Tryptophan alleviates chronic heat stress-induced impairment of antioxidant capacities, inflammatory response, and mitochondrial function in broilers.

The aim of this study was to investigate the effect of dietary tryptophan (Trp) supplementation on serum biochemical indices, antioxidant indices, cytokine levels, mitochondrial biosynthesis, and mitochondrial morphology of heat-stressed broilers. A total of 180 female Arbor Acres broilers (18-day-old) were randomly allocated into three groups with six replicates of 10 broilers each. Broilers in thermoneutral (TN) (23 ± 1 °C) group were fed a basal diet; the other two groups were fed the basal diet supplemented with 0 or 0.18% Trp under heat stress (HS) (34 ± 1 °C for 8 h/day (h/day) and 23 ± 1°C for the remaining time) condition. The heat stress lasted for 21 days (days 21 to 42). The results indicated that heat stress reduced serum total protein content (TP) and decreased the activities of serum superoxide dismutase (SOD) and total antioxidant capacity (T-AOC), but increased the levels of serum uric acid (UA), interleukin (IL)-1ß, IL-6, and IL-18 (P < 0.05) compared to the TN group. However, dietary supplementation with 0.18% Trp enhanced serum TP content, glutathione peroxidase (GSH-Px), SOD, catalase (CAT) activities, and T-AOC; decreased aspartate aminotransferase (AST) activities (P < 0.05); and lowered serum IL-1ß, IL-6, IL-18 contents (P < 0.05). Meanwhile, heat stress exposure downregulated the mRNA expression of mitochondrial transcription factor A (TFAM), cytochrome c oxidase subunit 1 (COX1), and cytochrome c oxidase subunit 5A (COX5A) in ileum (P < 0.05) as compared to the TN group. Dietary Trp supplementation enhanced the mitochondrial membrane potential (MMP) and the mRNA expression of TFAM, COX1 in ileum mucosa (P < 0.05) and ameliorated the damage of mitochondrial structure. Collectively, dietary supplementation with Trp could improve antioxidant capacity and mitochondrial structure and regulate mitochondrial function-related genes and decrease inflammatory response in heat-stressed broilers. Dietary Trp supplementation might be an effective nutritional strategy to protect against heat stress impairment.

Effects of dietary tryptophan supplementation on body temperature, hormone, and cytokine levels in broilers exposed to acute heat stress.

The present study was conducted to investigate the effects of tryptophan (Trp) supplementation on rectal temperature, hormone, and cytokine levels in broilers subjected to acute heat stress. A total of 300 18-day-old female Arbor Acres broilers were randomly allocated to five dietary treatment groups with six replicates per treatment group and ten birds per replicate. Broilers were fed a basal diet and in the thermoneutral conditions (TN, 23 ± 1 °C) was considered as the TN group. Broilers were fed a basal diet and exposed to acute heat stress (HS, 34 ± 1 °C) was regarded as the HS group, and other broilers exposed to acute heat stress (34 ± 1°C) were fed a basal diet supplemented with 0.09%, 0.18%, and 0.27% Trp. Results indicated that acute heat stress increased the rectal temperature (P < 0.05), enhanced the concentrations of corticosterone (CORT), dopamine (DA), adrenaline (Adr), adrenocorticotropic hormone (ACTH), and corticotropin-releasing hormone (CRH) in serum (P < 0.05), and elevated the levels of serum tryptophan hydroxylase (TPH)1, tryptophan 2, 3-dioxygenase (TDO), indoleamine 2, 3-dioxygenase (IDO), and kynurenic acid (P < 0.05), compared with the TN group. Meanwhile, acute heat stress increased the levels of serum Trp, hypothalamic Trp, 5-hydroxytryptophan (5-HT), and interleukin-22 (P < 0.05) relative to the TN group. However, compared with the heat stress group, Trp supplementation decreased the rectal temperature of heat-stressed broilers and dietary 0.09% Trp supplementation decreased the levels of serum CRH and TDO (P < 0.05), increased the levels of serum Trp and IL-22 (P < 0.05) in heat-stressed broilers. In addition, dietary supplemented with 0.18% Trp reduced the levels of serum DA, Adr, noradrenaline (NA), CRH, TDO, IDO, kynurenic acid, IL-1ß, and hypothalamic 5-HIAA/5-HT (P < 0.05), increased the levels of serum Trp, 5-HT, and IL-22, and upregulated the concentrations of hypothalamic Trp and 5-HT in heat-stressed broilers (P < 0.05). Moreover, dietary 0.27% Trp supplementation decreased the levels of serum DA, CRH, TDO, and hypothalamic 5-HIAA/5-HT (P < 0.05), and upregulated the levels of serum Trp, 5-HT, IL-22, hypothalamic Trp and 5-HT in heat-stressed broilers (P < 0.05). Taken together, dietary 0.18% Trp supplementation may be the optimal level for broilers reared under acute heat stress.

Effect of probiotic supplementation on the expression of tight junction proteins, innate immunity-associated genes, and microbiota composition of broilers subjected to cyclic heat stress.

This study investigated the effects of probiotic on intestinal innate immunity-associated gene expression and cecal microbiota in heat-stressed broilers. A total of 180 21-day-old male broilers were randomly assigned to three treatment groups with four replicates per group. The thermoneutral group (TN) (23 ± 1°C) received a basal diet, and another two heat-stressed groups (28-35-28°C for 12 h daily) were fed the basal diet (HS) or the basal diet supplemented with probiotic at a dose of 1.5 × 108 CFU/kg (HS_Pro) for 21 consecutive days. Compared with the TN group, the abundance of beneficial bacteria was decreased (p < 0.05) in the caecum of heat-stressed broilers. Heat stress downregulated (p < 0.05) the expression of Toll-like receptor (TLR)2 and upregulated (p < 0.05) the expressions of TLR5, TLR15, avian ß-defensin (AvBD)4, AvBD8, and AvBD14 in the ileum as compared with the TN group. Dietary supplementation of probiotic upregulated (p < 0.05) the occludin expression in the ileum, improved the microbiota balance in the caecum, and decreased (p < 0.05) the gene expressions of TLR5 and TLR15 in the ileum of heat-stressed broilers. Collectively, dietary probiotic supplementation could promote intestinal barrier function via improving gut microbiota community and regulating innate immunity-associated gene expressions in heat-stressed broilers.

Effect of probiotic supplementation on growth performance, intestinal morphology, barrier integrity, and inflammatory response in broilers subjected to cyclic heat stress.

This study investigated the protective effects of probiotic on heat stress-induced intestinal injury and inflammatory response in broilers. A total of 180 male broilers were randomly allocated to three treatments with four replicates each from 22 to 42 days of age. The broilers were either raised under thermoneutral (TN) conditions (23 ± 1°C) or subjected to cyclic heat stress (28-35-28°C for 12 hr daily). The broilers kept at TN conditions were fed a basal diet, and those exposed to heat stress were fed basal diets supplemented with or without probiotic at a dose of 1.5 × 108  cfu/kg. Compared with the TN group, heat stress decreased (p < .05) the growth performance, reduced (p < .05) villus height and villus height: crypt depth ratio in intestinal mucosa, increased (p < .05) serum levels of D-lactic acid on day 28 and endotoxin, TNF-α and IL-6 on day 42, and decreased (p < .05) serum IL-10 content on day 42. Dietary supplementation of probiotic reversed (p < .05) all these changes except for the growth performance in heat-stressed broilers. In conclusion, dietary inclusion of probiotic could improve intestinal morphology and barrier function, alleviate inflammatory response, but exert no ameliorative effect on growth performance of broilers under cyclic heat stress.

A Chinese Medicine Formula "Xian-Jia-Tang" for Treating Bladder Outlet Obstruction by Improving Urodynamics and Inhibiting Oxidative Stress through Potassium Channels.

The aim of this study is to investigate efficacy of a traditional Chinese medicine formula (named Xian-Jia-Tang, XJT) on bladder outlet obstruction (BOO) in rats and explore its mechanisms. Total 80 BOO model rats were established and randomly divided into 4 groups: physiological saline, XJT, Cesium Chloride (CC), and XJT and CC groups. Meanwhile, 12 rats were used as normal control. Bladder weight and urodynamics were measured. Oxidative stress level and mRNA expressions of potassium channels gene were detected in detrusor. The mRNA and protein levels of hypoxia inducible factor-α (HIF-α) in detrusor were detected by RT-PCR and Western blot. BOO model rats showed significantly higher bladder weight and abnormal urodynamics. XJT significantly improved the abnormal urodynamics and inhibited the oxidative stress and changes of mRNA levels of potassium channels genes in detrusor of BOO model rats. Moreover, KATP and SK2/3 mRNA were overexpressed in BOO model rats treated by XJT. Besides, the significantly increased levels of HIF-α mRNA and protein were also inhibited by XJT. However, these inhibition effects of XJT were weakened by CC. XJT could effectively improve the urodynamics and inhibit the oxidative stress caused by hypoxia through suppressing the role of potassium channels in BOO model rats.

[Bushen Huoxue Fang promotes the apoptosis of epithelial cells in the prostatic ductal system of rats with benign prostatic hyperplasia].

OBJECTIVE: To investigate the effects of Bushen Huoxue Fang (BSHX) on the apoptosis of epithelial cells in the prostatic ductal system of rats with benign prostatic hyperplasia (BPH) and its possible action mechanism. METHODS: One hundred 3- month-old male Wistar rats were randomly divided into four groups of equal number (control, castrated, BPH model, and BSHX). BPH models were made by subcutaneous injection of testosterone following castration; the rats in the BSHX group were treated intragastrically with BSHX at 2.34 g/ml after modeling, while those in the other two groups with equal volume of saline, all for 37 days. On the 38th day, all the rats were sacrificed and their prostates harvested for detection of the distribution of TGF-beta1 and alpha-actin and the count of positive cells in the prostatic ductal system by immunohistochemical staining. The apoptosis rate of epithelial cells in the prostatic ductal system was determined by TUNEL assay. RESULTS: The expression of TGF-beta1 was significantly increased in the rats of the BSHX group as compared with the BPH models in both the proximal prostatic duct ([15.28 +/- 4.30]% vs [36.42 +/- 8.10]%, P < 0.01) and the distal prostatic duct ([4.42 +/- 2.07]% vs [8.71 +/- 2.28 ]%, P < 0.05), while the expression of alpha-actin in the proximal duct was remarkably higher in the BSHX-treated rats than in the models ([28.14 +/- 7.43]% vs [18.28 +/- 4.07]%, P < 0.01), but lower than in the control animals ([33.57 +/- 6.85]%, P < 0.05). Compared with the control group, the BPH models and BSHX-treated rats both exhibited markedly decreased apoptosis of epithelial cells in the proximal prostatic duct ([39.42 +/- 9.20]% vs [3.86 +/- 1.34]%, P < 0.01, and [31.14 +/- 5.64]%, P < 0.01) and distal prostatic duct ([17.60 +/- 4.86]% vs [3.07 +/- 1.14]%, P < 0.01, and [12.37 +/- 2.25]%, P < 0.05). The apoptosis rate of epithelial cells in the prostatic ductal system was significantly higher in the BSHX-treated rats than in the BPH models (P < 0.01). CONCLUSION: By upregulating the expression of TGF-beta, BSHX can suppress the reduction of smooth muscle cells in the proximal prostatic duct, promote the apoptosis of prostatic epithelial cells, and thus effectively inhibit benign prostatic hyperplasia.

Simultaneous determination of nineteen major components in Qi She Pill by ultra-high-performance liquid chromatography-tandem mass spectrometry.

Qi She Pill (QSP) is a traditional Chinese medicine (TCM) prescription that has been used in treating cervical spondylosis radiculopathy for many years. In this study, a simple and sensitive method using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) on a reverse-phase C18 column was developed for the simultaneous determination of the 19 major components in QSP. We found that the optimum mobile phase for gradient elution was 0.1% formic acid and methanol. The correlation coefficients of all calibration curves were greater than 0.99. Recoveries measured at three concentration levels varied from 95.43% to 102.35%. Relative standard deviations of intra- and inter-day precisions were less than 4.45%. After successfully validating our method, we then applied it to the quantification of 19 components in QSP products to show that this method provides a new standard in quality assessment of TCM prescriptions containing multiple bioactive components.

Impact of intensive land-based fish culture in qingdao, china, on the bacterial communities in surrounding marine waters and sediments.

The impact of intensive land-based fish culture in Qingdao, China, on the bacterial communities in surrounding marine environment was analyzed. Culture-based studies showed that the highest counts of heterotrophic, ammonium-oxidizing, nitrifying, and nitrate-reducing bacteria were found in fish ponds and the effluent channel, with lower counts in the adjacent marine area and the lowest counts in the samples taken from 500 m off the effluent channel. Denaturing gradient gel electrophoresis (DGGE) analysis was used to assess total bacterial diversity. Fewer bands were observed from the samples taken from near the effluent channel compared with more distant sediment samples, suggesting that excess nutrients from the aquaculture facility may be reducing the diversity of bacterial communities in nearby sediments. Phylogenetic analysis of the sequenced DGGE bands indicated that the bacteria community of fish-culture-associated environments was mainly composed of Flavobacteriaceae, gamma- and deltaproteobacteria, including genera Gelidibacter, Psychroserpen, Lacinutrix, and Croceimarina.

[Effect of combination of bushen jianpi recipe and erythropoietin on serum tumor necrosis factor alpha in patients with anemia].

OBJECTIVE: To study the effect of treatment of renal anemia by combination of Bushen Jianpi Recipe (BJR, a Chinese experience recipe for supplementing Shen and supporting Pi) and low dosage erythropoietin (EPO), and the influence of treatment on change of serum tumor necrosis factor alpha (TNF-alpha) so as to explore the possible mechanism of integrative Chinese and western medicine (ICWM) in treating renal anemia. METHODS: Patients with renal anemia were randomly divided into two groups, the ICWM group and the control group, symptomatic and supporting treatment such as dialysis, supplementing of ferrous, foliac acid and vitamin B12, was given to both groups. Additionally, to the ICWM group, 50 IU/kg of EPO subcutaneous injection for twice every week, and oral intake of BJR, one dose per day taken in two parts, were given, and to the control group, EPO alone, 50 IU/kg by subcutaneous injecting, 3 times per week, was given. The therapeutic course for two groups was 3 months. Blood levels of hemoglobin (Hb), hematocrit (Hct), TNF-alpha were measured before treatment and the therapeutic effect was observed. RESULTS: After treatment, the levels of Hb and Hct increased significantly in both groups (P < 0.01), comparison between the two groups in Hb and Hct after being treated for 3 months showed significant difference (P < 0.05). The serum level of TNF-alpha was markedly higher than normal range in both groups before treatment, it significantly lowered after treatment in the ICWM group (P < 0.05), but unchanged in the control group. CONCLUSION: Combination of BJR and EPO could significantly inhibit the production of TNF-alpha, this may be an important factor for ICWM in effectively improving sensitivity to EPO.