[Establishment and validation of nomogram prediction model for complicated acute appendicitis].

Objective: To establish and internally validate a nomogram model for predicting complicated acute appendicitis (CA). Methods: The clinical data from 663 acute appendicitis patients from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from October 2015 to October 2022 were retrospectively analyzed. There were 411 males and 252 females, aged (M (IQR)) 41 (22) years (range: 18 to 84 years). There were 516 cases of CA and 147 cases of uncomplicated acute appendicitis. The minimum absolute contraction and selection operator regression model was used to screen the potential relative factors of CA, and the screened factors were included in the Logistic regression model for multivariate analysis. Software R was used to establish a preoperative CA nomogram prediction model, the receiver operating characteristic curve of the model was drawn, and the value of area under the curve (AUC) was compared to evaluate its identification ability, and the Bootstrap method was used for internal verification. Results: The elderly (age≥60 years) (OR=2.428, 95%CI: 1.295 to 4.549), abdominal pain time (every rise of 1 hour) (OR=1.089, 95%CI: 1.072 to 1.107), high fever (body temperature≥39 ℃) (OR=1.122, 95%CI: 1.078 to 1.168), total bilirubin (every rise of 1 µmol/L) (OR=2.629, 95%CI: 1.227 to 5.635) were independent relative factors of CA (all P<0.05). The AUC of this model was 0.935 (95%CI: 0.915 to 0.956). After internal verification using the Bootstrap method, the model still had a high discrimination ability (AUC=0.933), and the predicted CA curve was still in good agreement with the actual clinical CA curve. Conclusion: The clinical prediction model based on the elderly (age≥60 years), prolonged abdominal pain time, high fever (body temperature≥39 ℃), and increased total bilirubin can help clinicians effectively identify CA.

Exploration at the network pharmacology level of possible targeting mechanisms of Smilacis Glabrae Rhixoma for the treatment of osteoporosis.

OBJECTIVE: The aim of this study was to evaluate the therapeutic effect of Smilacis Glabrae Rhixoma (SGR) on osteoporosis at the level of network pharmacology, and to find new targets and mechanisms of SGR in the treatment of osteoporosis, to better find new drugs and their clinical applications. MATERIALS AND METHODS: In the original network pharmacology mode, we used an improved mode, such as screening the ingredients and targets of SGR through tools such as GEO database, Autodock Vina, and GROMACS. Through molecular docking, we conducted further screening for the targets acting on the effective ingredients of SGR, and finally we performed molecular dynamics simulation and consulted a large amount of related literature for the validation of the results. RESULTS: By screening and validating the data, we finally confirmed that there were mainly 10 active ingredients in SGR, which were isoeruboside b, smilagenin, diosgenin, stigmasterol, beta-sitosterol, sodium taurocholate, sitogluside, 4,7-dihydroxy-5-methoxy-6-methyl-8-formyl-flavan, simiglaside B, and simiglaside E, and mainly acted on eleven targets. These targets mainly exert therapeutic effects on osteoporosis by regulating 20 signaling pathways including Th17 cell differentiation, HIF-1 signaling pathway, apoptosis, inflammatory bowel disease, and osteoclast differentiation. CONCLUSIONS: Our study successfully explains the effective mechanism by which SGR ameliorates osteoporosis while predicting the potential targets NFKB1 and CTSK of SGR for the treatment of osteoporosis, which provides a novel basis for investigating the mechanism of action of new Traditional Chinese medicines (TCMs) at the network pharmacology level and a great support for subsequent studies on osteoporosis.

[Effects of Sidaxue, a Miao ethnomedicine recipe, on apoptosis and pyrolysis of human fibroblast-like synovial cells in rheumatoid arthritis].

OBJECTIVE: To investigate the effects of Sidaxue (SX), a recipe in Miao ethnomedicine, on apoptosis and pyrolysis of human fibroblast-like synovial cells in rheumatoid arthritis (RA-FLS). METHODS: The target proteins related with RA and those involved in cell apoptosis and pyroptosis were searched in different online databases, and Venny software was used to obtain apoptosis- and pyroptosis-related proteins in RA. RA-apoptosis-pyroptosis protein interaction (PPI) network was constructed to identity the key target proteins related with apoptosis and pyroptosis in RA. Autodock vina software was used to perform molecular docking to verify the binding ability of the main active ingredients in SX with the apoptosis- and pyroptosis-related proteins. In the cell experiment, MH7A cells were treated with 5 mg/L TGT (positive control) or 5, 10, 20, and 40 mg/L SX, and the changes in cell migration and invasion abilities and expressions of apoptosis- and pyroptosis-related proteins were examined using wound healing assay, Transwell assay, ELISA and Western blotting. RESULTS: We identified 9 RA-related apoptotic target proteins, 15 RA-related pyroptosis target proteins, and 4 overlapping target proteins related with RA, apoptosis and pyroptosis. The main active ingredients in SX had a high affinity with the target proteins including TNF-α, Fas, and Bax. In MH7A cells, SX treatment concentration-dependently increased the cell inhibition rate at 24, 48 and 72 h (P < 0.05), and significantly lowered the cell migration ability at 6, 12 and 24 h (P < 0.05); treatment with 20 and 40 mg/L SX for 24 h obviously suppressed MH7A cell invasion (P < 0.05). SX treatment (20 and 40 mg/L) and TGT treatment both significantly lowered the expression levels of TNF-α, IL-1ß, and IL-18 in the cells (P < 0.05). The Bax/Bcl-2 ratio and Fas and FasL expressions were increased significantly in cells treated with 20 and 40 mg/L SX (P < 0.05), and caspase-1 expression was decreased significantly in cells treated with 5 and 40 mg/L SX (P < 0.05). CONCLUSION: SX can induce apoptosis and pyroptosison in RA-FLSs possibly by down-regulating the expressions of TNF-α, IL-1ß and IL-18, up-regulating the expressions of Bax, Fas, and FasL, and inhibiting Bcl-2 and caspase-1 protein expressions.

[Genetic and clinical analysis of X-linked hypophosphatemic rickets].

Objective: To investigate the clinical and genetic features, and treatment of X-linked hypophosphatemic rickets (XLH). Methods: In this retrospective study, we reviewed the medical records of 25 pediatric patients with XLH who were admitted to Department of Endocrinology Genetics and Metabolism,Beijing Children's Hospital from January 2010 to January 2020. The clinical characteristics, PHEX gene variants, as well as clinical outcome of the patients were summarized. To analyze the correlation between genotype and phenotype, the patients were divided into different subgroups according to the location of the variants, including N-terminal-located vs. C-terminal-located variant, and Zn-binding domain exon 17 or 19 variant vs. non-exon 17 or 19 variant. The age at onset, height standard deviation score (HtSDS), intercondylar or intermalleolar distance, fasting serum phosphorus, and HtSDS and intercondylar or intermalleolar distance at the final follow-up were compared by rank sum test or t text. Results: Among the 25 children with XLH, 8 were boys and 17 were girls. The median age of onset was 1.2 (1.0, 1.8) years, and the median age of diagnosis was 2.5 (1.5, 4.3) years. The main clinical manifestations were abnormal gait and lower limb deformity. The HtSDS was -2.0(-3.2, -0.8), and the intercondylar or intermalleolar distance was 4.5 (3.0, 6.0) cm. The fasting serum phosphorus level was 0.8 (0.7, 0.9) mmol/L, while the serum alkaline phosphatase level was (721±41) U/L and the serum calcium level was (2.5±0.1) mmol/L. Three patients (12%) had parathyroid hormone levels above the upper limit of the normal range. Twenty-five patients (100%) showed radiographic changes of active rickets. Nephrocalcinosis was found in 2 cases (9%). Twenty-four different PHEX variations were detected in 25 patients, among whom 11 (44%) had not been reported previously. No hot spot variation was found. No statistical differences (all P>0.05) were identified in clinical features and outcomes either in comparing patients with N-terminal (21 cases) and C-terminal (4 cases) variants, or in comparing patients with variant located in exon 17 or 19 (4 cases) or not (21 cases). Twenty-four cases (96%) were treated regularly with phosphate supplements and active vitamin D. After 2.7 (1.6, 5.0) years of follow-up, clinical symptoms were relieved in 96% (24/25) of the patients. The HtSDS after treatment had no significant difference compared to that before treatment (-2.0(-3.2, -0.8) vs.-2.0(-2.8, -1.1),Z =-0.156, P>0.05), while the intercondylar or intermalleolar distance after treatment was significantly reduced compared to that before treatment (4.5(3.0, 6.0) vs. 1.5(0, 3.3) cm, Z =-3.043, P<0.05). Bone X-rays were reexamined in 17 cases after treatment, and radiographic signs of rickets were improved. Eighteen cases had secondary hyperparathyroidism and 7 cases had nephrocalcinosis. Conclusions: The main clinical manifestations of XLH are abnormal gait, lower limb deformity and short stature. A high proportion of novel variations of PHEX gene but no hot spot variation neither genotype-phenotype correlation are found. Regular treatment with phosphate supplements and active vitamin D can significantly improve the symptoms except for the height. However, the rate of adverse events including secondary hyperparathyroidism and nephrocalcinosis seems to be high.

Potential probiotics Lactobacillus casei K11 combined with plant extracts reduce markers of type 2 diabetes mellitus in mice.

AIMS: Probiotics and plant extracts have been used to prevent the development of type 2 diabetes mellitus (T2DM). The study aimed to explore the effect of the interaction between potential probiotics and bitter gourd extract (BGE) or mulberry leaf extract (MLE) on T2DM. METHODS AND RESULTS: Potential probiotics were tested for their gastrointestinal tract viability and growth situation combined with BGE and MLE in vitro. The diabetes model was constructed in C57BL/6 mice, and the potential effect and mechanism of regulating blood glucose were verified. Hematoxylin-eosin staining (HE), gas chromatography (GC), ELISA, and RT-PCR were also used for analysis. The results showed that Lactobacillus casei K11 had outstanding gastrointestinal tract viability and growth situation with plant extracts. Administration of L. casei K11 combined with BGE and MLE significantly reduced blood glucose levels and ameliorated insulin resistance in diabetic mice than the administration of Lactobacillus paracasei J5 combined with BGE and MLE. Moreover, in L. casei K11 combined with BGE and MLE groups, lipid metabolism, oxidative stress, and proinflammatory cytokine levels were regulated. Furthermore, the results indicated that L. casei K11 combined with BGE and MLE improved free fatty acid receptor 2 (FFAR2) upregulation, glucagon-like peptide-1 (GLP-1) secretion, and short-chain fatty acid (SCFA) levels. CONCLUSIONS: These findings showed that L. casei K11 combined with BGE and MLE modified the SCFA-FFAR2-GLP-1 pathway to improve T2DM. SIGNIFICANCE AND IMPACT OF THE STUDY: This study identified a new modality for evaluating interactions between potential probiotics and plant extracts. Our findings revealed that L. casei K11 combined with BGE and MLE significantly promoted the SCFA-FFAR2-GLP-1 pathway to inhibit T2DM.

An LCI-like protein APC2 protects ginseng root from Fusarium solani infection.

AIMS: We aimed to purify an antimicrobial protein from Bacillus amyloliquefaciens FS6 culture supernatant, verify its antimicrobial activity against Fusarium solani and evaluate its biocontrol potential for ginseng root rot. METHODS AND RESULTS: The antimicrobial protein was purified from FS6 culture supernatant using ammonium sulphate precipitation, anion exchange and gel chromatography. Based on mass spectrometry results, the purified protein was identified as an antimicrobial protein of the LCI family and was designated APC2 . The APC2 recombinant protein expressed in Escherichia coli (BL21) significantly inhibited F. solani and decreased the infection and spread of F. solani in ginseng root. An overexpressing APC2 strain FS6-APC2 was constructed and shown to have enhanced antimicrobial activity compared to the wild-type strain FS6. CONCLUSIONS: The APC2 protein shows strong antimicrobial activity against F. solani, reduces the incidence and severity of ginseng root rot caused by F. solani and exhibits a great biocontrol potential. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the inhibitory activity of APC2 protein (LCI family) against F. solani and its protective efficacy on ginseng root rot. These findings provide a scientific basis for future research on the biocontrol mechanism, as well as the development and application of FS6.

Plant protein diet-induced hypoimmunity by affecting the spiral valve intestinal microbiota and bile acid enterohepatic circulation in Amur sturgeon (Acipenser schrenckii).

An 8-week growth trial was conducted to study enterohepatic recirculation of bile acid metabolism and the intestinal microbiota of Amur sturgeon (Acipenser schrenckii) fed with three diets, including 540 g/kg, 270 g/kg or 0 g/kg fishmeal, which was correspondingly replaced by a plant protein blend (named P0, P50 and P100, respectively). The diets were designed to be isonitrogenous, isoenergetic and essential nutrients balanced. With rising levels of dietary plant protein, disruption of the spiral valve intestinal microbiota and more morbidity with liver disease were observed in the P100 group, although there were no haematological abnormalities observed. An obvious bile acids enterohepatic circulation disorder was found with phenotypes of increased liver bile acids compensatory synthesis, and reduced expression of bile acid receptors (FXR and TGR5), which induced BA accumulative toxicity. Accompanied by increased oxidative stress, it further induced hepatic lesions and hypoimmunity, which were non-negligible reasons for the high mortality and low utilization ability of plant protein by Amur sturgeon.

[A multicenter randomized prospective study of concurrent chemoradiation with 60 Gy versus 50 Gy for inoperable esophageal squamous cell carcinoma].

Objective: To determine whether 60 Gy is superior to standard 50 Gy for definitive concurrent chemoradiation(CCRT) in esophageal squamous cell carcinoma (ESCC) using modern radiation technology in a phase Ⅲ prospective randomized trial. Methods: From April 2013 to May 2017, 331 patients from 22 hospitals who were pathologically confirmed with stage ⅢA-ⅣA ESCC were randomized to 60 Gy or 50 Gy with random number table. Total of 305 patients were analyzed, including 152 in 60 Gy group and 153 in 50 Gy group. The median age was 63 years, 242(79.3%) males and 63(20.7%) females. The median length of primary tumor was 5.6 cm. The clinical characteristics between two groups were comparable. All patients were delivered 2 Gy per fraction, 5 fractions per week. Concurrent weekly chemotherapy with docetaxel (25 mg/m(2)) and cisplatin (25 mg/m(2)) and 2 cycles consolidation chemotherapy with docetaxel (70 mg/m(2)) and cisplatin (25 mg/m(2), d1-3) were administrated. The primary endpoint was local/regional progression-free survival (LRPFS). The data were compared with Pearson chi-square test or Fisher's exact test. Results: At a median follow-up of 27.3 months, the disease progression rate was 37.5% (57/152), 43.8% (67/153) in the high and standard-dose group, respectively (χ(2)=1.251, P=0.263). The 1, 2, 3-year LRPFS rate was 75.4%, 56.8%, 52.1% and 74.2%, 58.4%, 50.1%, respectively (HR: 0.95, 95%CI: 0.69-1.31, P=0.761). The 1, 2, 3-year overall survival rate was 84.1%, 64.8%, 54.1% and 85.4%, 62.9%, 54.0%, respectively (HR: 0.98, 95%CI: 0.71-1.38, P=0.927). The 1, 2, 3-year progression-free survival rate was 70.8%, 54.2%, 48.5% and 65.5%, 51.9%, 45.1%, respectively (HR: 0.93, 95%CI: 0.68-1.26, P=0.621). The incidence rates in toxicities between the two groups were similar except for higher rate of severe pneumonitis in high dose group (χ(2)=11.596, P=0.021). Conclusions: The efficacy in disease control is similar between 60 Gy and 50 Gy using modern radiation technology concurrent with chemotherapy for ESCC. The 50 Gy should be recommended as the regular radiation dose with CCRT for ESCC.

[Clinical characteristics of patients with delayed hepatolenticular degeneration].

Objective: To evaluate the clinical manifestations, metal metabolism, imaging characteristics and treatment response in patients with delayed Wilson disease (WD). Methods: Patients with untreated WD (40 with delayed onset and 40 with non-delayed onset) were enrolled. Twenty healthy people were included as normal controls. All patients were evaluated with modified Young scale neural symptom scores, grade of Child liver function and mental symptoms rating scale, magnetic resonance imaging (MRI) scan, magnetic sensitive imaging (susceptibility weighted imaging, SWI), metal metabolism. Corrected phase (CP) was measured at SWI. After 2 week treatment, neurologic symptoms, liver function, and metal metabolism were reviewed. Results: The total score of neurological symptoms in WD patients with delayed onset was lower than that of non-delayed onset (13.00±6.87 vs. 21.13±5.53, P=0.033). The scores of SCL-90 and HAMA depression scales in patients with delayed onset were lower than those of non-delayed onset. On T(2) weighted imaging, areas including substantia nigra and thalamus, the caudate nucleus, globus pallidus, putamen presented high signal rate in patients with delated onset than those with non-delayed (P=0.022, 0.037, 0.022, 0.037, 0.029 respectively). The SWI CP values of cangbai sphere and shell nucleus in patients with delayed onset were lower than those with non-delayed onset. Patients with delayed onset had higher urinary copper than those with non-delayed onset before and after treatment (P=0.040, 0.036). After treatment, the score of abnormal tremor and gait in patients with delayed onset was decreased (P=0.037, 0.044), while as the occurrence of neurological symptoms was increased by 10%, and the liver function level in patients with delayed WD was decreased in 3 cases. Conclusions: The brain of WD patients with delayed onset is mainly composed of metal deposits, however the cell damage is not apparent. Clinical symptoms are characterized by significant liver injury, but relatively mild neurological and psychiatric symptoms. Patients with delayed WD have higher urinary copper excretion than those with non-delayed WD. Chelating agents improves the neurological symptoms in patients with delayed onset.

Dietary N-Carbamylglutamate (NCG) alleviates liver metabolic disease and hepatocyte apoptosis by suppressing ERK1/2-mTOR-S6K1 signal pathway via promoting endogenous arginine synthesis in Japanese seabass (Lateolabrax japonicus).

N-Carbamylglutamate (NCG), an analogue of N-acetylglutamate (NAG), can promote the synthesis of endogenous Arginine (Arg) in mammals, but not well studied in fish. This study was conducted to investigate the capacity of Arg endogenous synthesis by NCG, and the effects of various dietary NCG doses on growth performance, hepatic health and underlying nutrient regulation metabolism on ERK1/2-mTOR-S6K1 signaling pathway in Japanese seabass (Lateolabrax japonicus). Four experimental diets were prepared with NCG supplement levels of 0 (N0), 360 (N360), 720 (N720) and 3600 (N3600) mg/kg, in which N360 was at the maximum recommended level authorized by MOA, China in fish feed, and the N720 and N3600 levels were 2 and 10-fold of N360, respectively. Each diet was fed to 6 replicates with 30 Japanese seabass (initial body weight, IBW = 11.67 ±â€¯0.02 g) in each tank. The results showed that the dietary NCG supplementation had no significant effects on the SGR and morphometric parameters of Japanese seabass, but 360-720 mg/kg NCG inclusion promoted PPV, while the 10-fold (3600 mg/kg) overdose of NCG had remarkably negative effects with significantly reduced feed efficiency, PPV and LPV. We found that Japanese seabass can utilize 360-720 mg/kg NCG to synthesis Arg to improve the amino acid metabolism by increasing plasma Arg and up-regulating intestinal ASL gene expression. Increased plasma GST and decreased MDA indicated the improved antioxidant response. Dietary NCG inclusion decreased plasma IgM and down-regulated the mRNA levels of inflammation (TNF-α and IL8), apoptosis (caspase family) and fibrosis (TGF-ß1) related genes in the liver. The immunofluorescence examination revealed significantly decreased hepatic apoptosis and necrosis signals in the NCG groups. The ameliorated liver function and histological structure were closely related to the improved lipid metabolism parameters with decreased plasma VLDL and hepatic TG and NEFA accumulation, down-regulated fatty acid and cholesterol synthesis and simultaneously increased lipolysis gene mRNA levels, which regulated by inhibiting phosphorylation of ERK1/2-mTOR-S6K1 signaling pathway. Consuming 3600 mg/kg of dietary NCG is not safe for Japanese seabass culturing with the significantly increased FCR and decreased protein and lipid retention, and reduced plasma ALB. Accordingly, the observed efficacy and safety level of dietary NCG in the diet of Japanese seabass is 720 mg/kg.

Baicalin inhibits Escherichia coli isolates in bovine mastitic milk and reduces antimicrobial resistance.

In this study, we aimed to evaluate the inhibitory effect of baicalin on Escherichia coli in vitro and the effects of baicalin treatment on antimicrobial resistance of the E. coli isolates. Through isolation, purification, and identification, a total of 56 E. coli strains were isolated from 341 mastitic milk samples. The study of inhibition effect of baicalin on the E. coli strains in vitro was focused on permeability and morphology of the isolates using an alkaline phosphatase kit and scanning electron microscopy. Furthermore, the resistance spectrum of the isolates to the common antimicrobial agents was tested at sub-minimum inhibitory concentrations of baicalin by the agar dilution method. Extended-spectrum ß-lactamase and plasmid-mediated quinolone resistance genes were amplified by PCR before and after incubation with baicalin. The results revealed that baicalin has certain inhibitory effects on the isolates in vitro. The alkaline phosphatase enzyme activity was significantly increased from 1.246 to 2.377 U/100 mL, and the surface of E. coli was concave and shriveled. Analysis of the resistance spectrum and PCR amplification showed that, after administration with baicalin, the sensitivity of most strains to the selected antimicrobial agents was enhanced. Strikingly, the drug-resistant genes from 71.43% (40/56) of these isolates were found to have drug-resistant genes to different extents. Altogether, the current study confirmed both the inhibitory effect on Escherichia coli in vitro and the reduction of antimicrobial resistance by baicalin. This is the first comprehensive study to report on baicalin, a traditional Chinese medicine that acts on E. coli isolated from the mastitic milk samples.

Dietary butylated hydroxytoluene improves lipid metabolism, antioxidant and anti-apoptotic response of largemouth bass (Micropterus salmoides).

A 10-week growth trail was conducted to investigate the efficacy and tolerance of dietary butylated hydroxytoluene (BHT) by evaluating inflammation, apoptosis and hepatic disease related to oxidative stress in largemouth bass (Micropterus salmoides). Four experimental diets were prepared with BHT supplement levels of 0 (B0), 150 (B150), 300 (B300) and 1500 (B1500) mg/kg, in which B150 was at the maximum recommended level established by European Union Regulation, and the B300 and B1500 levels were 2 and 10-fold of B150, respectively. Each diet was fed to 6 replicates with 30 largemouth bass (initial body weight, IBW = 6.20 ± 0.01 g) in each tank. The BHT inclusion level did not affect the specific growth rate, but fish in the B150 group showed the lowest feed conversion rate (P < 0.05). BHT inclusion significantly decreased the levels of plasma TC, TG, LDL, ALT and AKP, and increased the (HDL-C)/TC ratio (P < 0.05). Plasma MDA was significantly decreased in the B150 group and GSH-Px was extremely enhanced in each BHT inclusion group (P < 0.05). Hepatic T-AOC was significantly enhanced and O2- was significantly decreased in each BHT inclusion group compared to the B0 group (P < 0.05), as well as hepatic MDA was significantly decreased in B1500 group (P < 0.05). Dietary BHT inclusion down-regulated the hepatic mRNA levels of inflammation, apoptosis and fibrosis related genes, including TNFα, TGF-ß1, α-SMA, IL8, IL11ß and caspase-9. Moreover, BHT could improve hepatic lipid metabolism via up-regulating the mRNA levels of APOA1, CYP7A1, CYP8B1, and down-regulating the mRNA levels of PPAR-γ and APOB. Histological examination of the liver morphology with H&E and Sirius Red staining showed that BHT inclusion decreased necrotic degenerative changes and collagen deposition in largemouth bass. An immunofluorescence examination revealed significantly decreased cleaved caspase-3 signals in the BHT groups. In conclusion, the results demonstrated that ROS induces hepatic cell apoptosis and fibrosis via the intrinsic pathway of apoptosis by activating caspase-9 in the mitochondria and then initiates apoptosis by activating caspase-3. Consuming 2.32-23.80 mg/kg·bw/d (150-1500 mg/kg in diet) of BHT effectively improved the plasma and hepatic lipid metabolism, antioxidant response as well as reduced ROS production, protecting hepatic cells from injury. It is implied that even a 10-fold increase of the maximum level of BHT (150 mg/kg) is safe for the largemouth bass.

Effects of Chinese herbal medicine and cold exposure on plasma glucose, leucine and energy metabolism in sheep.

An experiment was carried out to assess the feeding effect of Chinese herbal medicine on N balance, ruminal fermentation characteristics, kinetics of plasma glucose, leucine and energy metabolism in sheep kept at thermoneutral environment (23°C) or exposed to cold (2-4°C). Four sheep were subjected to either mixed hay (MH-diet) or hay supplemented with 2% of a traditional nourishing Chinese herbal medicine mixture (Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet) over two 23-day periods using a crossover design. Cold exposure was conducted for 5 days. The isotope dilution of [U-13 C]glucose with open circuit calorimetry was used to determine the turnover and oxidation rates of plasma glucose and metabolic heat production. The rate of plasma leucine turnover was measured with an isotope dilution method using [1-13 C]leucine. N intake was higher, N excretion through faeces was lower and N digestibility was higher for the CHM-diet than the MH-diet. Rumen pH was lower, concentration of rumen NH3 was higher, concentrations of rumen total VFA and acetate tended to be higher and propionate was higher for the CHM-diet compared with the MH-diet. Turnover rate of plasma glucose was higher for the CHM-diet than the MH-diet and increased during cold exposure. Oxidation rate of plasma glucose did not differ between diets and also between environments. Turnover rate of plasma leucine was higher for the CHM-diet compared with the MH-diet but remained similar between environments. Heat production was greater for the CHM-diet than the MH-diet and increased during cold exposure. No significant diet × environment interaction was detected. The present results demonstrated that plasma glucose and energy metabolism were enhanced by both Chinese herbal medicine and cold exposure; plasma leucine metabolism was enhanced by Chinese herbal medicine but with lack of change in response to cold exposure in sheep under the conditions of the current experiment.

Growth performance, fatty-acid composition, lipid deposition and hepatic-lipid metabolism-related gene expression in juvenile pond loach Misgurnus anguillicaudatus fed diets with different dietary soybean oil levels.

A 10 week feeding trial was conducted to evaluate the effects of different dietary soybean oil (SO) levels on growth performance, fatty-acid composition and lipid deposition in viscera, histology and histochemistry of liver, intestine and hepatic-lipid metabolism-related gene expressions in pond loach Misgurnus anguillicaudatus juveniles. Misgurnus anguillicaudatus (mean ± s.d. mass 0·40 ± 0·01 g) were fed five experimental diets containing SO at different concentrations: 0, 20, 32, 56 and 100% SO and a diet containing 100% fish oil (100% FO). The mass gains and specific growth rates of M. anguillicaudatus fed 20% SO and 100% FO diets were significantly higher than those of the other groups (P < 0·05). The lipid content of viscera and the amount of cytoplasmic vacuolation in the liver increased with incremental dietary SO level. Meanwhile, increasing dietary SO levels up-regulated the messenger (m)RNA levels of lipogenic genes (such as Δ6fad, scd, pparγ, fas and srebp-1) and down-regulated the mRNA levels of the lipolytic genes (such as pparα, cpt1, atgl and hsl) in the liver. The percentage of 20:4n-6 significantly (P < 0·05) increased with increasing dietary SO level, which might be correlated with the up-regulation of the mRNA level of Δ6fad. The highest levels of dietary SO, however, had a negative effect on growth performance, lipid deposition of viscera and histology and histochemstry of liver and intestine. The increased lipid accumulation induced by incremental dietary SO level probably occurred through different strategies for lipid metabolism as a result of competition between lipolysis and lipogenesis and between export and import of lipids in this species.

Molecular clocks and the human condition: approaching their characterization in human physiology and disease.

Molecular clockworks knit together diverse biological networks and compelling evidence from model systems infers their importance in metabolism, immunological and cardiovascular function. Despite this and the diurnal variation in many aspects of human physiology and the phenotypic expression of disease, our understanding of the role and importance of clock function and dysfunction in humans is modest. There are tantalizing hints of connection across the translational divide and some correlative evidence of gene variation and human disease but most of what we know derives from forced desynchrony protocols in controlled environments. We now have the ability to monitor quantitatively ex vivo or in vivo the genome, metabolome, proteome and microbiome of humans in the wild. Combining this capability, with the power of mobile telephony and the evolution of remote sensing, affords a new opportunity for deep phenotyping, including the characterization of diurnal behaviour and the assessment of the impact of the clock on approved drug function.

Vancomycin serum trough concentration vs. clinical outcome in patients with gram-positive infection: a retrospective analysis.

WHAT IS KNOWN AND OBJECTIVE: Vancomycin is still the first-line treatment for resistant gram-positive infections, particularly for methicillin-resistant Staphylococcus aureus (MRSA) infections. The vancomycin treatment guideline is based on the association between vancomycin trough concentration and clinical outcome. We here present a retrospective analysis of whether the trough level of vancomycin is associated with clinical outcome in Chinese patients with gram-positive infections. METHODS: Clinical data were collected retrospectively from patients under vancomycin therapeutic drug monitoring in Huashan Hospital from March 2004 to September 2014. RESULTS AND DISCUSSION: A total of 148 inpatients with gram-positive infection were identified and data on their corresponding vancomycin serum trough concentration retrieved. A total of 113 strains of gram-positive bacteria were isolated from 111 patients, including 90 strains of MRSA. Vancomycin was used for 11 to 13 days on average. The overall bacterial eradication rate was 67·3% (76/113), including 61·1% (55/90) for MRSA and 91·3% (21/23) for Enterococcus. Multivariate logistic model analysis showed that vancomycin trough concentration was not associated with clinical outcome (OR: 1·0; 95% CI: 0·92, 1·08, P = 0·9613). The incidence of adverse drug reactions was low and not related to vancomycin trough concentration. WHAT IS NEW AND CONCLUSION: This retrospective analysis failed to demonstrate an association between vancomycin trough concentration and the clinical and microbiological response. Prospective controlled studies are necessary to further establish the need for the higher trough concentrations normally cited for clinical efficacy.

Effects of conservative laparoscopic endometrial cystectomy and use of kidney-reinforcing and blood activating traditional Chinese medicine on ovarian functions.

This study investigated functional changes in the ovaries of women who have had conservative laparoscopic endometrial cystectomy and the effects of traditional kidney-reinforcing and blood-activating Traditional Chinese Medicine (TCM) on ovarian function. Seventy female patients who underwent laparoscopic endometrial cystectomy were randomized into two groups: TCM group receiving kidney-reinforcing and blood-activating Chinese medicine (N = 35) and control group receiving only routine follow-up (N = 35). The serum levels of all study participants were measured for follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and inhibin B (INHB) prior to and after endometrial surgery. Postoperative menstruation conditions were also assessed. Compared to preoperative conditions, both LH and FSH levels during the postoperative 1st month increased in both groups, while E2 and INHB levels decreased (P < 0.05). In the TCM group, in contrast to the control group and the postoperative 1st month, we observed a decrease in LH and FSH levels during the postoperative 4th month, while E2 and INHB levels increased (P < 0.05). In this study, we found that certain TCM prescriptions lowered postoperative serum FSH and LH levels and increased the serum INHB and basal E2 levels, thereby improving the ovarian reserve.

Optimization of culture medium for anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl for microbial enhanced oil recovery.

UNLABELLED: Response surface methodology was employed to enhance the anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl. Glycerol is a promising carbon source used to anaerobically produce rhamnolipid. In a Plackett-Burman design, glycerol, KH2 PO4 and yeast extract were significant factors. The proposed optimized medium contained the following: 46·55 g l(-1) glycerol; 3 g l(-1) NaNO3 ; 5·25 g l(-1) K2 HPO4 ·3H2 O; 5·71 g l(-1) KH2 PO4 ; 0·40 g l(-1) MgSO4 ·7H2 O; 0·13 g l(-1) CaCl2 ; 1·0 g l(-1) KCl; 1·0 g l(-1) NaCl; and 2·69 g l(-1) yeast extract. Using this optimized medium, we obtained an anaerobic yield of rhamnolipid of 3·12 ± 0·11 g l(-1) with a 0·85-fold increase. Core flooding test results also revealed that Ps. stutzeri Rhl grown in an optimized medium enhanced the oil recovery efficiency by 15·7%, which was 6·6% higher than in the initial medium. Results suggested that the optimized medium is a promising nutrient source that could effectively mobilize oil by enhancing the in situ production of rhamnolipid. SIGNIFICANCE AND IMPACT OF THE STUDY: The ex situ application of rhamnolipid for microbial enhanced oil recovery (MEOR) is costly and complex in terms of rhamnolipid production, purification and transportation. Compared with ex situ applications, the in situ production of rhamnolipid in anaerobic oil reservoir is more advantageous for MEOR. This study is the first to report the anaerobic production optimization of rhamnolipid. Results showed that the optimized medium enhanced not only the anaerobic production of rhamnolipid but also crude oil recovery.

Effect of You Gui Wan on mouse sperm fertilising ability in vivo and in vitro.

This study is to explore whether YGW has an impact on sperm fertilising ability in mice. Twenty male mice were randomly divided into two groups. In vivo experiments, one group of animals were orally administrated with YGW decoction and another group administered with saline for 14 days. Afterwards, the animals were mated with their female partners. Percentages of retrieved zygotes were then compared. In vitro experiments, in vitro fertilisation (IVF) assay, sperm acrosome reaction and acrosin activity were used to compare sperm fertilising ability between the two groups. The YGW-treated group had a significantly higher percentage of zygotes than the saline controls (P = 0.005). The IVF rates induced by spermatozoa from the herb-treated mice were also significantly higher than those from the control animals (P = 0.015). The sperm acrosin activity of the herb-treated group was significantly higher than that of the saline-treated group (P = 0.048), although there was no significant difference in testicular weight, sperm count and sperm motility. These data suggest that YGW decoction has a significant effect on normal sperm fertilising ability both in vivo and in vitro, which may be due to, at least in part, increments in the sperm acrosin activity.

Mitigation of nutrient losses via surface runoff from rice cropping systems with alternate wetting and drying irrigation and site-specific nutrient management practices.

Resource-conserving irrigation and fertilizer management practices have been developed for rice systems which may help address water quality concerns by reducing N and P losses via surface runoff. Field experiments under three treatments, i.e., farmers' conventional practice (FCP), alternate wetting and drying (AWD), and AWD integrated with site-specific nutrient management (AWD + SSNM) were carried out during two rice seasons at two sites in the southwest Yangtze River delta region. Across site years, results indicated that under AWD irrigation (i.e., AWD and AWD + SSNM), water inputs were reduced by 13.4~27.5 % and surface runoff was reduced by 30.2~36.7 % compared to FCP. When AWD was implemented alone, total N and P loss masses via surface runoff were reduced by 23.3~30.4 % and 26.9~31.7 %, respectively, compared to FCP. However, nutrient concentrations of surface runoff did not decrease under AWD alone. Under AWD + SSNM, total N and P loss masses via surface runoff were reduced to a greater extent than AWD alone (39.4~47.6 % and 46.1~48.3 % compared to FCP, respectively), while fertilizer inputs and N surpluses significantly decreased and rice grain yields increased relative to FCP. Therefore, by more closely matching nutrient supply with crop demand and reducing both surface runoff and nutrient concentrations of surface runoff, our results demonstrate that integration of AWD and SSNM practices can mitigate N and P losses via surface runoff from rice fields while maintaining high yields.