Short-Chain Carboxylates Facilitate the Counting of Yeasts in Sub-High Temperature Daqu.

Sub-high temperature Daqu, a traditional solid fermenting agent used in Chinese strong-aroma Baijiu production, is abundant in diverse microorganisms, including bacteria, yeasts, molds, and actinomycetes. Among these, yeasts are pivotal for ethanol production and flavor formation. However, counting yeasts in Daqu is challenging due to interference from molds and bacteria. Antibiotics are employed to inhibit bacterial growth, but there is no practical way to suppress molds without affecting the growth of yeasts. In this study, short-chain carboxylates (C1-C6) were added to the culture medium at various pH conditions to investigate their effects on the growth of molds and yeasts. The results demonstrated distinct inhibitory effects of the short-chain carboxylates, depending on both pH and concentration. Several tested short-chain carboxylates effectively suppressed mold growth on agar plates while leaving yeast growth unaffected. This suggests a simple and feasible method for enhancing the efficiency of yeast isolation and counting in Daqu. Such an approach is valuable for studying yeasts in diverse and complex habitats.

Encoding Molecular Docking for Quantum Computers.

Molecular docking is important in drug discovery but is burdensome for classical computers. Here, we introduce Grid Point Matching (GPM) and Feature Atom Matching (FAM) to accelerate pose sampling in molecular docking by encoding the problem into quadratic unconstrained binary optimization (QUBO) models so that it could be solved by quantum computers like the coherent Ising machine (CIM). As a result, GPM shows a sampling power close to that of Glide SP, a method performing an extensive search. Moreover, it is estimated to be 1000 times faster on the CIM than on classical computers. Our methods could boost virtual drug screening of small molecules and peptides in future.

A fusion of VGG-16 and ViT models for improving bone tumor classification in computed tomography.

Background and Objective: Bone tumors present significant challenges in orthopedic medicine due to variations in clinical treatment approaches for different tumor types, which includes benign, malignant, and intermediate cases. Convolutional Neural Networks (CNNs) have emerged as prominent models for tumor classification. However, their limited perception ability hinders the acquisition of global structural information, potentially affecting classification accuracy. To address this limitation, we propose an optimized deep learning algorithm for precise classification of diverse bone tumors. Materials and Methods: Our dataset comprises 786 computed tomography (CT) images of bone tumors, featuring sections from two distinct bone species, namely the tibia and femur. Sourced from The Second Affiliated Hospital of Fujian Medical University, the dataset was meticulously preprocessed with noise reduction techniques. We introduce a novel fusion model, VGG16-ViT, leveraging the advantages of the VGG-16 network and the Vision Transformer (ViT) model. Specifically, we select 27 features from the third layer of VGG-16 and input them into the Vision Transformer encoder for comprehensive training. Furthermore, we evaluate the impact of secondary migration using CT images from Xiangya Hospital for validation. Results: The proposed fusion model demonstrates notable improvements in classification performance. It effectively reduces the training time while achieving an impressive classification accuracy rate of 97.6%, marking a significant enhancement of 8% in sensitivity and specificity optimization. Furthermore, the investigation into secondary migration's effects on experimental outcomes across the three models reveals its potential to enhance system performance. Conclusion: Our novel VGG-16 and Vision Transformer joint network exhibits robust classification performance on bone tumor datasets. The integration of these models enables precise and efficient classification, accommodating the diverse characteristics of different bone tumor types. This advancement holds great significance for the early detection and prognosis of bone tumor patients in the future.

The microRNA-4766/VEGFA axis mediates macrophage M2-type polarization to inhibit colorectal cancer proliferation and migration.

OBJECTIVE: To investigate the miR-4766/VEGFA axis in regulating M2-type macrophage polarization under hypoxia and its effect on the proliferation and migration of colorectal cancer (CRC) cells. METHODS: The differentially expressed genes (DEGs) in macrophages before and after hypoxia treatment in the dataset GSE154427 were analyzed. microRNA (miR)-4766 and VEGFA were selected as the research objects and then detected for mRNA expression and protein level using qRT-PCR and Western blot, respectively. The expression levels of M2 macrophage markers such as CD206, CD163, and ARG1 were detected to determine the M2-type macrophage polarization. The targeted binding of miR-4766 to VEGFA was verified using Dual-luciferase reporter gene assay. CCK-8 and Transwell assays were performed, respectively, to detect the capacity of cells to proliferate and migrate. IL-10 and TGF-ß levels in the conditioned medium were detected using ELISA. RESULTS: miR-4766 was upregulated, and VEGFA was downregulated in hypoxia-treated macrophages. miR-4766 inhibited, while VEGFA promoted the polarization of M2-type macrophages. miR-4766 targeted and negatively regulated VEGFA. miR-4766 inhibited the polarization of M2-type macrophages and then suppressed CRC cell proliferation and migration via targeting VEGFA. CONCLUSION: Restoring miR-4766 expression to inhibit VEGFA expression promised to be a potential strategy to suppress CRC development.

Brain image segmentation of the corpus callosum by combining Bi-Directional Convolutional LSTM and U-Net using multi-slice CT and MRI.

BACKGROUND AND OBJECTIVE: Traditional disease diagnosis is usually performed by experienced physicians, but misdiagnosis or missed diagnosis still exists. Exploring the relationship between changes in the corpus callosum and multiple brain infarcts requires extracting corpus callosum features from brain image data, which requires addressing three key issues. (1) automation, (2) completeness, and (3) accuracy. Residual learning can facilitate network training, Bi-Directional Convolutional LSTM (BDC-LSTM) can exploit interlayer spatial dependencies, and HDC can expand the receptive domain without losing resolution. METHODS: In this paper, we propose a segmentation method by combining BDC-LSTM and U-Net to segment the corpus callosum from multiple angles of brain images based on computed tomography (CT) and magnetic resonance imaging (MRI) in which two types of sequence, namely T2-weighted imaging as well as the Fluid Attenuated Inversion Recovery (Flair), were utilized. The two-dimensional slice sequences are segmented in the cross-sectional plane, and the segmentation results are combined to obtain the final results. Encoding, BDC- LSTM, and decoding include convolutional neural networks. The coding part uses asymmetric convolutional layers of different sizes and dilated convolutions to get multi-slice information and extend the convolutional layers' perceptual field. RESULTS: This paper uses BDC-LSTM between the encoding and decoding parts of the algorithm. On the image segmentation of the brain in multiple cerebral infarcts dataset, accuracy rates of 0.876, 0.881, 0.887, and 0.912 were attained for the intersection of union (IOU), dice similarity coefficient (DS), sensitivity (SE), and predictive positivity value (PPV). The experimental findings demonstrate that the algorithm outperforms its rivals in accuracy. CONCLUSION: This paper obtained segmentation results for three images using three models, ConvLSTM, Pyramid-LSTM, and BDC-LSTM, and compared them to verify that BDC-LSTM is the best method to perform the segmentation task for faster and more accurate detection of 3D medical images. We improve the convolutional neural network segmentation method to obtain medical images with high segmentation accuracy by solving the over-segmentation problem.

Lipofectamine 2000™ at transfection dose promotes EphA2 transcription in an HDAC4-dependent manner to reduce its cytotoxicity.

The cationic liposome is well-known as an efficient nucleic acid delivery tool; however, the stress responses induced by liposome per se have been rarely revealed. In this study, we found that Lipofectamine™ 2000 (lipo2000), a commonly used commercial cationic liposome transfection, could upregulate EphA2 mRNA expression in multiple cells at transfection dose. Furthermore, lipo2000 treatment could increase the level of EphA2 hnRNA (heterogeneous nuclear RNA). Lipo2000-induced EphA2 upregulation could be depleted upon global transcription inhibition, proving that lipo2000 upregulates EphA2 expression via activating its transcription. Moreover, HDAC4 depletion, a known EphA2 trans-acting regulatory factor, could eliminate the lipo2000-induced EphA2 upregulation, demonstrating that lipo2000 promotes EphA2 transcription in an HDAC4 dependent manner. Functionally, EphA2 knockdown did not affect GFP expression level and the interfering efficacy of siGAPDH, suggesting that EphA2 is unrelated to the nucleic acid delivery capacity of lipo2000. Nevertheless, EphA2 depletion significantly activated autophagy and apoptosis, increasing the cytotoxic effects of lipo2000, which could be rescued by EphA2 restoration, indicating that EphA2 is essential to overcome liposome-related cytotoxicity. Finally, we found that lipo2000 could activate EphA2 transcription in an HDAC4-dependent manner. EphA2 is not associated with the transfection efficiency of lipo2000, but it is vital to reduce lipo2000 cytotoxicity, suggesting that when conducting liposome-mediated gene function studies, especially for EphA2, the stress response of liposomes should be considered to obtain objective results.

Genetic Engineering Production of Ethyl Carbamate Hydrolase and Its Application in Degrading Ethyl Carbamate in Chinese Liquor.

Ethyl carbamate (EC), classified as a Group 2A carcinogen, is most abundant in the fermented foods, such as Cachaca, Shaoxing wine, and Chinese liquor (baijiu). Although biodegradation can reduce its concentration, a high ethanol concentration and acidic environment often limit its degradation. In the present study, a novel ethyl carbamate hydrolase (ECH) with high specificity to EC was isolated from Acinetobacter calcoaceticus, and its enzymatic properties and EC degradability were investigated. ECH was immobilized to resist extreme environmental conditions, and the flavor substance changes were explored by gas chromatography-mass spectrometry (GC/MS). The specific enzymatic activity of ECH was 68.31 U/mg. Notably, ECH exhibited excellent thermal stability and tolerance to sodium chloride and high ethanol concentration (remaining at 40% activity in 60% (v/v) ethanol, 1 h). The treatment of immobilized ECH for 12 h decreased the EC concentration in liquor by 71.6 µg/L. Furthermore, the immobilized ECH exerted less effect on its activity and on the flavor substances, which could be easily filtrated during industrial production.

Effects of beer, wine, and baijiu consumption on non-alcoholic fatty liver disease: Potential implications of the flavor compounds in the alcoholic beverages.

Non-alcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease and its global incidence is estimated to be 24%. Beer, wine, and Chinese baijiu have been consumed worldwide including by the NAFLD population. A better understanding of the effects of these alcoholic beverages on NAFLD would potentially improve management of patients with NAFLD and reduce the risks for progression to fibrosis, cirrhosis, and hepatocellular carcinoma. There is evidence suggesting some positive effects, such as the antioxidative effects of bioactive flavor compounds in beer, wine, and baijiu. These effects could potentially counteract the oxidative stress caused by the metabolism of ethanol contained in the beverages. In the current review, the aim is to evaluate and discuss the current human-based and laboratory-based study evidence of effects on hepatic lipid metabolism and NAFLD from ingested ethanol, the polyphenols in beer and wine, and the bioactive flavor compounds in baijiu, and their potential mechanism. It is concluded that for the potential beneficial effects of wine and beer on NAFLD, inconsistence and contrasting data exist suggesting the need for further studies. There is insufficient baijiu specific human-based study for the effects on NAFLD. Although laboratory-based studies on baijiu showed the antioxidative effects of the bioactive flavor compounds on the liver, it remains elusive whether the antioxidative effect from the relatively low abundance of the bioactivate compounds could outweigh the oxidative stress and toxic effects from the ethanol component of the beverages.

High-efficiency recovery, regeneration and recycling of 1-ethyl-3-methylimidazolium hydrogen sulfate for levulinic acid production from sugarcane bagasse with membrane-based techniques.

Ionic liquids have been proven efficient and environmental medium for producing platform chemical levulinic acid. Lack of high-efficiency, stable and low-cost recovery strategy with complex electrolyte form restricts the further scale-up of ionic liquids for platform chemicals production. Membrane-based techniques including ultrafiltration (UF) and bipolar membrane electrodialysis (BMED) were employed for the high-efficiency recovery, regeneration and recycling of 1-ethyl-3-methylimidazolium hydrogen sulfate [Emim][HSO4] for levulinic acid production from sugarcane bagasse. UF-BMED treatment works based on the interception of macromolecule biomass degradation products by UF treatment with regional recovery of Emim+ and SO42- by BMED treatment. Effect of major parameters on [Emim][HSO4] recovery performance was determined. Recovery ratio for Emim+ and SO42- approached 95.4% and 95.9%. Energy consumption of specific [Emim][HSO4] recovery was closed to 5.8 kWh/kg. Insight gained from this study suggests a high-efficiency and economical strategy for platform chemicals production with green solvent ionic liquids.

Integrated analysis of whole genome and transcriptome sequencing in a young patient with gastric cancer provides insights for precision therapy.

Gastric cancer is a leading cause of cancer-associated deaths worldwide and is considered to be an age-related disease. In younger patients, gastric cancer is biologically more aggressive, and prognosis is worse compared with that in elderly patients. In the present case report, the whole genome and transcriptome was sequenced in a 26-year-old patient with gastric cancer who presented with gastric cancer-related symptoms and was admitted to the First Affiliated Anhui Medical Hospital (Hefei, China) in December 2016. In total, 9 germline and 4 somatic mutations were identified in the patient, and there were more deleterious sites in the germline mutated genes. Genes with somatic mutations, such as MUC2, MUC4, SLC8A2, and with structural variations, including CCND3, FGFR2 and FGFR3, were found to be differentially expressed. Cancer-associated pathways, such as the 'calcium signaling pathway', 'cGMP-PKG signaling pathway' and 'transcriptional mis-regulation' were also enriched at both the genomic and transcriptomic levels. The genes found to have germline (SFRP4), somatic (MUC2, MUC4, SLC8A2) mutations, or structural variations (CCND3, FGFR2 and FGFR3) were differentially expressed in the patient and could be promising precision therapy targets.

The circular RNA 001971/miR-29c-3p axis modulates colorectal cancer growth, metastasis, and angiogenesis through VEGFA.

BACKGROUND: Colorectal cancer (CRC) is one of the most common malignant tumors globally. Angiogenesis is a key event maintaining tumor cell survival and aggressiveness. The expression of vascular endothelial growth factor A (VEGFA), one of the most significant tumor cell-secreted proangiogenic factors, is frequently upregulated in CRC. METHODS: The MTT assay was used to detect the viability of CRC cells. Transwell assays were performed to detect the invasion capacity of target cells. Relative protein levels were determined by immunoblotting. Pathological characteristics of tissues were detected by H&E staining and immunohistochemical (IHC) staining. A RIP assay was conducted to validate the predicted binding between genes. RESULTS: We observed that circ-001971 expression was dramatically increased in CRC tissue samples and cells. Circ-001971 knockdown suppressed the capacity of CRC cells to proliferate and invade and HUVEC tube formation in vitro, as well as tumor growth in mice bearing SW620 cell-derived tumors in vivo. The expression of circ-001971 and VEGFA was dramatically increased whereas the expression of miR-29c-3p was reduced in tumor tissue samples. Circ-001971 relieved miR-29c-3p-induced inhibition of VEGFA by acting as a ceRNA, thereby aggravating the proliferation, invasion and angiogenesis of CRC. Consistent with the above findings, the expression of VEGFA was increased, whereas the expression of miR-29c-3p was decreased in tumor tissue samples. miR-29c-3p had a negative correlation with both circ-001971 and VEGFA, while circ-001971 was positively correlated with VEGFA. CONCLUSIONS: In conclusion, the circ-001971/miR-29c-3p axis modulated CRC cell proliferation, invasion, and angiogenesis by targeting VEGFA.

Characterization of Microbial Diversity and Community Structure in Fermentation Pit Mud of Different Ages for Production of Strong-Aroma Baijiu.

In the traditional fermentation process of strong-aroma Baijiu, a fermentation pit mud (FPM) provides many genera of microorganisms for fermentation. However, the functional microorganisms that have an important effect on the quality of Baijiu and their changes with the age of fermentation pit (FP) are poorly understood. Herein, the Roche 454 pyrosequencing technique and a phospholipid fatty-acid analysis were employed to reveal the structure and diversity of prokaryotic communities in FPM samples that have been aged for 5, 30, and 100 years. The results revealed an increase in total prokaryotic biomass with an FP age; however, Shannon's diversity index decreased significantly (p < 0.01). These results suggested that a unique microbial community structure evolved with uninterrupted use of the FP. The number of functional microorganisms, which could produce the flavor compounds of strong-aroma Baijiu, increased with the FP age. Among them, Clostridium and Ruminococcaceae are microorganisms that directly produce caproic acid. The increase of their relative abundance in the FPM might have improved the quality of strong-aroma Baijiu. Syntrophomonas, Methanobacterium, and Methanocorpusculum might also be beneficial to caproic acid production. They are not directly involved but provide possible environmental factors for caproic acid production. Overall, our study results indicated that an uninterrupted use of the FP shapes the particular microbial community structure in the FPM. This research provides scientific support for the concept that the aged FP yields a high-quality Baijiu.In the traditional fermentation process of strong-aroma Baijiu, a fermentation pit mud (FPM) provides many genera of microorganisms for fermentation. However, the functional microorganisms that have an important effect on the quality of Baijiu and their changes with the age of fermentation pit (FP) are poorly understood. Herein, the Roche 454 pyrosequencing technique and a phospholipid fatty-acid analysis were employed to reveal the structure and diversity of prokaryotic communities in FPM samples that have been aged for 5, 30, and 100 years. The results revealed an increase in total prokaryotic biomass with an FP age; however, Shannon's diversity index decreased significantly (p < 0.01). These results suggested that a unique microbial community structure evolved with uninterrupted use of the FP. The number of functional microorganisms, which could produce the flavor compounds of strong-aroma Baijiu, increased with the FP age. Among them, Clostridium and Ruminococcaceae are microorganisms that directly produce caproic acid. The increase of their relative abundance in the FPM might have improved the quality of strong-aroma Baijiu. Syntrophomonas, Methanobacterium, and Methanocorpusculum might also be beneficial to caproic acid production. They are not directly involved but provide possible environmental factors for caproic acid production. Overall, our study results indicated that an uninterrupted use of the FP shapes the particular microbial community structure in the FPM. This research provides scientific support for the concept that the aged FP yields a high-quality Baijiu.

Changes in the gut microbiota during and after commercial helium-oxygen saturation diving in China.

OBJECTIVES: The influence of commercial helium-oxygen saturation diving on divers' gut microbiotas was assessed to provide dietary suggestion. METHODS: Faecal samples of 47 divers working offshore were collected before (T1), during (T2) and after (T3) saturation diving. Their living and excursion depths were 55-134 metres underwater with a saturation duration of 12-31 days and PaO2 of 38-65 kPa. The faecal samples were examined through 16S ribosomal DNA amplicon sequencing based on the Illumina sequencing platform to analyse changes in the bacteria composition in the divers' guts. RESULTS: Although the α and ß diversity of the gut microbiota did not change significantly, we found that living in a hyperbaric environment of helium-oxygen saturation decreased the abundance of the genus Bifidobacterium, an obligate anaerobe, from 2.43%±3.83% at T1 to 0.79%±1.23% at T2 and 0.59%±0.79% at T3. Additionally, the abundance of some short-chain fatty acid (SCFA)-producing bacteria, such as Fusicatenibacter, Faecalibacterium, rectale group and Anaerostipes, showed a decreased trend in the order of before, during and after diving. On the contrary, the abundance of species, such as Lactococcus garvieae, Actinomyces odontolyticus, Peptoclostridium difficile, Butyricimonas virosa, Streptococcus mutans, Porphyromonas asaccharolytica and A. graevenitzii, showed an increasing trend, but most of them were pathogens. CONCLUSIONS: Occupational exposure to high pressure in a helium-oxygen saturation environment decreased the abundance of Bifidobacterium and some SCFA-producing bacteria, and increased the risk of pathogenic bacterial infection. Supplementation of the diver diet with probiotics or prebiotics during saturation diving might prevent these undesirable changes.

Clinicopathological risk factors for gastric cancer: a retrospective cohort study in China.

OBJECTIVE: To examine the potential clinicopathological factors affecting the prognosis of patients with gastric cancer after surgical treatment in China. METHODS: Between 1 January 2001 and 31 December 2012, a total of 716 patients aged 22-84 years with gastric cancer were enrolled in the study. Survival analysis techniques including log rank test and Cox proportional hazard regression model were applied to evaluate the prognostic significance of clinicopathological characteristics in terms of survival time. RESULTS: Of the 24 demographic and pathological variables collected in the data, 16 prognostic factors of gastric cancer were found to have statistically significant influences on survival time from the unadjusted analyses. The adjusted analysis furtherly revealed that age, age square, lymph node metastasis rate group, tumour size group, surgical type II, number of cancer nodules, invasion depth group and the interaction between surgical type II and tumour size group were important prognosis and clinicopathological factors for gastric cancer in Chinese. CONCLUSION: Our study with relatively large sample size and many potential risk factors enable us to identify independent risk factors associated with the prognosis of gastric cancer. Findings from the current study can be used to assist clinical decision-making, and serve as a benchmark for the planning of future prognosis and therapy for patients with gastric carcinoma.

Long Non-Coding RNA MALAT1 Interacts With miR-204 to Modulate Human Hilar Cholangiocarcinoma Proliferation, Migration, and Invasion by Targeting CXCR4.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is involved in the development and progression of many types of tumors. An aberrant expression of MALAT1 was observed in many kinds of cancers. However, the exact effects and molecular mechanisms of MALAT1 in human hilar cholangiocarcinoma (HCCA) progression are still unknown. Here, we investigated the role of MALAT1 in human HCCA cell lines and clinical tumor samples in order to determine the function of this lncRNA. In our research, lncRNA-MALAT1 was specifically upregulated in HCCA tissues and cell lines, and was associated with pathological T stage, a larger tumor size, and perineural invasion. Knockdown of MALAT1 inhibited the proliferation, migration, and invasion of human HCCA cell. In addition, chemokine receptor-4 (CXCR4) was involved in MALAT1 induced human HCCA growth, migration, and invasion. By using online tools and a series of mechanistic analysis, we also demonstrated that miR-204-dependent CXCR4 regulation was required in MALAT1 modulating HCCA cell growth, migration and invasion. Taken together, our data indicated that MALAT1 might play an oncogenic role in HCCA through miR-204-dependent CXCR4 regulation, and could be regarded as a therapeutic target in HCCA. J. Cell. Biochem. 118: 3643-3653, 2017. © 2017 Wiley Periodicals, Inc.

Knockdown of Hypoxia-Inducible Factor 1α Improved the Efficacy of Low-Dose Metronomic Chemotherapy of Paclitaxel in Human Colon Cancer Xenografts.

Low-dose metronomic chemotherapy represents a new strategy for solid tumor treatments with a strong antiangiogenic activity and few side effects. However, low-dose metronomic therapy alone is not always as effective as traditional chemotherapy on eradication of tumor. On the contrary, low-dose metronomic in some cases could stimulate tumor growth due to hypoxia of tumor cells induced during therapy. Our study aimed to investigate whether knockdown of hypoxia-inducible factor-1α expression in tumor cell could facilitate low-dose metronomic therapy with paclitaxel for human colon cancer. Human colon cancer cell line (HT-29) stably transfected with specific short hairpin RNAs silencing hypoxia-inducible factor-1α exhibited marked attenuation of hypoxia-induced expression of the target genes such as vascular endothelial growth factor, glucose transporter 1, and P-glycoprotein. Compared with HT-29-c xenograft tumor model established by subcutaneous injection of HT-29 cells stably transfected with scrambled control short hairpin RNA, HT-29-ih xenograft tumor model showed more significant and long-lasting antitumor responses of empirical metronomic paclitaxel regimens, accompanied by drastic angiogenesis decrease and neglectable toxicity. All these data indicated that the combination of paclitaxel low-dose metronomic therapy with hypoxia-inducible factor-1α knockdown might provide a potent battle against colon cancer.

Developmental changes of the FAS and HSL mRNA expression and their effects on the content of intramuscular fat in Kazak and Xinjiang sheep.

Twenty-four male Kazak sheep and 30 Xinjiang fine wool sheep at different ages were selected to investigate the development-dependent expression levels of fatty acid synthase (FAS) gene and hormone-sensitive lipase (HSL) gene in muscle and their effects on the contents of intramuscular fat (IMF). Longissimus dorsal muscle was sampled to measure IMF and total RNA was extracted to determine FAS and HSL mRNA expression levels by real-time PCR. The results showed that: 1) The IMF content increased continuously with growth and showed significant differences (P < 0.05) between different age groups in male Kazak sheep, but in Xinjiang fine wool sheep there was no such difference observed. Furthermore, the IMF contents in Kazak were much higher (P < 0.01) than that of the other breed from day 30 to 90. 2) FAS mRNA expression level was the highest (P < 0.05) on day 0 in Kazak sheep and then declined with growth, in the other breed the gene showed a 'decline-rise-decline-rise' expression manner as the animals grew. HSL mRNA expression level had a similar model in two breeds, in Kazak sheep it was the highest on day 0 (P < 0.05) and in Xinjiang fine wool sheep on day 30 (P < 0.01), then both decreased after this term. 3) In male Kazak sheep, FAS and HSL mRNA expression level were both negatively related to IMF content (r = -0.485 (P = 0.02), r = -0.423 (P = 0.05)), and the ratio of FAS/HSL expression exhibited significantly negatively related IMF contents. In male Xinjiang sheep, there were no obvious relationship between FAS and HSL expression and IMF content (P > 0.05).

The tissue distribution and developmental changes of ghrelin mRNA expression in sheep.

Male Kazak sheep and Xinjiang fine wool sheep, six for each different age group (days 2, 30, 60, 90 and 120), were used in the present study to investigate the tissue distribution and developmental changes of ghrelin mRNA expression in abomasum; however, there was no 120-day-old Kazak sheep. After measurement of body weight, the tissues such as hypothalamus, pituitary, heart, liver, rumen, reticulum, omasum, abomasum, duodenum, and longissimus dorsi muscle were sampled. And the total RNA of different tissues was extracted to determine the abundance of ghrelin mRNA by RT-PCR and real-time PCR. The results showed that (1) for both breeds, body weight among different ages was significantly different (P<0.05). And from day 30 to 90, the body weight of Kazak was significantly higher than that of Xinjiang (P<0.01); (2) Ghrelin mRNA existed in all the above tissues and was significantly higher in the abomasum than in other tissues (P<0.05); (3) the temporal patterns of abomasum ghrelin mRNA expression in Kazak and Xinjiang were similar. From day 2 to 60 in Kazak and 2 to 90 in Xinjiang, there was a steady increase in the ghrelin mRNA level. By day 60 in Kazak and day 90 in Xinjiang, the level reached a plateau and remained steady. These results also demonstrated that from birth to day 90, ghrelin mRNA level was significantly higher in Kazak than in Xinjiang (P<0.01).