Rectifying cow infertility under heat stress by immunization against inhibin and supplementation of progesterone.

This study was undertaken for the development of novel techniques that are based on immunoneutralization of inhibin bioactivity to improve Holstein cow fertility. A series of 4 experiments were carried out on 2 farms that were located in subtropical or temperate regions, to test the effects of immunization against inhibin alpha subunit on cow fertility under varying degrees of heat stress conditions. Though immunization against inhibin alone improved conception rate (CR) after TAI moderately in cows under mild heat stress conditions, the treatment plus progesterone supplementation substantially enhanced CR in the range of 25 to 35 percentages from severe heat stress to comfortable weather conditions. There existed an additive effect between immunization against inhibin and progesterone supplementation that maximally enhanced CR. Further, immunization against inhibin increased both FSH and activin A concentrations in blood during both follicular and luteal phases. It also significantly increased blood concentrations of E2 in the follicular phase but decreased P4 concentrations during the early pregnancy. However, interferon-tau concentrations in blood around the time of pregnancy recognition were doubled in the inhibin immunized cows. In conclusion, immunization against inhibin plus P4 treatment enhances ovarian follicle and the subsequent early embryo developments that help to greatly improve the fertility of Holstein dairy cows.

Neuroprotective effect of gastrodin in methamphetamine-induced apoptosis through regulating cAMP/PKA/CREB pathway in cortical neuron.

OBJECTIVE: Methamphetamine (MA) abuse induces neurotoxicity and causes neuronal cell apoptosis. Gastrodin is a traditional Chinese herbal medicine used for the treatment of nerve injuries, spinal cord injuries, and some central nervous system diseases as well. The present study investigated the neuroprotective effects of gastrodin against MA-induced neurotoxicity in neuronal cells and its potential protective mechanism. METHODS: The primary cortex neuronal culture was divided into four groups (control group, MA group, MA + gastrodin group, and MA + gastrodin + small interfering RNA group). The neurotoxicity of MA was assessed by detecting apoptotic cells by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay and cell viability by cell counting kit 8 (CCK-8) method, the Tuj1-positive cells and the average axonal length were detected by immunofluorescence, and the expressions of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP-response element-binding (CREB), and brain-derived neurotrophic factor (BDNF) proteins were detected by Western blot. RESULTS: The results of CCK-8 assay showed that 0.5 mM MA was an optimal concentration that induced neurotoxicity (p < 0.01). Pretreatment with 25 mg/L gastrodin exerted maximum protective effects on neuronal cells. The expression levels of cAMP, PKA, phosphorylated PKA, CREB, phosphorylated CREB, and BDNF proteins were decreased in the MA group, and pretreatment with gastrodin upregulated the expression levels of these proteins (p < 0.01). The expressions of PKA and CREB proteins showed no significant changes in the control group, MA group, and gastrodin group. Compared the MA + gastrodin + small interfering RNA group with MA + gastrodin group, the Tuj1-positive cells and the average axonal length were decreased significantly, while the number of apoptotic cells was increased (p < 0.05). CONCLUSION: Gastrodin has neuroprotective effects against MA-induced neurotoxicity, which exerts neuroprotective effects via regulation of cAMP/PKA/CREB signaling pathway and upregulates the expression of BDNF.

In vitro skin penetration of petrolatum and soybean oil and effects of glyceryl monooleate.

OBJECTIVES: Petrolatum and soybean oil are common ingredients incorporated in topical skin formulations for skin protection and moisturization. However, the stratum corneum (SC) penetration kinetics of these two cosmetic ingredients has not been systematically studied. Glyceryl monooleate (GlyMOle) has been shown to enhance skin penetration of various compounds. It was hypothesized that GlyMOle could enhance skin penetration of petrolatum and soybean oil. This study aimed to examine the in vitro skin penetration of petrolatum and soybean oil in the presence or absence of GlyMOle. METHODS: Skin permeation experiments were conducted using the in vitro Franz diffusion cell model with split-thickness human skin and human epidermal membrane (HEM). The effect of permeant dose and the kinetics of permeant penetration were examined with and without GlyMOle in vitro. RESULTS: Petrolatum and soybean oil were found to permeate across HEM, and no effect of GlyMOle on skin permeation into the receptor chamber was observed. GlyMOle enhanced the penetration of petrolatum into the split-thickness skin at 50 µg dose (petrolatum:GlyMOle, 49 : 1, w/w). However, no effect of GlyMOle on petrolatum penetration was observed at 200 µg dose (of the same petrolatum:GlyMOle ratio), indicating a dose-dependent effect. GlyMOle at the level used in the study did not enhance the penetration of soybean oil with 50 and 200 µg doses at any timepoints. CONCLUSION: GlyMOle was a skin penetration enhancer for petrolatum under the in vitro conditions identified in this study.

Molecular cloning and expression analysis of tyr and tyrp1 genes in normal and albino yellow catfish Tachysurus fulvidraco.

The full-length complementary DNA of two genes related to vertebrate albinism, the tyrosinase gene tyr and tyrosinase-related protein 1 gene tyrp1, were cloned and analysed from normal and albino yellow catfish Tachysurus fulvidraco. The open reading frames (ORF) of tyr and tyrp1 encode putative peptides of 533 and 526 amino acids (amino-acid), both of which possess two conserved copper binding sites. The homologous identities of deduced amino-acid sequences showed that both Tyr and Tyrp1 of T. fulvidraco share considerable similarity with that of channel catfish Ictalurus punctatus. Both tyr and tyrp1 were expressed in a wide range of adult tissues. Tyr gene had the highest expression level in the brain of both normal and albino T. fulvidraco. Tyrp1 had the highest expression level in the skin of normal groups, and the fin of albino groups. The messenger (m)RNA expressions of tyr and tyrp1 were detectable at different early developmental stages and varied with embryonic and larval growth. Tyr and tyrp1 mRNA have obvious tissue specificity both in normal and albino T. fulvidraco and higher expression levels were detected in the normal group revealing that tyr and tyrp1 may have an important role in pigmentation. These results will provide useful data for understanding the molecular mechanism of melanin formation and the occurrence of albinism in T. fulvidraco.

DNMT3A-mediated down-regulation of microRNA-105 promotes gastric cancer cell proliferation.

OBJECTIVE: It has been well-established that microRNAs (miRNAs), a class of short non-coding RNA molecules, play an important role in the development of gastric cancer. In the present study, we focused on miR-105, a novel miRNA not previously linked to gastric cancer. PATIENTS AND METHODS: 36 paired surgically resected gastric cancer tissues and matched adjacent normal tissues were used to detect the expression of miR-105. AGS cells were used to overexpress or silence of miR-105 and to determine its effect on several tumorigenic properties. A cell proliferation enzyme-linked immunosorbent assay was used to analyze the incorporation of BrdU during DNA synthesis of AGS cells. Total cDNA from AGS cells was used to amplify the 3'-UTR of YY1 by PCR and luciferase activity was determined using the Dual-Luciferase Reporter Assay System RESULTS: We found that expression of miR-105 was reduced in gastric cancer tissues, compared with adjacent normal tissues, due to hypermethylation at its promoter region. Overexpression of miR-105 suppressed, whereas its inhibition promoted cell viability and proliferation. We further identified Yin Yang 1 (YY1) as a direct target of miR-105, by which miR-105 exerted its anti-proliferative role. Moreover, we found that DNMT3A was responsible for the down-regulation of miR-105 in gastric cancer cells. CONCLUSIONS: Our data demonstrate that miR-105 inhibits gastric cancer cell proliferation and progression, which might provide a therapeutical target for cancer therapy.

[Predictive and Prognostic significance of high-sensitivity modified Glasgow Prognostic Score (HS-mGPS) in advanced gastric cancer patients treated with neoadjuvant chemotherapy].

Objective: To study the predictive and prognostic significance of high-sensitivity modified Glasgow Prognostic Score (HS-mGPS) on the effect of neoadjuvant chemotherapy for advanced gastric cancer. Methods: 117 patients with advanced gastric cancer received neoadjuvant chemotherapy with SOX (oxaliplatin+ S1) or mFOLFOX 6(oxaliplatin+ CF+ 5-FU) regimen. HS-mGPS was calculated according to blood C-reactive protein (CRP) concentration and serum albumin (ALB) level. The correlation between HS-mGPS and clinicopathological characteristics was determined and the predictors of survival were analyzed. Results: 117 patients with stage ⅡB (43 cases), stage Ⅲ (60), and stage Ⅳ (14) received preoperative neoadjuvant chemotherapy. The overall response rate of neoadjuvant chemotherapy was 61.5%(72/117), and the tumor control rate was 88.0% (103/117), with a pathological response rate of 91.5% (107/117). The R0 resection rate was 81.2% (95/117). The median disease-free survival (DFS) was 21.0 (95% CI 6.4-35.6) months. The median overall survival (OS) was 39.0 (95% CI 21.4-56.6) months. Higher HS-mGPS was associated with higher T stage, local lymph-node metastasis, distant metastasis, lower chemotherapy overall response rate and lower pathological response rate (all P<0.05). The univariate analysis and multivariate analysis showed that higher HS-mGPS, presence of local lymph-node metastasis and non R0 resection were associated with poorer DFS and OS (P<0.05). Conclusion: HS-mGPS can be used to predict the benefits of neoadjuvant chemotherapy and as an independent prognostic factor for survival in patients with advanced gastric cancer.

[Effects of hyperbaric oxygen on mucosal inflammatory response after nasal surgery in rabbits].

Objective:To observe the effect of hyperbaric oxygen therapy for the inflammation of rabbit nasal in postoperation period, to investigate the clinical feasibility of hyperbaric oxygen therapy in reducing the inflammatory reaction and promoting mucosal healing in nasal recovery stage after surgery.Method:A total of 16 Japanese white rabbits were randomly assigned to hyperbaric oxygen group and non hyperbaric oxygen group, with 8 in each, and another 5 was divided into normal control group. The non hyperbaric oxygen group was in the normal atmospheric environment, the hyperbaric oxygen group was given hyperbaric oxygen treatment on the fifth day after the operation. All the animals were sacrificed to observe the morphological changes and pathological changes of the mucosa in the bilateral inferior turbinate surgery area at sixth weeks after operation. The normal control group was taken the same part of mucosa.Result:Specimen: hyperbaric oxygen group postoperative empyema in 2 side, 8 side of non hyperbaric oxygen group, hyperbaric oxygen group was significantly reduced compared with non hyperbaric oxygen group, the difference was statistically significant (P< 0.05). Pathological changes: in the hyperbaric oxygen group, the infiltration of inflammatory cells was mild in 12 sides and moderate in 4 sides; the non hyperbaric oxygen group was mild in 1 sides, moderate in 13 sides, and severe in 2 sides, the hyperbaric oxygen group was significantly lower than the non hyperbaric oxygen group, the difference was statistically significant (P< 0.01).Conclusion:Under the condition of this experiment, hyperbaric oxygen therapy can significantly reduce the inflammatory response of rabbit nasal mucosa after operation, and reduce the accumulation of purulent secretion.

Effects of conjugated linoleic acid or betaine on the growth performance and fatty acid composition in backfat and belly fat of finishing pigs fed dried distillers grains with solubles.

The objectives of this study were to determine the effects of conjugated linoleic acid (CLA) or betaine on the growth performance, carcass characteristics and fatty acid composition in backfat and belly fat of pigs fed distillers dried grains with solubles (DDGS). Thirty-two (60±2 kg) crossbred barrows (Duroc×Landrace×Yorkshine) were assigned to one of four diets randomly: (1) the control diet containing no corn DDGS (control group); (2) the diet containing 30% corn DDGS (DDGS-fed group); (3) the diet containing 30% corn DDGS and 10 g/kg CLA (CLA-fed group); (4) the diet containing 30% corn DDGS and 1 g/kg BET (BET-fed group). The pigs fed DDGS showed that the percentages of C18:2, polyunsaturated fatty acid (PUFA) and iodine value (IV) increased, while C18:1, saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA) decreased. Pigs fed the DDGS+CLA or DDGS+betaine diets showed the increased percentage of SFA, and the decreased percentage of C18:2, PUFA and IV. In conclusion, results confirmed that the diets containing 30% DDGS had no detrimental effects on growth performance, but increased the percentage of PUFA and IV and decreased the percentage of SFA and MUFA in the backfat and belly fat. However, supplementation with CLA or BET can part reverse these effects on carcass fat in finishing pigs.

Effects of isobutyrate supplementation on ruminal microflora, rumen enzyme activities and methane emissions in Simmental steers.

The objective of this study was to evaluate the effects of isobutyrate supplementation on rumen microflora, enzyme activities and methane emissions in Simmental steers consuming a corn stover-based diet. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. The treatments were control (without isobutyrate), low isobutyrate (LIB), moderate isobutyrate (MIB) and high isobutyrate (HIB) with 8.4, 16.8 and 25.2 g isobutyrate per steer per day respectively. Isobutyrate was hand-mixed into the concentrate portion. Diet consisted of 60% corn stover and 40% concentrate [dry matter (DM) basis]. Dry matter intake (averaged 9 kg/day) was restricted to a maximum of 90% of ad libitum intake. Population of total bacteria, cellulolytic bacteria and anaerobic fungi were linearly increased, whereas that of protozoa and total methanogens was linearly reduced with increasing isobutyrate supplementation. Real-time PCR quantification of population of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Fibrobacter succinogenes was linearly increased with increasing isobutyrate supplementation. Activities of carboxymethyl cellulase, xylanase and ß-glucosidase were linearly increased, whereas that of protease was linearly reduced. Methane production was linearly decreased with increasing isobutyrate supplementation. Effective degradabilities of cellulose and hemicellulose of corn stover were linearly increased, whereas that of crude protein in diet was linearly decreased with increasing isobutyrate supplementation. The present results indicate that isobutyrate supplemented improved microflora, rumen enzyme activities and methane emissions in steers. It was suggested that the isobutyrate stimulated the digestive micro-organisms or enzymes in a dose-dependent manner. In the experimental conditions of this trial, the optimum isobutyrate dose was approximately 16.8 g isobutyrate per steer per day.

Identification of novel kinase inhibitors by targeting a kinase-related apoptotic protein-protein interaction network in HeLa cells.

OBJECTIVES: Protein kinases orchestrate activation of signalling cascades in response to extra- and intracellular stimuli for regulation of cell proliferation. They are directly involved in a variety of diseases, particularly cancers. Systems biology approaches have become increasingly important in understanding regulatory frameworks in cancer, and thus may facilitate future anti-cancer discoveries. Moreover, it has been suggested and confirmed that high-throughput virtual screening provides a novel, effective way to reveal small molecule protein kinase inhibitors. Accordingly, we aimed to identify kinase targets and novel kinase inhibitors. MATERIALS AND METHODS: A series of bioinformatics methods, such as network construction, molecular docking and microarray analyses were performed. RESULTS: In this study, we computationally constructed the appropriate global human protein-protein interaction network with data from online databases, and then modified it into a kinase-related apoptotic protein-protein interaction network. Subsequently, we identified several kinases as potential drug targets according to their differential expression observed by microarray analyses. Then, we predicted relevant microRNAs, which could target the above-mentioned kinases. Ultimately, we virtually screened a number of small molecule natural products from Traditional Chinese Medicine (TCM)@Taiwan database and identified a number of compounds that are able to target polo-like kinase 1, cyclin-dependent kinase 1 and cyclin-dependent kinase 2 in HeLa cervical carcinoma cells. CONCLUSIONS: Taken together, all these findings might hopefully facilitate discovery of new kinase inhibitors that could be promising candidates for anti-cancer drug development.

Molecular mechanisms of Lycoris aurea agglutinin-induced apoptosis and G2 /M cell cycle arrest in human lung adenocarcinoma A549 cells, both in vitro and in vivo.

OBJECTIVES: Lycoris is aurea agglutinin (LAA) has attracted rising attention due to its remarkable bioactivities. Here, we aimed at investigating its anti-tumor activities. MATERIAL AND METHODS: In vitro methods including MTT, cellular morphology observation, FCM and immunoblotting were performed. In vivo methods like detection of tumor volume, body weight and survival ratio, as well as TUNEL staining were performed. RESULTS AND CONCLUSION: LAA triggers G2 /M phase cell cycle arrest via up-regulating p21expression as well as down-regulating cdk-1cyclinA singling pathway, and induces apoptotic cell death through inhibiting PI3K-Akt survival pathway in human lung adenocarcinoma A549 cells. While LAA has no significant cytotoxic effect toward normal human embryonic lung fibroblast HELF cells, and moreover, LAA could amplify the antineoplastic effects of cisplatin toward A549 cells. Lastly LAA also bears anti-cancer and apoptosis-inducing effects in vivo, and it could decrease the volume and weight of subcutaneous tumor mass obviously as well as expand lifespan of mice. These findings may provide a new perspective for elucidating the complicated molecular mechanisms of LAA-induced cancer cell growth-inhibition and death, providing a new opportunity of LAA as a potential candidate anti-neoplastic drug for future cancer therapeutics.

Virtual screening methods as tools for drug lead discovery from large chemical libraries.

Virtual screening methods have been developed and explored as useful tools for searching drug lead compounds from chemical libraries, including large libraries that have become publically available. In this review, we discussed the new developments in exploring virtual screening methods for enhanced performance in searching large chemical libraries, their applications in screening libraries of ~ 1 million or more compounds in the last five years, the difficulties in their applications, and the strategies for further improving these methods.

Cysteamine improves growth performance and gastric ghrelin expression in preweaning piglets.

The aim of the present study was to investigate the effect of cysteamine on growth performance of preweaning piglets and gastric expression of ghrelin mRNA in vivo and in vitro. Twelve litters of newborn piglets were allocated randomly to control and treatment groups. From 15 d of age, piglets in the control group were fed basal creep diet, whereas the treatment group received basal diet supplemented with 120 mg cysteamine per kg of diet until weaning on 35 d of age. Body weight gain, creep feed consumption, and diarrhea rates were recorded, and gastric mucosal tissues were collected for quantifying mRNA expression. To evaluate the direct effect of cysteamine on gastric ghrelin expression, primary cultures of gastric mucosal cells isolated from 35-d-old piglets were exposed to cysteamine for 20 h at 0, 1, 10, and 100 µg/mL, respectively. Dietary cysteamine increased (P < 0.05) average daily creep feed consumption and BW gain in preweaning pigs, which was accompanied by reduction in diarrhea rates. At 35 d of age, piglets treated with cysteamine showed increased (P < 0.05) ghrelin and gastrin and decreased (P < 0.05) somatostatin mRNA expression in gastric mucosa. Moreover, dietary cysteamine treatment increased serum concentration of gastrin (P < 0.05). In vitro, cysteamine significantly increased ghrelin mRNA expression in gastric mucosal cells at the concentration of 10 µg/mL. In conclusion, dietary cysteamine is effective in improving the growth performance and health condition of preweaning piglets, which is associated with its stimulatory effects on gastric ghrelin mRNA expression both in vivo and in vitro.

Vegetable-rich food pattern is related to obesity in China.

OBJECTIVE: To investigate the association between a vegetable-rich food pattern and obesity among Chinese adults. DESIGN: A food pattern rich in vegetables is associated with lower risk of obesity and non-communicable chronic disease in Western countries. A similar food pattern is found in the Chinese population but the cooking method is different. A cross-sectional household survey of 2849 men and women aged 20 years and over was undertaken in 2002 in Jiangsu Province (response rate, 89.0%). Food intake was assessed by food frequency questionnaire. Factor analysis was used to identify food patterns. Nutrient intake was measured by food weighing plus consecutive individual 3-day food records. Height, weight and waist circumference were measured. RESULTS: The prevalence of general obesity (BMI > or =28 kg m(-2)) was 8.0% in men and 12.7% in women, central obesity was 19.5% (> or =90 cm) and 38.2% (> or =80 cm), respectively. A four-factor solution explained 28.5% of the total variance in food frequency intake. The vegetable-rich food pattern (whole grains, fruits and vegetables) was positively associated with vegetable oil and energy intake. Prevalence of obesity/central obesity increased across the quartiles of vegetable-rich food pattern. After adjusting for sociodemographic factors and four distinct food patterns, the vegetable-rich pattern was independently associated with obesity. Compared with the lowest quartile of vegetable-rich pattern, the highest quartile had higher risk of general obesity (men, prevalence ratio (PR): 1.82, 95% confidence interval (CI): 1.05-3.14; women, PR: 2.25, 95% CI: 1.45-3.49). CONCLUSION: The vegetable-rich food pattern was associated with higher risk of obesity/central obesity in Chinese adults in both genders. This association can be linked to the high intake of energy due to generous use of oil for stir-frying the vegetables.

Allitridum mimics effect of ischemic preconditioning by activation of protein kinase C.

AIM: To investigate whether allitridum has the effect of pharmacological preconditioning and whether protein kinase C (PKC) plays a role in myocardial protection. METHODS: Thirty-four isolated rabbit hearts which subjected to 30 min of regional myocardial ischemia and 2 h reperfusion, were randomly divided into 5 groups: control group, ischemic preconditioning (PC) group, allitridum (A) group, polymyxin B (Poly B) group, allitridum + polymyxin B (A + Poly B) group. Infarct size was determined by triphenyltetrazolium staining. RESULTS: Pharmacological preconditioning in hearts with a 5 -min allitridum infusion 10 min before the prolonged regional ischemia resulted in significantly smaller infarcts (7 % +\- 6 % of risk area) than in control hearts (25 % +\- 7 %, P < 0.05). There is no significant difference in infarct size between (A+Poly B) group and control hearts (23 % +\- 5 % vs 25 % +\- 7 %, P > 0.05). CONCLUSION: These data indicate that allitridum can precondition rabbit ischemic myocardium and this protection can be effectively blocked by administration of Poly B, an inhibitor of PKC, implying that PKC has an important role in preconditioning.

Two hundred and seventeen cases of winter diseases treated with acupoint stimulation in summer.

217 cases of chronic bronchitis and asthma were clinically treated and analyzed for the effects of combining electric stimulation with topical application of drug on acupoints. The results suggested that the combined therapy was superior to unitary therapy (P < 0.05). It is indicated that the combined therapy has a good curative effect in both short- and long-terms.

Clinical observation on treatment of non-parvicellular carcinoma of the lung with jin fu kang oral liquid.

Jin Fu Kang Oral Liquid ([symbol: see text]), made of traditional Chinese drugs for supplementing qi and nourishing yin, was developed according to the common symptoms in lung carcinoma with deficiency of both qi and yin. Of the 96 cases in the Jin Fu Kang group, 1 case got complete remission (CR) after treatment, 8 cases partial remission (PR), 52 cases no change (NC), PR + NC covering 63.5%. Of the 52 cases in the group of Jin Fu Kang plus chemotherapy, 11 cases got PR after treatment, 26 cases NC, PR + NC covering 71.2%. Of the 25 cases in the chemotherapy group, 4 cases got PR after treatment, 11 cases NC, PR + NC covering 60.0%. The results show that the therapeutic effectiveness in the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy was better than that in the chemotherapy group. The one-year survival rate and the two-year survival rate after treatment in the Jin Fu Kang group were 67.3% and 67.3% respectively; 66.7% and 66.7% in the group of Jin Fu Kang plus chemotherapy; and 40.3% and 0.0% in the chemotherapy group. The improvement of clinical symptoms, increase of body weight and improvement of health situation (KPS marks) after treatment in both the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy were better than that in the chemotherapy group. Some indicators of immunology and hemogram after treatment were greatly improved in the Jin Fu Kang group, worse in the chemotherapy group, but no obvious improvement in the group of Jin Fu Kang plus chemotherapy.

ent-Kaurene diterpenoids from Isodon rubescens.

Two kaurenoids, taibairubescensins A and B, were isolated from the ethanol extract of the leaves and branches of Isodon rubescens. Their structures are designated as 2 beta, 3 beta-diacetoxy-11 beta, 13 alpha-dihydroxy-ent-kaur-16-en-15-one and 3 beta, 11 beta-diacetoxy-2 beta, 6 alpha-dihydroxy-ent-kaur-16-en-15-one, respectively, on the basis of detailed spectroscopic analyses.