Efficacy and safety of transanal opening of intersphincteric space in the treatment of high complex anal fistula: A meta­analysis.

The best treatment of high complex anal fistula (HCAF) is to avoid anal incontinence while improving the cure rate. On this basis, several surgical procedures for preserving the anal sphincter have been proposed. The purpose of the present study was to evaluate the efficacy and safety of transanal opening of intersphincteric space for treating HCAF. PubMed, Cochrane Library, China National Knowledge Infrastructure and the Wanfang databases were searched to collate all the articles on transanal opening of intersphincteric space for treating HCAF. A total of two researchers independently completed the whole process, from screening and inclusion to data extraction and the data was included in the RevMan 5.3 software for analysis. The main outcomes included the patients' essential characteristics, primary healing rate, management after recurrence, final healing rate, anal incontinence score before and after surgery, postoperative complication rate and types of complications. A total of six articles were included in this meta-analysis. The results showed that the weighted final healing rate of patients following transanal opening of intersphincteric space was 89% [risk differences (RD)=0.89; 95% confidence interval (CI)=0.86-0.92; I2=0%; P<0.00001]. The results of the anal incontinence score showed that there was no significant difference between the results before and after transanal opening of intersphincteric space surgery mean differences [(MD)=-0.04, Cl=-0.10-0.02, I2=0%; P=0.21]. Only 11 patients were reported to have complications, including urinary retention and bleeding following transanal opening of intersphincteric space with a complication rate of 8% (11/138) and the weighted average complication rate was 6% (RD=0.06,95% CI=0.02-0.10; I2=9%; P=0.003). Transanal opening of intersphincteric space has a high cure rate, a favorable anal incontinence score, fewer types of postoperative complications and a low complication rate; it can be used as a minimally invasive and sphincter-preserving surgical method for treating HCAF and is worthy of further promotion and research in clinical practice.

Lactococcus G423 improve growth performance and lipid metabolism of broilers through modulating the gut microbiota and metabolites.

This study aimed to explore whether Lactococcus G423 could improve growth performance and lipid metabolism of broilers by the modulation of gut microbiota and metabolites. A total of 640 1-day-old AA broilers were randomly divided into 4 groups [Control (CON), Lac_L, Lac_H, and ABX]. Average daily gain (ADG), average daily feed intake (ADFI), feed conversion ratio (FCR), breast muscle, thigh muscle, and abdominal fat pad were removed and weighed at 42 days of age. Serum was obtained by centrifuging blood sample from jugular vein (10 mL) for determining high-density lipoprotein (HDL), total cholesterol (TC), low-density lipoprotein (LDL), and triglyceride (TG) using ELISA. The ileal contents were harvested and immediately frozen in liquid nitrogen for 16S rRNA and LC-MS analyses. Then, the results of 16S rRNA analysis were confirmed by quantitative polymerase chain reaction (qPCR). Compared with the CON group, FCR significantly decreased in the Lac_H group (p < 0.05) in 1-21 days; ADG significantly increased and FCR significantly decreased in the Lac_H group (p < 0.05) in 22-42 days. 42 days weight body and ADG significantly increased in the Lac_H group (p < 0.05) in 42 days. Abdominal fat percentage was significantly decreased by Lactococcus G423 (p < 0.05), the high dose of Lactococcus G423 significantly decreased the serum of TG, TC, and LDL level (p < 0.05), and the low dose of Lactococcus G423 significantly decreased the serum of TG and TC level (p < 0.05). A significant difference in microbial diversity was found among the four groups. Compared with the CON group, the abundance rates of Firmicutes and Lactobacillus in the Lac_H group were significantly increased (p < 0.05). The global and overview maps and membrane transport in the Lac_L, Lac_H, and ABX groups significantly changed versus those in the CON group (p < 0.05). The results of LC-MS demonstrated that Lactococcus could significantly improve the levels of some metabolites (6-hydroxy-5-methoxyindole glucuronide, 9,10-DiHOME, N-Acetyl-l-phenylalanine, and kynurenine), and these metabolites were involved in four metabolic pathways. Among them, the pathways of linoleic acid metabolism, phenylalanine metabolism, and pentose and glucuronate interconversions significantly changed (p < 0.05). Lactococcus G423 could ameliorate growth performance and lipid metabolism of broilers by the modulation of gut microbiota and metabolites.

N-carbamylglutamate supplementation induces functional egg production in layers by modulating liver transcriptome profiles.

Eggs rich in polyunsaturated fatty acids (PUFA), known as functional eggs, are animal products deemed beneficial to human health and possess high economic value. The production of functional eggs involves supplementing exogenous additives with the ability to regulate lipid metabolism. As N-Carbamylglutamate (NCG) serves as an endogenous arginine synthesizer, and arginine acts as the substrate for the formation of nitric oxide (NO), the biological function of NCG is partially mediated by NO. NO is a key regulatory molecule in lipid metabolism, suggesting that NCG may also have the ability to modulate lipid metabolism. In order to assess the capacity of NCG in regulating liver lipid metabolism and its potential application in producing functional eggs, we conducted a study to investigate the effects of dietary supplementation of NCG on production performance, serum, and liver NO levels, yolk fatty acid composition, and the liver transcriptome of layers. In this study, we utilized 30 layers of the Jinghong No.1 breed, all aged 45 wk. All the birds were randomly divided into 2 groups. Each group had 5 replicates, and each replicate had 3 birds. We provided them with different diets: one group received the basic diet, and the other group's diet was supplemented with 0.08% NCG. The experiment lasted for 14 wk. The results did not reveal any positive impact of NCG on production performance. However, NCG supplementation elevated NO levels in serum and liver, along with an increase in yolk PUFA, ω-3, and ω-6 fatty acids. Liver transcriptome analysis identified 124 upregulated differentially expressed genes (DEGs) and 43 downregulated DEGs due to NCG supplementation. Functional annotation using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database highlighted 3 upregulated DEGs (CPT1A, MOGAT1, and CHKA) and 2 downregulated DEGs (FASN and ETNPPL) associated with lipid metabolism. Pathway enrichment analysis revealed that CPT1A was enriched in the AMPK signaling pathway and the PPAR signaling pathway, while FASN was enriched in the AMPK signaling pathway. Thus, CPT1A and FASN are potential functional genes related to lipid metabolism facilitated by NCG supplementation. In summary, our study suggests that NCG supplementation modulates liver lipid metabolism, leading to the production of functional eggs in layers.

N-carbamylglutamate supplementation regulates hindgut microbiota composition and short-chain fatty acid contents in Charollais and Small Tail Han crossbred sheep.

Introduction: The objective of this study was to investigate the effects of N-carbamylglutamate (NCG) supplementation on the growth performance, hindgut microbiota composition, and short-chain fatty acid (SCFA) contents in Charollais and Small Tail Han crossbred sheep. Methods: A total of 16 female crossbred mutton sheep (Charollais × Small Tail Han), aged 4 months, with an initial body weight of 30.03 ± 0.08 kg, were utilized in a 60 days experiment. The sheep were divided into two groups based on their initial body weight. Each group consisted of 8 replicates, with each individual sheep considered as a replicate. The dietary treatments comprised a basal diet supplemented with either 0.00% or 0.12% NCG. Results and discussion: Our findings indicate that NCG supplementation did not have a significant effect on the growth performance of mutton sheep. However, it did lead to changes in hindgut SCFA contents. Specifically, NCG supplementation increased the content of propanoic acid while decreasing acetic acid and hexanoic acid in the hindgut. Through microbiota analysis using the 16S rRNA technique, we identified Lachnospiraceae_NK3A20_group and Parasutterella as biomarkers for the hindgut microbiota in mutton sheep fed a diet containing NCG. Further analysis of the microbiota composition revealed that NCG supplementation significantly increased the abundance of Lachnospiraceae_NK3A20_group and Parasutterella, while decreasing unclassified_f_Lachnospiraceae and Lachnoclostridium. Correlation analysis between hindgut SCFA contents and microbiota composition revealed that the abundance of Lachnoclostridium was positively correlated with the contents of acetic acid and hexanoic acid, but negatively correlated with propanoic acid. Additionally, the abundance of Lachnospiraceae_NK3A20_group and Parasutterella was positively correlated with the content of propanoic acid, while being negatively correlated with acetic acid and hexanoic acid. Based on these findings, we conclude that dietary supplementation of 0.12% NCG can modulate hindgut SCFA contents in mutton sheep by regulating the composition of the hindgut microbiota.

Sodium dichloroisocyanurate: improving broiler health by reducing harmful microbial levels in the waterline.

Sodium dichloroisocyanurate (NaDCC) is commonly used for treating drinking water, industrial water, and wastewater. This study aimed to investigate the potential effects of NaDCC-treated waterline drinking water on the growth of AA+ broilers by reducing microbial levels in the waterline. A total of 480 healthy 1-day-old AA+ broilers (46.77 ± 0.50 g) were selected for the experiment and randomly divided into four groups with six replicates of 20 birds each. The control group received regular drinking water, while the test groups received drinking water with NaDCC concentrations of 10, 30, and 50 mg/L. The test groups consumed the treated water on specific days throughout the 42-day experimental period. Results showed that NaDCC treatment significantly reduced the levels of E. coli, Salmonella, S. aureus and Moulds in the drinking water at the waterline (p < 0.05). Drinking water with NaDCC also led to reduced broiler fecal emissions of NH3 and H2S, as well as reduced counts of E. coli, Salmonella, S. aureus and Moulds (p < 0.05), particularly at 30 mg/L and 50 mg/L concentrations. Broilers consuming NaDCC at 50 mg/L exhibited a significant increase in ADG from days 1-42 (p < 0.05). The levels of E. coli, Salmonella, S. aureus and Moulds in the drinking water at the waterline were significantly and positively correlated with the bacterial count in the feces (p < 0.05, R > 0.6). Additionally, bacterial levels in drinking water and broiler feces were negatively correlated with broiler production performance indicators, including ADG, ADFI, F/G and AWC. In conclusion, NaDCC can indirectly enhance broiler performance by reducing the levels of harmful bacteria in the waterline without affecting normal drinking water. The addition of 30 mg/L or 50 mg/L of NaDCC to the waterline in poultry production can effectively control harmful microorganisms and improve poultry health. Based on the experiment's results, it is recommended to preferentially use 30 mg/L NaDCC in the waterline to reduce farming costs.

Production performance, egg quality, and uterine gene expression for layers as affected by N-Carbamylglutamate supplementation.

Introduction: Supplementation of exogenous additives is a strategy to improve laying performance of layers by regulating uterine function. N-Carbamylglutamate (NCG) as an activator for endogenous arginine synthesis has the potential to regulate the laying performance of layers, but its effects have not been fully understood. Methods: This study investigated the effects of dietary supplementation of NCG on production performance, egg quality, and uterine gene expression in layers. A total of 360 45-week-old layers with a genetic line of Jinghong No. 1 were used in this study. The experimental period was 14 weeks. All birds were divided into 4 treatments with 6 replicates per treatment and 15 birds per replicate. Dietary treatments were based on a basal diet and supplemented with 0, 0.08, 0.12, or 0.16% NCG to form C, N1, N2, and N3 groups. Results and discussion: We found that layers in group N1 had higher egg production rate than those in group C. Egg weight was significantly reduced, while eggshell thickness was significantly improved, by treatment. However, the albumen height and Haugh unit were the lowest in group N3. Based on the above results, groups C and N1 were selected for further transcriptomics analysis of uterine tissue by RNA-seq. More than 7.4 Gb clean reads and 19,882 tentative genes were obtained using the Gallus gallus genome as a reference. Transcriptomics analysis in uterus tissue revealed that 95 differentially expressed genes (DEGs) were upregulated and 127 DEGs were downregulated. Functional annotation and pathway enrichment analysis showed that DEGs in uterine tissue were mainly enriched in glutathione metabolism, cholesterol metabolism, and glycerolipid metabolism, etc. Vitamin A metabolism-related gene, RBP1, nutrient transport-related gene, ALB, protein synthesis-related gene, METTL21C, and calcium transport-related gene, RYR2, CACNB2, RAMP3, and STAC, were significantly regulated by 0.08% NCG supplementation. Therefore, we concluded that NCG supplementation at a dose of 0.08% improved production performance and egg quality of layers by regulating uterus function.

Effects of dietary supplementation of N-carbamylglutamate on the haematology parameters, secondary sexual characteristics and testicular gene expression in roosters.

A total of 288 11-week-age roosters were used to evaluate the effects of dietary supplementation of N-carbamylglutamate (NCG) on reproductive traits and testicular gene expression. The experimental periods were 12 weeks. All birds were randomly assigned to 4 treatments with 6 replicates per treatment and 12 birds per replicate. Dietary conditions were based on a basal diet and supplemented with 0%, 0.08%, 0.12%, or 0.16% NCG to form C, N1, N2 and N3 groups respectively. Dietary supplementation of NCG had positive effects on the seminiferous tubule parameters, serum gonadotropin-releasing hormone and testosterone levels and the secondary sexual characteristics. Transcriptomics analysis was performed on the testicular tissues between C and N3 groups at the 16-week-age. Genes were mainly enriched in nine pathways, such as cytochrome P450 exogenous metabolism, drug metabolism, steroid hormone synthesis and glutathione metabolism, in which the ZP4 gene, cytochrome P450 family member 11A1 and other genes involved in the maintenance of gonadal function, steroid hormone biosynthesis and metabolism, and so forth, exist differences in expression levels. In summary, dietary supplementation of NCG had positive effects on the reproductive traits of roosters. NCG supplementation improved the development of reproductive traits of roosters by regulating the genes expression in testicular tissues and thus improved the synthesis of reproductive hormones in vivo.

Efficacy and safety of video-assisted anal fistula treatment in anorectal fistula: a meta-analysis.

INTRODUCTION: By searching relevant literature, the recurrence rate, complication rate after video-assisted anal fistula treatment (VAAFT), and efficacy and safety of the treatment were analyzed. EVIDENCE ACQUISITION: Articles that reported the outcomes of VAAFT up to December 2020 were searched in PubMed (Medline) and Cochrane Library, in accordance with the preferred reporting items for systematic review and meta-analysis (PRISMA) screening guidelines. Two researchers independently completed the whole process from screening and inclusion to quality evaluation and bias risk assessment, and the data was included in the RevMan 5.3 software for analysis. The main outcomes were demographic data of patients, detection rate, classification of internal opening of anorectal fistula, postoperative recurrence rate, and incidence of complications. EVIDENCE SYNTHESIS: A total of 10 articles were included (779 patients). The average age of the patients was 44 years old, average operation time was 60 min, and the average follow-up time was 22 months. The ratio of male to female was 2.4:1, the ratio of high anorectal fistula to low anorectal fistula was 6.6:1, the detection rate of internal openings was 98%, the weighted recurrence rate was 24%, and the weighted complication rate was 1%. CONCLUSIONS: VAAFT is effective and safe in the treatment of anorectal fistulas.

A pivotal role of selective autophagy in mitochondrial quality control: Implications for zinc oxide nanoparticles induced neurotoxicity.

Excessive occupational, medical, and environmental exposure of zinc oxide nanoparticles (ZnONPs) caused its accumulation in the nervous system and raised global concerns over its detrimental effects. However, very few researches had been conducted on the impact of mitochondrial quality control process on central nervous system (CNS) after ZnONPs administration, including mitochondrial fission, fusion, biogenesis, and autophagy. In present study, mitochondrial dysfunction and apoptosis were triggered in ZnONPs-exposed human neuroblastoma SH-SY5Y cells. Upregulation of mitochondrial biogenesis regulator (PGC-1α) and fission proteins (Drp1) and downregulation of fusion proteins (OPA1 and Mfn2) were observed in 3 and 6 µg/mL ZnONPs-treated cells. Meanwhile, loss of mitochondrial dynamics and biogenesis was observed in the severe impaired cells (treated with 12 µg/mL ZnONPs). More, autophagy and mitophagy were significantly activated in ZnONPs-treated cells. The increased Beclin1 and LC3 II proteins, decreases of p62 protein, and activated PINK1/Parkin signaling were quantified. The autophagy agonist (Rapamycin), inhibitor (3-MA), and mitophagy inhibitor (Cyclosporine A, CsA) were employed to verify the roles of autophagy and mitophagy in ZnONPs-treated cells. Consequently, mitochondrial dysfunction and apoptosis were aggravated by the blockage of autophagy and mitophagy. Our research could be used to evaluate the risk assessment of ZnONPs exposure in CNS neurons so as to provide a crucial guideline for their future biological applications.

Hypothalamic and pituitary transcriptome profiling using RNA-sequencing in high-yielding and low-yielding laying hens.

The reproductive physiology and laying performance of laying hens are regulated by the hypothalamus and pituitary. To understand the mechanism of egg laying regulation, we sequenced and analysed the hypothalamus and pituitary expression profiles in high- and low-yielding laying Chinese Dagu Chickens (CDC) using RNA-seq. More than 46 million clean reads and 24,873 tentative genes were obtained using the Gallus gallus genome as a reference. Transcriptome analysis in hypothalamus and pituitary revealed seven and 39 differentially expressed genes (DEGs) between high- and low-yielding CDC hens, respectively. A total of 24 and 22 DEGs were up-regulated and down-regulated, respectively, and 13 novel genes were identified. Functional annotation and pathway enrichment analysis showed that DEGs in the hypothalamus were mainly enriched in glycosaminoglycan biosynthesis. DEGs significantly enriched in the pituitary primarily affected the extracellular matrix, the protein extracellular matrix, and the extracellular space. Pathways involving phenylalanine metabolism, 2-oxocarboxylic acid metabolism, the glycosphingolipid biosynthesis-ganglion series, and local adhesion were significantly enriched in the pituitary. Eight DEGs, PRDX6, TRIB2, OVCH2, CFD, Peptidase M20, SLC7A10, and two other amino acid transporters, are involved in the metabolism and transport of amino acids. To our knowledge, this is the first study comparing the hypothalamus and pituitary transcriptomes of high- and low-yielding laying hens. Our findings suggest that putative differences in gene expression can provide a base for further research in this field. Moreover, we identified increased expression of genes involved in amino acid metabolism, glycosaminoglycan biosynthesis, and oestrogen negative feedback systems in low-yielding laying hens, highlighting their potential as biomarkers of egg production.

Inhibition of alpha-hemolysin expression by resveratrol attenuates Staphylococcus aureus virulence.

Staphylococcus aureus is an important zoonotic pathogen that causes a variety of life-threatening diseases. The increasing emergence of drug resistance further complicates the treatment of S. aureus infections. The critical role of alpha-hemolysin (Hla) in virulence renders this toxin an ideal target for the development of anti-infective agents for S. aureus. Here, We found that resveratrol, a natural compound widely found in fruits without antibacterial activity, could effectively inhibit Hla expression via down-regulating the transcription of hla, the gene that encodes Hla, and RNAIII, the effector molecule of the agr system. The addition of resveratrol to a co-culture system of S. aureus and A549 cells significantly alleviated bacteria-mediated cellular injury. Furthermore, treatment with resveratrol effectively protected mice from S. aureus pneumonia. Our results established resveratrol as an effective Hla inhibitor that reduces Hla expression without antimicrobial activity and can be further developed into novel therapeutics against S. aureus infections.

Extraction, preliminary characterization and antioxidant properties of polysaccharides from the testa of Salicornia herbacea.

Response surface methodology and central composite design based on single-factor experiments were used to optimize the extraction parameters (i.e., extraction time, extraction temperature, and solid-liquid ratio) of polysaccharides from the testa of Salicornia herbacea (STP). The optimal conditions included a liquid-solid ratio of 57:1 (volume/mass), an extraction time of 300min and an extraction temperature of 87°C. Under these conditions, the maximal yield of crude STP was 1.30±0.06%, which agreed with model predictions. Preliminary characterization of the purified acidic polysaccharide (STP II-1) by high performance liquid chromatography indicated that it was composed mainly of d-mannose, d-rhamnose, d-glucose, d-galactose, d-arabinose, glucuronic acid and galacturonic acid. The antioxidant capacity of STP II-1 was analyzed by monitoring both the scavenging rate of the 2,2-diphenyl-1-picrylhydrazyl free radical and inhibition rate of H2O2-induced erythrocyte hemolysis.

Expression of nerve growth factor and its receptor, tyrosine kinase receptor A, in rooster testes.

Nerve growth factor (NGF), which is required for the survival and differentiation of the nervous system, is also thought to play an important role in the development of mammalian reproductive tissues. To explore the function of NGF in the male reproductive system of non-mammalian animals, we determined the presence of NGF and its receptor, tyrosine kinase receptor A (TrkA), in rooster testes and investigated the regulation of NGF and TrkA expression by follicle-stimulating hormone (FSH). The mRNA and protein levels of NGF and TrkA in 6-week-old rooster testes were lower than those in 12-, 16- or 20-week age groups; levels were highest in the 16-week group. Immunohistochemistry showed that NGF and TrkA were both detected in spermatogonia, spermatocytes and spermatids. NGF immunoreactivity was observed in Leydig cells and strong TrkA signals were present in Sertoli cells. Meanwhile, FSH increased TrkA transcript levels in rooster testes in a dose-dependent manner. We present novel evidence for the developmental and FSH-regulated expression of the NGF/TrkA system, and our findings suggest that the NGF/TrkA system may play a prominent role in chicken spermatogenesis.

Association of inhibin-α gene polymorphisms with follicular cysts in large white sows.

Ovarian follicular cysts are anovulatory follicular structures that have been identified in sows and are known to cause infertility. The pathogenesis of follicular cysts remains poorly understood. Hormones play key roles in the formation and persistence of cysts. The hormone inhibin is a member of the TGF-ß superfamily and is named for its negative regulation of FSH, another hormone that controls follicular recruitment and growth. In the present study, 48 sows with follicular cysts and 60 normal sows with no cysts were screened for mutations in the inhibin-α gene to examine the association of inhibin-α gene polymorphisms with the presence of follicular cysts. The results show that the c.-42G>A and c.3222G>A polymorphisms are significantly associated with follicular cysts and that sows with c.-42GG and c.3222GG genotypes have lower risk of developing cysts. Our findings may provide novel biological biomarkers and promising gene therapy candidates for follicular cyst formation in sows, which would greatly benefit pig breeding programs.

Downregulation of the expression of inhibin α subunit and betaglycan in porcine cystic follicles.

Inhibins, as members of the transforming growth factor beta (TGF-ß) superfamily, downregulate the synthesis and secretion of follicle-stimulating hormone (FSH) in an endocrine manner. The role of inhibin/betaglycan in the ovary regulation recently gained attention. To date, no data exist on the function of inhibin α subunit and betaglycan in cystic follicles. In this study, the expressions of inhibin α subunit and betaglycan in cystic follicles were investigated using immunohistochemistry, real-time PCR and Western blot analysis. Both inhibin α subunit and betaglycan immunoreactivities were mainly localized in the granulosa cells of follicles. Expression of inhibin α subunit and betaglycan was inferior in cystic follicles compared with that in normal large follicles. However, the result of enzyme-linked immunosorbent assay showed no significant difference in the decreasing in concentration of inhibin α subunit in cystic follicular fluid compared with the control (P>0.05). In this study, we explored the effects of FSH on betaglycan expression in granulosa cells in vitro. As expected, a significant increase in the expressions of betaglycan mRNA and protein in granulosa cells was observed in response to exogenous FSH (30 ng/ml) (P<0.05) compared with the control. Consequently, this study provides evidence that the expressions of inhibin α subunit and betaglycan are inferior in cystic follicles, and this may be caused by the decrease in FSH in the presence of a cystic follicle.

The interactions between nerve growth factor and gonadotrophins in bovine oviduct.

Nerve growth factor promotes the survival and differentiation of nervous cells and is thought to play an important role in the development of reproductive tissues. The aims of this work were to detect the presence of NGF and its receptor NTRK1 in bovine oviduct samples, and to investigate the regulatory interactions between NGF/NTRK1 and gonadotrophins in bovine oviduct epithelial cells. Both transcripts and proteins of NGF and NTRK1 were detected by RT-PCR and Western blotting, and the corresponding proteins were specifically immunolocalized in oviduct epithelial cells. In addition, real-time PCR experiments revealed that the levels of NGF and NTRK1 mRNA in oviduct epithelial cells treated with exogenous FSH or LH were greater than those in negative control cells (P<0.05). Similarly, treatment with NGF significantly increased the expression of FSHR and LHR in oviduct epithelial cells via its effects on NTRK1 (P<0.05). This process was suppressed by treatment with the NTRK1 inhibitor K252α. We conclude that NGF/NTRK1 may have a role in regulating the function of bovine oviducts via its interactions with gonadotrophins.

NF-κB pathway contributes to cadmium-induced apoptosis of porcine granulosa cells.

To better understand the mechanism of cadmium (Cd)-induced apoptosis of porcine granulosa cells, we examined the nuclear factor-kappa B (NF-κB) p65 subunits intracellular translocation and the expression of some downstream apoptotic-related genes. Apoptosis and reactive oxygen species (ROS) production in porcine granulosa cells exposed to cadmium chloride (CdCl2) were determined by acridine orange/ethidium bromide double staining and 2,7-dichlorodihydro-fluorescein-diacetate oxidation staining, respectively. The results showed that the apoptosis of porcine granulosa cells induced by CdCl2 significantly increased in a time- and dose-dependent manner along with the increasing of ROS production, and 10 µM parthenolide, an inhibitor NF-κB, can accelerate the process of apoptosis. Moreover, immunofluorescence and western blot results showed that CdCl2 could stimulate the translocation of p65 into nucleus in porcine granulosa cells. Furthermore, CdCl2 also significantly stimulate the expression of Bcl-2 proteins in porcine granulosa cells than that in the control. In contrast, we did not find any change of Bax expression in granulosa cells upon exposure of cadmium. Taken together, these results demonstrate that the activation of NF-κB pathway may play a crucial role in cadmium-induced apoptosis of porcine granulosa cells.

Interaction between neurotrophin 4 and gonadotrophin in bovine oviducts.

The expression and localization of neurotrophin 4 (NT4) and its receptor, tyrosine kinase B (TRKB), in the bovine oviduct, and their interaction with gonadotrophins in bovine oviduct epithelial cells (BOECs), were examined. Transcripts for NT4 and TRKB were detected by reverse transcription polymerase chain reaction (RT-PCR) in bovine oviducts in the follicular and luteal phases, and their proteins were immunolocalized in BOECs. Based on real time PCR, NT4 mRNA did not differ significantly between the two phases of the cycle, although TRKB mRNA expression was higher (P < 0.05) in the luteal phase than that in follicular phase. The BOECs were treated with various concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in vitro; for NT4, mRNA and protein were higher (P < 0.05) than those in the control (based on real time PCR and enzyme-linked immunosorbent assay (ELISA) assays). The effects of NT4 and the TRKB inhibitor (K252a) on the expression of LH receptor (LHR) and FSH receptor (FSHR) in the oviduct epithelial cells were also studied using a monolayer culture model. Expression levels of LHR and FSHR mRNA in BOECs treated with various concentrations of NT4 were higher (P < 0.05) than those in the control. However, these expressions were blocked by treatment with K252α. We concluded that neurotrophin 4 may have a role in regulating the function of bovine oviducts by interacting with gonadotrophins.

Absence of nerve growth factor and comparison of tyrosine kinase receptor A levels in mature spermatozoa from oligoasthenozoospermic, asthenozoospermic and fertile men.

BACKGROUND: Nerve growth factor (NGF) and the Tyrosine kinase receptor (TrkA) are expressed in mammalian ovaries and testes, where they are involved in oocyte maturation and spermatogenesis. We assessed the possibility that NGF and TrkA gene products are present in human spermatozoa, and explored differences in levels of NGF and TrkA in spermatozoa from oligoasthenozoospermic, asthenozoospermic and fertile men. METHODS: The presence of NGF and TrkA in human spermatozoa was investigated using RT-PCR, immunofluorescence and Western blotting. Real-time PCR and ELISA were used to determine expression levels of NGF and TrkA. RESULTS: No NGF transcripts were detected in human spermatozoa. The levels of TrkA mRNA in spermatozoa from oligoasthenozoospermic samples were significantly lower than those in spermatozoa from fertile and asthenozoospermic samples (P<0.05). NGF concentrations in seminal plasma samples from oligoasthenozoospermic men were lower than those in seminal plasma from fertile and asthenozoospermic men, but these differences were not statistically significant (P>0.05). CONCLUSIONS: The levels of both NGF protein in seminal plasma and TrkA mRNA in spermatozoa are low in samples from oligoasthenozoospermic men.