zn-ZCP or ZnSO4 was higher (P < 0.05) than that in the control group. Dietary treatments did not affect the mRNA expression of Zn transporter 1 (ZnT-1) or metal response element-binding transcription factor-1 (MTF-1). In conclusion, as a Zn supplement, Zn-ZCP was comparable to ZnSO4 for enhancing Zn accumulation and growth performance of broilers during 1 to 21 d. Broilers fed the diet with 80 mg/kg Zn from Zn-ZCP had different expressions of ZnT-2 and ZnT-5 in the duodenum compared with those fed the ZnSO4 diet.

Proliferation and osteo/odontoblastic differentiation of stem cells from dental apical papilla in mineralization-inducing medium containing additional KH(2)PO(4).

OBJECTIVES: Stem cells from the dental apical papilla (SCAPs) can be induced to differentiate along both osteoblast and odontoblast lineages. However, little knowledge is available concerning their differentiation efficiency in osteogenic media containing additional KH2 PO4 . MATERIALS AND METHODS: Stem cells from the dental apical papilla were isolated from apical papillae of immature third molars and treated with two kinds of mineralization-inducing media, MM1 and MM2, differing in KH2 PO4 concentration. Proliferation and osteo/odontogenic differentiation capacity of MM1/MM2-treated SCAPs were investigated and compared both in vitro and in vivo. RESULTS: Cell counting and flow cytometry demonstrated that MM2 containing 1.8 mm additional KH2 PO4 significantly enhanced proliferative potential of SCAPs, compared to MM1. Osteo/odontogenic capacity of SCAPs was much better in MM2 medium than in MM1, as indicated by elevated alkaline phosphatase activity, increased calcium deposition and upregulated expression of osteo/odontoblast-specific genes/proteins (for example, runt-related transcription factor 2, osterix, osteocalcin, dentin sialoprotein, and dentin sialophosphoprotein). In vivo transplantation findings proved that SCAPs in MM2 group generated more mineralized tissues, and presented higher expression of osteo/odontoblast-specific proteins (osteocalcin and dentin sialoprotein) than those in the MM1 group. CONCLUSION: Mineralization-inducing media supplemented with 1.8 mm additional KH2 PO4 significantly enhanced cell proliferation and improved differentiation capacity of SCAPs along osteo/odontogenic cell lineages, compared to counterparts lacking additional KH2 PO4 .

Effects of feeding an herbal preparation to sows on immunological performance of offspring.

The objective of this study was to determine the effects of feeding Chinese herbal ultra-fine (CHU) powder to sows during the last week of gestation and during the lactation period on immunological performance of the offspring. In this experiment, 15 pregnant sows (mean BW = 235.6 ± 3.7 kg) were randomly assigned to 1 of 3 treatments including no additive (Control), 0.75% CHU powder (Group A), or 1.5% CHU powder (Group B) added to a maize- and soybean meal-based diet. Blood from 10 piglets per group was collected at d 7, d 14, or d 21 of age to measure serum metabolites, lymphocyte proliferating activity, and serum antibody and cytokine concentrations. Dietary supplementation of sows with CHU powder increased (P < 0.05) serum concentrations of total protein, albumin, and triglycerides of offspring, whereas the concentration of glucose was reduced (P < 0.05) compared with Controls. The CHU powder enhanced (P < 0.05) serum concentrations of IgG in Group B offspring on d 7 and IgM in Group A offspring on d 7 and d 14, increased IL-10 in Group A offspring on d 7, as well as IL-2 in offspring from Groups A and B on all days of determination. The CHU powder increased interferon gamma in Group A offspring on d 14 and in Group B offspring on d 14 and d 21, and increased tumor necrosis factor alpha in offspring of Group A on d 14 and in Group B on all days surveyed. Compared with Controls, a greater number (P < 0.05) of T lymphocyte subpopulations were detected in Group A and B offspring including CD4+ cells in Group A on d 7 and d 21, CD4+ cells in Group B on d 14 and d 21, and CD8+ cells in Group A on d 7 and d 14. Collectively, these findings indicate a beneficial effect of CHU powder treatment of sows in later gestation and during lactation on serum metabolism and cellular and humoral immune responses of their offspring.

Effects of the preoperative administration of Yunnan Baiyao capsules on intraoperative blood loss in bimaxillary orthognathic surgery: a prospective, randomized, double-blind, placebo-controlled study.

Yunnan Baiyao is a well-known Chinese herbal medicine that has been used as a haemostatic drug for nearly 100 years. The aim of this clinical trial was to evaluate the efficacy and safety of Yunan Baiyao capsules on the reduction of blood loss in bimaxillary orthognathic surgery. 87 consecutive patients scheduled for simultaneous maxillary Le Fort I osteotomies and bilateral sagittal split ramus osteotomies (BSSRO) were enrolled in a prospective, randomized, double-blind, placebo-controlled clinical trial. Patients were administered Yunnan Baiyao capsules or placebo capsules, orally for 3 days before surgery. Intraoperative blood loss was estimated and the safety of Yunnan Baiyao capsules was evaluated. The total blood loss in the Yunnan Baiyao group (mean, 330.5+/-134.4 ml) was significantly lower than in the control group (mean, 420.3+/-175.9 ml). No allergic reactions, thromboembolic events or other side effects were recorded in this trial. It can be concluded that the preoperative use of Yunnan Baiyao capsules, in combination with hypotension anaesthesia, results in a reduction in intraoperative blood loss in bimaxillary orthognathic surgery. Yunnan Baiyao capsules are an effective and safe haemostatic Chinese medicine.

Synthesis of Drosophila melanogaster acetylcholinesterase gene using yeast preferred codons and its expression in Pichia pastoris.

To improve the expression level of recombinant Drosophila melanogaster AChE (R-DmAChE) in Pichia pastoris, the cDNA of DmAChE was first optimized and synthesized based on the preferred codon usage of P. pastoris. The synthesized AChE cDNA without glycosylphosphatidylinositol (GPI) signal peptide sequence was then ligated to the P. pastoris expression vector, generating the plasmid pPIC9K/DmAChE. The linearized plasmid was homologously integrated into the genome of P. pastoris GS115 via electrotransformation. Finally seven transformants with high expression level of R-DmAChE activity were obtained. The highest production of R-DmAChE in shake-flask culture after 5-day induction by methanol was 718.50 units/mL, which was about three times higher than our previous expression level of native DmAChE gene in P. pastoris. Thus, these new strains with the ability to secret R-DmAChE in the medium could be used for production of R-DmAChE to decrease the cost of the enzyme expense for rapid detection of organophosphate and carbamate insecticide residues.