Chimeric TIM-4 receptor-modified T cells targeting phosphatidylserine mediates both cytotoxic anti-tumor responses and phagocytic uptake of tumor-associated antigen for T cell cross-presentation.

To leverage complementary mechanisms for cancer cell removal, we developed a novel cell engineering and therapeutic strategy co-opting phagocytic clearance and antigen presentation activity into T cells. We engineered a chimeric engulfment receptor (CER)-1236, which combines the extracellular domain of TIM-4, a phagocytic receptor recognizing the "eat me" signal phosphatidylserine, with intracellular signaling domains (TLR2/TIR, CD28, and CD3ζ) to enhance both TIM-4-mediated phagocytosis and T cell cytotoxic function. CER-1236 T cells demonstrate target-dependent phagocytic function and induce transcriptional signatures of key regulators responsible for phagocytic recognition and uptake, along with cytotoxic mediators. Pre-clinical models of mantle cell lymphoma (MCL) and EGFR mutation-positive non-small cell lung cancer (NSCLC) demonstrate collaborative innate-adaptive anti-tumor immune responses both in vitro and in vivo. Treatment with BTK (MCL) and EGFR (NSCLC) inhibitors increased target ligand, conditionally driving CER-1236 function to augment anti-tumor responses. We also show that activated CER-1236 T cells exhibit superior cross-presentation ability compared with conventional T cells, triggering E7-specific TCR T responses in an HLA class I- and TLR-2-dependent manner, thereby overcoming the limited antigen presentation capacity of conventional T cells. Therefore, CER-1236 T cells have the potential to achieve tumor control by eliciting both direct cytotoxic effects and indirect-mediated cross-priming.

[Content analysis and quality evaluation of main active components and mineral elements of Cynomorium songaricum in different habitats].

To clarify the content characteristics of the main active components and mineral elements of Cynomorium songaricum under different habitat conditions, and further explore the relationship between the quality of C. songaricum and habitats, this study took C. songaricum from 25 different habitats in China as the research object, and measured the contents of 8 main active components and 12 mineral elements separately. Diversity analysis, correlation analysis, principal component analysis and cluster analysis were carried out. The results showed that the genetic diversity of total flavonoids, ursolic acid, ether extract, potassium(K), phosphorus(P) and zinc(Zn) in C. songaricum was high. The coefficient of variation of crude polysaccharide, ether extract, gallic acid, protocatechuic aldehyde, catechin, epicatechin, calcium(Ca), sodium(Na), magnesium(Mg), sulfur(S), iron(Fe), manganese(Mn), selenium(Se) and nickel(Ni) were all over 36%, indicating that the quality of C. songaricum was significantly affected by habitats. There were strong synergistic and weak antagonistic effects among the contents of the 8 active components, and complex antagonistic and synergistic effects among the contents of the 12 mineral elements. Principal component analysis revealed that crude polysaccharide, ursolic acid, catechin, epicatechin and total flavonoids could be used as the characteristic components to evaluate the quality of C. songaricum, and Na, copper(Cu), Mn and Ni were the characteristic elements to evaluate the quality of C. songaricum. In cluster ana-lysis, the second group with the main active components as cluster center had better quality in terms of the content of active substances, and the second group with the mineral elements as cluster center had higher utilization potential in the exploitation of mineral elements. This study could provide a basis for resource evaluation and breeding of excellent varieties of C. songaricum in different habitats, and provide a reference for cultivation and identification of C. songaricum.

Content analysis and quality evaluation of main active components and mineral elements of Cynomorium songaricum in different habitats / 中国中药杂志

To clarify the content characteristics of the main active components and mineral elements of Cynomorium songaricum under different habitat conditions, and further explore the relationship between the quality of C. songaricum and habitats, this study took C. songaricum from 25 different habitats in China as the research object, and measured the contents of 8 main active components and 12 mineral elements separately. Diversity analysis, correlation analysis, principal component analysis and cluster analysis were carried out. The results showed that the genetic diversity of total flavonoids, ursolic acid, ether extract, potassium(K), phosphorus(P) and zinc(Zn) in C. songaricum was high. The coefficient of variation of crude polysaccharide, ether extract, gallic acid, protocatechuic aldehyde, catechin, epicatechin, calcium(Ca), sodium(Na), magnesium(Mg), sulfur(S), iron(Fe), manganese(Mn), selenium(Se) and nickel(Ni) were all over 36%, indicating that the quality of C. songaricum was significantly affected by habitats. There were strong synergistic and weak antagonistic effects among the contents of the 8 active components, and complex antagonistic and synergistic effects among the contents of the 12 mineral elements. Principal component analysis revealed that crude polysaccharide, ursolic acid, catechin, epicatechin and total flavonoids could be used as the characteristic components to evaluate the quality of C. songaricum, and Na, copper(Cu), Mn and Ni were the characteristic elements to evaluate the quality of C. songaricum. In cluster ana-lysis, the second group with the main active components as cluster center had better quality in terms of the content of active substances, and the second group with the mineral elements as cluster center had higher utilization potential in the exploitation of mineral elements. This study could provide a basis for resource evaluation and breeding of excellent varieties of C. songaricum in different habitats, and provide a reference for cultivation and identification of C. songaricum.

The effects of the supplementation of multi-strain probiotics on intestinal microbiota, metabolites and inflammation of young SPF chickens challenged with Salmonella enterica subsp. enterica.

This study assessed the effect of probiotics on cecal microbiota, cecal short-chain fatty acids (SCFAs), and the gene expression of cytokines in young specific-pathogen-free (SPF) chickens infected with S. enterica subsp. enterica. One-day-old SPF chickens (n = 105) were randomly assigned to one of the three treatment groups: control (Cont) group, Salmonella-infected (Sal) group, and a Salmonella-infected group treated with multi-strain probiotics (ProSal group). All chickens except those in the Cont group were challenged orally with 1 × 108  cfu/ml of Salmonella 4 days after hatching. Chickens in the Sal group exhibited more abundance of Proteobacteria than those in the Cont and ProSal groups. At the genus level, chickens in ProSal group exhibited increased numbers of Lactobacillus and Oscillospira compared with those in the other groups. Chickens in the ProSal group exhibited a significant increase of cecal SCFAs compared with chickens in the Sal group. Chickens in the ProSal group exhibited increased gene expression of anti-inflammatory cytokines, IL-10 and TGF-ß4, and decreased expression of the proinflammatory cytokine, IFN-γ, in the cecal tonsil compared with those in the Sal group. The results of this study indicated that the administration of probiotics can modulate microbiota, SCFAs, and immunomodulatory activity in SPF chickens.

Effects of Multi-Strain Probiotics Combined with Gardeniae fructus on Intestinal Microbiota, Metabolites, and Morphology in Broilers.

This study was conducted to investigate the effects of a multi-strain probiotic combined with Gardeniae fructus on the growth performance, intestinal microbiota composition and metabolites, and intestinal morphology of broiler chickens. The dietary treatments included the basal diet without any antimicrobials (C), the basal diet supplemented with 10 ppm avilamycin (A), the basal diet supplemented with 0.1% multi-strain probiotics powder containing Lactobacillus acidophilus LAP5, L. fermentum P2, L. casei L21, and Pediococcus acidophilus LS (1×107 CFU/g) (P), and the basal diet supplemented with a mixture of 0.1% multi-strain probiotics and 0.05% herbal medicine G. fructus (PH). The results showed no significant differences in growth performance across all groups. A denaturing gradient gel electrophoresis analysis indicated that the groups PH, P, and A exhibited an increase in the similarity coefficients of their intestinal microbial populations. The real-time polymerase chain reaction (PCR) analysis showed that the relative concentrations of Firmicutes and Lactobacillus in the cecum and Bifidobacterium spp. in the ileum were higher in the groups PH, P, and A than in group C, and the diet supplemented with multi-strain probiotics combined with G. fructus decreased the concentrations of cecal Escherichia spp. and Clostridium perfringens. The broilers fed with multi-strain probiotics combined with G. fructus showed a significant increase (P<0.05) in the cecal short-chain fatty acids (total SCFA, acetic acid, and butyric acid) compared to the other groups. The treatment with antibiotics, multi-strain probiotics, or multi-strain probiotics combined with G. fructus increased the villus height/crypt depth ratio in the ileum of broilers. In conclusion, the supplementation of multi-strain probiotics combined with G. fructus was beneficial to the intestinal microflora composition, metabolites, and morphology in broilers.

Xuezhikang contributes to greater triglyceride reduction than simvastatin in hypertriglyceridemia rats by up-regulating apolipoprotein A5 via the PPARα signaling pathway.

Xuezhikang (XZK), an extract of Chinese red yeast rice, is recommended as an optimal choice for patients with coronary heart disease (CHD) with markedly elevated triglyceride (TG) levels. This study was designed to compare the hypotriglyceridemic effects between XZK and simvastatin. The role of apolipoprotein A5 (apoA5), a key regulator of TG metabolism and a target gene of peroxisome proliferator-activated receptor α (PPARα), was to be identified in XZK-related hypotriglyceridemic actions. For these goals, hypertriglyceridemia of rats was induced by a high-fructose diet. In order to investigate the hypotriglyceridemic effects of XZK and simvastatin on these animals based on an equivalent low-density lipoprotein cholesterol (LDL-C) lowering power, we titrated their doses (XZK 80 mg/kg/d versus simvastatin 1 mg/kg/d) according to plasma LDL-C reduction of rats. Similarly, we titrated the target doses of the two agents (XZK 500 µg/ml versus simvastatin 10 µM) according to hepatocyte LDL receptor expressions, and then compared the effects of the two agents on TG and apoA5 of hepatocytes in vitro. Our results showed that XZK (80 mg/kg/d) had higher hypotriglyceridemic performance than simvastatin (1 mg/kg/d) on these animals albeit their equivalent LDL-C lowering power. Higher plasma apoA5 levels and hepatic apoA5 expressions were observed in rats treated with XZK (80 mg/kg/d) than simvastatin (1 mg/kg/d). Further, XZK (80 mg/kg/d) contributed to higher hepatic PPARα expressions of rats than simvastatin (1 mg/kg/d). Although the two agents led to an equivalent up-regulation of LDL receptors of hepatocytes, more TG reduction and apoA5 elevation were detected in hepatocytes treated with XZK (500 µg/ml) than simvastatin (10 µM). However, PPARα knockdown eliminated the above effects of XZK on hepatocytes. Therefore, our study indicates that XZK has greater hypotriglyceridemic performance than simvastatin in the setting of an equivalent LDL-C lowering power, which is attributed to more apoA5 up-regulation by this agent via the PPARα signaling pathway.

[Determination and principal component analysis of mineral elements based on ICP-OES in Nitraria roborowskii fruits from different regions].

The content of elements in fifteen different regions of Nitraria roborowskii samples were determined by inductively coupled plasma-atomic emission spectrometry(ICP-OES), and its elemental characteristics were analyzed by principal component analysis. The results indicated that 18 mineral elements were detected in N. roborowskii of which V cannot be detected. In addition, contents of Na, K and Ca showed high concentration. Ti showed maximum content variance, while K is minimum. Four principal components were gained from the original data. The cumulative variance contribution rate is 81.542% and the variance contribution of the first principal component was 44.997%, indicating that Cr, Fe, P and Ca were the characteristic elements of N. roborowskii.Thus, the established method was simple, precise and can be used for determination of mineral elements in N.roborowskii Kom. fruits. The elemental distribution characteristics among N.roborowskii fruits are related to geographical origins which were clearly revealed by PCA. All the results will provide good basis for comprehensive utilization of N.roborowskii.

Effects of recombinant lycopene dietary supplement on the egg quality and blood characteristics of laying quails.

This study was conducted to determine the effect of dietary supplement of bacterial lycopene (BL) produced by Escherichia coli on the egg quality and blood characteristics of laying quails. The antioxidant activity measurement showed that BL exhibited 100% 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging capacity at a concentration of 4.65 µg/ml, which was more effective than butylated hydroxytoluene (BHT) and commercial lycopene (CL). Moreover, seven dietary groups of laying quails consisting of 10 100-day-old quails (Coturnix coturnix japonica) each were fed with the basal diet supplemented with BL, CL or canthaxanthin (CA) for 4 weeks. Consequently, the triglyceride content of yolk was significantly lower in the group with BL and CL supplement. The serum malondialdehyde (MDA) level of the BL- and CA-supplemented groups at 18 mg/kg was lower than the control group. In conclusion, BL has a high antioxidant activity and is promising as a feed additive in the diet of laying quails.

Coptidis rhizome and Si Jun Zi Tang can prevent Salmonella enterica serovar Typhimurium infection in mice.

Salmonella, a common zoonotic pathogen, causes gastroenteritis in both humans and animals. Traditional Chinese Medicine (TCM) has been used to improve gastrointestinal dysfunction and to modify the immune response to inflammation for centuries. This study used six herbal plants and four TCM formulae to rate their efficacy in preventing S. Typhimurium infection via mouse model. Minimum bactericidal concentration (MBC) of Coptidis rhizome (CR) against the reference strain tallied 12.5 mg/ml and against clinical isolate ST21 was 25 mg/ml. MBCs of other herbal extracts and formulae on Salmonella Typhimurium strains were above 50 mg/ml. In the mice model, CR and Si Jun Zi Tang (SJZT) could significantly decrease the bacterial load in organs and blood after being challenged, along with body weight loss due to the infection. CR and SJZT alleviated infection-induced interferon-gamma levels in the serum and tissues, and tumor necrosis factor-alpha (TNF-α) levels in intestinal tissues. CR and SJZT serum metabolites could suppress S. Typhimurium invasion and TNF-α expression in RAW264.7 cells. The therapeutic activity of CR and SJZT may involve berberine, ginsenoside Rb1, and glycyrrhizin, interfering with Salmonella when invading macrophages. CR and SJZT has shown potential in preventing S. Typhimurium infection through the regulation of the immune response.

The protective role of MnTBAP in oxidant-mediated injury and inflammation in a rat model of lung contusion.

BACKGROUND: Lung contusion (LC) is a unique direct and focal insult that is considered a major risk factor for the initiation of acute lung injury and acute respiratory distress syndrome. We have shown recently that consumption of nitric oxide (due to excess superoxide) resulting in peroxynitrite formation leads to decreased vascular reactivity after LC. In this study, we set out to determine whether the superoxide scavenger Mn (III) tetrakis (4-benzoic acid) porphyrin chloride (MnTBAP) plays a protective role in alleviating acute inflammatory response and injury in LC. METHODS: Nonlethal, closed-chest, bilateral LC was induced in a rodent model. Administration of the superoxide dismutase mimetic MnTBAP concurrently in LC in rats was performed, and bronchoalveolar lavage (BAL) and lung samples were analyzed for degree of injury and inflammation at 5 and 24 h after the insult. The extent of injury was assessed by the measurement of cells and albumin with cytokine levels in the BAL and lungs. Lung samples were subjected to H&E and superoxide staining with dihydro-ethidium. Protein-bound dityrosine and nitrotyrosine levels were quantified in lung tissue by tandem mass spectrometry. RESULTS: The degrees of lung injury after LC as determined by BAL albumin levels were significantly decreased in the MnTBAP-administered rats at all the time points when compared to the corresponding controls. The release of proinflammatory cytokines and BAL neutrophils was significantly less in the rats administered MnTBAP after LC. Administration of MnTBAP decreased tissue damage and decreased necrosis and neutrophil-rich exudate at the 24-h time point. Staining for superoxide anions showed significantly greater intensity in the lung samples from the LC group compared to the LC+ MnTBAP group. High-performance liquid chromatography/tandem mass spectrometry revealed that MnTBAP treatment significantly attenuated dityrosine and nitrotyrosine levels, consistent with decreased oxidant injury. CONCLUSION: Superoxide dismutase mimetic-MnTBAP reduced permeability and oxidative injury in LC and may have a therapeutic role in diminishing inflammation in LC.

Application of Scutellariae radix, Gardeniae fructus, and Probiotics to Prevent Salmonella enterica Serovar Choleraesuis Infection in Swine.

Salmonella enterica serovar Choleraesuis, a host-adapted pathogen of swine, usually causes septicemia. Lactic acid bacteria (LAB) strains have been widely studied in recent years for their probiotic properties. In this study, a mouse infection model first screened for potential agents against infection, then a pig infection model evaluated effects of LAB strains and herbal plants against infection. Scutellariae radix (SR) and Gardeniae fructus (GF) showed abilities to reduce bacteria shedding and suppressing serum level of TNF- α induced by infection in swine. Bioactivities of SR and GF were enhanced by combining with LAB strains, which alone could speed up the bacteria elimination time in feces and boost immunity of infected pigs. Baicalein and genipin exhibited stronger cytotoxicity than baicalin and geniposide did, as well as prevent Salmonella from invading macrophages. Our study suggests LAB strains as exhibiting multiple functions: preventing infection, enhancing immunity to prepare host defenses against further infection, and adjusting intestinal microbes' enzymatic activity in order to convert herbal compounds to active compounds. The SR/GF-LAB strain mixture holds potential infection-prevention agents supplied as feed additives.

Effect of Echinacea purpurea L. on oxidative status and meat quality in Arbor Acres broilers.

BACKGROUND: Echinacea purpurea L. (EP) is a popular herbal antioxidant and immunomodulator. The present study was conducted to evaluate the effects of EP on meat quality and oxidative status in broilers. Two hundred and fifty (1-day-old) male broilers (Arbor Acres) were randomly allocated to five groups including the control (corn-soybean meal diet) and 0.1, 0.5, 1.0 and 2.0% EP powder groups, with two replicates per treatment group. RESULTS: The results indicated that the addition of 0.5% and 1.0% EP significantly increased water-holding capacity and decreased storage loss of breast and thigh fillets at 35 days old. For fillet colour, L* (lightness) values were lower, and a* (redness) and b* (yellowness) values were higher with EP supplementation. Lower crude fat contents were observed in EP groups in comparison with control at 35 days of age in breast and thigh fillets, respectively. Production of malondialdehyde was slightly reduced in serum of EP supplemented birds compared to the control group. Results for Trolox equivalent antioxidant capacity, catalase and superoxide dismutase were significantly higher for the 0.5, 1.0 and 2.0% EP supplemental groups than control group in serum. Liver and spleen tissues results showed that the antioxidative enzymes activities were higher with EP powder at 35 days of age. CONCLUSION: Dried EP can be used as a feed additive to improve the meat quality and oxidative status in Arbor Acres broilers.

Starch characteristics and their influences on in vitro and pig prececal starch digestion.

The main objective of this research was to study the characteristics of starch granules and their influences on in vitro and pig prececal starch digestion of corn, dehulled barley, wheat, and potato. Scanning electron microscopy was used to study the starch endosperm structure in the parent material as well as in vitro starch digestion. The results showed that corn starch granules were polyhedral, with a diameter ranging from 2 to 10 µm, whereas those of dehulled barley and wheat were spherical, with a diameter ranging from 5 to 20 µm. Potato had the largest starch granules among starch sources reported herein, with oval spheres of 10-50 µm in diameter. In vitro starch hydrolysis showed that starch granules of corn degraded faster than the starch of dehulled barley and wheat, with the potato starch being degraded the slowest. The in vivo digestibility trial using ileal-cannulated pigs confirmed the starch degradation of grains. The in vitro (x, %) and in vivo (y, %) digestibility were highly correlated [y = 6.5304x - 538.48 (R(2) = 0.9924)]. On the basis of the results, in vitro starch hydrolysis might be useful in predicting in vivo prececal starch digestibility. The digestion kinetic characteristics of different starch sources might be employed to evaluate the starch digestive rate at the pig ileum.

[Effects of climate change on potato growth in semi-arid region of Loess Plateau, China].

Based on the 1988-2008 located observation and 2007-2008 encrypted observation of potato growth and the 1957-2008 meteorological observation in semi-arid region of Loess Plateau, this paper studied the effects of climate change on the potato growth in this region. In 1957-2008, the annual precipitation in this region had a descending trend, with a linear fitting rate of the annual precipitation change curves being - 13.359 mm x (10 a)(-1), while the annual mean temperature displayed an ascending trend, with a linear fitting rate of the annual mean temperature change curves being 0.239 degrees C x (10 a)(-1). During potato growth period, the aridity index displayed a marked ascending trend, and the linear fitting rate of the aridity index change curves was 0.102 x (10 a)(-1). The growth rate of potato tuber became faster from the 96th day after sowing, reached the maximum on the 110th day, and turned slower from the 124th day. The interval from sowing to seedling emergence was shortened by 1-2 d x (10 a)(-1), and that from inflorescence formation to reaping and of whole growth period was lengthened by 9-10 d x (10 a) (-1). In the study region, climate warming shortened the vegetative growth stage, but lengthened the reproductive growth stage and whole growth period of potato.

Porcine lactoferrin expression in transgenic rice and its effects as a feed additive on early weaned piglets.

The purpose of this study is to determine the growth performance and immune characteristics of early weaned piglets receiving rice bran expressing porcine lactoferrin as a feed additive. Full-length cDNA encoding porcine lactoferrin (LF) driven by a rice actin promoter was transformed into rice plants, and its integration into the rice genome was verified by Southern blot analysis. The expression of recombinant LF (rLF) in whole grains and rice bran was also confirmed, and the amount of rLF accumulated in rice bran was estimated by immunoblot assay to be approximately 0.1% of rice bran weight. An iron-binding assay showed that the rLF retained iron-binding activity and the binding capacity of 1 mg/mL rLF would be saturated by 100 microM of FeCl(3). Thirty-six early weaned piglets at 21 days old were randomly selected into two groups and fed a diet containing 5% transgenic rice bran containing 50 mg/kg rLF (rLF group) and 5% rice bran (control group) to investigate the piglets' growth performance and immune characteristics. The results showed no significant difference in growth performance between the groups during the feeding period. However, the aerobic bacteria, anaerobic bacteria, and coliform counts in the cecal contents of the rLF-fed group were significantly lower than those of the control group. Additional immune characteristics such as the IgG concentration in the rLF group was higher than the control group at the 28th day, but leukocyte counts and the peripheral lymphocyte ratio remained similar. In summary, porcine LF expressed in rice bran, a byproduct of rice, can be used as a functional additive to improve antimicrobial capabilities and IgG concentration of early weaned piglets.

[Traditional Chinese and Western medicine treatment of mycoplasmal pneumonia in children and the serum cytokine changes].

OBJECTIVE: To observe the therapeutic effect of traditional Chinese medicine (TCM) and Western medicine (WM) treatments on mycoplasmal pneumonia in children and the changes in the serum cytokines. METHODS: Ninety children with mycoplasmal pneumonia were randomly divided into the treatment group and the control group. TCM was given orally and azithromycin at the daily dose of 10 mg/kg was administered intravenously in the treatment group. In the control group, only intravenous azithromycin was given. After a 7-day treatment, the response rate, time of symptom disappearance, and serum levels of IL-6, IL-8 and TNF-alpha were observed. RESULTS: The total response rate was 93.33% in the treatment group and 73.33% in the control group, showing a significantly better therapeutic effect in the treatment group (P<0.05). The combined treatments also showed better effects in alleviating fever, coughing and rales (P<0.05), and resulted in more obvious reduction in the serum levels of cytokines (P<0.05). CONCLUSION: Combined treatment with TCM and WM produce good therapeutic effects in children with mycoplasmal pneumonia.

Use of PCR primers and probes based on the 23S rRNA and internal transcription spacer (ITS) gene sequence for the detection and enumerization of Lactobacillus acidophilus and Lactobacillus plantarum in feed supplements.

Novel polymerase chain reaction (PCR) primers designed from the 16S-23S internal transcription spacer (ITS) rRNA and 23S rRNA genes, respectively, were used for the specific detection of Lactobacillus acidophilus and Lactobacillus plantarum. Molecular weights of the PCR products were 221 and 599 bp, respectively. Strains of L. acidophilus and L. plantarum obtained from the culture center, dairy products, infant stool and other samples, could be identified with these PCR primers. DNAs from other lactic acid bacteria (LAB) species including strains of Lactobacillus pentosus which was closely related to L. plantarum, and bacteria species other than LAB, would not generate the false positive results. When this PCR primer set was used for the detection of L. acidophilus and L. plantarum in feed supplement or feed starter samples, reliable results were obtained. Furthermore, when these L. acidophilus or L. plantarum specific primers were used as DNA probes for the colony hybridization of L. acidophilus and L. plantarum, the viable cells of these LAB species in culture and feed supplements or starter products could be identified and enumerized. The method described here thus offers a rapid and economic way to inspect and assure the quality of the feed supplements or fermentation starters.