IFN-γ affects pancreatic cancer properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway

BackgroundMetastasis-related in colon cancer 1 (MACC1) is highly expressed in a variety of solid tumours, but its role in pancreatic cancer (PC) remains unknown. Interferon gamma (IFN-γ) affecting MACC1 expression was explored as the potential mechanism following its intervention.MethodsExpressions of MACC1 treated with IFN-γ gradient were confirmed by quantitative real-time PCR (qRT-PCR) and western blot (WB). Proliferation, migration, and invasion abilities of PC cells treated with IFN-γ were analysed by CCK8, EDU, colony formation, Transwell (with or without matrix gel) and wound-healing assays. Expression of antisense long non-coding RNA of MACC1, MACC1-AS1, and proteins of AKT/mTOR pathway, (pho-)AKT, and (pho-)mTOR was also assessed by qRT-PCR and WB. SiRNA kit and lentiviral fluid were conducted for transient expression of MACC1 and stable expression of MACC1-AS1, respectively. Rescue assays of cells overexpressing MACC1-AS1 and of cells silencing MACC1 were performed and cellular properties and proteins were assessed by the above-mentioned assays as well.ResultsIFN-γ inhibited MACC1 expression in a time- and dose-dependent manner; 100 ng/mL IFN-γ generally caused downregulation of most significant (p ≤ 0.05). In vitro experiments revealed that IFN-γ decreased cellular proliferation, migration, and invasion abilities and downregulated the expression of pho-AKT and pho-mTOR (p ≤ 0.05). Conversely, overexpression of MACC1-AS1 upregulated pho-AKT and pho-mTOR proteins, and reversed cellular properties (p ≤ 0.05). Rescue assays alleviated the above changes of pho-AKT/ mTOR and cellular properties.ConclusionIFN-γ affected PC properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway, which provides novel insight for candidate targets for treating PC.

IFN-γ affects pancreatic cancer properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway.

BACKGROUND: Metastasis-related in colon cancer 1 (MACC1) is highly expressed in a variety of solid tumours, but its role in pancreatic cancer (PC) remains unknown. Interferon gamma (IFN-γ) affecting MACC1 expression was explored as the potential mechanism following its intervention. METHODS: Expressions of MACC1 treated with IFN-γ gradient were confirmed by quantitative real-time PCR (qRT-PCR) and western blot (WB). Proliferation, migration, and invasion abilities of PC cells treated with IFN-γ were analysed by CCK8, EDU, colony formation, Transwell (with or without matrix gel) and wound-healing assays. Expression of antisense long non-coding RNA of MACC1, MACC1-AS1, and proteins of AKT/mTOR pathway, (pho-)AKT, and (pho-)mTOR was also assessed by qRT-PCR and WB. SiRNA kit and lentiviral fluid were conducted for transient expression of MACC1 and stable expression of MACC1-AS1, respectively. Rescue assays of cells overexpressing MACC1-AS1 and of cells silencing MACC1 were performed and cellular properties and proteins were assessed by the above-mentioned assays as well. RESULTS: IFN-γ inhibited MACC1 expression in a time- and dose-dependent manner; 100 ng/mL IFN-γ generally caused downregulation of most significant (p ≤ 0.05). In vitro experiments revealed that IFN-γ decreased cellular proliferation, migration, and invasion abilities and downregulated the expression of pho-AKT and pho-mTOR (p ≤ 0.05). Conversely, overexpression of MACC1-AS1 upregulated pho-AKT and pho-mTOR proteins, and reversed cellular properties (p ≤ 0.05). Rescue assays alleviated the above changes of pho-AKT/ mTOR and cellular properties. CONCLUSION: IFN-γ affected PC properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway, which provides novel insight for candidate targets for treating PC.

[Determination of Three Types of New Psychoactive Tryptamines in Blood by QuEChERS Combined with UPLC-MS/MS].

ABSTRACT: Objective To establish a method combining QuEChERS and ultra-high liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for rapid screening and testing of three types of new psychoactive tryptamines in human blood： 5-MeO-DALT, 5-MeO-MiPT and 5-MeO-DiPT. Methods The effects of the type of extractant, the type and dosage of salting-out agent, and the dosage of adsorbent on the test results of the three tryptamines were investigated. Blood samples were processed by QuEChERS method and then determined by UPLC-MS/MS. Results The linear relationships of 5-MeO-DALT, 5-MeO-MiPT and 5-MeO-DiPT in human blood were good in the range of 0.5-100, 0.5-100 and 0.2-100 ng/mL, respectively, with their coefficients higher than 0.99. The limits of detection （LODs） were 0.1-0.2 ng/mg. The recoveries ranged from 84.86% to 94.57%. Intra-day and inter-day precisions were good. Conclusion The method is simple, rapid, easy to operate and has a high recovery. It is suitable for the qualitative and quantitative study of tryptamines in blood and can provide the reference for public security organs to deal with related cases.

Effects of dietary Lactobacillus rhamnosus CF supplementation on growth, meat quality, and microenvironment in specific pathogen-free chickens.

The probiotic effects of Lactobacillus rhamnosus strain CF (Chen Fu) on growth performance, meat quality, and microenvironment in specific pathogen-free (SPF) chickens were investigated and compared with Enterococcus faecium. One-hundred-eighty 7-day-old SPF chickens were randomly assigned into 3 groups with 3 replicate pens of 20 chickens each. Group 1 served as a control that was fed a basal diet without probiotics supplementation. Groups 2 and 3 were fed the basal diet supplemented with L. rhamnosus CF and E. faecium, respectively. On d 12 and 24, BW, ADG, feed conversion ratio (FCR), dressing percentage (DP), and apparent digestibility of crude protein (AD-CP) were calculated. Meat color, fat content, shear force, water content, and pH value of breast and thigh muscles; ammonia, urea nitrogen, and uric acid content in plasma; pH value, Enterococcus, Lactobacillus, and E. coli in ceca; and ammonia emission were determined. Compared with group 1, group 2 exhibited higher BW, ADG, AD-CP, DP, cecal Lactobacilli, and muscle fat content (P < 0.05) as well as lower FCR, muscle water content, plasma ammonia, pH value, E. coli, and Enterococcus in ceca, and ammonia emission (P < 0.05), and group 3 exhibited higher BW, ADG, AD-CP, DP, and muscle fat content (P < 0.05), as well as lower FCR, meat color, plasma ammonia, E. coli and Enterococcus in ceca, and ammonia emission (d 24) (P < 0.05). Compared with group 3, group 2 exhibited lower plasma ammonia level, E. coli, and pH value in ceca and ammonia emission (P < 0.05) and higher AD-CP, meat color, pH value in thigh muscles, fat content in breast muscles, and number of Lactobacillus in ceca (P < 0.05). Thus, L. rhamnosus CF improves growth performance, meat quality, and microenvironment and is a potential probiotic additive in chickens.

[Regulatory role of calcium activated chloride channel in pulmonary vascular structural remodeling in rats with pulmonary arterial hypertension induced by high pulmonary blood flow].

OBJECTIVE: To explore the regulatory role of calcium activated chloride channel (CaCC) in vascular structural remodeling in pathogenesis of pulmonary arterial hypertension (PAH) induced by high pulmonary blood flow. METHOD: An abdominal aorta and inferior vena cava shunting operation was used to induce high pulmonary blood flow and establish a PAH rat model.Seventy-five SD rats were randomly divided into normal, sham, shunt, niflumic acid (NFA) 1(0.2 mg/(kg·d))and NFA 2 (0.4 mg/(kg·d)) groups. There were 15 rats in each group. Pulmonary artery pressure and vascular structural remodeling were measured, arteriole contraction ratio among these groups were compared using vascular tone analysis system, and the electrophysiology of pulmonary artery smooth muscle cell (PASMC) was recorded using patch clamp technology. Differences between multiple groups were compared through variance analysis and that between groups with q test. RESULT: Compared with normal ((14.4±1.3 ) mmHg, 1 mmHg=0.133 kPa)and sham groups ((13.5±2.3 ) mmHg), mean pulmonary artery pressure in shunt group ((27.4±2.4 ) mmHg) increased significantly (P<0.05). Compared with shunt group, mean pulmonary artery pressure in NFA 1 group ((21.2±2.0) mmHg) and NFA 2 group ((22.3±2.0) mmHg) decreased significantly (P<0.05). Pulmonary vascular structural remodeling including pulmonary artery stenosis presented in shunt group. Compared with normal ((114.3±1.2)%) and sham ((115.5±1.1)%) groups, arteriole contraction ratio to 10(-5) mol/L phenylephrine in shunt group ((132.6±1.4)%) increased significantly (P<0.05). Compared with shunt group, pulmonary vascular structural remodeling alleviated in NFA 1 and NFA 2 groups. Arteriole contraction ratio in NFA 1 group ((126.4±1.3)%) and NFA 2 group ((124.6±1.0)%) decreased significantly compared with shunt group (P<0.05). Patch clamp technique recorded typical CaCC currents. Compared with normal ((32.3±2.3 ) pA/pF) and sham groups ((35.3±1.2) pA/pF), the CaCC current density of PASMC in shunt group ((51.3±2.7) pA/pF) increased significantly (P<0.05). Compared with shunt group, the CaCC current density of PASMC in NFA 1 group ((40.2±1.5 ) pA/pF) and NFA 2 group ((42.7±2.2) pA/pF) decreased significantly (P<0.05). CONCLUSION: CaCC is involved in pulmonary arterial hypertension induced by high pulmonary blood flow through regulating membrane potential. NFA attenuate pulmonary vascular structural remodeling and pulmonary pressure through decreasing CaCC current density of PASMC membrane.

Effect of weaning age on cortisol release in piglets.

The effect of weaning age on the adrenal cortex, which plays a vital role in the stress response, is currently unknown. Therefore, plasma adrenocorticotropic hormone (ACTH) and cortisol levels, weights and relative weights of adrenal glands, and steroidogenesis-related protein and enzyme expression levels in piglets weaned on different days were determined. Piglets weaned at 35 days had significantly lower ACTH levels than those weaned at 14 or 21 days, and cortisol levels of piglets weaned at 21, 28, and 35 days were significantly lower than those of piglets weaned on day 14. Adrenal gland weights of piglets weaned at 28 and 35 days and relative adrenal gland weights of piglets weaned at 35 days were significantly lower than those of piglets weaned at 14 days. However, no significant difference was detected in the expression of melanocortin-type 2 receptor mRNA, which is associated with weaning age. Steroidogenic acute-regulatory (StAR) mRNA and cholesterol side-chain cleavage cytochrome P450 mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 or 21 days, and P450 11ß mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 days. Therefore, early-weaned piglets exhibited increased adrenal gland weights and StAR and steroidogenic enzyme expression, all of which contributed to high cortisol levels. The high plasma ACTH and cortisol levels in early-weaned piglets indicate that these animals would be greatly affected by stress.

Sequence variation in the Toxoplasma gondii ROP20 gene among strains from different hosts and geographical locations.

Toxoplasma gondii, an opportunistic protozoan parasite, infects almost all warm-blooded animals. In this study, we examined the sequence variation in rhoptry protein 20 (ROP20) genes among 18 T. gondii isolates collected from different hosts and geographical regions. Full length ROP20 genes were amplified and sequenced. The results showed that the genes were 1659 bp in length and contained only a single exon, and that the A+T content varied from 46.68 to 47.20% among the 18 strains. The results of sequence alignment indicated that there were 30 variable nucleotide positions (0-1.40%) in the 18 T. gondii strains containing 18 transitions and 11 transversions, representing 1.81% overall sequence variation. Phylogenetic analysis of the ROP20 sequences showed that ROP20 variation could differentiate between the clonal lineage genotypes I and ToxoDB #9, indicating that ROP20 exhibits a relatively marked degree of sequence diversity and might represent a novel genetic marker for intraspecies phylogenetic analyses of T. gondii.

Seroprevalence, risk factors and genetic characterization of Toxoplasma gondii in free-range white yaks (Bos grunniens) in China.

The objective of this study was to determine the seroprevalence, risk factors and genotypes of Toxoplasma gondii in white yaks (Bos grunniens) in China. A total of 974 serum samples were collected from white yaks in Tianzhu Tibetan Autonomous County (TTAC), Gansu province, northwest China for detecting T. gondii antibodies by the modified agglutination test (MAT), and 414 tissues belonging to 138 white yaks were collected for detecting T. gondii DNA by amplification of B1 gene with a semi-nested PCR. A total of 155 serum samples (15.91%) were seropositive for T. gondii antibodies at a 1:100 cut-off, and 10 DNA samples (7.25%) were positive for the T. gondii B1 gene, which were genetically characterized using multilocus PCR-RFLP. Only one genotype (ToxoDB#9) was identified from two samples with complete genotyping results. Statistically significant differences were not observed between T. gondii seroprevalence and gender, season or pregnancy in the logistic regression analysis (P>0.05). Ages of white yaks was considered as a main risk factor associated with T. gondii infection. Our results indicated a widespread exposure to T. gondii among white yaks, and revealed the genotype ToxoDB#9 of T. gondii in white yaks for the first time in China.

First serologic survey of Q fever in free-range yaks in China.

The objective of the present investigation was to determine the seroprevalence of Coxiella burnetii infection in free-range yaks in China. A total of 552 serum samples were collected from yaks in Gansu province, northwest China between April 2013 and January 2014, and antibodies against C. burnetii were evaluated using enzyme-linked immunosorbent assay (ELISA). Overall, 13.59% (75/552, 95% CI: 10.73-16.45) of the examined animals were positive for C. burnetii antibodies. There was no significant difference in C. burnetii seroprevalence between female yaks (13.78%, 95% CI: 10.36-17.19) and male yaks (13.13%, 95% CI: 7.89-18.36) (P>0.05). Coxiella burnetii seroprevalence in yaks in different age groups ranged from 10.88% to 15.26%, but the difference was not statistically significant (P>0.05). Coxiella burnetii seroprevalence in yaks sampled in different seasons ranged from 12.06% (autumn) to 18.33% (summer), but the difference was not statistically significant (P>0.05). This is the first report of C. burnetii seroprevalence in free-range yaks in China, indicating the need for measures to be taken to control C. burnetii infection in free-range yaks in China.

Detection of common deafness mutation by maternal plasma cell-free DNA.

OBJECTIVES: The aim is to investigate the use of the ligase detection reaction (LDR) microarray to examine the difference of the single nucleotide between the pregnant woman and the fetus by cell-free DNA in the maternal plasma in congenital deafness. MATERIALS AND METHODS: The proband and the couples' venous blood samples and the amniotic fluid/ chorionic villi collected from seven deafness families for prenatal diagnosis were analyzed. The cell-free DNA from maternal plasma was examined to determine if they carried the mutations of GJB2 235delC. RESULTS: Three samples were found to carry the mutation of GJB2 235delC. It is in agreement with the sequencing results. The affected fetuses were suggested to take invasive procedure for confirmation. CONCLUSIONS: The chip may be a potential method to screen for congenital deafness based on maternal plasma DNA.

Prevalence and genetic characterization of Toxoplasma gondii infection in bats in southern China.

Toxoplasma gondii can infect a wide variety of warm-blooded animals, including bats. Limited information on T. gondii infection in bats is available in China. The objective of the present study was to determine prevalence and genetic diversity of T. gondii infection in bats in southern China. A total of 608 bats representing 12 species, including 120 Aselliscus stoliczkanus, 59 Myotis chinensis, 11 Miniopterus schreibersii, 53 Rhinolophus affinis, 32 Rhinolophus pusillus, 81 Hipposideros armiger, 28 Hipposideros fulvus, 32 Cynopterus brachyotis, 14 Cynopterus sphinx, 45 Eonycteris spelaea, 109 Hipposideros larvatus, and 24 Taphozous melanopogon, were collected from Yunnan and Guangxi provinces, southern China. They were examined for the presence of T. gondii DNA by amplification of the B1 gene using a nested PCR, and the positive samples were genotyped at 11 genetic loci (SAG1, 5'- and 3'-SAG2, alternative SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) using multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology. Fifty-nine (9.7%) of these bats were detected positive by PCR but only five of these positive DNA samples were completely typed at all loci; of which 4 samples, 2 from A. stoliczkanus, and 2 from H. larvatus, belonged to ToxoDB Genotype #10, and the other one from H. larvatus was identified as ToxoDB Genotype #9 (http://toxodb.org/toxo/). To our knowledge, this is the first report of molecular detection and genetic characterization of T. gondii infection in bats in China. The results show that these bats are potential reservoirs for T. gondii transmission, which may pose a threat to human health.

Diagnostic value of magnetic resonance cholangiopancreatography for biliary complications in orthotopic liver transplantation: a meta-analysis.

BACKGROUND AND OBJECTIVE: Magnetic resonance cholangiopancreatography (MRCP) is a noninvasive procedure to diagnose biliary complications. The aim of the present meta-analysis was to establish the overall diagnostic accuracy of MRCP to diagnose biliary complications post-orthotopic liver transplantation (OLT). METHODS: A systematic review was performed by searching electronic bibliographic databases prior to May 2012. Sensitivity, specificity, and other measures of the accuracy of MRCP for diagnosis of post-OLT were summarized using a random-effects or a fixed-effects model. Receiver operating characteristic curves were used to summarize overall test performance. RESULTS: Fourteen studies, which involved 892 subjects were eligible for the analysis. The summary estimates of sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and area under cure of MRCP for diagnosis of biliary complications were as follows: 0.95, 0.92, 10.23, 0.08, 206.59, and 0.979, respectively. The results for biliary strictures in four studies involving 177 subjects were 0.94, 0.95, 0.96, 0.09, 178.33, and 0.973 respectively. CONCLUSIONS: MRCP is a sensitive and specific technique to diagnose biliary complications.

Unbalanced translocation in an adult patient with premature ovarian failure and mental retardation detected by spectral karyotyping and array-comparative genomic hybridization.

BACKGROUND: There are only three cases of unbalanced translocation (X;1) reported in childhood in the literature, while no such phenotypic information is available in adults. MATERIALS AND METHODS: To delineate the phenotype-genotype relationship of unbalanced translocation (X;1) in adulthood, we reported here a 20-year-old female with an unbalanced translocation (X;1) which was determined by spectral karyotyping, array-comparative genomic hybridization and subtelomeric fluorescence in situ hybridization (FISH). RESULTS: The phenotype of partial trisomy 1 and partial monosomy X of the present case was much attenuated, including premature ovarian failure, mental retardation, class I obesity, mild dysmorphism and delayed secondary sexual characteristics. The breakpoints of the unbalanced translocation were accurately located at Xq28 and 1q32.1. The large amplification on Chromosome 1 q arm was found to involve 312 genes and the deletion on Chromosome X q arm also involved 141 genes. Overall, genes associated with physiological process (47 genes), cellular process (33), development (23), response to stimulus (1) and reproduction (1) were observed in the amplification on Chromosome 1 q arm. In addition, genes related to physiological process (23 genes), cellular process (13), development (6) and response to stimulus (2) were observed in the large deletion on chromosome X q arm. Late-replication studies revealed the existence of skewed X inactivation in the derivative X chromosome. CONCLUSIONS: The phenotype of partial monosomy X and partial trisomy 1q is much attenuated in case of unbalanced translocation (X;1) in adulthood probably owing to skewed X inactivation in derivative X chromosome.

[Pattern recognition of stereoscopic features of the leaves epidermis of medicinal Curcuma plants in China by image analysis].

OBJECTIVE: To provide new ways for classifying and identifying medicinal curcuma plants. METHOD: Based on classical taxonomy, the microscopic features of the epidermis of Curcuma plants in China were scored quantitatively by stereology and image analysis. RESULT: It showed that there were a lot of differences in density, size and shape of the epidermic cells among the medicinal Curcuma plants in China; that the average perimeter of the upper epidermic cells, average sectional area of the lower epidermis, stoma density and trichoma distribution of the upper and lower epidermis, etc. could be considered to be the main evidences for the microscopic identification of leaves of Curcuma. The 21 materials which belong to 11 species of Curcuma in China were systematized into 6 species, 1 species complex, 2 cultivated varieties. CONCLUSION: It can be concluded that stereology, and image analysis are advanced and feasible in pharmacognosy and taxonomy especially in the authentication of the relative and easily confused species.

[Culture techniques of Asarum sieboldii Miq].

Asarum sieboldii was domesticated in Sichuan, China. The rational planting density was selected according to this study. In the deep, loose and fertile soil, it grows well under semisunlight. Seedlings are raised in the seedling bed for two or three years, and then transplanted to the field, Harvest is started after three of four years of growth. Fertilizer is applied at the beginning of March and the beginning or middle of July.

[Preliminary studies on medicinal plants of Corydalis in Sichuan].

This paper reports the species, distribution and medical uses of Corydalis in Sichuan Province, of which 3 species e.g., Corydalis linstowiana and C. moupinensis, are found in China for the first time, 5 species C. balansae, C. trifoliolata e. g. are found in Sichuan for the first time. Medica effects of 10 species are described.

[A primary survey of the resources of Chinese materia medica in the Ganzi District].

This paper gives an account of the classification, distribution and resources of medicinal plants, traditional Zang folk medicine and historical changes of some rare crude drugs in Ganzi district. Rational suggestions have been made with regard to the preservation, development and utilization of the natural resources that are medicinally and economically important.