Effect of age at first egg on reproduction performance and characterization of the hypothalamo-pituitary-gonadal axis in chickens.

As an important indicator of sexual maturity of hens, age at first egg (AFE) is significantly associated with reproduction performance. In this study, 400 hens were divided into 6 groups based on AFE to analyze the difference of reproduction performance, reproduction hormone levels and the characterization of the hypothalamo-pituitary-gonadal axis in chickens. The results showed that the egg production of the hens in the late-maturing groups was significantly lower than that of the ones in other groups and the precocious hens had a lower egg production rate. The hens in late-maturing group had a lower fertility rate, LH levels and shorter duration of peak of egg production (PEP), the precocious hens had lower PRL levels. In addition, the characterization of the hypothalamo-pituitary-gonadal axis showed that the individuals with normal AFE had higher GNRH, GNRHR, ESR1, KITLG, and CYP11A1 expression levels than late-maturing and precocious individuals, which indicated that the chickens with normal AFE advantages on reproduction regulation system.

RFamide-related peptides' gene expression, polymorphism, and their association with reproductive traits in chickens.

RFamide-related peptides (RFRP) are synthesized by the hypothalamus and have a regulatory role in gonad development. The goal of this study was to investigate the association between SNP of the RFRP gene and the reproductive traits and hormone levels of Zhenning yellow chickens. The mRNA expression levels were detected based on different tissues, ages, and genotypes. Eleven mutation sites were detected in the RFRP gene, 4 of which were significantly related to reproductive traits and hormone levels. Association analysis revealed that A276G was associated with egg production at 300 d of age (EP300) and amount of prehierarchical follicles (P < 0.05). G1396A was associated with egg weight at 300 d of age and luteinizing hormone (LH) and prolactin levels (P < 0.05). G1694A showed significant associations with fertilization rate and LH levels (P < 0.05), and A2659G was associated with EP300 (P < 0.05). The results of expression analysis showed that the RFRP mRNA expression levels in the hypothalamus were higher than those in other tissues (P < 0.01). The expression in immature individuals was higher than that in mature ones (P < 0.01). There were also differences in mRNA expression levels between different genotypes (P < 0.05). In summary, the results of this study might provide potential markers and a theoretical basis for the improvement of chicken reproductive traits.

H-FABP gene expression and genetic association with meat quality traits in domestic pigeons (Columba livia).

1. H-FABP (heart-type fatty acid-binding protein) is an important fatty acid-binding protein that participates in the metabolism and intracellular transportation of lipids. This study was designed to study the H-FABP gene expression and genetic association with meat quality traits in domestic pigeons.2. The spatio-temporal expression patterns showed that H-FABP was widely expressed in all eleven tissues in birds from hatching to 4 weeks of age, and the expression level in the liver was the highest, and spleen and subcutaneous fat showed relatively high levels at all time points, and increased markedly as squabs became older.3. Nine novel SNPs were found; three of them (g.42 C > T and g.123 C > T in exon 1 and g.3128 C > T in exon 2) were synonymous mutations, and six of them (g.3304 T > C, g.3305 G > A, g.3313A>G, g.3343 C > T, g.3372A>C, g.3410 T > C in exon 3) were located in the 3' untranslated region (UTR). In addition, four haplotypes and six diplotypes were formed and showed weak linkage disequilibrium. Association analysis showed that in the SNP of g.3305 G > A, the AB genotype showed higher (P < 0.05) inosinic acid concentrations in breast muscle than the BB genotype. In the SNP of g.3313A>G, the AA and AB genotypes showed significantly higher (P < 0.01) inosinic acid concentrations than the BB genotype, AA showed the highest intramuscular fat content, and the relative mRNA expression level of the AA genotype was the highest (P < 0.05).4. Association analysis of diplotypes showed no correlation with meat quality traits. Moreover, correlation analysis indicated that the H-FABP mRNA expression level was tightly related to intramuscular fat content (P < 0.01).5. This study suggested that the H-FABP gene may be a candidate gene in marker-assisted selection of pigeons for high-quality meat.

[Multimodality navigation for liver resection of complicated alveolar echinococcosis].

Objective: To investigate the clinical application of multimodality navigation for liver resection in the treatment of complicated alveolar echinococcosis (AE). Methods: From October 2019 to February 2020, the clinical data and perioperative results of patients with AE treated by surgery in our department were retrospectively studied. Hepatic parenchyma disconnection plane and liver resection were navigated and performed with three-dimensional reconstruction and HITACHI real-time multi-image fusion interventional navigation system (RVS). Results: All of six patients were successful performed radical liver resection without mortality. The operation time was (301±106)min and the median blood loss was 200 ml. Two patients needed blood transfusion intraoperative (33.33%). The postoperative hospital stay was (10.8±2.8) day, and the cost of hospitalization was (82 584±995.61) yuan. Clavien-Dindo grade Ⅲ complication occurred in one patient. Conclusions: Multimodality navigation might provide precise intraoperative navigation of the surgical plane and effectively assist liver resection for the treatment of complicated AE.

POMC gene expression, polymorphism, and the association with reproduction traits in chickens.

Reproduction trait is one of the most important economic traits in poultry industry. This study was aimed to investigate the mRNA expression levels, single nucleotide polymorphisms (SNP) of POMC gene, and the association with reproduction traits in chickens. Five SNP (g.958 G > A, g.1374 G > C, g.1393 G > A, g.1817 C > T, and g.1918G > A) were detected in introns of POMC gene in 317 Zhenning yellow chickens. Association analysis revealed that g.958 G > A and g.1817 C > T showed significantly associations with fertilization rate, hatching rate of hatching eggs, and hatching rate of fertilized eggs in chickens. Simultaneously, g.1374 G > C and g.1918G > A were both associated with egg weight at 300 D of age (P < 0.05). The SNP of g.958 G > A, g.1393 G > A, and g.1817 C > T were all associated with E2 hormone levels (P < 0.05). The result of mRNA expression levels in different tissues showed that POMC mRNA expression level in the pituitary was higher than those in the other tissues and varied in different genotypes. In conclusion, the results in this study provided new evidences that polymorphisms of the POMC gene have potential effects on reproduction traits in chickens. The 5 SNP detected in this study could be potential markers for improving reproduction traits in chickens.

Expression of MSTN gene and its correlation with pectoralis muscle fiber traits in the domestic pigeons (Columba livia).

Myostatin (MSTN) is a negative regulator of skeletal muscle growth and plays an important role in muscle development. In this research, we constructed a tissue expression profile of the pigeon MSTN gene in eight tissues and a spatio-temporal expression profile in the pectoral muscle using qRT-PCR method. And the pectoralis muscle fiber traits during pigeon post-hatching stages at 1, 7, 14, 21, and 28 D were analyzed through the paraffin sections. Then the correlations between the muscle fiber diameter, cross-sectional area, density, and the expression of MSTN in the pectoral muscle were analyzed. Results showed that MSTN mRNA was mainly expressed in breast muscle, heart, spleen, and kidney and it was almost unexpressed in the liver and lungs. Moreover, the MSTN mRNA expression level in breast muscle was significantly higher than that in other tissues (P < 0.05), and showed an interesting trend that it decreased in the first week and then increased with age. Meanwhile, decrease of myostatin transcripts was accompanied by the down-regulation of Myf5 and the up-regulation of MyoG during the first week post-hatching. In addition, the paraffin sections analysis results revealed that the diameter and cross-sectional area of pectoralis muscle fiber significantly increased with age (P < 0.05), and a significant positive correlation was shown between the MSTN gene expression level and muscle fiber diameter (P < 0.05). These fundamental researches might contribute to further understanding of the roles MSTN played in the post-hatching muscle fiber development in pigeon.

Association of ADSL gene polymorphisms with meat quality and carcass traits in domestic pigeons (Columba livia).

1. Adenylosuccinate lyase (ADSL) plays an important role in the synthesis of inosine monophosphate (IMP). In this trial, a total of 200 pigeons were sampled and slaughtered. Seven meat quality traits and 11 carcass traits were measured. DNA sequencing was used to detect nucleotide mutations, and associations between ADSL gene polymorphisms and meat quality and carcass traits were analysed. 2. Sequencing results showed that 9 nucleotide mutations were found in the exons of the ADSL gene. All the mutations were synonymous except C13065G, which caused a change in amino acids (Ser to Arg). In addition, two of the detected single nucleotide polymorphisms (SNPs) had significant associations with meat quality and carcass traits. 3. For the C13065G SNP located in exon11, the IMP content of breast muscle in AA and AB genotype was higher than in the BB genotype (P < 0.01). The SNP G10696A located in exon10 was significantly associated with carcass rate, in which AA and AB genotype were higher than the BB genotype (P < 0.01). 4. The results indicated that the ADSL gene has a close association with meat quality and carcass traits in domestic pigeons, and G10696A and C13065G SNPs could be applied as genetic markers in molecular marker-assisted breeding of pigeons.

Effects of rearing systems on reproductive hormones secretion and their receptors gene expression in Xianju chickens under summer conditions.

Previous study in our lab showed that indigenous Xianju chickens from free-range system (FRS) under summer conditions had lower egg production than those from conventional cage rearing system (CRS). The objective of this study was to preliminarily determine the FRS-dependent mechanism of depressing laying performance according to determining the effect of rearing systems on reproductive hormones secretion and their receptors mRNA expression in Xianju chickens reared under summer conditions. A total of 360 indigenous Xianju chickens were randomly allocated to CRS and FRS groups, each of which included 5 replicates of 36 hens. The experiment lasted between 21 and 29 wk of age. We found that the ovarian weight, numbers of small yellow follicles, and large white follicles in the FRS group were lower than those in the CRS group (P < 0.05). Changing from CRS to FRS increased serum concentrations of prolactin and decreased serum-luteinizing hormone and progesterone levels (P < 0.05). Gene expressions in the preovulatory follicles from FRS hens were upregulated for prolactin receptor and downregulated for luteinizing hormone receptor and progesterone receptor, compared to those from CRS hens (P < 0.05). It can be concluded that changing from CRS to FRS in the current experimental conditions depressed egg production traits in Xianju chickens by inducing a synergistic activity of reproductive hormones and the gene expressions of their receptors.

Association of DGAT2 gene polymorphisms with carcass and meat quality traits in domestic pigeons (Columba livia).

1. Diacylglycerol acyltransferase (DGAT) plays an important role in the synthesis of triacylglycerol, but its effects on meat quality and carcass composition in pigeons are unclear. In this study, single-nucleotide polymorphisms (SNPs) in the exons of the DGAT2 gene were identified and analysed by using DNA sequencing methods in 200 domestic pigeons (Columba livia). The associations between DGAT2 polymorphisms and carcass and meat quality traits were also analysed. 2. Sequencing results showed that 5 nucleotide mutations were detected in exons 3, 4, 5 and 6 of the DGAT2 gene. The analysis revealed three genotypes (AA, AB and BB) in G18398T and G22484C, in which the AA genotype and A allele had the highest frequency. 3. In the SNP of G18398T located in exon 5, individuals with genotype BB had significantly higher meat quality and lower abdominal fat content than those with AA or AB genotype. In the SNP of G22484C located in exon 6, the genotype AA showed highest carcass trait values, while the genotype BB represented better meat quality, compared to AA and AB genotypes. 4. The results imply that DGAT2 gene has a close relationship with carcass and meat quality traits in pigeons, and the SNPs of G18398T and G22484C can be used as genetic markers for marker-assisted breeding in pigeon.

Effects of rearing systems on laying performance, egg quality, and serum biochemistry of Xianju chickens in summer.

The objective of this study was to compare the laying performance, egg quality, and serum biochemistry of hens maintained in conventional cage rearing system (CRS), flat net-rearing system (NRS), and free range system (FRS) under summer conditions. Indigenous Xianju chickens (n = 540) were randomly allocated into cages or pens of rearing system groups, within each system there were 5 replicates with 36 hens in each replicate. The experiment lasted between 21 and 29 wk of age. Hen-day egg production (P = 0.00) and egg mass (P = 0.00) were higher in the CRS but were similar in the NRS and FRS. Lowest egg weight (P = 0.02), yolk weight (P = 0.00) and yolk ratio (P = 0.01), and feed intake (P = 0.01) were observed from the FRS, whereas lowest feed conversion ratio (FCR) was recorded from the CRS (P = 0.01). Rearing systems had negligible effect on egg quality. Serum Ca (P = 0.04) and total protein (P = 0.03) levels were found to be higher in the CRS but were lower in the FRS. Serum levels of glucose (P = 0.01), cholesterol (P = 0.00), and triglyceride (P = 0.00) in the CRS increased compared with the NRS and FRS groups, whereas serum levels of high density lipoprotein cholesterol (HDL-C; P = 0.01) in the CRS decreased. It can be concluded that under summer conditions, Xianju chickens from CRS had an advantage in terms of productivity parameters, but exhibited higher levels of serum lipids and glucose.

[The lesser omentum approach for laparoscopic pancreatic enucleation].

Objective: To evaluate the feasibility and strategy of the lesser omentum approach for laparoscopic pancreatic enucleation. Methods: Between June 2011 and October 2013, 6 laparoscopic pancreatic enucleations were performed by lesser omentum approach.The average age was 42 years, including 1 male and 5 female.The preoperation diagnosis of 6 cases were pancreatic islet cell tumors determined by abdominal CT/MRI, ultrasound and digital subtraction angiography.The tumors of 3 cases located in pancreatic neck, 2 tumors located in neck and body of pancreas, and 1 tumor located in pancreatic body.Their biggest tumor diameter were 0.8-2.5 cm. Results: Among the 6 cases, all laparoscopic pancreatic enucleations were successfully performed.None of the patients were converted to open operation.Eestimated blood loss was (26.7±18.6)ml, operating time was (82.5±19.4)minutes, and postoperative length of stay was (5.17±1.17)days.Additionally, postoperative complication included grade A pancreatic fistula in 1 case.After 36-64 months followed-up, there was no tumor recurrence and clinical symptom disappeared. Conclusion: For the islet cell tumors located in pancreatic neck and body, the lesser omentumapproach may contribute to good surgical view and operative space, which can make pancreatectomy safer and easier for clinical application.

Association of MYF5 and KLF15 gene polymorphisms with carcass traits in domestic pigeons (Columba livia).

Single nucleotide polymorphisms (SNPs) in the exons of the myogenic factor 5 (MYF5) and Kruppel-like factor 15 (KLF15) genes were identified and analysed by using DNA sequencing methods in 60 female domestic pigeons (Columba livia). Five SNPs (T5067A, C5084T, C5101T, T5127A and C5154G) were detected in exon 3 of MYF5 and 6 SNPs (C1398T, C1464T, G1542A, C1929T, G1965A and A2355G) were found in exon 2 of KLF15, respectively. The analysis revealed three genotypes, in which the AA genotype was dominant and the A allele showed a dominant advantage. For the MYF5 gene, the C5084T and T5127A SNP genotypes were significantly associated with carcass traits of pigeons. Within those two SNPs, the BB genotype showed relatively higher trait association values than those of AA or AB genotypes. No significant association was observed between the KLF15 SNP genotypes and carcass traits. These results indicated that the MYF5 gene is a potential major gene affecting carcass traits in domestic pigeons. The BB genotype of the C5084T and T5127A SNPs could be a potential candidate genetic marker for marker-assisted selection in pigeon.

Moxibustion attenuates inflammatory response to chronic exhaustive exercise in rats.

Exercise is recognized as an activator to elicit an inflammatory response whilst moxibustion in traditional Chinese medicine has been previously found to modulate immune functioning. However, whether moxibustion can alleviate the inflammatory cytokines response to chronic exhaustive exercise remains unknown. In the present study, rats were randomly assigned to a sedentary control group (Sed), a sedentary moxibustion group (Sed + Moxa), and 2 trained groups- one submitted to a 3-week exhaustive swimming (Trained), and the other a trained moxibustion group (Trained + Moxa). We found that chronic exhaustive exercise significantly increased the serum levels of pro-inflammatory cytokines (IL-1ß, TNF-α, IFN-γ) and the IFN-γ/IL-4 ratio, and decreased the anti-inflammatory cytokines (IL-4, IL-10). Moxibustion treatment markedly reduced the serum levels of IL-1ß, IFN-γ and the IFN-γ/IL-4 ratio, while elevated the IL-4 and IL-10 productions in trained rats. However, TNF-α level was not significantly affected. Our results suggested that an excessive inflammatory response and a potential inflammatory damage may be involved during chronic exhaustive exercise. Moxibustion could attenuate the inflammatory impairment and have an anti-inflammatory effect. The beneficial effects of moxibustion might be mediated by reducing the pro-inflammatory cytokines, increasing the anti-inflammatory cytokines, and modulating the balance between pro- and anti-inflammatory cytokines.

Sertoli cells induce xenolymphocyte apoptosis in vitro.

BACKGROUND: Testicular Sertoli cells can protect pancreatic islet grafts from allo- and autoimmune destruction; however, the mechanisms underlying immune privilege of the testicle are not well understood, especially in xenotransplantation. The purpose of this study was to investigate whether rat Sertoli cells could induce mouse lymphocyte apoptosis in vitro. METHODS: Testis was isolated from 2- to 4-week-old Sprague Dawley (SD) rats. Sertoli cells were successfully prepared by digestion with collagenase type V, trypsin, and DNase I, and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. We examined the apoptosis rates of Balb/c mouse lymphocytes, which were cocultured with SD rat Sertoli cells by FACS. The expression of Fas ligand (Fasl), transforming growth factor (TGF)-beta1 and clusterin on Sertoli cells were detected by immunocytochemistry. RESULTS: In the cocultured system, Sertoli cells accounted for more than 93%. With our isolation method, the viability of Sertoli cells was more than 95% and the apoptosis rate was 10.87% +/- 3.87%. The lymphocyte apoptosis ratio was 15.52 +/- 0.17 (P < .01, compared with the control groups). SABC immunochemistry staining showed that the sertoli cells could express FasL, TGF-beta, and clusterin. CONCLUSIONS: In our coculture in vitro, rat Sertoli cells expressed FasL and TGF-beta1 as well as induced the apoptosis of mouse lymphocytes. These results indicated that the expression of FasL and TGF-beta1 on Sertoli cells might relate to immune privilege in xenotransplantation.

Iridoid compounds from Boschniakia rossica.

Four iridoid compounds were isolated from methanol extract of Boschniakia rossica by repeated column chromatography. Their structures were determined as boschnaloside (1), boschnarol (2), bosnarol methylether (3), and 7-deoxy 8-epiloganic acid (4), respectively. Compound 2, 3, and 4 were isolated for the first time from this plant.

[Inhibitory effect of methanol extract of Boschniakia rossica Fedtsch. et Flerov on rat hepatic preneoplastic lesions induced by diethylnitrosamine].

OBJECTIVE: To investigate the inhibitory effect of Boschniakia rossica(BR) on hepatocarcinogenesis in rats. METHOD: Based on immunohistochemistry techniques, the expression of placental form glutathione S-transferase(GST-P), mutant p53 and p21 protein were investigated in hepatic preneoplastic lesions induced by Solt-Farber protocol in the liver of rats that had been treated with the above method, administered with BR extract and of control group. RESULT: The extract of BR(500 mg/kg) has inhibitory effect on the formation of diethylnitrosamine-induced GST-P-positive foci in F344 rat and the expression of mutant p53 and p21 protein was lower than that of hepatic preneoplastic lesions, and the increasing gamma-glutamyltranspeptidase(gamma-GT) activity in rat liver treated with Solt-Farber protocol was decreased by the extract of BR. CONCLUSION: These results indicate that BR has inhibitory effect on DEN induced hepatic preneoplastic lesions in F344 rat.

Effect of 3,6-dimethamidodibenzopyriodonium citrate on Ca2+ in rabbit platelet in vitro.

AIM: To study the effect of 3,6-dimethamidodibenzopyriodonium citrate (I-65) on the cytoplasmic free Ca2+ ([Ca2+]i) concentration in rabbit platelet. METHODS: Measurement of the cytosolic Ca2+ of platelets in vitro by using Quin 2-AM fluorescence technique. RESULTS: In the presence of CaCl2 1 mmol.L-1, I-65 (10, 20, and 30 mumol.L-1) reduced the rise in [Ca2+]i induced by thrombin and calcimycin from 142 +/- 22 nmol.L-1 and 124 +/- 18 nmol.L-1 to 118 +/- 20, 78 +/- 12, 40 +/- 10 nmol.L-1, respectively and 108 +/- 15, 77 +/- 14, 37 +/- 14 nmol.L-1, respectively. In the presence of egtazic acid 2 mmol.L-1, I-65 (10, 20, and 30 mumol.L-1), reduced the Ca2+ release induced by thrombin from 52 +/- 11 nmol.L-1 to 34 +/- 9, 19 +/- 6, and 11 +/- 5 nmol.L-1, respectively. In addition, I-65 (10, 20, and 30 mumol.L-1) also reduced the Ca2+ influx induced by thrombin from 91 +/- 13 nmol.L-1 to 84 +/- 15, 58 +/- 15, and 28 +/- 19 nmol.L-1, respectively. CONCLUSION: I-65 inhibited not only the Ca2+ release, but also the influx of Ca2+ in activation platelet.

[Value of immunohistochemical investigation of anti-GST-pi antibody in the early diagnosis of gastric carcinoma and precancerous lesion].

Human placental form of glutathione S-transferase (GST-pi) was detected in 38 human gastric carcinoma and 28 precancerous lesions by Avidin biotin peroxidase complex (ABC) method using anti-GST-pi antibody. Of 22 normal gastric mucosa, 91% were negative for GST-pi stain, 95% of 21 well and moderately differentiated gastric adenocarcinomas, all of 5 poorly differentiated gastric adenocarcinomas, 75% of undifferentiated gastric carcinomas, 85% of intestinal metaplasia and 100% of atypical hyperplasia were positive for GST-pi stain. These results indicate that GST-pi is a new sensitive marker for immunohistochemical detection of gastric carcinoma and precancerous lesion.