Selection Signal Analysis Reveals Hainan Yellow Cattle Are Being Selectively Bred for Heat Tolerance.

Hainan yellow cattle are indigenous Zebu cattle from southern China known for their tolerance of heat and strong resistance to disease. Generations of adaptation to the tropical environment of southern China and decades of artificial breeding have left identifiable selection signals in their genomic makeup. However, information on the selection signatures of Hainan yellow cattle is scarce. Herein, we compared the genomes of Hainan yellow cattle with those of Zebu, Qinchuan, Nanyang, and Yanbian cattle breeds by the composite likelihood ratio method (CLR), Tajima's D method, and identifying runs of homozygosity (ROHs), each of which may provide evidence of the genes responsible for heat tolerance in Hainan yellow cattle. The results showed that 5210, 1972, and 1290 single nucleotide polymorphisms (SNPs) were screened by the CLR method, Tajima's D method, and ROH method, respectively. A total of 453, 450, and 325 genes, respectively, were identified near these SNPs. These genes were significantly enriched in 65 Gene Ontology (GO) functional terms and 11 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (corrected p < 0.05). Five genes-Adenosylhomocysteinase-like 2, DnaJ heat shock protein family (Hsp40) member C3, heat shock protein family A (Hsp70) member 1A, CD53 molecule, and zinc finger and BTB domain containing 12-were recognized as candidate genes associated with heat tolerance. After further functional verification of these genes, the research results may benefit the understanding of the genetic mechanism of the heat tolerance in Hainan yellow cattle, which lay the foundation for subsequent studies on heat stress in this breed.

Knowledge, attitudes and practices towards advance directives among clinical nurses: Multicenter cross-sectional survey.

AIMS AND OBJECTIVES: This study aimed to describe the knowledge, attitudes and practices (KAP) of nurses in implementing advance directives (ADs) for older patients and analyze the influencing factors before the establishment of the first advance directives act in China. DESIGN: Multicenter cross-sectional survey. The standards for reporting the STROBE checklist are used. METHODS: This cross-sectional study developed a self-designed structured questionnaire to assess nurses' knowledge, attitudes and practices about ADs. Nurses were recruited by stratified random sampling through the Nursing Departments of 12 hospitals in southwest China and were asked to fill out the questionnaire face to face about knowledge, attitudes and practices. Data were analyzed following descriptive statistics, rank-sum test and multiple linear regression. RESULTS: This study included 950 nurses. The study found that nurses were extremely supportive of ADs. Unmarried nurses had better knowledge of ADs than married ones. Nevertheless, there was a discrepancy between the participants' knowledge, attitude and practice. The participants' practice was lower (4.3%) compared with their attitude (81.9%) and knowledge (42.2%). Knowledge on, attitudes towards and standardized procedures for ADs in the workplace affected nursing practice. CONCLUSIONS: The study recommends that courses on ADs and appropriate support from medical institutions should be provided to nurses to increase their knowledge and confidence in implementing ADs. Healthcare professionals should be sufficiently equipped to implement ADs and handle their execution appropriately to provide adequate end-of-life care corresponding to patients' wishes. RELEVANCE TO CLINICAL PRACTICE: The study results inform rich insights as it discusses the numerous interrelating factors influencing these three fundamental aspects that affect the success of any AD policy by surveying the knowledge, attitudes and practices of clinical nurses. Furthermore, our results hint at distinct areas of improvement in the nursing practice to facilitate the wider implementation and acceptance of ADs in China. PATIENT OR PUBLIC CONTRIBUTION: This study involved no patient.

Association of nitric oxide synthase gene polymorphism with asthma: A systematic review and meta-analysis.

INTRODUCTION: This study examines the associations between asthma and nitric oxide (NO) synthase (NOS) gene polymorphisms. METHODS: After a systematic literature search in electronic databases, studies were selected based on eligibility criteria. Data were extracted from research articles and were synthesized and tabulated. Where a particular polymorphism data were reported by multiple studies, meta-analyses of odds ratios were performed, or odds ratios reported by individual studies were pooled. RESULTS: Twenty studies (4450 asthma patients and 5306 non-asthmatic individuals) were identified. Many studies did not find any association between CCTTT repeat polymorphism in NOS2 gene and asthma. However, a study reported that pretreatment mean exhaled NO levels in asthmatics were found to be significantly higher in genotypes with higher number of CCTTT repeats. Also, alleles with <11 CCTTT repeats were associated with poor asthma treatment outcomes. A single nucleotide polymorphism, G894T, in NOS3 gene was not found to be significantly associated with asthma by at least four studies. However, a T allele at this locus was associated with lower NO levels. Also, G894T frequency was significantly higher in asthmatic children who responded to inhaled corticosteroids along with long-lasting beta2-agonists. A T allele of NOS3 786C/T polymorphism increased the probability of bronchial asthma with comorbid essential hypertension in asthma patients. Asthma severity also differed for different Ser608Leu exon 16 variants of NOS2 gene. CONCLUSIONS: Several polymorph NOS gene variants are identified, some of which appear to have influence on asthma prevalence or outcomes. However, data are varying depending on the nature of variant, ethnicity, study design, and disease parameters.

Dynamic Changes in the Gut Microbiota and Metabolites during the Growth of Hainan Wenchang Chickens.

Gut microbiota and their metabolites play important roles in animal growth by influencing the host's intake, storage, absorption, and utilization of nutrients. In addition to environmental factors, mainly diet, chicken breed and growth stage also affect changes in the gut microbiota. However, little research has been conducted on the development of gut microbiota and its metabolites in local chickens. In this study, the cecal microbiota and metabolites in different developmental stages of Hainan Wenchang chickens (a native breed of Bantam) were investigated using 16S rRNA sequencing and untargeted metabolomics. With aging, the structure of gut microbiota tended to be more stable. The relative proportions of dominant bacteria phyla Firmicutes, Bacteroidetes, and Proteobacteria showed stage changes with the development. With aging, gut microbiota and their metabolites may have structural and functional changes in response to nutrient metabolism and immune requirements in different physiological states. Several microbial and metabolic biomarkers with statistical differences were detected in different development stages. The bacteria that form networks with their significant related metabolites were different in various growth stages, including uncultured_bacterium_f_Ruminococcaceae, Ruminococcaceae_UCG-014, Faecalibacterium, uncultured_bacterium_o_Bacteroidales, and uncultured_bacterium_f_Lachnospiraceae. Partially differential bacteria were significantly correlated with short-chain fatty acids such as butyric acid. These findings may provide new insights into the physiological and molecular mechanisms of developmental changes of local chicken breeds, as well as resources for microbial and metabolic biomarker identification to improve growth efficiency.

Effect of altitude hypoxia on blood-brain barrier after subarachnoid hemorrhage based on phosphatidylinositol 3-kinase/protein kinase B/nuclear factor κB pathway in rats / 解剖学报

Objective To investigate the effect of plateau hypoxia on the blood-brain barrier after subarachnoid hemorrhage (SAH) in rats. Methods Adult male SD rats (n = 78) were randomly divided into 4 groups: sham group (sham), SAH model group (SAH), plateau hypoxia sham group (Hp sham) and plateau hypoxia SAH model group (Hp SAH). The rat model of plateau hypoxia was established through low-pressure simulation chamber (altitude 5000 m), and the SAH model was established by endovascular perforation method. At 24 hours after SAH, neurobehavior score and SAH grade were assessed. The morphological changes of neurons and apoptosis of nerve cells in the CA1 region of hippocampal were observed by the staining of Nissl and TUNEL. The expression of phosphorylated PI3K (p-PI3K), PI3K, phosphorylated Akt (p-Akt), Akt, phosphorylated nuclear factor κB (p-NF-κB), NF-κB, matrix metalloproteinase-9 (MMP-9), occludin and claudin-5 in hippocampal were detected by the method of Western blotting. The expression of occludin and claudin-5 proteins in the CA1 region of hippocampal were observed by immunofluorescent staining. Results At 24 hours after SAH, the neurobehavior score decreased significantly and SAH grade increased significantly in the SAH and Hp SAH group (P< 0.05). Neurobehavior score decreased significantly in the Hp SAH group compared with the SAH group (P < 0.05). In the SAH group, neurons in the CA1 region of hippocampus were atrophied and deformed, the arrangement were disordered, the number of neurons decreased significantly, and the apoptosis of nerve cells increased significantly(P< 0.05). Plateau hypoxia could aggravate the morphological damage of neurons and apoptosis of nerve cells. The expression of p-PI3K/PI3K, p-Akt/Akt, occludin and claudin-5 proteins decreased significantly, while the expression of p-NF-κB/NF-κB and MMP-9 proteins increased significantly in the SAH and Hp SAH group (P< 0.05). The expression of p-PI3K/PI3K and MMP-9 proteins increased significantly in Hp SAH group compared with the SAH group. The expression of claudin-5 protein increased significantly in Hp sham group compared with the sham group (P < 0.05). Immunofluorescent staining showed that the expression of occludin and claudin-5 proteins in the CA1 region of hippocampus decreased in the SAH group. Plateau hypoxia could further decreased the expression of occludin and claudin-5 proteins. Conclusion Plateau hypoxia aggravates blood-brain barrier disruption after subarachnoid hemorrhage in rats through inhibiting PI3K/Akt/NF-κB pathway.

Mammary myofibroblastoma: a clinicopathological analysis of fifteen cases / 中华病理学杂志

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of breast myofibroblastoma. Methods: The clinicopathological data and prognostic information of 15 patients with breast myofibroblastoma diagnosed at the Department of Pathology of the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China from 2014 to 2022 were collected. Their clinical characteristics, histological subtypes, immunophenotypes and molecular characteristics were analyzed. Results: There were 12 female and 3 male patients, ranging in age from 18 to 78 years, with a median and average age of 52 years. There were 6 cases in the left breast and 9 cases in the right breast, including 12 cases in outer upper quadrant, 2 cases in inner upper quadrant and 1 case in outer lower quadrant. Most of the cases showed a well-defined nodule grossly, including pushing growth under the microscope in 13 cases, being completely separated from the surrounding breast tissue in 1 case, and infiltrating growth in 1 case. Among them, 12 cases were classic subtype and composed of occasional spindle cells with varying intervals of collagen fiber bundles; eight cases had a small amount of fat; one case had focal cartilage differentiation; one case was epithelioid subtype, in which epithelioid tumor cells were scattered in single filing or small clusters; one case was schwannoma-like subtype, and the tumor cells were arranged in a significant palisade shape, resembling schwannoma, and one case was invasive leiomyoma-like subtype, in which the tumor cells had eosinophilic cytoplasm and were arranged in bundles, and infiltrating into the surrounding mammary lobules like leiomyoma. Immunohistochemical studies showed that the tumor cells expressed desmin (14/15) and CD34 (14/15), as well as ER (15/15) and PR (15/15). Three cases with histologic subtypes of epithelioid subtype, schwannoma-like subtype and infiltrating leiomyoma-like subtype showed RB1 negative immunohistochemistry. Then FISH was performed to detect RB1/13q14 gene deletion, and identified RB1 gene deletion in all three cases. Fifteen cases were followed up for 2-100 months, and no recurrence was noted. Conclusions: Myofibroblastoma is a rare benign mesenchymal tumor of the breast. In addition to the classic type, there are many histological variants, among which the epithelioid subtype is easily confused with invasive lobular carcinoma. The schwannoma-like subtype is similar to schwannoma, while the invasive subtype is easily misdiagnosed as fibromatosis-like or spindle cell metaplastic carcinoma. Therefore, it is important to recognize the various histological subtypes and clinicopathological features of the tumor for making correct pathological diagnosis and rational clinical treatment.

Value of Contrast-enhanced Ultrasound in Sentinel Lymph Node after Primary Breast Cancer Resection / 中山大学学报(医学科学版)

ObjectiveTo investigate the value of percutaneous and intravenous contrast-enhanced ultrasound（P-Ⅳ-CEUS） in sentinel lymph nodes（SLNs） after resection of early-stage primary breast cancer. MethodsA retrospective analysis was done on the clinical and imaging data of 42 early breast cancer patients. Following primary tumor resection， all these patients underwent reoperation in our hospital. SLNs were examined by preoperative P-Ⅳ-CEUS and intraoperative sentinel lymph node biopsy（SLNB） was performed by using Methylene blue as a tracer. Then we analyzed the detection and false-negative rate in CEUS and SLNB respectively. By using the surgical pathological results as the gold standard， the diagnostic efficacy of CEUS for SLNs was explored. ResultsThe detection rate and false negative rate of SLNs in percutaneous contrast-enhanced ultrasound （P-CEUS） were 92.9% （39/42） and 7.1% （3/42）， respectively. The detection rate in methylene blue staining was 100% （41/41） and one patient underwent neoadjuvant therapy due to biopsy-confirmed metastasis. The sensitivity， specificity， positive predictive value， negative predictive value and accuracy of P-Ⅳ-CEUS were 66.7% （2/3）， 100% （37/37）， 100% （2/2）， 97.3% （36/37） and 97.4% （38/39）， respectively. ConclusionsP-Ⅳ-CEUS after resection of early-stage primary breast cancer can accurately detect SLNs and characterize their status， which is a reliable clinical basis for reducing invasive SLNB.

Prevalence and clinical implications of polypharmacy and potentially inappropriate medication in elderly patients with heart failure: results of six months' follow-up / 老年心脏病学杂志（英文版）

OBJECTIVES@#To investigate the prevalence of polypharmacy and potentially inappropriate medication (PIM) in elderly patients with heart failure (HF) and their impact on readmission and mortality.@*METHODS@#We conducted a study of 274 participants aged 60 years or older with HF. The prevalence of polypharmacy (defined as the use of five or more medications) was calculated, and the 2019 American Geriatrics Society Beers criteria were applied to access PIMs. Medications and PIMs were characterized at admission and discharge, and changes in prescriptions during hospitalization were compared. The impact of polypharmacy and PIM on readmission and mortality were investigated.@*RESULTS@#The median age of this study population was 68 years old. The median number of prescribed drugs was 7 at admission and 10 at discharge. At discharge, 99.27% of all patients were taking five or more drugs. The incidence of composite endpoint and cardiovascular readmission increased with the number of polypharmacy within 6 months. The use of guideline-directed medical therapy reduced the incidence of composite endpoint events and cardiovascular readmission, while the use of non-cardiovascular medications increased the composite endpoint events. The frequency of PIMs was 93.79% at discharge. The incidence of composite endpoint events increased with the number of PIMs. "PIMs in older adults with caution" increased cardiovascular readmission and "PIMs based on kidney function" increased cardiovascular mortality. Several comorbidities were associated with cardiovascular mortality or non-cardiovascular readmission.@*CONCLUSIONS@#Polypharmacy and PIM were highly prevalent in elderly patients with HF, and their use was associated with an increased risk of composite endpoint events, readmission and mortality. Non-cardiovascular medications, "PIMs in older adults with caution", "PIMs based on kidney function" and several comorbidities were important factors associated with hospital readmission and mortality. Our findings highlight the importance of medication optimization in the management of HF in elderly patients.

Characterization of a novel recombinant halophilic ß-glucosidase of Trichoderma harzianum derived from Hainan mangrove.

BACKGROUND: ß-glucosidase is an important biomass-degrading enzyme and plays a vital role in generating renewable biofuels through enzymatic saccharification. In this study, we analyzed the transcriptome of Trichoderma harzianum HTASA derived from Hainan mangrove and identified a new gene encoding ß-glucosidase Bgl3HB. And the biochemically characterization of ß-glucosidase activity was performed. RESULTS: Bgl3HB showed substantial catalytic activity in the pH range of 3.0-5.0 and at temperatures of 40 ℃-60 ℃. The enzyme was found quite stable at 50 ℃ with a loss of only 33.4% relative activity after 240 min of heat exposure. In addition, all tested metal ions were found to promote the enzyme activity. The ß-glucosidase activity of Bgl3HB was enhanced by 2.12-fold of its original activity in the presence of 5 M NaCl. Surprisingly, Bgl3HB also showed a remarkable ability to hydrolyze laminarin compared to other measured substrates. Enzyme efficiency was examined in the sugarcane bagasse saccharification processes, in which Bgl3HB with 5 M NaCl worked better supplementing Celluclast 1.5L than the commercial Novozyme 188 ascertained it as an admirably suited biocatalyst for the utilization of agricultural waste. In this work, this is the first report of a halophilic ß-glucosidase from Trichoderma harzianum, and represents the ß-glucosidase with the highest known NaCl activation concentration. And adding 5 M NaCl could enhance saccharification performance even better than commercial cellulase. CONCLUSIONS: These results show that Bgl3HB has great promise as a highly stable and highly efficient cellulase with important future applications in the industrial production of biofuels.

Protective effect of orange essential oil on the formation of non-alcoholic fatty liver disease caused by high-fat diet.

The purpose of this study was to investigate the protective effect of sniffing orange essential oil (OEO) on the formation of non-alcoholic fatty liver disease (NAFLD) caused by a high-fat diet. The results confirmed that sniffing OEO could reduce obesity caused by a high-fat diet (HFD) by reducing the levels of triglycerides (TGs), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C). In addition, the observation of liver tissue sections showed that sniffing OEO could reduce lipid accumulation in liver cells. Further analysis by western blot analysis showed that OEO treatment made the expression levels of acetyl-CoA carboxylase (ACC) and Cytochrome P450 2E1 (CYP2E1) down-regulated and the expression levels of peroxisome proliferator-activated receptor-α (PPAR-α) and carnitine palmitoyltransferase-1 (CPT-1) up-regulated. These results indicate that the treatment of sniffing OEO could enhance the antioxidant capacity of mice and reduce liver damage caused by a high-fat diet. Furthermore, sniffing OEO could inhibit lipid synthesis and oxidative stress stimulated by a high-fat diet. Overall, OEO treatment had a certain protective effect on NAFLD-related diseases caused by a high-fat diet. Therefore, aromatherapy may be introduced as a treatment of long-term chronic diseases.

Correlation between serum cystatin C and left ventricular geometry in patients with obstructive sleep apnea syndrome complicated with prehypertension / 中华超声影像学杂志

Objective:To investigate the correlation between serum cystatin C (Cys C) and left ventricular geometry in patients with obstructive sleep apnea syndrome (OSAS) complicated with prehypertension(PH).Methods:A total of 408 patients with PH and OSAS diagnosed by polysonogram monitoring in the sleep monitoring room of Respiratory Department, the First Hospital of Shanxi Medical University from July 2018 to June 2021 were collected. Serum Cys C concentration and echocardiography were performed in all patients. According to the classification of left ventricular geometry, all patients were divided into four groups: normal configuration (NG) group( n=297), concentric remodeling (CR) group( n=49), eccentric hypertrophy (EH) group( n=33), and concentric hypertrophy (CH) group( n=29). General clinical data, sleep parameters, blood biochemical parameters, Cys C concentration and echocardiographic parameters were compared among the four groups, and the correlation between serum Cys C and left ventricular geometry was analyzed. Results:①The serum Cys C concentration increased successively from NG group, CR group, EH group to CH group, and the increase in CH group was the most obvious ( P<0.05). In addition, there were statistically significant differences in body mass index (BMI), waist circumference, systolic blood pressure (SBP), diastolic blood pressure (DBP), severity of OSAS, the percentage of the time that the blood oxygen saturation was less than 90% of the total sleep time (T90), lowest oxygen saturation (Lowest-SaO 2), mean oxygen saturation (Mean-SaO 2), left ventricular end diastolic diameter (LVEDD), inter-ventricular septal diameter (IVST), left ventricular posterior wall thickness diameter (LVPWT), left ventricular mass index (LVMI), relative wall thickness (RWT), left ventricular ejection fraction (LVEF) and E/A among all groups (all P<0.05). ②Multivariate Logistic regression analysis showed that Cys C was independently correlated with CR (β=0.721, OR=2.057, P=0.047), EH(β=0.961, OR=2.614, P=0.017) and CH (β=1.180, OR=3.254, P=0.010). Conclusions:There is a correlation between serum Cys C and left ventricular geometry in OSAS patients with PH, suggesting that serum Cys C might be involved in the change of left ventricular geometry.

Effects of different doses of fluoride on expression of IRE1α-ASK1-JNK proteins in rat renal cortex / 中华地方病学杂志

Objective:To investigate the expression and significance of endoplasmic reticulum stress apoptosis pathway related proteins in renal cortex of rats with chronic fluorosis.Methods:Twenty four healthy SD rats were divided into 4 groups (6 rats/group, half male and half female) according to their body mass (100 - 120 g) by random number table method, rats in control group drank tap water (fluoride content < 0.5 mg/L), and in low, medium and high fluoride groups drank tap water with fluoride content (sodium fluoride) of 10, 50 and 100 mg/L, respectively. After 180 days of feeding, dental fluorosis was examined, 24-hour urine sample was collected and the content of fluoride in urine was detected by fluoride ion selective electrode method. Renal tissue was taken after anesthesia, and the pathological changes of renal cortex were observed by hematoxylin-eosin (HE) staining. The expressions of endoplasmic reticulum stress apoptosis pathway related proteins [inositol-requiring enzyme 1α (IRE1α), apoptosis signal-regulating kinase 1 (ASK1), C-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK)] were determined by immunohistochemical staining in rat renal cortex.Results:No dental fluorosis was found in control group. The incidence of dental fluorosis in low, medium and high fluoride groups were 2/6, 5/6 and 6/6, respectively. Compared with control group [(5.707 ± 1.190) mg/L], the urinary fluoride in low, medium and high fluoride groups [(17.028 ± 3.006), (34.378 ± 12.045), (94.759 ± 31.773) mg/L] was significantly higher ( P < 0.05), and the urinary fluoride in high fluoride group was higher than that in low and medium fluoride groups ( P < 0.05). HE staining showed that, compared with control group, the cell volume of renal tubules and glomeruli in medium and high fluoride groups increased, the cells arranged closely, and the eosinophilia of the cytoplasm increased. The immunohistochemical staining results showed that there was no significant difference in the expression of JNK protein in rat renal cortex between control group and low, medium and high fluoride groups ( F = 0.07, P > 0.05). The expressions of IRE1α, ASK1 and P-JNK proteins in rat renal cortex in high fluoride group were higher than those of control, low and medium fluoride groups ( P < 0.05), and the expressions of IRE1α and ASK1 proteins in medium fluoride group were significantly higher than those in control and low fluoride groups ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to renal cortex injury in rats, and the mechanism of injury may be related to the activation of IRE1α-ASK1-JNK endoplasmic reticulum stress apoptosis pathway.

The Relationship between MicroRNA Expression Profiling in Imatinib-Resistant Cell Line K562/G and Potential Mechanism through FOXO3/Bcl-6 Signaling Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the drug resistant related FOXO3/Bcl-6 signaling pathway in K562/G cell line and its related microRNA(miRNA) mechanisms.@*METHODS@#The drug resistance potency of imatinib on K562/G was detected by MTT assay. The expression of FOXO3 and Bcl-6 proteins in K562 and K562/G cells was detected by Western blot. Real-time PCR (RT-PCR) was used to detect the expression of FOXO3 and Bcl-6 mRNA. The miRNA expression profiling in K562 and K562/G cells was analyzed by microarray technique, and the miRNA targeted to FOXO/Bcl-6 signaling pathway was identified.@*RESULTS@#The expression of FOXO3 and Bcl-6 protein was significantly increased in K562/G cells as compared with that in K562 cells (P<0.01), the expression level of Bcl-6 mRNA showed no increase in K562/G cells. However, FOXO3 mRNA was up-regulated in K562/G cells (P<0.05). MiRNA microarray results showed that 109 miRNAs were expressed differentially in K562 and K562/G cells. The expression of 81 miRNAs were up-regulated while 28 miRNAs were down-regulated. Through reverse prediction by bioinformatics, miR-6718-5p, miR-5195-5p, miR-4711-3p, miR-4763-5p, miR-4664-5p and miR-3176 were related to FOXO/Bcl-6 signaling pathway.@*CONCLUSION@#The FOXO3/Bcl-6 signaling pathway contributes to imatinib resistance in K562/G cell line, and the miRNA expression profiles showed significant differences between K562/G and K562 cells.

Effects of heart rate control on short-term prognosis of heart failure in hospitalized elderly patients / 中华老年医学杂志

Objective:To examine the effects of heart rate control during hospitalization on short-term prognosis of heart failure in elderly patients.Methods:As a prospective study, 150 elderly patients with heart failure were selected from the Department of Geriatrics, Qilu Hospital of Shandong University.The subjects were divided into an experimental group and a control group by digitally generated random numbers, with 75 individuals in each group.Both groups received conventional anti-heart failure therapy during hospitalization, but patients from the control group had doses of heart rate control drugs adjusted every 2-4 weeks, with no special requirement for the heart rate before hospital discharge.In contrast, patients from the experimental group were given heart rate control drugs with timely dose adjustment to achieve more proactive heart rate control, aiming for a rate <70 beat/min, as long as heart failure symptom improvement and good volume management could be maintained.Values of cardiac function indexes were compared between the two groups at discharge and 6 months after discharge.Heart failure readmission rates within 6 months, cardiovascular disease mortality rates and the incidences of composite endpoint events after readmission due to heart failure aggravation were compared between the two groups.Treatment safety was also evaluated.Results:There was no statistical difference in blood pressure, heart rate, N-terminal pro-B-type natriuretic peptide(NT-pro-BNP), left ventricular ejection fraction(LVEF), left ventricular end systolic diameter(LVESD), or left ventricular end diastolic diameter(LVEDD)between the two groups at admission( P>0.05), and there was no statistical difference in the average length of hospitalization between the two groups( P>0.05). The experimental group had a lower average heart rate and diastolic pressure than the control group at discharge and 6 months latter[at discharge: (61.6±4.2)beat/min(1 mmHg=0.133 kPa) vs.(78.0±7.1)beat/min, (62.1±10.4)mmHg vs.(66.1±10.2)mmHg; at 6 months: (64.7±12.1)beat/min vs.(71.8±11.2)beat/min, (62.8±11.2)mmHg vs.(68.6±10.2)mmHg; P<0.05 or P<0.01]. NT-pro-BNP in the experimental group was significantly lower than that in the control group at discharge[(1 706±1 408)ng/L vs.(2 806±3 812)ng/L, P<0.05]. The absolute values of changes in LVEF(ΔLVEF), LVESD(ΔLVESD)and LVEDD(ΔLVEDD)after 6 months in the experimental group were significantly higher than those in the control group[ΔLVEF: (0.08±0.09) vs.(0.02±0.09), P<0.05; ΔLVESD: (-5.82±7.44)mm vs.(-1.63±6.07)mm, P<0.01; ΔLVEDD: (-2.76±5.52)mm vs.(-0.86±4.44)mm, P<0.05]. The rate of readmission and the incidence of composite endpoint events within 6 months in the experimental group were significantly lower than those in the control group[21.3%(16 cases) vs.36.0%(27 cases), P<0.05]; 25.3%(19 cases) vs.44.0%(33 cases), P<0.05.There was no significant difference in all-cause mortality between the two groups( P>0.05). Conclusions:For elderly patients with heart failure, proactive active heart rate control during hospitalization and a rate <70 beat/min before discharge will improve cardiac function indexes and lower the rate of readmission with exacerbation of heart failure, cardiovascular disease mortality and the incidence of composite end-point events after readmission.This strategy has good safety and is beneficial for short-term prognosis.

Screening of Reference Genes for RT-qPCR in Chicken Adipose Tissue and Adipocytes.

Reverse transcription quantitative real-time PCR is the most commonly used method to detect gene expression levels. In experiments, it is often necessary to correct and standardize the expression level of target genes with reference genes. Therefore, it is very important to select stable reference genes to obtain accurate quantitative results. Although application examples of reference genes in mammals have been reported, no studies have investigated the use of reference genes in studying the growth and development of adipose tissue and the proliferation and differentiation of preadipocytes in chickens. In this study, GeNorm, a reference gene stability statistical algorithm, was used to analyze the expression stability of 14 candidate reference genes in the abdominal adipose tissue of broilers at 1, 4, and 7 weeks of age, the proliferation and differentiation of primary preadipocytes, as well as directly isolated preadipocytes and mature adipocytes. The results showed that the expression of the TATA box binding protein (TBP) and hydroxymethylbilane synthase (HMBS) genes was most stable during the growth and development of abdominal adipose tissue of broilers, the expression of the peptidylprolyl isomerase A (PPIA) and HMBS genes was most stable during the proliferation of primary preadipocytes, the expression of the TBP and RPL13 genes was most stable during the differentiation of primary preadipocytes, and the expression of the TBP and HMBS genes was most stable in directly isolated preadipocytes and mature adipocytes. These results provide reference bases for accurately detecting the mRNA expression of functional genes in adipose tissue and adipocytes of chickens.

Multi-Omics Association Reveals the Effects of Intestinal Microbiome-Host Interactions on Fat Deposition in Broilers.

Growing evidence indicates that gut microbiota factors cannot be viewed as independent in the occurrence of obesity. Because the gut microbiome is highly dimensional and complex, studies on interactions between gut microbiome and host in obesity are still rare. To explore the relationship of gut microbiome-host interactions with obesity, we performed multi-omics associations of gut metagenome, intestinal transcriptome, and host obesity phenotypes in divergently selected obese-lean broiler lines. Metagenomic shotgun sequencing generated a total of 450 gigabases of clean data from 80 intestinal segment contents of 20 broilers (10 of each line). The microbiome comparison showed that microbial diversity and composition in the duodenum, jejunum, ileum, and ceca were altered variously between the lean- and fat-line broilers. We identified two jejunal microbes (Escherichia coli and Candidatus Acetothermia bacterium) and four cecal microbes (Alistipes sp. CHKCI003, Ruminococcaceae bacterium CPB6, Clostridiales bacterium, and Anaeromassilibacillus sp. An200), which were significantly different between the two lines (FDR < 0.05). When comparing functional metagenome, the fat-line broilers had an intensive microbial metabolism in the duodenum and jejunum but degenerative microbial activities in the ileum and ceca. mRNA-sequencing identified a total of 1,667 differentially expressed genes (DEG) in the four intestinal compartments between the two lines (| log2FC| > 1.5 and FDR < 0.05). Multi-omics associations showed that the 14 microbial species with abundances that were significantly related with abdominal fat relevant traits (AFRT) also have significant correlations with 155 AFRT-correlated DEG (p < 0.05). These DEG were mainly involved in lipid metabolism, immune system, transport and catabolism, and cell growth-related pathways. The present study constructed a gut microbial gene catalog of the obese-lean broiler lines. Intestinal transcriptome and metagenome comparison between the two lines identified candidate DEG and differential microbes for obesity, respectively. Multi-omics associations suggest that abdominal fat deposition may be influenced by the interactions of specific gut microbiota abundance and the expression of host genes in the intestinal compartments in which the microbes reside. Our study explored the interactions between gut microbiome and host intestinal gene expression in lean and obese broilers, which may expand knowledge on the relationships between obesity and gut microbiome.

Protective Effect of Huangqi Guizhi Wuwutang Combined with Shengmaiyin on Cardiac Function of Diabetic Cardiomyopathy / 中国实验方剂学杂志

Objective:To observe the protective effect of Huangqi Guizhi Wuwutang combined with Shengmaiyin on the heart function in patients with diabetic cardiomyopathy and explore its anti-myocardial fibrosis and anti-inflammatory effects. Method:The 96 patients were randomly divided into observation group （48 cases） and control group （48 cases）. Both groups were given comprehensive measures to control blood sugar， blood lipids， blood pressure and heart failure. Patients in control group took Tongmai Jiangtang capsule， 3 granules/time， 3 times/day. Patients in observation group took Huangqi Guizhi Wuwutang combined with modified Shengmaiyin， 1 dose/day. The treatment courses were three months in both groups. Left ventricular ejection fraction （LVEF）， early diastolic peak velocity E peak/late diastolic peak velocity A peak （E/A）， left ventricular end diastolic diameter （LVEDd） and cardiac output per stroke （SV） through echocardiography were recorded before and after therapy. Cardiac troponin-I （cTn I）， troponin T （cTn-T）， creatine kinase isoenzyme -MB （CK-MB）， lactate dehydrogenase （LDH）， transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁） before and after treatment ， matrix metalloproteinase-2 （MMP-2）， insulin-like growth factor-1 （IGF-1）， interleukin-6 （IL-6）， IL-1， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， N-terminal pro-B-type natriuretic peptide （NT-proBNP）， soluble ST2 （sST2） and galectin-3 （Gal-3） levels were detected. Symptom of cardiac insufficiency and traditional Chinese medicine （TCM） syndrome score were evaluated before and after treatment. Result:The LVEF and E/A data in observation group were higher than those in control group （<italic>P</italic><0.01）. The levels of cTn-I， cTn-T， LDH and CK-MB in the observation group were lower than those in the control group （<italic>P</italic><0.01）. After treatment， the levels of TGF-<italic>β</italic>₁， MMP-2， IGF-1， IL-6， IL-1， TNF-<italic>α</italic>， NT-proBNP， sST2 and Gal-3 in the observation group decreased and were lower than those in the control group （<italic>P</italic><0.01）. The clinical efficacy of the observation group was better than that of the control group （<italic>Z</italic>=1.974，<italic>P</italic><0.05）. Conclusion:On the basis of conventional intervention of western medicine， Huangqi Guizhi Wuwutang combined with modified Shengmaiyin has anti-inflammatory and anti-myocardial fibrosis effects， with inhibitory effect on myocardial remodeling， and can reduce myocardial tissue damage to improve ventricular diastolic function and protect heart function. With such high clinical efficacy， it is worthy of clinical use.

Effect of Danggui Shaoyaosan on Diminished Ovarian Reserve in Model Rats： An Exploration Based on TGF-β1/ Smads Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the improving effect of Danggui Shaoyaosan on diminished ovarian reserve （DOR） in rats triggered by Tripterygia wilfordii polyglycoside tablet combined with stress， and to explore the role of transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁）/Smads signaling pathway in such improvement. Method:Forty-eight female SD rats with normal sexual cycle were selected and randomly divided into a normal group （<italic>n</italic>=8） and a modeling group （<italic>n</italic>=40）， and the ones in the modeling group were given Tripterygium wilfordii polyglycoside tablets （50 mg·kg^-1） combined with random stress for 15 d. After successful modeling， the rats were randomized into the model group， low-， medium-， and high-dose （3.96， 7.92， 15.84 g·kg^-1） Danggui Shaoyaosan groups， and estradiol valerate group （0.09 mg·kg^-1）， with eight in each group. Under the premise of stress exposure， they were separately gavaged with the normal saline， low-， medium- and high-dose Danggui Shaoyaosan， and estradiol valerate for 15 successive days. The estrous cycle of rats in each group was observed daily. After intervention， the rats were sacrificed and the ovarian visceral index was calculated. The pathological changes in ovarian tissues were observed by hematoxylin eosin （HE） staining. The protein expression levels of TGF-<italic>β</italic>₁ and TGF-<italic>β</italic>₁receptor （TGF-<italic>β</italic>₁R） in the ovarian tissues of rats were measured by immunohistochemistry （IHC）， and the mRNA expression levels of Smad2， Smad3， and Smad7 in the ovarian tissues by real-time polymerase chain reaction （Real-time PCR）. Result:Compared with the normal group， the model group exhibited disordered estrus cycle （<italic>P</italic><0.05）， reduced visceral index （<italic>P</italic><0.01）， and down-regulated TGF-<italic>β</italic>₁ and TGF-<italic>β</italic>₁R protein and Smad2 and Smad3 mRNA expression in the ovarian tissues （<italic>P</italic><0.01）， and up-regulated Smad7 mRNA expression （<italic>P</italic><0.01）. Compared with the model group， Danggui Shaoyaosan at the low， medium， and high doses and estradiol valerate improved the estrus cycle of rats to varying degrees （<italic>P</italic><0.05） and increased the visceral index， with better effects observed in the medium-group and high-dose Danggui Shaoyaosan groups （<italic>P</italic><0.05，<italic>P</italic><0.01）. Besides， the protein expression levels of TGF-<italic>β</italic>₁ and TGF-<italic>β</italic>₁R and the mRNA expression levels of Smad2 and Smad3 in the ovarian tissues were elevated to varying degrees （<italic>P</italic><0.01）， and the Smad7 mRNA expression declined （<italic>P</italic><0.01）. The improvements in TGF-<italic>β</italic>₁ and TGF-<italic>β</italic>₁R protein expression of the medium-dose Danggui Shaoyaosan group and estradiol valerate group were more obvious. Conclusion:Danggui Shaoyaosan significantly improves ovarian reserve in DOR rats， which is closely related to the regulation of TGF-<italic>β</italic>₁/Smads signaling pathway.

Study on Network Pharmacology Mechanism of Liangfu Pills in the Treatment of Gastric Ulcer / 中国药房

OBJECTIVE：To study the m echanism of Liangfu pil ls in the tre atment of gastric ulcer. METHODS ：Based on network pharmacology ，the active ingredients ，targets and gastric ulcer related targets of Liangfu pill were screened by searching TCMSP，GeneCards，OMIM and DisGeNET databases ，and the common targets of Liangfu pill and gastric ulcer were screened by Venny 2.1 software. The protein interaction network of Liangfu pill in the treatment of gastric ulcer was constructed by STRING database. Cytoscape 3.7.2 software was used to construct topological analysis of the network.The function of GO and the enrichment of KEGG pathway were analyzed by DAVID database. Active component-gastric ulcer target-KEGG pathway network was constructed for Liangfu pill. In animal experiment ，mice were randomly divided into blank group ，model group ，ranitidine group （positive control ，39 mg/kg），Liangfu pill low- ，medium- and high- dose groups （0.78，1.56，3.12 g/kg），and were given corresponding drugs intragastrically ，once a day ，for consecutive 7 days. After last administration ，except for blank group ，the other groups were given absolute ethanol （0.01 mL/g）intragastrically to induce the gastric ulcer model. After modeling ，the pathological changes of gastric tissue were observed and the gastric ulcer index was calculated ；the expression of TP 53，c-Jun，p38 MAPK，Akt，p-Akt and TNF-α protein in gastric tissues of mice were detected. RESULTS：A total of 9 active ingredients （i.g. isorhamnetin ，β-gsterol，kaempferol）and 166 common targets were screened in Liangfu pills. Liangfu pill can treat gastric ulcer through GO functions such as cell response to chemical stimulation ，protein binding and extracellular space ，and KEGG pathways such as cancer signaling pathway ， hepatitis B ， Δ 基金项目：国家自然科学基金资助项目（No.81660649）；贵州省 bladder cancer ，pancreatic cancer ，TNF signaling pathway and 优秀青年科技人才项目（No.黔科合平台人才〔2019〕5658）；贵州中医 prostate. The results of animal experiments showed that 药大学博士启动基金项目 （No.2019-38）；贵州中医药大学科研项目 compared with model group ，Liangfu pills could significantly （No.贵中医科院内〔2019〕39号） ＊讲师，博士。研究方向：中药药效物质基础及作用机理。电话： reduce the pathological changes of gastric tissue and gastric 0851-88308060。E-mail：997845460@qq.com ulcer index ，as well as the protein expression of TP 53， # 通信作者：教授，博士。研究方向：中药药效物质基础及作用机 c-Jun，p38 MAPK，Akt，p-Akt and TNF-α（P＜0.05 or P＜ 理。E-mail：weina-0613@163.com 0.01）. CONCLUSIONS ：The 9 active components and 166 中国药房 2021年第32卷第9期 China Pharmacy 2021Vol. 32 No. 9 ·1063· targets of Liangfu pill were screened out. Liangfu pill can inhibit the expression of inflammatory factors by inhibiting MAPK ，NF-κ B，TNF and PI 3K/Akt signaling pathway ，and thus play an important role in improving gastric ulcer.

[Over-expression of circNR4A1/Hsa-circ-0026352 inhibits proliferation and induces apoptosis in MCF-7 cells].

Objective To investigate the expression of circNR4A1/Hsa-circ-0026352 in breast tumor tissue, peripheral blood, cell lines and its effect on the proliferation and apoptosis of MCF-7 breast cancer cells. Methods Using circRNA microarray, different expression of human circRNAs was found out between the breast cancer tissues and adjacent normal tissues. The expression of circNR4A1 in breast cancer tissues, peripheral blood and breast cancer cell lines including MCF-7, HCC-1937, MDA-MB-231, T-47D and MCF-10A were detected by quantitative real-time PCR (qRT-PCR). Circular RNA interactome software and KEGG software were used to predict its target miRNA and bio-function. The circNR4A1 over-expression vector was constructed and transfected into MCF-7 cells. The effect of circNR4A1 on the proliferation of MCF-7 cells was detected by CCK-8 assay and clone formation assay. The cell apoptosis and cell cycle distribution were analyzed by flow cytometry and Hoechst 33258 stainning. The qRT-PCR was performed to detect the mRNA expression levels of MEK/ERK signal pathway genes (K-RAS, MEK, ERK) and apoptosis-related genes (Bcl2, BAX, BAD, caspase-3). Western blotting was used to detect the expression of MEK, ERK, Bcl2, BAD and caspase-3 proteins. Results The expression of circNR4A1(Hsa-circ-0026352) was lower in breast cancer tissues, peripheral blood and breast cancer cell lines than in adjacent normal tissues, healthy individuals' peripheral blood and human breast epithelial cells. Over-expression of circNR4A1 in MCF-7 cells inhibited the cell proliferation, promoted cell apoptosis and blocked the cell cycle in G1/S phase. The mRNA and protein expression of MEK, ERK, Bcl2 were significantly depressed, while the expression of BAD, caspase-3 were enhanced in the MCF-7 cells over-expressing circNR4A1. Conclusion The expression of circNR4A1/Hsa-circ-0026352 is lower in breast cancer tissues, peripheral blood and breast cancer cells. Over-expression of circNR4A1 can inhibit cell proliferation and induce cell apoptosis in MCF-7 cells.