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Traumatic brain injury (TBI) induced neuroinflammation is featured as excessive glial inflammatory activation and violent neurologic destruction and dysfunction. Massive microglia activation in situ and disrupt of blood-brain barrier contribute to severely collapsed nervous system. Tizoxanide (TIZ), a synthetic thiazolide derivative agent possessing a broad-spectrum anti-infective effect, currently shows a potential resistance against pathogens like bacteria, virus and parasites, while its underlying role in neuroinflammation is elusive. The study aimed to explore the effect of TIZ on neuroinflammation in vitro microglia.Primary microglia were accepted to neuroinflammatory activation via lipopolysaccharide (LPS) administration. TIZ was conducted to pretreatment of microglia. Cell viability, inflammatory cytokines, chemotaxis, nitric oxide release, inflammation-related enzymes, and mitogen-activated protein kinase (MAPK) pathway activation in microglia were investigated respectively.We demonstrated that TIZ administration attenuates inflammatory cytokines and chemokines through quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) of medium supernatant. In addition, TIZ reduces pro-inflammatory mediators and nitric oxide release in microglia. Furtherly, TIZ inhibits the level of p38/MAPK pathway in LPS stimuli, indicating that TIZ negatively regulates neuroinflammation via inhibiting p38/MAPK pathway.TIZ is verified to be an anti-inflammation effect on neuroinflammation in microglia via downregulation of p38/MAPK pathway, which restrains inflammation by reduced inflammatory cytokines, chemokines and mediators and decreased nitric oxide release. To summarize, TIZ is considered to be a promising reagent to alleviate neuroinflammation targeting microglia in nervous system injury.

Adipogenesis plays a key role in increasing fat mass, which is a main characteristic for obesity, and involves preadipocyte proliferation and differentiation. Recently, more and more evidences suggested that microRNAs (miRNAs) is an important member of the regulatory network of adipogenesis. In this study, miR-143a-3p was highly expressed in adipose tissues of obese mice, and was up-regulated at the middle and last stage of 3T3-L1 adipocyte differentiation. Using mouse 3T3-L1 cells line, which is an ideal model in vitro for the study of adipogenesis, we observed that overexpression of miR-143a-3p inhibited the preadipocyte proliferation, and enhanced the preadipocyte differentiation. In contrast, the inhibition of miR-143a-3p expression promoted the preadipocyte proliferation, and inhibited the preadipocyte differentiation. Further analysis suggested that miR-143a-3p mediating preadipocyte differentiation might be involved in fatty acid metabolism. In addition, we found that miR-143-3p and PPARγ, an activator of miR-143a-3p transcription, could regulate each other. Compared with miR-143a-3p, MAPK7 played an opposite role in the proliferation and differentiation of adipocyte. Further analysis indicated that MAPK7 is a target gene of miR-143a-3p in 3T3-L1 cells, and inhibition of MAPK7 recede the effect of miR-143a-3p on preadipocyte proliferation and differentiation. Taken together, these results indicated that as a regulator of PPARγ, miR-143a-3p play an important role in adipogenesis via regulating MAPK7 and fatty acid.

Green leaf volatiles (GLVs) have been reported to play an important role in the host-locating behavior of several folivores that feed on angiosperms. However, next to nothing is known about how the green leafhopper, Empoasca vitis, chooses suitable host plants and whether it detects differing emission levels of GLV components among genetically different tea varieties. Here we found that the constitutive transcript level of the tea hydroperoxide lyase (HPL) gene CsiHPL1, and the amounts of (Z)-3-hexenyl acetate and of total GLV components are significantly higher in tea varieties that are susceptible to E. vitis (Enbiao (EB) and Banzhuyuan (BZY)) than in varieties that are resistant to E. vitis (Changxingzisun (CX) and Juyan (JY)). Moreover, the results of a Y-tube olfactometer bioassay and an oviposition preference assay suggest that (Z)-3-hexenyl acetate and (Z)-3-hexenol offer host and oviposition cues for E. vitis female adults. Taken together, the two GLV components, (Z)-3-hexenol and especially (Z)-3-hexenyl acetate, provide a plausible mechanism by which tea green leafhoppers distinguish among resistant and susceptible varieties. Future research should be carried out to obtain the threshold of the above indices and then assess their reasonableness. The development of practical detection indices would greatly improve our ability to screen and develop tea varieties that are resistant to E. vitis.

The aim of the study was to investigate the influences of micro ribonucleic acid (miR)-21 and downstream Toll-like receptor 4 (TLR4)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway on myocardial apoptosis induced by myocardial ischemia-reperfusion (I/R) injury in rats.Recombinant adeno-associated virus rAAV9-ZsGreen-pre-miR-21 and blank control virus were constructed. A total of 48 Sprague-Dawley (SD) rats were randomly divided into S1 group (open chest only), S2 group (transfection with blank virus + open chest), I/R1 group (transfection with blank virus + 6 d of myocardial I/R), and I/R2 group (transfection with miR-21 + 6 d of myocardial I/R). The cardiac function and myocardial infarct size of rats were evaluated in each group. Quantitative Polymerase Chain Reaction (qPCR) was applied to measure the expression level of miR-21 in the myocardium. The level of myocardial apoptosis in each group was detected through terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) staining. Western blotting was performed to determine the protein expression levels of B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (Bax), Caspase-3, TLR4, and NF-κB in the myocardium. The content of interleukin-6 (IL-6) and IL-10 was measured using enzyme-linked immunosorbent assay (ELISA) kits.The cardiac function of rats in I/R1 and I/R2 groups was significantly lower than that in S1 and S2 groups (p0.01). Rats in I/R2 group had better cardiac function than those in I/R1 group (p0.01). In I/R1 group, the level of myocardial apoptosis of rats was overtly increased compared with that in S1, S2, and I/R2 groups (p0.01), while the expression level of miR-21 in myocardium was evidently lower than that in S1, S2, and I/R2 groups (p0.01). Compared with S1, S2, and I/R2 groups, I/R1 group had markedly decreased Bcl-2/Bax expression level and IL-10 content and overtly elevated expression levels of Caspase-3, p-TLR4, p-NF-κB, and IL-6 content in the myocardium (p0.01).Myocardial I/R injury in rats leads to decreased expression of miR-21. The overexpression of miR-21 is able to effectively inhibit the TLR4/NF-κB pathway and reduce the level of myocardial apoptosis of rats and the release of inflammatory factors.

Chemo-resistance of colon cancer remains a major problem in therapy. The role of miR-215-3p in the chemo-sensitivity of colon cancer remains unidentified.Here, we constructed a 5-Fluoracil (5-Fu) resistant HCT116 cell line (HCT116/5-Fu) and miR-215-3p expression levels were measured in 56 cases of colon cancer tissues and 23 cases of normal tissues by quantitative real-time polymerase chain reaction (qRT-PCR). The effects of miR-215-3p on colon cancer cell growth and apoptosis were investigated using cell counting kit-8 (CCK-8) and apoptosis assay, respectively. In addition, CXC-chemokine receptor type1 (CXCR1) was identified as a target of miR-215-3p by using luciferase reporter assay.miR-215-3p was down-expressed in the 5-FU resistant cell compared to the parent cell. The level of miR-215-3p was correlated with the 5-Fu sensibility of colorectal cancer cell and the alteration of miR-215-3p affected the sensibility of colorectal cancer cells toward 5-Fu. Furthermore, miR-215-3p accelerated the apoptosis of colorectal cancer cell which was treated with 5-Fu. Mechanically, miR-215-3p regulated the level of endogenous CXCR1 in HCT116 cell and alternation of CXCR1 affected the 5-Fu sensibility mediated by miR-215-3p. Finally, overexpression of miR-215-3p restrained the growth of HCT116/5-Fu cells in the xenograft model.MiR-215-3p improved the 5-Fu sensibility via regulating the expression of CXCR1 in the colorectal cancer cell.

To investigate the relationship between microRNA-203 (miR-203) and diabetic nephropathy and its potential mechanism.The expression of microRNA-203 in mice with diabetic nephropathy and M4200 cells cultured with high glucose was detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Toll-like receptor 4 (TLR4), the target gene of microRNA-203, was predicted and screened by bioinformatics method. Real-time quantitative PCR and Western blot were used to detect the endogenous TLR4 level in renal cortex of db/db mice with diabetic nephropathy and glomerular mesangial cells cultured in high glucose or low glucose. The expression of microRNA-203 and TLR4 mRNA were evaluated by RT-PCR after treatment of miR-203 mimics and inhibitor. The protein of TLR4 level was detected by Western blot. Additionally, the proliferation ability of cells was evaluated by Cell Counting Kit-8 (CCK8). The target relationship between microRNA-203 and TLR4 3' UTR was confirmed by luciferase reporter assay RESULTS: The expression of miR-203 was significantly decreased in the kidney of mice with diabetic nephropathy and M4200 cells cultured in high glucose. On the contrary, TLR4 expression was significantly increased. Results of in vitro experiments showed that miR-203 could bind to 3'UTR region of TLR4. Overexpression of microRNA-203 significantly decreased the levels of TLR4 mRNA and protein. Meanwhile, low expression of miR-203 leaded to increased TLR4 expression, resulting in an enhanced proliferation of M4200 cells.The downregulation of microRNA-203 leaded to an increased level of TLR4, thus promoting proliferation of M4200 cells in the pathogenesis of diabetic nephropathy.

N^(6)-methyladenosine (m^(6)A) has been identified as a conserved epitranscriptomic modification of eukaryotic mRNAs, and plays important biological roles in the regulation of cellular metabolic processes. However, its role in myogenic differentiation is unclear. Here, we altered the m^(6)A RNA methylation level by overexpression of METTL3, and explored the effect of m^(6)A RNA methylation on myogenic differentiation of murine myoblasts in vitro. The m6A RNA methylation level is regulated by exogenous methylation inhibitor cycloleucine (Cyc) and methyl donor betaine (Bet). Therefore, chemical reagents of Cyc and Bet were used to test the regulatory effect of m^(6)A RNA methylation on myogenic differentiation. Results showed that METTL3 and Bet positively regulated the m^(6)A RNA methylation levels, and Cyc negatively regulated m^(6)A RNA methylation levels. In addition, m^(6)A methylation positively regulated myogenic differentiation in murine myoblasts. These findings provide insight in the mechanisms underlying the effect of m^(6)A RNA methylation on myogenesis.

MiR-222-3р has been implicated in tumor cell proliferation and has an important role in the differentiation and maturation of myogenic cells. However, its role in skeletal myoblast proliferation is still unclear. In this study, we found that miR-222-3р expression increases initially and then decreases during C2C12 myoblast proliferation. Using synthetic miRNA mimics and inhibitors in gain- or loss-of-function experiments, we snowed that miR-222-3р overexpression in C2C12 cells promotes myoblast proliferation and represses myofiber formation, while miR-222-3р downregulation has the opposite effect. Using a prediction program, BTG2 was identified as a possible target gene of miR-222-3р. During myogenesis, miR-222-3р mimics repress BTG2 expression, while miR-222-3р inhibitors promote BTG2 expression. Using dual-luciferase reporter assay, we further demonstrated that miR-222-3р specifically targets BTG2. Additionally, we show that siRNA-mediated downregulation of BTG2 expression in C2C12 myoblasts promotes the proliferation and suppresses differentiation. In conclusion, we provide a novel insight into the mechanism by which miR-222-3р regulates the proliferation and differentiation of C2C12 myoblasts by targeting BTG2. This information contributes to our understanding of the role of miRNAs in skeletal muscle development.

The aim of this study was to analyze the results of two crossing systems between wild boars and different domesticated pig breeds. Hybrid wild boars were produced by crossing captured wild boars with Meishan pigs and LY sows according to the traditional production system. The resultant commercial hybrids were black and white in coat color, respectively. Significant differences were found in the carcass and meat quality traits and nutritional values between these two hybrid wild boars. Compared with the white hybrid wild boars, at the age of 300 days, the body weight of black hybrid wild boars was 9.41 kg lower, while percent lean was 2.51% less and percent fat 2.45% higher (P < 0.05). The black hybrid wild boars had higher pH2 (6.17 vs 6.09) and intramuscular fat (3.34 vs 2.52%), lower drip loss (2.21 vs 2.68%) and shear force (44.00 vs 52.23) (P < 0.05), and more unsaturated fatty acids and essential amino acids (P < 0.05). In conclusion, cross breeding was shown to be an effective method to improve the overall production performance of wild boars, but crossing with different dam line breeds caused different responses. Compared with the white hybrid wild boars, the black hybrid wild boars had worse growth rate and carcass traits, but better meat quality traits and nutritional values.

DNA methylation is an important component of the epigenetic machinery and plays a critical role in transcriptional regulation. It mostly occurs in CpG abundant regions, known as CpG islands (CGIs). G protein-coupled receptor 120 (GPR120) functions as an omega-3 fatty acid receptor and is involved in multiple-biological processes, including lipogenesis. Herein, we show that GPR120 is highly expressed in porcine mature adipose tissue and is positively associated with adipose tissue development (r = 0.86, P < 0.01). We also predicted 5 CGIs across the GPR120 genomic sequence and investigated their methylation status using the MassArray approach. Our results show that these CGIs exhibit significantly different methylation states (P(CGI) < 0.01), and that the DNA methylation of GPR120 5ꞌ-untranslated and first exon regions can negatively regulate its expression levels. This study will aid further investigations on the epigenetic mechanism regulating GPR120 expression.

The myosin heavy chain (MyHC) composition, glycolytic potential, mitochondrial content, and gene expression related to energy metabolism were analyzed in eight muscles from Tibetan pigs, to study how meat quality develops in different muscle tissues. The muscles were classified into three clusters, based on MyHC composition: masseter, trapezius, and latissimus dorsi as 'slow-oxidative-type'; psoas major and semimembranosus as 'intermediate-type'; and longissimus dorsi, obliquus externus abdominis, and semitendinosus as 'fast-glycolytic-type'. The 'slow-oxidative-type' muscles had the highest MyHC I and MyHC IIA content (P < 0.01); 'intermediate-type' muscles, the highest MyHC IIx content (P < 0.01); and 'fast-glycolytic-type' muscles, the highest MyHC IIb content (P < 0.01). The pH values measured in 'slow-oxidative-type' muscles were higher than those in the other clusters were; however, the color of 'fast-glycolytic-type' muscles was palest (P < 0.01). Mitochondrial content increased in the order: fast-glycolytic-type < intermediate-type < slow-oxidative-type. In the 'slow-oxidative-type' muscles, the expression levels of genes related to ATP synthesis were higher, but were lower for those related to glycogen synthesis and glycolysis. Mitochondrial content was significantly positively correlated with MyHC I content, but negatively correlated with MyHC IIb content. MyHC I and mitochondrial content were both negatively correlated with glycolytic potential. Overall, muscles used frequently in exercise had a higher proportion of type I fibers. 'Slow-oxidative-type' muscles, rich in type I fibers with higher mitochondrial and lower glycogen and glucose contents, had a higher ATP synthesis efficiency and lower glycolytic capacity, which contributed to their superior meat quality.

DNA methylation is a stable epigenetic mark mediating gene expression. Methylation is crucial for diverse biological processes, including aging and embryo development. FASN (fatty acid synthase) plays an important role in de novo lipogenesis, through catalyzing the reductive synthesis of long-chain fatty acids. In this study, we investigated the FASN gene expression pattern and corresponding DNA methylation status in the inner layer of backfat from Jinhua pigs at different developmental stages. Our results showed that FASN gene expression increases with age and is positively associated with adipocyte volume (r = 0.98, P < 0.01). In addition, the DNA methylation level for the first exon (0.11, CGI 3) of the FASN gene is approximately 8-fold lower than levels for its promoter (0.94, CGI 1&2) (two-way ANOVA, PCGI < 0.01). The association analysis revealed that both promoter (r = -0.944, P < 0.01) and first exon methylation (r = -0.774, P < 0.01) are significantly and negatively correlated with FASN gene expression. Our results will benefit future investigations of the epigenetic mechanism underlying FASN gene expression.

Pseudorabies virus (PRV) early protein UL31 is a homologue of herpes simplex virus 1 (HSV-1) UL31, which is a multifunctional protein important for HSV-1 infection. However, the precise roles of PRV UL31 in virus life cycle are still poorly understood. A relatively crucial tool for uncovering the function of UL31 is an antiserum that specifically detects UL31 in the PRV-infected cells. For this purpose, a recombinant UL31 protein consisting of N-terminal 27 aa of UL31 fused to EYFP and His-tag was expressed, purified and used for the preparation of antiserum in BALB/c mice. Our results show that Western blot analysis and immunofluorescence assay showed that this antiserum could specifically detect the purified recombinant UL31 as well as full-length UL31 in the PRV infected cells. These results demonstrate that the prepared antiserum could serve as a valuable tool for further studies of UL31 functions in PRV infection.

Inflammatory bowel disease (IBD) is a common disease in human resulted from a various of factors including genetic background, immune system and environment factors.Recent studies suggest pro-inflammatory cytokine IL-17 producing cell subset was involved in the disease development and the maintenance of IBD. And the differentiated and activation of IL-17 producing cells were mostly dependent on the cytokines profile secreted by innate cells in intestinal tissues. In this study, we examined the functions of IL-6 signal in regulatory of IL-17 production in acute IBD model.Wildtype mice were treated with anti-IL-6 neutralizing antibodies to block IL-6 signal And then treated with DSS to induce acute IBD.Mice treated with anti-IL-6 neutralizing antibodies show severe colitis and high level of pro-inflammatory cytokine IL-17 production in DSS-induced acute IBD model when compared with control group. Our research suggested blockade of IL-6 signal pathways in acute colitus model resulted in specific activation of IL-17 producing cell population. Furthermore, CD44+ activated Th17 cell population and CD44- IL-17 producing T cells exhibited different susceptibility to IL-6 signal in our model.Blockade of IL-6 signal in DSS-induced acuted IBD model increased IL-17 production level specifically in CD44- T cells and reduced CD44+ Th17 cell population.

To study the impact of cold ischemia on tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) expression after liver transplantation, a stable model of partial liver transplantation in rats was established. The experimental animals were divided into the following groups: a partial hepatectomy control group, a group that received partial liver transplantation after 30 min of cold ischemia (experimental group A), and a group that received a partial liver transplantation after 10 h of cold ischemia (experimental group B). The survival rate was observed in each group. The liver tissue was sampled 1, 2, and 4 days after surgery, and immunohistochemical detection of proliferating cell nuclear antigen TNF-α and IL-10 was performed. The correlation between liver regeneration and TNF-α and IL-10 expression was analyzed, and the impact of the 2 cytokines on rat liver regeneration after liver transplantation was evaluated. The survival rates of rats in the partial hepatectomy control group, in the group that received a partial liver transplantation after 30 min of cold ischemia, and the group that received a partial liver transplantation after 10 h of cold ischemia were 100, 70, and 33.3%, respectively. The expression of proliferating cell nuclear antigen and TNF-α was decreased (P < 0.05), and IL-10 expression was increased (P < 0.05) in animals that received a partial liver transplant after 10 h of cold ischemia compared with that in the animals that received a partial liver transplant after 30 min of cold ischemia. We conclude that with the extension of cold ischemic time, liver regeneration and survival rate after liver transplantation decreased. TNF-α and IL-10 play important regulatory roles in the regeneration process of transplanted livers.

This study analyzed the effect of muscle-fiber type composition on glycogenin-1 (GYG) gene expression and its impact on pH. The longissimus dorsi (LD) muscle contains more type IIB fibers (75.10%) than does the psoas major (PM) muscle (41.58%), while the PM has more type I (3.65 vs 0.94%), type IIA (34.15 vs 10.63%), and type IIX (20.62 vs 13.33%) fibers. Compared with PM, glycolytic potential (GP), pH45 min, and ΔpH from 45 min to 24 h post-mortem were all relatively higher in LD. Glycogen metabolites (lactate and GP) were negatively correlated with pH24 h and positively correlated with ΔpH. Expression of GYG was generally higher in LD. GYG expression was positively correlated with glycogen metabolite (lactate and GP) content and ΔpH, and was negatively correlated with pH24 h. These data confirm that the muscle-fiber type and GP have significant effects on ultimate pH and pH decline, and suggest that expression of GYG in muscles is related to the metabolism of glycogen and may impact GP, ΔpH, and ultimate pH. High expression of GYG was associated with a high glycogen content, large pH decline, and low ultimate pH in muscles post-mortem.

DNA methylation may be involved in regulating the expression of protein-coding genes, resulting in different fat and muscle phenotypes. Using a methylation-sensitive amplified polymorphism approach, we obtained 7423 bands by selective amplification of genomic DNA from six different fat depots and two heterogeneous muscle types from Duroc/Landrace/Yorkshire cross-bred pigs. The degrees of DNA methylation, determined by the percentages of hemi- and fully methylated sites relative to the total number of CCGG sites, were similar in male and female pigs for each specific tissue [χ(2) test; P (two-tailed) > 0.05]. Gender bias was therefore ignored. There were significant differences in the degree of DNA methylation among the eight tissue types [χ(2) test; P(total) (two-tailed) = 0.009]. However, similar degrees of methylation were observed among the six fat depots [χ(2) test; P(fat) (two-tailed) = 0.24 > 0.05]and between the two muscle types [χ(2) test; P(muscle) (two-tailed) = 0.76 > 0.05]. We conclude that the degree of DNA methylation differs between porcine fat and muscle tissue, but that the methylation status of a particular tissue type is similar, despite being deposited at different body sites.

Malate dehydrogenases 1 and 2 (MDH1 and MDH2), and malic enzyme 1 (ME1) play important roles in the Krebs cycle for energy metabolism. The mRNA abundance changes of MDH1, MDH2 and ME1 genes were measured across six different adipose tissues from the leaner Landrace and fatty Rongchang pig breeds using quantitative real-time PCR. The mRNA of MDH1, MDH2 and ME1 was more abundant in fatty Rongchang pigs than in leaner Landrace pigs. In both breeds, females exhibited higher adipocyte volume and mRNA abundance of MDH1, MDH2 and ME1 compared with males. These values were higher in the subcutaneous adipose tissue compared with visceral adipose tissue. Furthermore, mRNA abundance changes of MDH1, MDH2 and ME1 have the remarked significant positive correlation with adipocyte volume across the six adipose tissue types. We conclude that there are breed-, gender- and tissue-specific expression patterns of ME1, MDH1 and MDH2, which highlight their potential as candidate genes for selecting for fat volume in pigs.

The Yanan (YN) pig is a traditional Chinese indigenous breed that is raised in southwest China in the Sichuan Province, but there is little data on the germplasm characteristics of this breed. To evaluate carcass characteristics and meat quality of the YN pig, we compared carcass and meat quality of YN pigs and Landrace × Yanan (CY) hybrid pigs; 30 YN pigs and 30 CY pigs weighing 20 ± 2 kg were reared and slaughtered at the normal slaughter weight (100-120 kg). The carcasses were chilled and the left carcass side was dissected into bone, lean meat, fat, and skin; meat quality parameters were measured. Carcasses of YN pigs were lighter (88.85 vs 90.05 kg, P < 0.05) and shorter (71.88 vs 77.61 cm, P < 0.001); they contained less lean meat (41.60 vs 49.25%, P < 0.001), less ham and breech (25.93 vs 27.53%, P < 0.001) and less carcass bone (9.83 vs 10.53%, P < 0.01) than did carcasses of CY pigs. On the other hand, YN pigs had more carcass subcutaneous fat and skin (48.58 vs 40.23%, P < 0.001), thicker backfat (3.67 vs 3.43 cm, P < 0.001) and smaller loin muscle area (9.83 vs 26.91 cm(2), P < 0.001) compared with CY pigs. Among meat quality parameters, YN pigs had higher pH(1) (6.41 vs 6.17, P < 0.001), higher color score(u) (3.86 vs 3.36, P < 0.001) and lower Minolta L(u) values (40.89 vs 45.32, P < 0.01) than CY pigs. On the other hand, YN pigs had lower drip loss (1.31 vs 2.26%, P < 0.05) and lesser fiber area (2351.34 vs 3025.43 μm(2), P < 0.01) than CY pigs. Both breeds had high intramuscular fat (4.46% in YN and 4.45% in CY). No significant differences in other carcass traits and meat quality were found in the two populations. We conclude that YN pigs could be used in commercial pig production to provide good tasting and high-quality niche products.

To normalize a set of quantitative real-time PCR (q-PCR) data, it is essential to determine an optimal number/set of housekeeping genes, as the abundance of housekeeping genes can vary across tissues or cells during different developmental stages, or even under certain environmental conditions. In this study, of the 20 commonly used endogenous control genes, 13, 18 and 17 genes exhibited credible stability in 56 different tissues, 10 types of adipose tissue and five types of muscle tissue, respectively. Our analysis clearly showed that three optimal housekeeping genes are adequate for an accurate normalization, which correlated well with the theoretical optimal number (r ≥ 0.94). In terms of economical and experimental feasibility, we recommend the use of the three most stable housekeeping genes for calculating the normalization factor. Based on our results, the three most stable housekeeping genes in all analysed samples (TOP2B, HSPCB and YWHAZ) are recommended for accurate normalization of q-PCR data. We also suggest that two different sets of housekeeping genes are appropriate for 10 types of adipose tissue (the HSPCB, ALDOA and GAPDH genes) and five types of muscle tissue (the TOP2B, HSPCB and YWHAZ genes), respectively. Our report will serve as a valuable reference for other studies aimed at measuring tissue-specific mRNA abundance in porcine samples.

Chronic cholangitis caused by hepatolithiasis is a common disease in Southeast Asia. Few studies have addressed the effects of chronic cholangitis on cyclic activity of the sphincter of Oddi (SO). In this study, we investigated the changes of myoelectric activity in rabbits with chronic cholangitis in vivo and in vitro.Chronic cholangitis was induced in rabbits by initially introducing three pieces of 2-0 silk suture and sequentially injecting E. coli into the choledochus through the tube in ductus cysticus. In in vivo experiments, myoelectric activity of SO was recorded by a circular electrode through the jejunum stump in conscious animals. In in vitro experiments, the SO was completely isolated and the myoelectric activity was recorded by a circular electrode in a 10-mL organ bath filled with Krebs solution, with or without addition of cholecystokinin-8 (CCK-8), KCl, ionomycin or induction of capacitative calcium entry (CCE).In comparison with control and non-infected rabbits, the rabbits with chronic cholangitis showed higher levels of alkaline phosphatase and gamma-glutamyltransferase and significant pathological changes including increased inflammatory infiltration and collagen deposition in mucosae or muscular layer. Cyclic myoelectric activity of SO at phases 2 and 3 of migrating motor complex and the excitatory response to CCK-8 were dramatically decreased in animals with chronic cholangitis. Myoelectric activity of SO was also significantly decreased in vitro with or without agonists or with induction of CCE.Myoelectric activity of SO and its response to agonists are decreased in rabbits with chronic cholangitis both in vivo and in vitro.

The microbicidal activity of a complex glutaraldehyde disinfectant was observed. The result showed that when the temperature of the test was 20 degrees C and 10% calf serum were added to the bacterial suspensions 2% diluted solution of the disinfectant for a two-minute contact could kill 99.99% of Staphylococcus aureus, and 1% diluted solution for two minutes could kill 99.99% of E. coli. The undiluted disinfectant with five-minute contact time could destroy the antigenicity of HBsAg. The undiluted disinfectant with thirty-five minute contact and 50% diluted solution with forty-five minute contact time could completely kill the spores of Bacillus subtilis var. niger under 50 degrees C. It was discovered that the germicidal concentration of the disinfectant was four times as high as the inhibition concentration and the used concentration determinal with capacity test was four times higher than as germicidal concentration when the three concentrations were compared. This result indicated that selection of suitable inactivator was very important. The practical concentration of disinfectant should be decided under the basis of the result from capacity test.

Electrophoretic techniques have been used to survey five different field-collected isolates (i.e., Anhui, Hubei, Guangxi, Sichuan and Yunnan) of S. japonicum from the mainland of China for genetic variation at 7 isozymes markers representing 9 loci. Polymorphic loci were observed for LDH-1, LDH-2, MDH and PGM, respectively, the proportion of polymorphic loci being 44.4%. Average heterozygosity varied between 0.223 and 0.425, with a mean of 0.332. Nei's genetic distance (D) among the populations of 5 isolates gave values between 0.001 and 0.039 with an average of 0.023, indicating that these isolates are very closely related.

Five different field-collected isolates (i.e., Anhui, Hubei, Guangxi, Sichuan and Yunnan) of Schistosoma japonicum from the mainland of China were compared by means of multilocus enzyme electrophoresis in polyacrylamide gels of individual male worm extracts. The enzyme systems used in the study included GDH, G6PD, LDH, MDH, PGI, PGM and SOD. Of 9 loci examined in the 7 isozyme systems, 4 were found to be polymorphic. These were LDH-1, LDH-2, MDH and PGM, the remainder of GDH, G6PD-1, G6PD-2, PGI and SOD being monomorphic. Our results indicated that the genetic polymorphism occurring in the natural population of different isolates of S. japonicum in the mainland of China. The allele frequencies at polymorphic loci coding for enzyme as well as the genetic identity and genetic distance in different isolates of S. japonicum will be reported elsewhere.