Rs13405728 is associated with slow ovarian response in assisted reproductive technology.

OBJECTIVE: To explore the association between rs13405728 with slow ovarian response in assisted reproductive technology (ART). PATIENTS AND METHODS: 236 women, aged 21 to 35 years, were enrolled and grouped according to their genotypes, The polymorphism of rs13405728 was genotyped by DNA sequencing. RESULTS: There was no evidence of any difference in anti-Müllerian hormone (AMH), antral follicle count (AFC) among the three genotypes (p>0.05). The occurrence of slow response in genotype GG was lower than those in the other two genotypes (p<0.05). There were independent correlations between slow ovarian response with the dose of luteinizing hormone (LH) required and the genotypes of rs13405728 (p<0.05). CONCLUSIONS: There were significant independent correlations between slow ovarian response with the dose of LH required and the genotypes of rs13405728. Different mechanisms may be involved in poor response and slow response.

Impaired p65 degradation by decreased chaperone-mediated autophagy activity facilitates epithelial-to-mesenchymal transition.

Aberrant activation of nuclear factor-κB (NF-κB) has been observed in a wide range of human cancers and is thought to promote tumorigenesis and metastasis. As a central component of NF-κB pathway, p65 protein level is tightly regulated and could be subjected to proteasome degradation. Here we demonstrated that p65 can bind to HSC70 with four consensus recognition motif in its RHD domain and be constitutively transported to the lysosome membrane to bind with lysosome-associated membrane protein type 2A and degraded within the lysosome in two epithelial cell lines, proposing that p65 can be degraded by chaperone-mediated autophagy (CMA). Of great importance, there is a decreased CMA activity together with impaired degradation of p65 in a process of epithelial-mesenchymal transition (EMT). The resulted accumulation of p65 leads to higher NF-κB activity and contributes to the progression and maintenance of the EMT program. Taken together, our results define a novel regulatory mechanism for the important transcription factor p65, and these findings would shed new light on the inhibition of EMT, as well as metastasis of cancer cells.

Effect of alpha-lipoic acid on relieving ammonia stress and hepatic proteomic analyses of broilers.

Ammonia in poultry houses not only affects worker health but also induces a variety of poultry diseases. Alpha-lipoic acid (LA) is an effective antioxidant that protects cells against oxidative injury during various toxic and pathological processes. This study was designed to evaluate the mitigating effects of LA supplementation on ammonia stress and hepatic proteome changes in broilers. Male broilers (22 d old) were allocated to 3 groups: (1) a control group without ammonia stress (CTRL); (2) exposure to 70 ppm ammonia (AM); and (3) exposure to 70 ppm ammonia and dietary administration of 300 mg/kg LA (AM+LA). Ammonia exposure significantly decreased broiler growth performance and plasma glutathione peroxidase activity (P < 0.05), and increased plasma malondialdehyde content and glutamic-pyruvic transaminase activity (P < 0.05). These negative effects were eliminated by LA supplementation. Comparative proteomic analyses revealed 291 differentially expressed proteins in the AM group compared to the CTRL and AM+LA groups. A total of 30 proteins were differentially expressed between the AM/CTRL and (AM+LA)/AM groups. The addition of LA restored 24 of these proteins to control levels; these proteins were mainly related to transcription regulation, detoxification, protein translation and degradation, and immune and stress responses. The differentially expressed proteins included the high mobility group box (HMGB) and glutathione S-transferase (GST), which is closely related to immune response and oxidative stress, and collagens, which are implicated in liver injury. The addition of LA to broiler diet may reduce ammonia toxicity by maintaining the antioxidant system, xenobiotic metabolism, and metabolic pathways.

HIF-1α downregulates miR-17/20a directly targeting p21 and STAT3: a role in myeloid leukemic cell differentiation.

Hypoxia-inducible factor 1 (HIF-1) is a crucial transcription factor for the cellular adaptive response to hypoxia, which contributes to multiple events in cancer biology. MicroRNAs (miRNAs) are involved in almost all cellular activities such as differentiation, proliferation, and apoptosis. In this work, we use miRNA microarrays to profile miRNA expression in acute myeloid leukemia (AML) cells with inducible HIF-1α expression, and identify 19 differentially expressed miRNAs. Our study shows that HIF-1α represses the expression of miR-17 and miR-20a by downregulating c-Myc expression. These two miRNAs alleviate hypoxia and HIF-1α-induced differentiation of AML cells. More intriguingly, miR-17 and miR-20a directly inhibit the p21 and STAT3 (signal transducer and activator of transcription 3) expression, both of which can reverse miR-17/miR-20a-mediated abrogation of HIF-1α-induced differentiation. Moreover, we show in vivo that miR-20a contributes to HIF-1α-induced differentiation of leukemic cells. Taken together, our results suggest that HIF-1α regulates the miRNA network to interfere with AML cell differentiation, representing a novel molecular mechanism for HIF-1-mediated anti-leukemic action.

Long-lasting infiltration anaesthesia by lidocaine-loaded biodegradable nanoparticles in hydrogel in rats.

BACKGROUND: Infiltration of a long-lasting anaesthetic is helpful during the post-operative period. The recently developed local drug delivery system, biodegradable nanoparticles in a thermo-sensitive hydrogel (nanogel system), may possibly provide an extended duration of drugs. Therefore, we evaluated whether prolonged infiltration anaesthesia could be achieved by loading lidocaine into this delivery system. METHODS: Thirty male rats were randomized into five groups of six rats each: saline; 2% hydrochloride lidocaine solution; lidocaine-loaded nanogel system and its compositing formulations, namely lido-nano gel; lido-nano; and lidogel. Durations of local anaesthesia with subcutaneously injected agents were measured by tail flick latency tests in a randomized, blind fashion. RESULTS: Lido-nano gel produced effective anaesthesia for 360+/-113 min, compared with 150+/-33 min by lidogel, 180+/-37 min by lido-nano, and 110+/-45 min by lidocaine solution (P<0.001, means+/-SD), and elicited complete sensory blockade for 300+/-114 min, compared with 75+/-37 min by lidogel, 105+/-53 min by lido-nano, and 60+/-33 min by lidocaine solution (P<0.001, means+/-SD) without severe skin/systemic toxicity. CONCLUSION: Lidocaine-loaded biodegradable nanoparticles in hydrogel produced prolonged infiltration anaesthesia in rats without severe toxicity, indicating a possible way to develop long-lasting local anaesthetics.

Effect of soybean peptides on growth performance, intestinal structure and mucosal immunity of broilers.

Five-hundred 1-day-old broilers were randomly assigned to five groups, 100 chickens for each group. From group 1 to 3, 80, 120 and 200 mg/kg soybean peptides was added to the diets respectively; in the fourth group, 3.2 mg/kg genramycin was added; and the fifth group was the control without soybean peptides and antibiotics. At the age of 28 and 49 days, the number of goblet cells (GC), intestine intraepithelial lymphocyte, immunoglobulin A-forming cells, the ratio of villous height and crypt depth (V/C) of broiler's duodenum, jejunum and cecum were observed by the application of haematoxylin and eosin or histochemistry staining. The results indicated that soybean peptides added with 80-120 mg/kg could increase daily weight gain, the number of GC and V/C. Soybean peptides could modulate intestinal mucosal immunity of broilers.

Multi-sites cleavage of leukemogenic AML1-ETO fusion protein by caspase-3 and its contribution to increased apoptotic sensitivity.

Leukemia-associated fusion protein AML1-ETO is a product of the chromosome translocation (8;21) frequently occurred in acute myeloid leukemia (AML). The fusion oncoprotein blocks leukemic cell differentiation, and it also induces growth arrest with increased sensitivity to apoptosis induction. Such dichotomous functions make it difficult to clarify the role of AML1-ETO in leukemogenesis. Here, we systematically showed that constitutively and overexpressed AML1-ETO protein was cleaved to four fragments of 70, 49, 40 and 25 kDa by activated caspase-3 during apoptosis induction by extrinsic mitochondrial and death receptor signaling pathways. The in vitro proteolytic system combined with MALDI-TOF/TOF mass spectrometer confirmed that AML1-ETO and wild-type ETO but not RUNX1 (AML1) proteins were direct substrates of apoptosis executioner caspase-3. Site-directed mutagenesis analyses identified two nonclassical aspartates (TMPD188 and LLLD368) as caspase-3-targeted sites in the AML1-ETO sequence. When these two aspartates were mutated into alanines, more intriguingly, the apoptosis-amplified action of AML1-ETO induction completely disappeared, while inducible expression of the caspase-3-cleaved 70 kDa fragment of AML1-ETO after tetracycline removal is sufficient to enhance apoptotic sensitivity. Further investigations on the potential in vivo effects of such a cleavage and its possible role in leukemogenesis would provide new insights for understanding the biology and treatment of AML1-ETO-associated leukemia.

Comparison of blood counts in splenic, renal and mesenteric arterial and venous blood in post-pubertal rabbits.

OBJECTIVES: To compare blood counts between arterial and venous blood to and from visceral organs and indirectly look into the function of the organs. METHODS: Splenic, renal and superior mesenteric arterial and venous blood samples were obtained from the arteries and veins in 38 post-pubertal rabbits and blood profile, including complete and differential blood counts, haemoglobin concentration and haematocrit, were measured with an automatic haematology analyser. RESULTS AND CONCLUSIONS: The rabbit spleen released a large amount of leucocytes (both lymphocytes and granulocytes) into the splenic venous blood (a venous increase of 33% in total leucocyte count), and also received more leucocytes (36-58% more in terms of concentration) from the artery than the kidney or intestine. Significantly fewer red blood cells were present in the renal venous blood than in the arterial blood (a venous reduction of 5% in erythrocyte count), but it remains to be clarified why and how the reduction was induced. More than 3-4% of water might be taken into the mesenteric venous blood during microcirculation (a venous reduction of 3-4% in erythrocyte-related parameters) and a significant number of leucocytes (mainly large leucocytes) in the mesenteric blood capillaries might migrate into the surrounding intestinal tissue (a venous reduction of 13% in leucocyte count).

Evidence for paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov.

Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prevalent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville Reservoir, the mat-forming filamentous cyanobacterium L. wollei, a species that had recently invaded from other areas of the southern United States, was studied to determine if it could produce any of the known cyanotoxins. Of the 91 field samples collected at 10 locations at Guntersville Reservoir, Ala., on the Tennessee River, over a 3-year period, 72.5% were toxic. The minimum 100% lethal doses of the toxic samples ranged from 150 to 1,500 mg kg of lyophilized L. wollei cells-1, with the majority of samples being toxic at 500 mg kg-1. Samples bioassayed for paralytic shellfish toxins by the Association of Official Analytical Chemists method exhibited saxitoxin equivalents ranging from 0 to 58 micrograms g (dry weight)-1. Characteristics of the neurotoxic compound(s), such as the lack of adsorption by C18 solid-phase extraction columns, the short retention times on C18 high-performance liquid chromatography (HPLC) columns, the interaction of the neurotoxins with saxiphilin (a soluble saxitoxin-binding protein), and external blockage of voltage-sensitive sodium channels, led to our discovery that this neurotoxin(s) is related to the saxitoxins, the compounds responsible for paralytic shellfish poisonings. The major saxitoxin compounds thus far identified by comparison of HPLC fluorescence retention times are decarbamoyl gonyautoxins 2 and 3. There was no evidence of paralytic shellfish poison C toxins being produced by L. wollei. Fifty field samples were placed in unialgal culture and grown under defined culture conditions. Toxicity and signs of poisoning for these laboratory-grown strains of L. wollei were similar to those of the field collection samples.