[Mechanism of Qingluo Tongbi Formula for regulating immune-bone erosion in rheumatoid arthritis].

OBJECTIVE: To explore the mechanism of Qingluo Tongbi formula for regulating "immune-bone erosion" in rheumatoid arthritis (RA). METHODS: Sixty-four RA patients were randomized into two groups to receive treatment with oral methotrexate or Qingluo Tongbi Formula for 12 weeks. Flow cytometry was used to analyze the changes in the percentages of CD3-CD19+, CD19+CD27 and CD19+BAFFR+B cell subpopulations in peripheral blood of the patients, and serum levels of B cell activating factor (BAFF), RANKL, RANK and osteoprotegerin (OPG) levels were detected using ELISA. Before and after the treatment, serum levels of ß-CTX, TRACP-5b, BGP, BALP, and PINP were measured with ELISA, and bone mineral density was determined with DXEA dual-energy X-ray absorptiometry. In the cell experiment, RAW264.7 cells were induced to differentiated into osteoclasts and treated with Qingluo Tongbi Formula at low-, moderate and high doses (125, 250 and 500 µg/mL, respectively) or with methotrexate (2 µg/mL) for 48 h, and the changes in the expression levels of RANKL, RANK, OPG and c-Fos were detected using Western blotting. RESULTS: The B cell subgroups in RA patients were correlated with the RANKL/RANK/OPG system. Treatment with Qingluo Tongbi Formula obviously down-regulated the percentages of the B cell subgroups, lowered serum levels of BAFF, ß-CTX and TRACP-5b, increased the levels of BGP, BALP and PINP, and improved lumbar bone density of RA patients (P<0.05); All these changes were significantly correlated with the regulation of B cell expressions (P<0.05). In RAW264.7 cells-derived osteoclasts, Qingluo Tongbi Formula significantly decreased the expressions of RANKL, RANK and c-Fos and increased the expression of OPG (P<0.05). CONCLUSION: Qingluo Tongbi Formula inhibits bone erosion in RA possibly by regulating B cell subset percentages and BAFF expression and inhibiting osteoclast differentiation via the RANKL/RANK/OPG pathway.

[The distribution of musk and river deer in local chronicles in Guizhou].

Some information and records about musk (She, ) and river deer (Zhang, /) can be found in local chronicles or documents in Guizhou. They were taken as the same species in terms of a medicinal animal. The records for their living areas in Guizhou were neither clear nor in detail in the Ming Dynasty, but were specific in the Qing Dynasty and more concise in the period of the Republic of China. The living areas for musk and river deer reduced from the Ming Dynasty to the Republic of China. Such change was believed to be the result of the natural environment and the social factors as well, such as the reclamation of mountain areas in Guizhou, the reduction of forests, and the demand and resulting exorbitant prices that led to excessive hunting.

Treatment of secondary lower limb lymphedema after gynecologic cancer with complex decongestive therapy.

Secondary lower extremity lymphedema is a common complication of treatment for gynecological cancers. Conservative therapy plays an important role in the treatment of patients with secondary lower extremity lymphedema; in particular, complex decongestive therapy (CDT) has been recognized as an effective nonoperative technique for these patients. But CDT therapy for secondary lower extremity lymphedema remains a problem in China because this technique and its effectiveness have not achieved widespread use and popularity. Our goal was to assess effects of CDT in patients with secondary lower limb lymphedema after treatment for gynecological cancers. The retrospective study consisted of 60 patients who were treated with 20 sessions of CDT. Assessments included objective changes in limb circumference, degree of LE, imaging features, and incidence of erysipelas before and after CDT treatment. We found that CDT can effectively improve lymph stasis and promote backflow, and decrease circumference, interstitial fluid content, and incidence of erysipelas of lymphedematous lower limb. Our results demonstrate that CDT is an effective treatment method for patients with secondary lower limb lymphedema following treatment for gynecologic cancers. This technique should be more widely utilized and popularized in China to improve the quality of life of millions of patients with secondary lower limb lymphedema.

[Association of vitamin D nutritional status with calcaneal bone mineral density in school-age children: a prospective cohort study].

Objective: To investigate the relationships between vitamin D nutritional status and the calcaneal bone mineral density (BMD) in children. Methods: Data were obtained from School-based Cardiovascular and Bone Health Promotion Program. In 2017, a total of 15 391 children aged 6-16 years in Beijing selected through stratified cluster sampling were included in the baseline survey. A follow-up investigation was conducted in 2019. The questionnaire survey, detection of serum 25-hydroxyvitamin D [25(OH)D] level and ultrasound measurement of calcaneal BMD were conducted. Multivariable linear and logistic regression models were used to analyze the relationships between baseline vitamin D nutritional status and the follow-up calcaneal BMD. Results: A total of 10 914 children aged (11.5±3.3) years (boys accounting for 49.6%) were included in the analysis. The average 25(OH)D level was (35.4±12.0) nmol/L, and the deficiency rate was 36.1%. After the adjustment for age, gender, body mass index, smoking status, alcohol use status, dairy products intake, vitamin D supplement, calcium supplement, physical activity, pubertal development, and baseline calcaneal BMD Z-score, for per 10 nmol/L increase in 25(OH)D, the follow-up calcaneal BMD Z-score increased by 0.01(P=0.041), and the OR(95%CI) of decreased calcaneal BMD Z-score after 2 years was 0.96 (0.93-1.00)(P=0.030). Compared with vitamin D adequacy, the follow-up calcaneal BMD Z-score of children with vitamin D insufficiency and deficiency decreased by 0.03(P=0.307) and 0.06 (P=0.046), and the risk of decreased calcaneal BMD Z-score after 2 years increased by 15%(P=0.037) and 21%(P=0.006), respectively (P for trend<0.05). Conclusions: Vitamin D nutritional status was closely related to calcaneal BMD, and children with adequate vitamin D nutritional status tended to obtain higher BMD. Children and adolescents are encouraged to maintain sufficient vitamin D levels, strengthen nutrition and exercise to promote bone health.

[Association of vitamin D nutritional status with body muscle mass in school-age children adolescents].

Objective: To investigate the association between vitamin D nutritional status and the body muscle mass in children. Methods: Data were obtained from School-based Cardiovascular and Bone Health Promotion Program. In 2017, a total of 15 391 children aged 6-16 years in Beijing were selected through stratified cluster sampling in baseline survey. A follow-up investigation was conducted in 2019. The questionnaire survey and the detection of serum 25-hydroxyvitamin D [25(OH)D] level were conducted. The bioelectrical impedance analysis (BIA) apparatus was used to measure body muscle mass, and muscle mass index (MMI) was calculated. Multivariable linear models were used to analyze the association of vitamin D nutritional status with the baseline and follow-up MMI measures. Results: A total of 10 890 children aged (11.5±3.3) years(boys accounting for 49.6%) were included in the analysis. The average 25(OH)D level was (35.4±12.0) nmol/L, with an adequacy ratio of 11.1%. After multivariate linear regression adjustment for age, sex, body fat mass, smoking status, alcohol use status, dairy supplement, calcium supplement, physical activity, and pubertal development, no statistically significant association between vitamin D nutritional status and baseline MMI level was observed (P>0.05). For the follow-up MMI, the Z-score increased by 0.008 (P=0.058) for per 10 nmol/L increase in 25(OH)D, which were 0.002 (P=0.815) and 0.037 (P=0.031) higher in children with insufficient and adequate vitamin D than those with vitamin D deficiency, respectively (P for trend =0.089). Subgroup analysis showed that in the normal BMI group, for per 10 nmol/L increase in 25 (OH) D, the MMI at baseline survey and MMI Z-score at follow-up of children with adequate vitamin D and increased by 0.019 and 0.014, respectively (both P<0.05). Conclusions: Vitamin D nutritional status was related to muscle mass in children, and children with adequate vitamin D tended to obtain higher MMI. Children and adolescents are encouraged to maintain sufficient vitamin D levels, strengthen nutrition and exercise to promote body health.

Supplementation with Yucca schidigera improves antioxidant capability and immune function and decreases fecal score of dairy calves before weaning.

Yucca schidigera (YS) is a species of plant rich in antimicrobials, antioxidants, and immunomodulators. It has been used as feed additive to improve animal performance and decrease methane emissions in cattle. However, few studies have evaluated YS in dairy calves. In this study, we evaluated the effects of YS on the growth performance, antioxidant capacity, and immune function in dairy calves before weaning. We randomly assigned 40 newborn female Holstein calves (4 d old; 40 ± 5 kg of body weight) to 1 of 4 treatments (n = 10 per treatment), which were fed 0, 3, 6, or 9 g/d of YS powder. The YS allowance was mixed into milk or milk replacer and fed twice daily. Dry matter intake (both liquid and starter feed) and fecal score were recorded daily, and body weight, withers height, body length, and heart girth were measured at 4, 14, 28, 42, and 60 d of age. Blood was sampled from the jugular vein at 14, 42, and 60 d of age after the afternoon feeding for analysis of serum antioxidant capacity and immune function. Feeding YS did not affect dry matter intake, but decreased the feed-to-gain ratio with a quadratic dose effect. Over the whole study period, the average daily gain tended to linearly increase with the increasing YS doses, and it was 6.8% higher in diets supplemented with 9 g/d of YS than in the basal control diet without YS. The YS supplementation linearly decreased fecal score in a dose-dependent manner, and the frequency of diarrhea was significantly decreased as the YS supplementation increased throughout the whole study period. The YS supplementation also linearly decreased maleic dialdehyde concentration in the serum compared with the control group. The activity of catalase tended to linearly and quadratically increase, and that of glutathione peroxidase increased linearly with the increased YS supplementation. Serum concentrations of IgA and IgG increased linearly with the increased YS supplementation, and that of IgG tended to increase quadratically. To the best of our knowledge, this is the first study that demonstrated that feeding YS to young calves could improve growth, feed efficiency, and immunity, and decrease fecal score and diarrhea. The results of this study indicated that feeding YS at 9 g/d may be recommended to benefit dairy calves before weaning.

[The change in nutritional status of pancreatic cancer patients undergoing neoadjuvant therapy and its influence on these patients' postoperative outcomes as well as prognosis].

Neoadjuvant therapy has been one of the standard therapies for borderline resectable pancreatic cancer and locally advanced pancreatic cancer, but it may deteriorate these patients' nutritional status while controlling the progress of cancer, which inevitably influences these patients' postoperative outcomes and prognosis.In this article, we tried to answer this problem by reviewing other studies.And we found that neoadjuvant therapy would influence patients' postoperative outcomes and prognosis by deteriorating their nutritional status.But effective nutritional support can prevent it.It indicates that there is a need for these patients to receive nutritional support as soon as possible.However, in consideration of the limited number of relevant studies and their limitations, there is a need for more studies with strict design to answer this problem.And it can provide evidence for early nutritional support in pancreatic cancer patients who is going to undergo neoadjuvant therapy.

Dietary supplementation of total flavonoids from Rhizoma Drynariae improves bone health in older caged laying hens.

Caged layer osteoporosis (CLO) is a common bone metabolism diseases and poses a great threat to the production of laying hens. So far, there is no effective nutrition intervention to prevent CLO. The objective of this study was to evaluate the effects of dietary total flavonoids from Rhizoma Drynariae (TFRD), a Chinese herbal, on bone health, egg quality, and serum antioxidant capacity of caged laying hens. A total of two hundred sixteen, 54-wk-old Lohmann Pink-shell laying hens at were allocated to 3 groups with 6 replicates of 12 hens per replicate. The control group was fed a basal diet (BD) and 2 treatment groups additionally supplied with 0.5 or 2.0 g/kg TFRD, respectively. Results showed that supplying 2.0 g/kg TFRD enhanced the activities of serum total antioxidant capacity (P < 0.01) and glutathione peroxidase (P < 0.05) and had higher femur and tibia bone mineral density (both P < 0.05) compared with the control group. Dietary 2.0 g/kg TFRD also reduced the activities of serum alkaline phosphatase (P < 0.01), tartrate resistant acid phosphatase (P < 0.01), and the contents of osteocalcin (P < 0.01). Furthermore, tibia histomorphology observation showed that the microstructure of bone tissue was improved after TFRD treatment. Egg quality was not affected by TFRD while the egg weight significantly increased (P < 0.01). These findings suggested that TFRD has beneficial effects on bone health in older caged laying hens.

Diversity of sugarcane root-associated endophytic Bacillus and their activities in enhancing plant growth.

AIMS: Plant tissues are the reservoirs of beneficial and harmful microbes that regulates plant growth. In the present study, we investigated the diversity, function and colonization of sugarcane roots associated with Bacillus spp. METHODS AND RESULTS: A total of 20 Bacillus strains were isolated and identified by 16S rRNA gene sequencing, and their genetic diversity was examined by BOX, ERIC, REP, (GTG)5 PCR techniques. Among all Bacillus isolates, 65% showed indole acetic acid-like compounds production, 50% solubilized phosphorus and 25% of the isolates were able to secrete siderophore. Moreover, all 20 Bacillus isolates showed antifungal activity against eight fungal pathogens and 11 of them (55%) antagonized tomato grey mold. Based on the plant growth-promoting traits and antifungal potential, isolate Y8 was selected for root and plant tissue colonization assays and a greenhouse-level sugarcane growth promotion study. Fluorescence microscopy results confirmed that isolate Y8 has a strong ability to colonize in the sugarcane root and leaves, and the root surface association of Y8 was confirmed by scanning electron microscopy. Furthermore, greenhouse experimental results demonstrated that Y8 has a significant effect on enhancing sugarcane biomass and root length. CONCLUSIONS: Endophytic Bacillus strains have growth-promoting properties and anti-fungal ability that can enhance plant fitness in an eco-friendly manner. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic Bacillus strains would be a potential alternative to chemical fertilizer as well as a biocontrol agent in the future.

[The association between body fat distribution and calcaneal bone mineral density in children and adolescents aged 6-16 years in Beijing].

Objective: To understand the association between body fat distribution and calcaneal bone mineral density (BMD) in children and adolescents aged 6-16 years in Beijing. Methods: Children and adolescents were selected in 30 schools (8 primary schools, 21 middle schools and one 12-year education school) from Dongcheng, Tongzhou, Fangshan and Miyun districts of Beijing by using a stratified cluster sampling method from November 2017 to January 2018. A total of 15 030 students in grade 1 to 4 of primary school, grade 1 of junior and senior middle school were enrolled after excluding subjects who were not able to participate into this study due to trauma or other uncomfortable physical conditions or with missing key values or with diabetes and kidney diseases. Questionnaire survey, body composition and calcaneus bone mineral density were conducted. The relation of fat mass percentage (FMP), trunk to total fat ratio (TrTFR), trunk to limb fat ratio (TrLFR), limb to total fat ratio (LTFR) and viscera to total fat ratio (VTFR) with calcaneus BMD were assessed using the multivariate linear regression models after adjusting for possible confounding effects. The central fat distribution types were divided into 4 groups including type 1: both trunk fat and visceral fat greater than the sex-and age-specific internal P(75); type 2: barely trunk fat greater than P(75); type 3: barely visceral fat greater than P(75); type 4: both trunk fat and visceral fat greater than P(75). The central fat distribution types were included into the model in the form of dummy variables to analyze its relationship with calcaneal BMD. The sex-and age-specific z-scores of fat distribution indicators and BMD were calculated. Results: A total of 15 030 participants aged (11.4±3.3) years (50.2% boys) were involved in the analysis. In both genders, after adjusting for age, height, lean mass index, smoking, drinking, physical activity, milk intake, vitamin D and calcium supplementation, FMP, TrTFR, TrLFR and VTFR were negatively correlated with calcaneal BMD (all P value<0.05), while LTFR was positively associated with calcaneal BMD (all P values<0.05). Compared to the central fat distribution type 1, the regression coefficients (95% CI) of type 2, 3 and 4 were -0.253 (-0.418, -0.087), -0.385 (-0.567, -0.204) and -0.428 (-0.487, -0.369) in boys, respectively; the regression coefficients (95% CI) of type 3 and 4 were -0.158 (-0.301, -0.015) and -0.226 (-0.290, -0.163), respectively. Conclusion: Body fat distribution and central fat distribution in children and adolescents were correlated with calcaneus bone mineral density.

[The association between blood lipids and calcaneus bone mineral density in children and adolescents aged 6-16 years in Beijing].

Objective: To investigate the association between blood lipid and calcaneus bone mineral density (BMD) in children and adolescents aged 6-16 years in Beijing. Methods: Children and adolescents were selected in 30 schools (8 primary schools, 21 middle schools and one 12-year education school) from Dongcheng, Tongzhou, Fangshan and Miyun districts of Beijing by using a stratified cluster sampling method from November 2017 to January 2018. A total of 14 303 students in grade 1 to 4 of primary school, grade 1 of junior and senior middle school were enrolled after excluding subjects who were not able to participate into this study due to trauma or other uncomfortable physical conditions or with missing key values or with diabetes and kidney diseases. Questionnaire survey, blood lipid and calcaneus BMD were conducted. Multivariate linear regression was applied to quantify the association between calcaneal BMD as a dependent variable and blood lipid level as an independent variable after adjusting for the potential confounding factors. Furthermore, quantile regression was used to analyze the association between blood lipid level and different percentiles (P(25), P(50) and P(75)) of ultrasonic velocity values of bone mineral density, and parallel test was conducted for regression coefficients of different percentiles. Results: A total of 14 303 participants aged (11.4±3.3) years (49.9% boys) were involved in the analysis. The mean age of 14 303 participants was (11.0±3.3) years. 7 142 boys accounted for 49.9%. The mean±SD of calcaneal BMD, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), and high density lipoprotein-cholesterol (HDL-C) were (1 540.9±33.8) m/s, (3.90±0.76), (2.18±0.62), and (1.40±0.32) mmol/L, respectively. The P(5)0 (P(25), P(75)) of triglyceride (TG) was 0.69 (0.49-0.94) mmol/L. After the adjustment of age, height, smoking, drinking, vitamin D and calcium supplementation, dairy intake, physical activity, FMI, and MMI, a significantly inverse association (P<0.05) between TG level and calcaneus BMD was observed in both genders, which the regression coefficients (95%CI) in boys and girls were -0.064 (-0.085, -0.044) and -0.073 (-0.094, -0.053), respectively. Conclusion: The level of BMD was associated with TG in boys and girls. Therefore, it is important to prevent children from hypertriglyceridemia for the bone health promotion.

Electro-acupuncture at ST37 and ST25 induce different effects on colonic motility via the enteric nervous system by affecting excitatory and inhibitory neurons.

BACKGROUND: On the basis of the importance of the enteric nervous system (ENS) in gastrointestinal motility, we hypothesized that the ENS may mediate the therapeutic efficacy of electro-acupuncture (EA) in constipation by regulating the mechanisms underlying the effects of EA on gastrointestinal function. METHODS: Model mice with constipation were generated by gastric instillation of 0-4°C normal saline. Defecation time and stool (form and wet and dry weight) were assessed. The effect of EA at ST37 or ST25 on colorectal motility and proximal colonic motility was assessed using a water-filled balloon. The expression of protein gene product 9.5 (PGP9.5), the cholinergic neuron marker acetyltransferase (ChAT) and the anticholinergic neuron marker nitric oxide synthase (nNOS) was detected by immunohistochemistry, real-time quantitative polymerase chain reaction (qPCR) and western blot analysis. KEY RESULTS: ST37 and ST25 improved colorectal pressure; however, ST37 but not ST25 improved proximal colonic pressure. In the proximal colon, the expression of PGP9.5 returned to normal after EA at ST 37, while EA at ST25 did not have this effect. In addition, qPCR and western blot analysis showed that ST37 could downregulate the expression of nNOS and upregulate the expression of ChAT to normal levels, while ST25 could only downregulate the expression of nNOS to normal levels. CONCLUSIONS AND INFERENCES: Electro-acupuncture at specific acupoints can improve intestinal motility in constipation by altering the ENS and differentially affecting excitatory and inhibitory neurons, restoring the coordination between contraction and relaxation muscles, and working in concert with the central nervous system and peripheral neural pathways.

[Clinical effect of alanyl glutamine in the treatment of patients with gastrointestinal function obstacle caused by severe phorate poisoning].

Objective: To observe the therapeutic efficacy of alanyl glutamine injection on patients with gastrointestinal function obstacle caused by severe phorate poisoning. Methods: A total of 80 eligible patients with gastrointestinal function obstacle caused by severe phorate poisoning were randomly divided into the control group (n=40) and treatment group (n=40) . The control group was treated with the conventional therapy, which included forbidden diet, atropine, pralidoxime iodide, anti-inflammatory, albumin infusion, ω-3 fish oil fat emulsion, protection of organs function, blood perfusion, and Fat Emulsion, Amino Acids (17) and Glucose Injection. The treatment group was treated with alanyl glutamine injection plus the conventional therapy. To observe the time of recovering to normal of gastrointestinal function between the two groups, compared the AChE activity and changes of prealbumin, albumin and total protein of the two groups respectively. Furthermore, the total atropine dosage, the total pralidoxime iodide dosage and ICU stay time between the two groups were also compared. Results: The gastrointestinal function recovery time of patients in the treatment group was less than the control group, the difference was statistically significant (P<0.05) . From the third day of treatment, the serum cholinesterase activity of the treatment group was higher than the control group, the difference was statistically significant (P<0.05) . On the 5th day and 10th day of the treatment, the prealbumin, albumin and total protein of the treatment group were significantly higher than these indexes of the control group in the same period, the difference were statistically significant (P<0.05) . The total atropine dosage, the total pralidoxime iodide dosage and ICU stay time in the treatment group were lower than the control group, the difference were statistically significant (P<0.05) . Conclusion: Alanyl glutamine injection has a great therapeutic effect for gastrointestinal function obstacle patients caused by severe phorate poisoning.

[Systematic review on orofacial myofunctional therapy to treat obstructive sleep apnea-hypopnea syndrome].

Obstructive sleep apnea-hypopnea syndrome (OSAHS) is an important risk factor for cardiovascular and cerebrovascular diseases. The widespread collapse of the airway, which can't maintain its normal position and structure, is an important cause of OSAHS. Noninvasive ventilator-assisted ventilation is the preferred treatment for OSAHS, but there are shortcomings of poor compliance. Orofacial myofunctional therapy (OMT) is a noninvasive, simple, low-cost, low-risk therapy that is expected to replace noninvasive ventilator-assisted ventilation to a certain extent. So far, there is little Chinese literature on OMT, especially no training methods in Chinese version. Therefore, this paper systematically reviewed anatomical abnormalities and pathology of OSAHS, possible treatment mechanism of OMT and related research progress. And we translated two kinds of training method of OMT (Guimarase's and Hemmat Baz's) into Chinese for researchers and clinicians' reference, which is widely cited in foreign literature.

Protective effect of quercetin on acute lung injury in rats with sepsis and its influence on ICAM-1 and MIP-2 expression.

This study aimed to explore the protective effect of quercetin on acute lung injury (ALI) in rats with sepsis and the related mechanism. Rats were administered different doses of quercetin intraperitoneally, and blood samples and lung tissue were collected at 24 h after treatment. Arterial blood gases, lung water content, protein content, and cell counts in bronchoalveolar lavage fluid (BALF) were measured. Morphological changes in lung tissue pathology were observed under a light microscope. Serum intercellular adhesion molecule (ICAM)-1 and macrophage inflammatory protein 2 (MIP-2) levels were detected and ICAM-1 and MIP-2 mRNA expression in lung tissue was determined. Compared with that in the control model group, arterial blood gases, lung water content, protein content, and cell counts in BALF improved in the high- and low-dose quercetin groups (P < 0.05), with maximal improvement observed for the high-dose quercetin (P < 0.05). Lesions on the lungs improved in the high- and low-dose quercetin groups than those in the control model group, and the high-dose quercetin group showed better improvement than the low-dose group (P < 0.05). Compared with that in the sham-operated group, both serum and lung tissue ICAM-1 and MIP-2 expression increased significantly in the model group (P < 0.05). The quercetin groups presented lower ICAM-1 and MIP-2 expression than the control model group, with the lowest expression observed in the high-dose group (P < 0.05). Quercetin may protect against ALI in rats with sepsis by inhibiting ICAM-1 and MIP-2 expression.

Characterization of the Microbial Communities in the Ant Lion Euroleon coreanus (Okamoto) (Neuroptera: Myrmeleontidae).

Euroleon coreanus (Okamoto) is widely distributed in China, and the larval stage can be treated as traditional Chinese medicine. However, the host-bacterium relationship remains unexplored, as there is a lack of knowledge on the microbial community of ant lions. Hence, in the current study, we explored the microbial community of the larval ant lion E. coreanus using Illumina MiSeq sequencing. Results indicated that a total of 10 phyla, 126 genera, and 145 species were characterized from the second instars of E. coreanus, and most of the microbes were classified in the phylum Proteobacteria. Cronobacter muytjensii was the most abundant species characterized in the whole body and gut of E. coreanus, and the unclassified species in the genera Brevundimonas and Lactobacillus were relatively more abundant in the head and carcass. In addition, no Wolbachia-like bacteria were detected, whereas bacteria like Francisella tularensis subsp. Holarctica OSU18 and unclassified Rickettsiella were first identified in ant lion E. coreanus.

Effects of garlic oil, nitrate, saponin and their combinations supplemented to different substrates on in vitro fermentation, ruminal methanogenesis, and abundance and diversity of microbial populations.

AIMS: To investigate the effect of garlic oil (G), nitrate (N), saponin (S) and their combinations supplemented to different forage to concentrate substrates on methanogenesis, fermentation, diversity and abundances of bacteria and Archaea in vitro. METHODS AND RESULTS: The study was conducted in an 8 × 2 factorial design with eight treatments and two substrates using mixed ruminal batch cultures obtained. Quillaja S (0·6 g l(-1) ), N (5 mmol l(-1) ) and G (0·27 g l(-1) ) were used separately or in binary and tertiary combinations. The two substrates contained grass hay and a dairy concentrate mixture at a 70 : 30 (high-forage substrate) ratio or a 30 : 70 (high-concentrate substrate) ratio. Ruminal fermentation and cellulolytic bacterial populations were affected by interaction between substrate and anti-methanogenic compounds. The inhibitor combinations decreased the methane production additively regardless of substrate. For the high-concentrate substrate, S decreased methane production to a greater extent, so did G and N individually for the high-forage substrate. Feed degradability and total volatile fatty acid (VFA) concentrations were not decreased by any of the treatments. Fibre degradability was actually improved by N+S for the high-forage substrate. VFA concentrations and profiles were affected differently by different anti-methanogenic inhibitors and their combinations. All treatments inhibited the growth of Archaea, but the effect on Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens varied. CONCLUSIONS: The results suggest that substrate influences the efficacy of these inhibitors when they are used separately, but in combinations, they can lower methanogenesis additively without much influence from the substrate. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented research provided evidence that binary and tertiary combination of garlic oil, nitrate and saponin can lower the methane production additively without adversely impacting rumen fermentation and degradability, and forage to concentrate ratio does not change the above effects. These anti-methanogenic inhibitors in combination may have practical application to mitigate methane emission from ruminants.

Chronopharmacology of analgesic effect and tolerance induced by six narcotic analgesics in mice.

Narcotic analgesics, especially morphine, exert significantly different effects depending on the time within one day. The objective of this study was to observe whether the dosing time of 6 narcotic analgesics in mice affected their efficacy, pain tolerance and recovery of tolerance. The chronopharmacology of these 6 narcotics was evaluated using a hot-plate model. Maximum possible effect (MPE) of morphine showed a significant 24 h rhythm, which was higher during the dark phase and lower during the light phase (P<0.05). Conversely, MPEs of fentanyl and bucinnazine groups during the light phase exceeded those during the dark phase (P<0.05). Pain tolerance developed after drug administration at 9:00 am or 9:00 pm for 5 days, of which bucinnazine produced lower tolerance at 9:00 am. After a 2-day washout period, the mice rapidly recovered from tolerance at 3:00 pm for 5-day morphine dosing at 9:00 pm, and for fentanyl dosing at 9:00 am. Not all narcotic analgesics displayed significant circadian variations, and the dosing time-dependent effects also depended on the types of narcotics. Therefore, the time of administration is crucial in clinical pain treatment. Chronotherapy may be more effective to relieve pain while reducing side effects.

Prebiotic Oligosaccharides: Comparative Evaluation Using In Vitro Cultures of Infants' Fecal Microbiomes.

The objective of this study was to systematically assess the bifidogenic effect of three commonly used prebiotic products using in vitro cultures of infant fecal samples. Fresh stool samples collected from six term infants, each exclusively fed human milk (n = 3) or infant formula (n = 3), at 28 days of age were used as inocula. The following prebiotic products were added at concentrations applicable to infant formula: Vivinal GOS 15 (containing 28.5% galacto-oligosaccharide [GOS]) at 7.2 g/liter, Beneo HP (99.5% long-chain inulin [IN]) at 0.8 g/liter, Beneo Synergy 1 (enriched oligofructose and inulin [OF-IN]) at 4 g/liter, and a combination of Vivinal GOS 15 (7.2 g/liter) and Beneo HP (0.8 g/liter) (GOS-IN). The growth of total bacteria, Bifidobacterium, Bacteroides, Bifidobacterium longum, and Escherichia coli was quantified using specific quantitative PCR (qPCR). Bifidobacterium was also enumerated on selective Beerens agar plates, with representative colonies identified by sequencing of their 16S rRNA genes. Volatile fatty acids (VFA) and pH in the cultures were also determined. Irrespective of the feeding methods, the GOS product, either alone or in combination with Beneo HP, resulted in substantially higher growth of total bifidobacteria, and much of this growth was attributed to growth of B. longum. Beneo Synergy 1 also increased the abundance of total bifidobacteria and B. longum. Corresponding to the increases in these two bacterial groups, acetic acid concentrations were higher, while there was a trend of lower E. coli levels and pH. The lower pH and higher acetic acid concentration might be directly responsible for the lower E. coli population. At the concentrations studied, the GOS product was more bifidogenic and potent in inhibiting E. coli than the other products tested. These results suggest that supplementation of infant formula with GOS may increase intestinal bifidobacteria and benefit infant health.

Investigation of ruminal bacterial diversity in dairy cattle fed supplementary monensin alone and in combination with fat, using pyrosequencing analysis.

The objective of this study was to examine and compare the effects of monensin, both alone and together with dietary fat, on ruminal bacterial communities in dairy cattle fed the following 3 diets: a control diet, the control diet supplemented with monensin, and the control diet supplemented with both monensin and fat. Bacterial communities in the liquid and the adherent fractions of rumen content were analyzed using 454 pyrosequencing analysis of 16S rRNA gene amplicons. Most sequences were assigned to phyla Firmicutes and Bacteroidetes, irrespective of diets and fractions. Prevotella was the most dominant genus, but most sequences could not be classified at the genus level. The proportion of Gram-positive Firmicutes was reduced by 4.5% in response to monensin but increased by 12.8% by combination of monensin and fat, compared with the control diet. Some of the operational taxonomic units in Firmicutes and Bacteroidetes were also affected by monensin or by the combination of monensin with fat. The proportion of numerous bacteria potentially involved in lipolysis and (or) biohydrogenation was increased by both monensin and fat. The Shannon diversity index was decreased in the control diet supplemented with both monensin and fat, compared with the other 2 diet groups. Supplementary fats hinder bacterial attachment to plant particles and then result in decreased bacterial diversity in the rumen. The finding of this study may help in understanding the effect of monensin and fat on ruminant nutrition and the adverse effect of monensin and fat, such as milk fat depression and decreased feed digestibility.