A more accurate relationship between serum androgen and metabolism among healthy, nonobese, reproductive-age women based on liquid chromatography-tandem mass spectrometry.

This study explored a more precise association between androgens and glycolipid metabolism in healthy women of different ages. Body mass index (BMI), waist circumference (WC), and waist-to-hip ratio were used as body fat indicators. High-density lipoprotein (HDL), low-density lipoprotein, triglycerides, and total cholesterol were used as lipid markers. Fasting blood glucose (FBG), fasting insulin, and the homeostatic model assessment of insulin resistance were used to assess insulin resistance and glucose metabolism. Liquid chromatography-tandem mass spectrometry was used to measure androgen indicators, including testosterone, sex hormone-binding globulin (SHBG), free testosterone (FT), dihydrotestosterone (DHT), androstenedione (A4), dehydroepiandrosterone (DHEA), and dehydroepiandrosterone sulfate (DHEAS). DHEAS levels varied across age groups. Correlation analyses with Spearman's coefficient showed that the free androgen index correlated positively with WC (p = 0.040), FT correlated positively with BMI (p = 0.033) and WC (p = 0.049), SHBG correlated positively with HDL (p = 0.013), and A4 correlated positively with FBG (p = 0.017). Multiple linear regression analysis showed that among healthful women aged 36-40 years, A4 increased with FBG, and SHBG increased with HDL. Even within healthy, nonobese women, lipid and glucose metabolism were robustly correlated with androgens. Yearly metabolic assessments are necessary, particularly for FBG and HDL, since these markers can predict the likelihood of hyperandrogenemia, enabling timely interventions.

A missed diagnosis of sputum crust with fiberoptic bronchoscope causing extubation failure: a case report.

BACKGROUND: Fiberoptic bronchoscopy (FOB) and bronchoscopic biopsy are the established methods for diagnosing and treating sputum crust. However, sputum crust in concealed locations can sometimes be missed or undiagnosed, even with bronchoscopy. CASE PRESENTATION: We present the case of a 44-year-old female patient who experienced initial extubation failure and postoperative pulmonary complications (PPCs) due to the missed diagnosis of sputum crust by FOB and low-resolution bedside chest X-ray. The FOB examination showed no apparent abnormalities prior to the first extubation, and the patient underwent tracheal extubation 2 h after aortic valve replacement (AVR). However, she was reintubated 13 h after the first extubation due to a persistent irritating cough and severe hypoxemia, and a bedside chest radiograph revealed pneumonia and atelectasis. Upon performing a repeat FOB examination prior to the second extubation, we serendipitously discovered the presence of sputum crust at the end of the endotracheal tube. Subsequently, we found that the sputum crust was mainly located on the tracheal wall between the subglottis and the end of the endotracheal tube during the "Tracheobronchial Sputum Crust Removal" procedure, and most of the crust was obscured by the retained endotracheal tube. The patient was discharged on the 20th day following therapeutic FOB. CONCLUSION: FOB examination may miss specific areas in endotracheal intubation (ETI) patients, particularly the tracheal wall between the subglottis and distal end of the tracheal catheter, where sputum crust can be concealed. When diagnostic examinations with FOB are inconclusive, high-resolution chest CT can be helpful in identifying hidden sputum crust.

Effect of lidocaine perioperative infusion on chronic postsurgical pain in patients undergoing thoracoscopic radical pneumonectomy.

BACKGROUND: Thoracoscopic radical pneumonectomy is associated with a high incidence of postoperative chronic pain. Studies on the benefits of lidocaine intravenous infusion during the perioperative period were still controversial in thoracoscopic surgery. METHODS: Sixty-four lung cancer patients scheduled for thoracoscopic radical pneumonectomy were randomly divided into two groups: normal saline group (control group) or lidocaine group. In the lidocaine group, 1.5 mg/kg lidocaine was administered during the anesthesia induction, and 2 mg·kg-1·h-1 lidocaine was continuously intravenous infused until the end of the surgery. After the surgery, a mixture of 2 µg/kg sufentanil and 10 mg/kg lidocaine was continuously intravenous infused by postoperative patient-controlled intravenous analgesia pump (100 ml). In the control group, the same volume of normal saline was administered according to the calculation of lidocaine during anesthesia induction, maintenance and postoperative patient-controlled intravenous analgesia. The primary outcome was the incidence of chronic postoperative pain at 3 months after the surgery. The secondary outcomes include the incidence of chronic postoperative pain at 6 months after the surgery; the effect of lidocaine on postoperative pain within the first 24 and 48 h; total amount of sufentanil administered during entire procedure and the number of PCA triggers within 48 h after surgery. RESULTS: Compared with the control group, the incidence of chronic pain at 3 months after the surgery was significantly lower (13 cases, 46.4% vs. 6 cases, 20.7%, p < 0.05), but no significant difference at 6 months between two group. The cumulative dosage of sufentanil in perioperative period was significantly lower (149.64 ± 18.20 µg vs. 139.47 ± 16.75 µg) (p < 0.05), and the number of PCA triggers (8.21 ± 4.37 vs. 5.83 ± 4.12, p < 0.05) was significantly greater in the control group. The NRS pain scores at 24 h (1.68 ± 0.72 vs. 1.90 ± 0.86) and 48 h (1.21 ± 0.42 vs. 1.20 ± 0.41) after the operation were no significant difference. CONCLUSION: Perioperative infusion lidocaine significantly reduced the number of PCA triggers and the incidence of chronic postoperative pain at 3 months after the thoracoscopic radical pneumonectomy. TRIAL REGISTRATION: http://www.chictr.org.cn : ChiCTR1900024759, frist registration date 26/07/2019.

A combined in vitro and in silico study of the inhibitory mechanism of angiotensin-converting enzyme with peanut peptides.

Food-derived peptides with low molecular weight, high bioavailability, and good absorptivity have been exploited as angiotensin-converting enzyme (ACE) inhibitors. In the present study, in-vitro inhibition kinetics of peanut peptides, in silico screening, validation of ACE inhibitory activity, molecular dynamics (MD) simulations, and HUVEC cells were performed to systematically identify the inhibitory mechanism of ACE interacting with peanut peptides. The results indicate that FPHPP, FPHY, and FPHFD peptides have good thermal, pH, and digestive stability. MD trajectories elucidate the dynamic correlation between peptides and ACE and verify the specific binding interaction. Noteworthily, FPHPP is the best inhibitor with a strongest binding affinity and significantly increases NO, SOD production, and AT2R expression, and decreases ROS, MDA, ET-1 levels, ACE, and AT1R accumulation in Ang II-injury HUVEC cells.

Application and discoveries of metabolomics and proteomics in the study of female infertility.

Introduction: Female infertility is defined as the absence of clinical pregnancy after 12 months of regular unprotected sexual intercourse. Methods: This study employed metabolomics and proteomics approaches to investigate the relationship between metabolites and proteins and female infertility. The study used metabolomics and proteomics data from the UK Biobank to identify metabolites and proteins linked to infertility. Results: The results showed that GRAM domain-containing protein 1C and metabolites fibrinogen cleavage peptides ADpSGEGDFXAEGGGVR and 3-Hydroxybutyrate had a positive correlation with infertility, whereas proteins such as Interleukin-3 receptor subunit alpha, Thrombospondin type-1 domain-containing protein 1, Intestinal-type alkaline phosphatase, and platelet and endothelial cell adhesion molecule 1 exhibited a negative correlation. These findings provide new clues and targets for infertility diagnosis and treatment. However, further research is required to validate these results and gain a deeper understanding of the specific roles of these metabolites and proteins in infertility pathogenesis. Discussion: In conclusion, metabolomics and proteomics techniques have significant application value in the study of infertility, allowing for a better understanding of the biological mechanisms underlying infertility and providing new insights and strategies for its diagnosis and treatment. These research findings provide a crucial biological mechanistic basis for early infertility screening, prevention, and treatment.

Postoperative jaundice related to UGT1A1 and ABCB11 gene mutations: A case report and literature review.

BACKGROUND: Patients with obstructive jaundice caused by intrahepatic bile duct stones can be effectively managed by surgery. However, some patients may develop postoperative complications, liver failure, and other life-threatening situations. Here, we report a patient with mutations in the uridine 5'-diphospho-glucuronosyltransferase 1A1 (UGT1A1) and bile salt export pump (adenosine triphosphate-binding cassette subfamily B member 11, ABCB11) genes who presented multiple intrahepatic bile duct stones and cholestasis, and the jaundice of the patient increased after partial hepatectomy. CASE SUMMARY: A 52-year-old male patient admitted to the hospital on October 23, 2021, with a progressive exacerbation of jaundice, was found to have multiple intrahepatic bile duct stones with the diagnoses of obstructive jaundice and acute cholecystitis. Subsequently, the patient underwent left hepatectomy with biliary exploration, stone extraction, T-tube drainage, and cholecystectomy without developing any intraoperative complications. The patient had a dark urine color with worsening jaundice postoperatively and did not respond well to plasma exchange and other symptomatic and supportive treatments. Since the progressive increase in postoperative bilirubin could not be clinically explained with any potential reason, including, if not at all, viral infection, cholangitis, autoimmune liver disease, and other causes, the patient underwent whole-exon screening for any genetic diseases, which surprisingly identified UGT1A1 and ABCB11 gene mutations related to glucuronidation of indirect bilirubin as well as bile acid transport in hepatocytes, respectively. Thus, we hypothesized that postoperative refractory cholestasis might result from UGT1A1 and ABCB11 gene mutations and further recommended liver transplantation to the patient, who eventually declined it and died from liver failure six months later. CONCLUSION: Surgery may aggravate cholestasis in patients with multiple intrahepatic bile duct stones and cholestasis associated with UGT1A1 and ABCB11 gene mutations. A liver transplant may be the best option if active medical treatment fails.

Clinical Application of the Classical Theory of Traditional Chinese Medicine in Diabetic Nephropathy.

Objective: To explore the clinical application of the classical theory of traditional Chinese medicine (TCM) in diabetic nephropathy (DN). Methods: A total of 100 patients with DN treated in our hospital from May 2019 to June 2021 were included. The patients were randomly assigned to the control group and the study group. The control group received routine treatment, and the study group was treated with the classical theory of TCM. The efficacy, TCM syndrome score, urine proteinuria (UTP), urine albumin-creatinine ratio (UACR), plasma albumin (ALB), hemoglobin A1c (HbA1C), fasting blood glucose (FBG), blood urea nitrogen (BUN), creatinine (Cr), and treatment safety were compared between the two groups. Results: In comparison to the curative effect, the study group was significantly effective in 34 cases, effective in 12 cases, and ineffective in 4 cases, and the effective rate was 92.00%; the control group was significantly effective in 16 cases, effective in 18 cases, and ineffective in 16 cases, and the effective rate was 68.00%. The effective rate in the study group was higher compared to the control (P < 0.05). In comparison to the TCM syndrome scores, there exhibited no significant difference before treatment (P > 0.05), but after treatment, the TCM syndrome scores of the two groups decreased, and the TCM syndrome scores of the study group were lower compared to the control at 6 weeks, 12 weeks, 24 weeks, and 36 weeks of treatment (P < 0.05). There exhibited no significant difference in the indexes of UTP and UACR before treatment, but the indexes of UTP and UACR in the two groups decreased after treatment, and the indexes of UTP and UACR in the study group were lower compared to the control at 6 and 12 weeks after treatment. There was no significant difference in the indexes of ALB, HbA1C, and FBG before treatment, but after treatment, the indexes of ALB increased, the indexes of HbA1C and FBG decreased in both groups, and the indexes of HbA1C and FBG i4n the study group were lower compared to the control, while the index of ALB in the study group was higher. The indexes of BUN and Cr were compared, and there was no significant difference before treatment, but after treatment, the indexes of BUN and Cr in the two groups decreased, and the indexes of BUN and Cr in the study group were lower compared to the control (P < 0.05). In terms of the treatment safety of the two groups, there was no abnormality in blood, urine, stool routine, and liver and kidney function examination in the study group. No obvious adverse reactions were found in all patients. There were 1 case of abnormal liver function and 2 cases of rash in the control group, and there exhibited no significant difference (P > 0.05). Conclusion: Under the guidance of classical theory, the optimization scheme of comprehensive treatment of TCM may improve renal function by improving metabolic disorders, vascular lesions, neurotrophic disorders, antioxidant stress, and other ways to repair nerve injury, improving the changes of TCM syndromes, signs, and indicators of patients, and delay the progress of DN.

Mechanisms by Which Traditional Chinese Medicines Influence the Intestinal Flora and Intestinal Barrier.

The effect of a drug on the intestinal flora and the intestinal barrier is an important evaluation index for drug safety and efficacy. Chemical synthetic drugs are widely used due to their advantages of fast efficacy and low doses, but they are prone to cause drug resistance and inhibit proton pumps, which may harm intestinal health. Traditional Chinese medicine (TCM) has been applied clinically for thousands of years, and how TCMs regulate intestinal health to achieve their effects of disease treatment has become a hot research topic that needs to be resolved. This paper reviews the recent research on the effects of TCMs on intestinal microorganisms and the intestinal mucosal barrier after entering the intestine, discusses the interaction mechanisms between TCMs and intestinal flora, and details the repair effect of TCMs on the intestinal mucosal barrier to provide a reference for the development, utilization, and modernization of TCM.

Application of traditional Chinese medicine in film drug delivery system.

Film drug delivery systems have the advantages of precise administration, simple process and easy portability, compared with other traditional drug delivery systems such as tablets, capsules, syrups, ointments, etc. The traditional Chinese medicine (TCM) are normally developed in four categories of film agent like patch film, coating, spray film and gel film, which are applied to the treatment of oral ulcers, chronic diseases of lower limbs, burns, scalds, gynecological disease and body care. So the TCM film has great research value and prominent market prospect. In this review, we summarized the research progress of the material composition, pharmaceutical production, clinical application and pharmacology mechanism of various TCM film agents. It may provide a comprehensive reference for further development and utilization of TCM film agents.

Transcriptome and Metabonomics Analysis Revealed the Molecular Mechanism of Differential Metabolite Production of Dendrobium nobile Under Different Epiphytic Patterns.

The cultivation medium of Dendrobium nobile has an effect on the contents of its main medicinal components, but the specific mechanism is still unclear. In this study, the callus, seedlings, rhizomes, and leaves of D. nobile were sequenced for the PacBio SMRT. The 2-year-old stems were selected for the Illumina sequencing and metabolome sequencing to analyze the genetic mechanism of metabolic differences under different epiphytic patterns. As a result, a total of 387 differential genes were obtained, corresponding to 66 differential metabolites. Different epiphytic patterns can induce a series of metabolic changes at the metabolome and transcriptome levels of D. nobile, including flavonoid metabolism, purine metabolism, terpenoid backbone biosynthesis, amino acid metabolism, and alpha-linolenic acid metabolic, and related regulatory genes include ALDH2B7, ADC, EPSPS-1, SHKA, DHAPS-1, GES, ACS1, SAHH, ACS2, CHLP, LOX2, LOX2.3, and CYP74B2. The results showed that the genetic mechanism of D. nobile under various epiphytic patterns was different. In theory, the content of metabolites under the epiphytic patterns of Danxia stone is higher, which is more suitable for field cultivation.

Genome-Wide Identification of Genes Related to Biosynthesis of Phenolic Acid Derivatives in Bletilla striata at Different Suspension Culture Stages.

To screen the genes regulating the biosynthesis of phenolic acid derivatives from the genome of Bletilla striata, we designed a suspension culture system to sample the cells for the following experiments. The contents of four phenolic acid derivatives were determined by high-performance liquid chromatography, and several full-length transcriptome sequencings of RNA samples at 10 time points were performed for bioinformatics analysis. The correlation analysis was used to identify and verify the key DEGs involved in the biosynthesis of the four phenolic acid derivatives. The results showed that the contents of p-hydroxybenzylalcohol (HBA), Dactylorhin A, Militarine, and Coelonin peaked at 33 days postinoculation (Dpi), 18 Dpi, 39 Dpi, and 39 Dpi of the culture system, respectively. Based on transcriptome data, 80 DEGs involved in the biosynthesis of phenolic acid derivatives were obtained. The KEGG pathway enrichment analysis classified them mostly into five metabolic pathways: phenylpropane biosynthesis, starch and sucrose metabolic, cyanoamino acid metabolism, gluconeogenesis and glycolysis, and phenylalanine metabolism. qPCR analysis revealed that the relative gene expression levels were consistent with the overall trend of transcriptome sequencing results. Among them, 14, 18, 23, and 41 unigenes were found to be involved in the synthesis of HBA, Dactylorhin A, Coelonin, and Militarine, respectively. These unigenes laid a solid foundation for elucidating the biosynthesis mechanism of phenolic acid derivatives in suspension cells of B. striata.

Impact of Auricularia cornea var. Li polysaccharides on the physicochemical, textual, flavor, and antioxidant properties of set yogurt.

This study evaluated quality attributes and in vivo antioxidant activity of Auricularia cornea var. Li polysaccharide (ACP)-fortified set yogurt during 21 days of storage (4 °C). Set yogurt was manufactured using a commercial yogurt culture, and 3% (w/v) ACP was added. Physicochemical (pH, titratable acidity, and water-holding capacity), textural, rheological, microstructural, flavor, and antioxidant properties of set yogurt were investigated. The results showed that the addition of ACP significantly enhanced WHC, viscosity, firmness, and cohesiveness, while inhibiting post-acidification of set yogurt during storage. The yogurt supplemented with ACP showed a larger hysteresis area and higher G' and G″ values, formed a porous, dense, mesh-like structure and exhibited a unique mushroom flavor. Antioxidant results showed that administration of ACP-fortified yogurt significantly decreased serum alanine aminotransferase and aspartate aminotransferase enzyme activities and malondialdehyde levels, while increasing superoxide dismutase, catalase, phospholipid hydroperoxide glutathione peroxidase, and total antioxidant capacity in the liver and hippocampus of the mice. ACP-fortified yogurt might alleviate hepatic damage and hippocampal neuroinflammation induced by d-galactose. Additionally, ACP-fortified yogurt downregulated the expression of Keap1 and upregulated the expression of Nrf2 and HO-1 in the liver. In conclusion, ACP may be used as an ingredient to produce yogurt with desired properties.

Breast tubular carcinoma at parasternal and retro-nipple area: Two case reports.

INTRODUCTION: Breast tubular carcinoma (TC) is a well-differentiated infiltrating ductal carcinoma, common in postmenopausal women. PATIENT CONCERNS: Two patients concerned their abnormality of their breasts, one at deep parasternal higher chest wall in a 74-year-old female and the other behind the nipple in a 39- year-old female. DIAGNOSIS: These masses were detected by mammography, ultrasound and magnetic resonance imaging (MRI) examinations. The parasternal mass identified by mammography showed long spicules along the edges of the mass. Ultrasound examination revealed that these masses had solid irregular hypoechoic nodules. The color Doppler ultrasound of the retro-nipple mass presented with increased blood flow resistance index. The dynamic contrast-enhanced MRI examination of the retro-nipple nodule demonstrated an intensely enhancing mass with a plateau-type time-signal intensity curve. INTERVENTIONS: Two cases were surgically removed by local resection of foci under ultrasound guidance. OUTCOMES: These imaging examinations strongly suggest possible breast tubular carcinoma, which was confirmed by the pathological evaluation of frozen sections from surgically removed masses. CONCLUSION: Although uncommon, breast tubular carcinoma may be considered in the differential diagnosis of small solid masses with long spicules at parasternal breast or behind the nipples in adult women.

Is there a correlation between miR-301a expression and neoadjuvant chemotherapy efficacy in breast cancer tissue?

Neoadjuvant chemotherapy (NAC) is the standard therapeutic regimen for locally advanced breast cancer. However, clinical physical examination and imaging results fail to accurately assess the treatment response, and postoperative pathological examination has a time lag in response to therapeutic effect which is not conducive to the timely adjustment of treatment strategies. A previous study has shown that miR-301a was associated with invasion and metastasis in breast cancer, and was found to be involved in endocrine therapy resistance; however, evidence regarding the correlation between miR-301a expression and NAC efficacy remains scarce. In this study, 101 patients with locally advanced breast cancer were included. All patients received anthracycline based chemotherapy. The expression level of miR-301a in pretreatment core needle biopsy tissues was determined by real-time polymerase chain reaction analysis. Relevant clinicopathological data were collected, and the correlation between miR-301a expression and NAC efficacy was assessed. Based on our data, miR-301a cannot be used to identify whether breast cancer benefits from NAC, and no correlation was observed between miR-301a expression and clinicopathological characteristics. In conclusion, miR-301a may not be a potential prognostic biomarker of NAC efficacy in breast cancer.

Common apolipoprotein E gene mutations contribute to lipoprotein glomerulopathy in China.

BACKGROUND: Lipoprotein glomerulopathy (LPG) is a unique disease characterized by thrombus-like lipoprotein deposition in glomeruli and an increased serum apolipoprotein E level (ApoE protein or APOE gene). Several APOE mutations contribute to the occurring of LPG. METHODS: We confirmed LPG in 7 individuals by renal biopsy, and investigated families of 2 patients with urinalysis, serum creatinine and serum lipid examination. Exons of APOE of all individuals as well as their relatives were amplified and sequenced directly. RESULTS: Two types of APOE mutations were identified in the 7 patients and their relatives. APOE Maebashi (Arg142-Leu144-->0) heterozygotes were found in 5 individuals who were from 4 different families. APOE Kyoto (Arg25-Cys) was confirmed heterogeneous in another 2 individuals. Both mutations present incomplete penetrance. CONCLUSION: Our research indicates that APOE Maebashi (Arg142-Leu144-->0) is a common mutation in Chinese LPG. However, not all carriers of the 2 mutations have LPG, although hyperlipidemia and high serum ApoE level are tested. There are likely other reasons, such as a local mechanism in the glomeruli, which participated in the renal injury.

Mechanism of the feedback-inhibition resistance in aspartate kinase of Corynebacterium pekinense: from experiment to MD simulations.

In microorganisms and plants, aspartate kinase (AK) is the initial committed enzyme of the biosynthesis of the aspartate acid family amino acids and is inhibited by end products. In the paper, we mutated the key allosteric regulatory site A380 around the binding site of the Lys inhibitor in Corynebacterium pekinense AK (CpAK). A single-mutant A380C was obtained with 12.35-fold higher enzyme activity through high-throughput screening. On this basis, T379 as another key allosteric regulatory site was further modified, and the double-mutant T379N/A380C with 22.79-fold higher enzyme activity was obtained. Molecular dynamics (MD) simulations were used to investigate the mechanism of allosteric inhibition by Lys. The results indicated that the binding of Lys with CpAK resulted in conformational changes and a larger distance between the phosphorus atom of ATP and the oxygen atom of Asp, which was detrimental for the catalytic reaction. However, the mutation of allosteric sites opens the "switch" of allosteric regulation and can prevent the conformational transformation. Some key residues such as G168, R203, and D193 play an important role in maintaining the substrate binding with CpAK and further enhance the enzyme activity.

Peptides from walnut (Juglans mandshurica Maxim.) protect hepatic HepG2 cells from high glucose-induced insulin resistance and oxidative stress.

Oxidative stress is an important factor in the pathogenesis of insulin resistance (IR) in type 2 diabetes mellitus (T2DM). Bioactive peptides from nuts have been shown to alleviate oxidative stress and IR. However, the specific mechanisms underlying their activity remain unclear. This study investigated the protective effects of three novel peptides derived from Juglans mandshurica Maxim., LVRL, LRYL, and VLLALVLLR, against high glucose-induced IR and oxidative stress in HepG2 cells. The possible mechanisms underlying these effects were also elucidated. The walnut-derived peptides improved glucose consumption, glucose uptake, and glucose transporter type 4 (GLUT4) translocation, and elevated the phosphorylation of insulin receptor substrate 1 (IRS-1), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt). They also increased the activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD), the nuclear transport of nuclear factor E2-related factor 2 (Nrf2), and the protein expression of heme oxygenase-1 (HO-1). Furthermore, the walnut-derived peptides reduced high glucose-induced ROS overproduction and the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. These results suggested that walnut-derived peptides protect HepG2 cells from high glucose-induced IR and oxidative stress by activating IRS-1/PI3 K/Akt and Nrf2/HO-1 signaling pathways.

Full Transcriptome Analysis of Callus Suspension Culture System of Bletilla striata.

BACKGROUND: Bletilla striata has been widely used in the pharmacology industry. To effectively produce the secondary metabolites through suspension cultured cells of B. striata, it is important to exploring the full-length transcriptome data and the genes related to cell growth and chemical producing of all culture stages. We applied a combination of Real-Time Sequencing of Single Molecule (SMRT) and second-generation sequencing (SGS) to generate the complete and full-length transcriptome of B. striata suspension cultured cells. METHODS: The B. striata transcriptome was formed in de novo way by using PacBio isoform sequencing (Iso-Seq) on a pooled RNA sample derived from 23 samples of 10 culture stages, to explore the potential for capturing full-length transcript isoforms. All unigenes were obtained after splicing, assembling, and clustering, and corrected by the SGS results. The obtained unigenes were compared with the databases, and the functions were annotated and classified. RESULTS AND CONCLUSIONS: A total of 100,276 high-quality full-length transcripts were obtained, with an average length of 2530 bp and an N50 of 3302 bp. About 52% of total sequences were annotated against the Gene Ontology, 53,316 unigenes were hit by KOG annotations and divided into 26 functional categories, 80,020 unigenes were mapped by KEGG annotations and clustered into 363 pathways. Furthermore, 15,133 long-chain non-coding RNAs (lncRNAs) were detected. And 68,996 coding sequences were identified based on SSR analysis, among which 31 pairs of primers selected at random were amplified and obtained stable bands. In conclusion, our results provide new full-length transcriptome data and genetic resources for identifying growth and metabolism-related genes, which provide a solid foundation for further research on its growth regulation mechanisms and genetic engineering breeding mechanisms of B. striata.

Lipid Catabolism in Starved Yak Is Inhibited by Intravenous Infusion of ß-Hydroxybutyrate.

Lipid is the chief energy source for starved animals. ß-hydroxybutyrate (BHBA) is the main ketone body produced by lipid decomposition. In Chinese hamster ovary (CHO) cell experiment, it was found that BHBA could be used not only as an energy substance, but also as a ligand of GPR109A for regulating lipid metabolism. However, whether BHBA can regulate lipid metabolism of yaks, and its effective concentration and signal pathway are not clear. This study investigated the effects and mechanism of starvation and BHBA on the lipid metabolism of yak. Eighteen male Jiulong yaks were selected and then randomly divided into three groups: normal feeding group (NG), starvation group (SG), and starvation with BHBA infusion group (SBG). The yaks in the NG group were freely fed during the trial, while the yaks in the SG and SBG groups fasted; from 7th to 9th days of the experiment, the NG and SG were infused continuous with 0.9% normal saline and SBG was infused 1.7 mmol/L BHBA solution respectively. The blood samples were collected on the 0th, 1st, 3rd, 5th, 7th, and 9th day of experiment. The subcutaneous adipose tissue of all the yaks in this study were taken from live bodies after infusion. Serum glucose, lipid metabolites, hormone concentrations, and mRNA and protein expressions of key factors of lipid metabolism and signaling pathway in subcutaneous adipose tissue were measured. The results showed that, as compared with NG, starvation significantly reduced the body weight of yak in SG, and significantly increased the concentration of BHBA in serum and the mRNA expression of PKA and CREB1 in subcutaneous adipose tissue, while the mRNA expression of MEK, PKC, ERK1/2, the area of adipocytes, and the proportion of saturated fatty acid were decreased. Whereas, further increase of BHBA concentration through infusion promoted the mRNA expression of GPR109A receptor in the subcutaneous adipose tissue of SBG, inhibited the mRNA expression of AC and PKA, and decreased the phosphorylation protein abundance of CREB1, and significantly increased the diameter and area of adipocytes. These findings suggest that starvation led to enhanced lipid catabolism in yaks. An increasing BHBA concentration could increase the mRNA expression of GPR109A receptor in subcutaneous adipose tissue and inhibit the cAMP/PKA/CREB signaling pathway and lipid decomposition.

Effects of Nutritional Deprivation and Re-Alimentation on the Feed Efficiency, Blood Biochemistry, and Rumen Microflora in Yaks (Bos grunniens).

Yak suffers severe starvation and body weight reduction in the cold season and recovers relatively rapid growth in the warm season every year. Herein, we investigated the effects of starvation and refeeding on the growth, feed efficiency, blood biochemistry and rumen microbial community as well as functions of yaks. The results showed that starvation significantly reduced the body weight of yaks. Serum glucose and triglyceride concentrations significantly decreased, and ß-hydroxybutyric acid and non-esterified fatty acid levels were significantly increased during the starvation period. Starvation also dramatically inhibited rumen microbial fermentations. Whereas, refeeding with the same diet significantly increased the feed efficiency, nutrient digestibility together with rumen acetate, propionate and microbial protein productions compared with those before starvation. The 16S rDNA sequencing results showed that starvation mainly decreased the ruminal protein-degrading bacteria Prevotella and propionate-producing bacteria Succiniclasticum populations and dramatically increased the denitrifying bacteria Thauera populations. Refeeding reduced the Euryarchaeota population and increased propionate-producing bacteria Succinivibrionaceae UCG-002 and starch-degrading bacteria Ruminobacter populations when compared with those before starvation. The predicted microbial metabolic pathways, related to amino acid and starch metabolisms, were also significantly altered during the starvation and refeeding. The results indicated that the rumen microorganisms and their metabolism pathways changed with feed supply, and these alterations in part contributed to yak adaption to starvation and re-alimentation. This study is helpful for enhancing the understanding and utilization of this natural character of yaks to explore and improve their growth potential.