First report of Alternaria alternata causing leaf yellow spot on Heteropanax fragrans in China.

Heteropanax fragrans (Roxb.) Seem is a common garden landscape tree in China. In December 2020, a leaf disease on H. fragrans was observed in a 2 ha field in Zhanjiang (20.85° N, 109.28° E), Guangdong province, China. Early symptoms were small yellow spots on leaves. Later, the spots gradually expanded and turned into necrotic tissues with a clear yellow halo and a white center. The disease incidence on plants was 100%. Twenty diseased leaves were collected from the field. The margin of the diseased tissues was cut into 2 mm × 2 mm pieces, surface disinfected with 75% ethanol and 2% sodium hypochlorite for 30 and 60 s, respectively, and rinsed thrice with sterile water before isolation. The tissues were plated onto potato dextrose agar (PDA) medium and incubated at 28 ℃. After 2-day incubation, grayish fungal colonies appeared on the PDA, then pure cultures were produced by transferring hyphal tips to new PDA plates. Single-spore isolation method was used to recover pure cultures for three isolates (HFA-1, HFA-2, and HFA-3). The colonies first produced a light-grayish aerial mycelia, which turned dark grayish upon maturity. Conidiophores were branched. Conidia numbered from two to four in chains, were dark brown, ovoid, or ellipsoid and mostly beakless; had 1-4 transverse and 0-3 longitudinal septa; measured within 7.2-17.8 (average = 10.2) × 2.5-7.5 (average = 4.3) µm (n = 30). Molecular identification was performed using the colony polymerase chain reaction method with MightyAmp DNA Polymerase (Takara-Bio, Dalian, China) (Lu et al. 2012) to amplify the large subunit (LSU), internal transcribed spacer (ITS) region, translation elongation factor (TEF) , and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) with NL1/LR3, ITS1/ITS4, EF-1α-F/EF-1α-R, and GDF1/GDR1 (Walther et al. 2013;Woudenberg et al. 2015; Nishikawa and Nakashima. 2020). Amplicons of the isolates were sequenced and submitted to GenBank (LSU, ON088978-ON088980; ITS, MW629797, ON417005 and ON417006; TEF, MW654167, ON497264,and ON497265;GAPDH, MW654166, ON497262,and ON497263). The obtained sequences were 100% identical with those of Alternaria alternata strain CBS 102600 upon BLAST analysis . The sequences were also concatenated for phylogenetic analysis by maximum likelihood. The isolates clustered with A. alternata (CBS 102600, CBS 102598, CBS 118814, CBS 918.96,CBS 106.24, CBS 119543, CBS 916.96). The fungus associated with leaf yellow spot on H. fragrans was thus identified as A. alternata. Pathogenicity tests were conducted in a greenhouse at 24 â„ƒ-30 â„ƒ with 80% relative humidity. Individual plants were grown in pots (n = 5, 1 month old). The unwounded leaflets were inoculated with 5 mm-diameter mycelial plugs of the isolates or agar plugs (as control). The test was performed thrice. Disease symptoms were found on the leaves after 7 days, whereas the controls remained healthy. The pathogen was re-isolated from infected leaves and phenotypically identical to the original isolates to fulfill Koch's postulates. To our knowledge, this report is the first one on A. alternata causing leaf yellow spot on H. fragrans. Thus, this work provides an important reference for the control of this disease in the future.

First Report of Fusarium proliferatum Associated with Leaf Yellow on Alternanthera philoxeroides in China.

Alternanthera philoxeroides (Mart.) Griseb is a highly invasive weed commonly found in rice fields in China. In May 2021, leaf yellowing was observed on this weed (about 10 ha) in Zhanjiang (21°19'N, 110°20'E), Guangdong Province, China. Disease incidence was approximately 20% (n = 100 investigated plants). Ten yellow leaves from 10 plants were sampled, surface-sterilized with 75% ethanol for 30 s, followed by 2% NaClO for 5 min. The leaves were rinsed three times in sterile distilled water and four sections of each leaf were placed onto potato dextrose agar (PDA). Pure cultures were obtained by transferring hyphal tips to new PDA plates. Twenty-two isolates of Fusarium ssp. (69% of the isolates) were obtained from 55% of the leaf samples. Three representative single-spore isolates (APF-1, APF-2, and APF-3) were used for further study. Colonies were white to pink on PDA. Conidiogenous cells were monophialidic or polyphialidic. Macroconidia were slightly curved, tapering apically with three to five septa, and measured from 32.5-55.8 µm × 2.5-5.1 µm in size (n=50). The morphological features of these fungi were noted to be in line with those of Fusarium proliferatum (Leslie and Summerell, 2006). For molecular identification, a colony PCR method (Lu et al. 2012) was used to amplify the internal transcribed spacer (ITS) and portions of elongation factor 1-α (EF1-α), RNA polymerase II largest subunit (RPB1), and RNA polymerase II second largest subunit (RPB2) genes using primers ITS1/ITS4, EF1-728F/EF1-986R, RPB1-R8/RPB1-F5, and RPB2-7CF/fRPB2-11aR, respectively (O'Donnell et al. 1998; O'Donnell et al. 2010). The sequences were submitted to GenBank under accession numbers MZ026797-MZ026799 (ITS) and MZ032209-MZ032217 (RPB1, RPB2, EF1-α). The sequences of the three isolates were 100% identical (ITS, 537/537 bp; RPB1, 1606/1606 bp; RPB2, 770/770 bp and EF1-α, 683/683 bp) with those of F. proliferatum (accession nos. MT378328, MN193921, MH582196, and MH582344) through BLAST analysis. Analysis of the sequences revealed a 99.87 - 100% identity with the isolates of the F. proliferatum (F. fujikuroi species complex, Asian clade) by polyphasic identification using the FUSARIUM-ID database (Yilmaz et al. 2021). The sequences were also concatenated for phylogenetic analysis by the maximum likelihood method. The isolates clustered with F. proliferatum. Pathogenicity was tested through in vivo experiments. The inoculated and control plants (n = 5, 30 days old) were sprayed with a spore suspension (1 × 105 per mL) of the three isolates individually and sterile distilled water, respectively, until run-off (Feng and Li. 2019). The test was performed three times. The plants were grown in pots in a greenhouse at 25 °C to 28 °C, with relative humidity of approximately 80%. Yellowing was observed on the inoculated plants after 7 days, while the control plants remained healthy. The pathogen re-isolated from all the inoculated plants was identical to the inoculated isolates in terms of morphology and ITS sequences. No fungi were isolated from the control plants. To the best of our knowledge, this study is the first to report F. proliferatum causing yellow symptoms on A. philoxeroides. The fungus has some potential biological control properties, but its host range needs to be further determined.

Pseudocercospora rhododendricola Causing Leaf dark Spot on Rhododendron pulchrum by the first phylogenetic analyses.

Rhododendron pulchrum Sweet is a famous ornamental flower in China. In December 2020, a leaf spot disease was observed on cv. Maojuan in Zhanjiang (21.17 N, 110.18 E), Guangdong, China. The spots were irregular and distributed on both sides of the main vein. They were dark to black, and their borders were obvious. The coalescence of the spots eventually led to leaf wilt. The disease incidence was 100% (n = 100, about 50 ha ). Thirty infected leaves were collected from the field, and the margin of the diseased tissues was cut into 2 mm × 2 mm pieces. Samples were surface disinfected with 75% ethanol and 2% sodium hypochlorite for 30 and 60 s, respectively. They were rinsed thrice with sterile water before isolation. The tissues were plated on potato dextrose agar (PDA) medium and incubated at 28 ℃. After 5 days, fungal colonies appeared on the PDA. Pure cultures were produced by transferring hyphal tips to new PDA plates. Three isolates (RSP-1, RSP-2, and RSP-3) were obtained and the colonies of isolates were preserved in glycerol (15%) at -80 °C deposited at the Museum of Guangdong Ocean University. The morphology of these three isolates was consistent, and their sequences showed 100% homology according to ITS, TEF1, and ACT analysis results. The colonies grew to approximately 5 cm in diameter after 10 days. They showed olive green with off-white aerial mycelia. Stromata and conidia were observed on leaf lesions. Stromata were olivaceous brown. Conidia were solitary, cylindrical to narrowly obclavate, mildly curved, obtuse to rounded at the apex, and 1- to 3-septate; they had dimensions of 20 to 60 × 2.0 to 3.0 µm (n = 30). These morphological characteristics were not different from the description of Pseudocercospora rhododendricola (J.M. Yen) Deighton (Liu et al. 1998). For molecular identification, the colony PCR method with MightyAmp DNA Polymerase (Takara-Bio, Dalian, China) (Lu et al. 2012) was used to amplify the internal transcribed spacer (ITS), translation elongation factor 1-α gene (TEF1), and actin (ACT) loci of the isolates using primer pairs ITS4/ITS5, EF1/EF2, and ACT-512F/ACT-783R, respectively (White et al., 1990; O'Donnell et al. 1997). The sequences of the isolate RSP-1 were deposited in the GenBank (ITS, MW629798; TEF1, MW654168; and ACT, MW654170). BLAST analysis showed that the sequences of P. rhododendricola were submitted to GenBank for the first time by the author of this paper. A phylogenetic tree was generated based on the concatenated data of ITS, TEF1, and ACT sequences from GenBank by the Maximum Likelihood method. The isolates were closest to Pseudocercospora sp. CPC 14711 (Crous et al., 2013). Phylogenetic and morphological analyses identified the isolates as P. rhododendricola. Pathogenicity tests were conducted in a greenhouse at 24 °C-30 â„ƒ with 80% relative humidity. Healthy cv. Maojuan were grown in pots. Unwounded leaflets were inoculated with 5 mm-diameter mycelial plugs of the isolates or agar plugs (as control) (5 leaflets per plant, 3 plants, 2-month-old plants). The test was performed thrice. Disease symptoms were found on the leaves after 2 weeks, whereas the control plants remained healthy. The fungus was re-isolated from the infected leaves and confirmed as the same isolates by morphological and ITS analyses. P. rhododendricola was the cause of leaf spot of Rhododendron sp. from Singapore (Liu et al., 1998). For the first time, this pathogen was identified by combining phylogenetic and morphological analyses. The sequences in this study would be used as the reference sequences for further studies.

Efficacy of intrauterine perfusion of peripheral blood mononuclear cells (PBMC) for infertile women before embryo transfer: meta-analysis.

This meta-analysis was intended to evaluate the effects of intrauterine perfusion of peripheral blood mononuclear cells (PBMC) on the pregnancy outcomes including clinical pregnancy rates, embryo implantation rates, live birth rates and miscarriage rates of infertile women who were undergoing in vitro fertilisation (IVF) treatment. By searching Pubmed, Embase database, five articles meeting the inclusion criteria were included, and 1173 women were enrolled (intrauterine PBMC group: n = 514; NO-PBMC group: n = 659). For the entire IVF/ICSI population and one or two embryo transfer failure patients, there was no significant difference in endometrial thickness, embryo implantation rates, live birth rates, and miscarriage rates between the PBMC group and NO-PBMC group. Although the clinical pregnancy rates of the PBMC group were higher than that of the NO-PBMC group, the confidence interval was close to the line of unity. As for the patients with three or more implantation failures, the clinical pregnancy rates, embryo implantation rates and live birth rates were much higher in the PBMC group than that of the NO-PBMC group. In summary, current evidence suggests that intrauterine perfusion of PBMC can significantly improve pregnancy outcomes in patients who have three or more implantation failures.Impact statementWhat is already known on this subject? An increasing number of studies have shown that immune cells play an important role in embryo transfer. There is no reliable evidence to confirm the clinical efficacy of intrauterine perfusion of PBMC.What do the results of this study add? The current evidence suggests that intrauterine perfusion of PBMC can significantly improve pregnancy outcomes in patients who have three or more implantation failures.What are the implications of these findings for clinical practice and/or further research? To the best of our knowledge, this meta-analysis is the first to evaluate the effect of intrauterine perfusion of PBMC on pregnancy outcomes before embryo transfer. Our study indicated that intrauterine perfusion of PBMC significantly increased clinical pregnancy rates, embryo implantation rates, and live birth rates in patients who failed more than three implants.

[Detection and analysis of FAM19A4 promoter methylation in cervical exfoliated cells].

Objective: To investigate the cinical value of FAM19A4promoter methylation in cervicalexfoliated cells for triage of cervical cancer. Methods: A total of 162 high-risk HPV-infected patients who were pathologically confirmed as different cervical lesions from August 2017 to December 2017 were collected in Guangdong Women and Children Hospital. Taqman probe-based quantitative PCR (qPCR) was used to detect the methylation of FAM19A4 promoterin different grades of cervical lesions, and the value of FAM19A4 methylation in predicting cervical HSIL and the above lesions was calculated by diagnostic test. Results: (1)The positive rates of FAM19A4 methylation in cervical exfoliated cells increased with the severity of cervical lesions, which were 7.69% (4/52) , 34.62% (9/26) , 55.56% (20/36) , 95.83% (46/48) in normal cervix/cervicitis, cervical LSIL, HSIL, and cervical cancer, respectively(P<0.05).(2)There was no significant difference in the detection rates of FAM19A4 methylation between different age groups, pathological types, clinical stage, tumor size and lymph node metastasis status (P>0.05). (3) The specificity and positive predictive value of FAM19A4 methylation in detecting cervical HSIL alone and ≥HSIL lesions were the optimal, with the AUC of 0.69 and 0.84, respectively. When combined with HPV16/18 genotyping, the sensitivity was significantly improved. Conclusions: The detection of FAM19A4 promoter methylation in cervical exfoliated cells has a high clinical value of discriminating ≥HSIL lesions; and the cotest of methylated FAM19A4 and HPV16/18 genotyping can identify ≥HSIL lesions more sensitively.

Oral administration of short chain fatty acids could attenuate fat deposition of pigs.

Short chain fatty acids (SCFAs) are the main products of indigestible carbohydrates that are fermented by microbiota in the hindgut. This study was designed to investigate the effects of oral SCFAs administration on the lipid metabolism of weaned pigs. A total of 21 barrows were randomly allocated into three groups, including control group (orally infused with 200 mL physiological saline per day), low dose SCFAs group (orally infused with 200 mL SCFAs containing acetic acid 20.04 mM, propionic acid 7.71 mM and butyric acid 4.89 mM per day), and high dose SCFAs group (orally infused with 200 mL SCFAs containing acetic acid 40.08 mM, propionic acid 15.42 mM and butyric acid 9.78 mM per day). The results showed that the average daily feed intake of SCFAs groups were lower than that of control group (P<0.05). Oral administration of SCFAs decreased the concentrations of triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol and insulin (P<0.05), and increased the leptin concentration in serum (P<0.05). The total fat, as well as TC and TG levels in liver, was decreased by oral SCFAs administration (P<0.05). In addition, SCFAs down-regulated the mRNA expressions of fatty acid synthase (FAS) and sterol regulatory element binding protein 1c (P<0.05), and enhanced the mRNA expression of carnitine palmitoyltransferase-1α (CPT-1α) in liver (P<0.05). SCFAs also decreased FAS, acetyl-CoA carboxylase (ACC) and peroxisome proliferator activated receptor σ mRNA expressions in longissimus dorsi (P<0.05). And in abdominal fat, SCFAs reduced FAS and ACC mRNA expressions (P<0.05), and increased CPT-1α mRNA expression (P<0.05). These results suggested that oral administration of SCFAs could attenuate fat deposition in weaned pigs via reducing lipogenesis and enhancing lipolysis of different tissues.

Harnessing low-density lipoprotein receptor protein 6 (LRP6) genetic variation and Wnt signaling for innovative diagnostics in complex diseases.

Wnt signaling regulates a broad variety of processes in both embryonic development and various diseases. Recent studies indicated that some genetic variants in Wnt signaling pathway may serve as predictors of diseases. Low-density lipoprotein receptor protein 6 (LRP6) is a Wnt co-receptor with essential functions in the Wnt/ß-catenin pathway, and mutations in LRP6 gene are linked to many complex human diseases, including metabolic syndrome, cancer, Alzheimer's disease and osteoporosis. Therefore, we focus on the role of LRP6 genetic polymorphisms and Wnt signaling in complex diseases, and the mechanisms from mouse models and cell lines. It is also highly anticipated that LRP6 variants will be applied clinically in the future. The brief review provided here could be a useful resource for future research and may contribute to a more accurate diagnosis in complex diseases.

Excess of dietary montmorillonite impairs growth performance, liver function, and antioxidant capacity in starter pigs.

Montmorillonite (MMT) is widely used as a mycotoxin adsorbent in animal feeds, but its safety remains unclear. This study was conducted to investigate the safety of MMT supplementation in diets fed to starter pigs. A total of 120 32-d-old piglets (initial weight, 8.0 ± 0.9 kg) were randomly allotted into dietary treatments with graded MMT levels (0 [FS 0], 0.5% [FS 0.5], 1.0% [FS 1.0], 2.5% [FS 2.5], and 5.0% [FS 5.0]) with 6 replicate pens per treatment and 4 pigs per pen. All diets were fed for 28 d. As the MMT level increased, ADG and G:F changed in a linear and quadratic manner, while ADFI was linearly decreased ( > 0.05). Compared with FS 0, ADG, ADFI, and G:F of pigs in FS 1.0 increased ( < 0.05). However, the ADFI in pigs of FS 5.0 was lower than that in pigs of FS 0 ( < 0.05). The relative liver weight activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) changed in a linear and quadratic manner ( < 0.05). Compared with FS 0, pigs in FS 2.5 and FS 5.0 had a greater serum ALT ( < 0.05), while AST activity significantly increased in pigs of FS 5.0 ( < 0.05). Dietary MMT supplementation decreased serum Mg content in a linear and quadratic manner ( < 0.05), while Zn and Cu contents were linearly decreased ( < 0.05). Serum Zn and Cu contents of pigs in FS 0.5, FS 2.5, and FS 5.0 groups were lower than those in the control. Pigs fed with 2.5% and 5% MMT showed hepatic histopathological changes, including swelling, granular and vesicular degeneration, and apparent vacuolar degeneration. In addition, the content of serum total antioxidant capacity (T-AOC) and activity of glutathione peroxidase (GSH-PX) decreased in a linear and quadratic manner ( < 0.05). Compared to the control, 5.0% MMT significantly increased piglets' serum malondialdehyde (MDA) concentration and decreased GSH-PX activity ( < 0.05). T-AOC concentration in the pigs fed 2.5% and 5.0% MMT was lower than that in the control group ( < 0.05). Serum superoxide dismutase (SOD) activity changed in a quadratic manner ( < 0.05). Piglets in FS 1.0 showed a higher SOD activity when compared with the control ( < 0.05). These results indicate that supplementation of MMT higher than 1.0% can negatively affect liver structure and serum mineral content, and 5.0% MMT supplementation would also decrease feed intake, aggravate liver damage, and reduce the antioxidant capacity of starter pigs. Therefore, excess supplementation of MMT is not safe in starter pigs.

MicroRNA-499-5p regulates porcine myofiber specification by controlling Sox6 expression.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that negatively regulate gene expression of target messenger RNAs (mRNAs) and miRNAs have been proven to play vital roles in skeletal muscle development. The miRNA-499-5p has been reported to be negatively related with the expression of Sox6, a critical transcription factor for the maintenance of fast-twitch skeletal muscle. In this study, we amplified a length of 2012-bp mRNA that contains a 1512-bp porcine Sox6 (pSox6) 3'UTR from skeletal muscle of a Duroc×Landrace×Yorkshire pig. By luciferase reporter assay we verified that pSox6 is a target of miR-499-5p. In extensor digitorum longus and Soleus muscles of pigs, the expression levels of miR-499-5p and pSox6 mRNA were also inversely correlated. Besides, overexpression of miR-499-5p in porcine satellite cells promoted the expression of MyHC I and MyHC IIa mRNA, along with a reduction of pSox6 mRNA. Taken together, these results indicate that miR-499-5p may facilitate the oxidative myofibers formation by downregulating pSox6 expression.

Benzoic acid beneficially affects growth performance of weaned pigs which was associated with changes in gut bacterial populations, morphology indices and growth factor gene expression.

This study was conducted to investigate the effects of benzoic acid (BA) on growth performance, intestinal development and intestinal barrier function in weaned pigs. Ninety weaned pigs were randomly assigned to one of three treatments: a basal diet (CON), the basal diet supplemented with 2000 mg/kg benzoic acid (BA1) and 5000 mg/kg benzoic acid (BA2). At the end of days 14 and 42, six pigs per treatment were randomly selected to collect plasma and intestinal samples. Results showed that BA supplementation not only improved final body weight, daily growth and feed conversion ratio from days 15 to 42 and days 1 to 42, but also decreased the activity of plasma diamine oxidase (day 42) and the pH values of jejunal contents (day 14) (p < 0.05). Ileal Bacillus populations (day 14) were increased by BA, while Escherichia coli counts in the ileum and caecum (day 42) were decreased (p < 0.05). Higher Lactobacillus counts occurred in the ileum (day 14, 42) of BA1-fed piglets as compared to CON and BA2-fed piglets (p < 0.05). In addition, BA supplementation increased the ratio of villus height to crypt depth (day 14, 42) and decreased the crypt depth (day 14) (p < 0.05). Growth-stimulating factors (insulin-like growth factor-1, day 42; insulin-like growth factor-1 receptor, day 14, 42) and tight junction protein (occludin, day 14, 42; zonula occludens-1, day 42)-related gene mRNA levels were upregulated in the jejunum of piglets fed BA diets (p < 0.05). In conclusion, this study provides the first evidence that BA has beneficial effects on intestinal development and intestinal barrier function of weaned pigs, which can partly explain why growth performance of pigs was improved by dietary BA supplementation.

Dietary spray-dried chicken plasma improves intestinal barrier function and modulates immune status in weaning piglets.

The objective of this study was to evaluate the effects of dietary addition of spray-dried chicken plasma (SDCP) as a replacement for spray-dried porcine plasma (SDPP) on serum biochemistry, intestinal barrier function, immune parameters, and the expression of intestinal development-related genes in weaning pigs. One hundred and forty-four 25-d-old weaning piglets with BW of 6.43 ± 0.39 kg were randomly allotted to 1 of 4 dietary treatments: 1) CON (basal diet; control), 2) SDPP (containing 5% SDPP), 3) SDPP + SDCP (containing 2.5% SDPP and 2.5% SDCP), and 4) SDCP (containing 5% SDCP). After a 28-d trial, 6 pigs from each treatment were randomly selected to collect serum and intestinal samples. On d 14 after the initiation of the trial, pigs in the SDPP, SDPP + SDCP, and SDCP groups had an increase ( < 0.05) in serum concentrations of total protein and IgG and a decrease ( < 0.05) in activities of alanine aminotransferase and diamine oxidase compared with the CON group. In the jejunum, supplementation with SDPP and SDCP reduced ( < 0.05) the concentration of tumor necrosis factor-α (TNF-α) and upregulated ( < 0.05) the mRNA levels of zonula occludens 1 (ZO-1), zonula occludens 2 (ZO-2), occludin (OCLN), Toll-like receptor 2 (TLR2), glucagon-like peptide 2 (GLP2), and IGF-1 compared with the CON group. In the ileum, feeding SDPP, SDPP + SDCP, and SDCP decreased ( < 0.05) the concentrations of TNF-α and secretory IgA (sIgA) and upregulated ( < 0.05) the mRNA levels of claudin 1 (CLDN-1) and TLR2 compared with feeding CON. However, there were no differences among the SDPP, SDPP + SDCP, and SDCP groups. Furthermore, supplementation with SDCP reduced ( < 0.05) the concentration of IL-10 and upregulated ( < 0.05) the mRNA levels of GLP-2, mucin 2 (MUC2), and trefoil factor family 3 (TFF3) in the ileum compared with feeding CON. Collectively, the current results indicate that dietary addition of SDCP has a beneficial influence on the health condition of weaning pigs by alleviating liver damage, promoting intestinal development, improving intestinal barrier function, and reducing overstimulation of immune response. The efficacy of SDCP is comparable to that of SDPP.

Spray-dried chicken plasma improves intestinal digestive function and regulates intestinal selected microflora in weaning piglets.

One hundred forty-four 25-d-old weaning piglets with BW of 6.43 ± 0.39 kg were used in a 28-d trail to evaluate the effects of dietary addition of spray-dried chicken plasma (SDCP) as a replacement for spray-dried porcine plasma (SDPP) on growth performance, nutrient digestibility, diarrhea incidence, small intestinal morphology, digestive enzyme activity, and microflora. Pigs were randomly allotted to 1 of 4 dietary treatments: 1) CON (control; a basal diet), 2) SDPP (containing 5% SDPP), 3) SDPP + SDCP (containing 2.5% SDPP and 2.5% SDCP), and 4) SDCP (containing 5% SDCP). Six pigs from each treatment were randomly selected to collect serum and intestinal samples. Compared with the CON group, both the SDPP and the SDPP + SDCP groups improved final BW of pigs (P < 0.05), but there were no differences among the SDPP, SDPP + SDCP, and SDCP groups. From d 1 to 14 and d 15 to 28, pigs fed the SDPP and SDPP + SDCP diets had a greater (P < 0.05) ADG than pigs fed the CON diet. During the overall period, both ADG and ADFI of pigs in the SDPP and SDPP + SDCP groups were improved (P < 0.05) compared with pigs in the CON group. Furthermore, pigs fed diets containing SDPP or SDCP had a greater (P < 0.05) apparent total tract digestibility (ATTD) of CP, ether extract, Ca, and ash and less (P < 0.05) incidence of diarrhea than pigs fed the CON diet. However, no differences were observed for ATTD and diarrhea incidence between the SDPP and SDCP groups. Compared with the CON group, duodenal villus height and the ratio of villi to crypt were increased (P < 0.05) in the SDPP, SDPP + SDCP, and SDCP groups and jejunal crypt depth was decreased in the SDPP + SDCP and SDCP groups (P < 0.05). Pigs in the SDPP group had greater (P < 0.05) activities of amylase, maltase, and trypsin than pigs in the CON group. However, no significant differences were observed between the SDCP and SDPP groups. Additionally, inclusion of SDCP in diet decreased (P < 0.05) the population of Escherichia coli. In conclusion, these results demonstrated that the addition of SDCP in pigs' diet had an effect similar to SDPP on improving growth performance through the promotion of the small intestinal development, increasing digestive enzymes activities, enhancing ATTD of nutrients, and decreasing diarrhea incidence.

Differential expression of lipid metabolism-related genes and myosin heavy chain isoform genes in pig muscle tissue leading to different meat quality.

The aim of this study was to investigate the variations in meat quality, lipid metabolism-related genes, myosin heavy chain (MyHC) isoform genes and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) gene mRNA expressions in longissimus dorsi muscle (LM) of two different pig breeds. Six Rongchang and six Landrace barrows were slaughtered at 161 days of age. Subsequently, meat quality traits and gene expression levels in LM were observed. Results showed that Rongchang pigs not only exhibited greater pH, CIE a*24 h and intramuscular fat content but also exhibited lower body weight, carcass weight, dressing percentage, LM area and CIE b*24 h compared with Landrace pigs (P<0.05). Meanwhile, the mRNA expression levels of the lipogenesis (peroxisome proliferator-activated receptor gamma, acetyl-CoA carboxylase and fatty acid synthase) and fatty acid uptake (lipoprotein lipase)-related genes were greater in the Rongchang (P<0.05), whereas the lipolysis (adipose triglyceride lipase and hormone sensitive lipase) and fatty acid oxidation (carnitine palmitoyltransferase-1B)-related genes were better expressed in the Landrace. Moreover, compared with the Landrace, the mRNA expression levels of MyHCI, MyHCIIa and MyHCIIx were greater, whereas the mRNA expression levels of MyHCIIb were lower in the Rongchang pigs (P<0.05). In addition, the mRNA expression levels of PGC-1α were greater in Rongchang pigs than in the Landrace (P<0.05), which can partly explain the differences in MyHC isoform gene expressions between Rongchang and Landrace pigs. Although the small number of samples does not allow to obtain a definitive conclusion, we can suggest that Rongchang pigs possess better meat quality, and the underlying molecular mechanisms responsible for the better meat quality in fatty pigs may be partly due to the higher mRNA expression levels of lipogenesis and fatty acid uptake-related genes, as well as the oxidative and intermediate muscle fibers, and due to the lower mRNA expression levels of lipolysis and fatty acid oxidation-related genes, as well as the glycolytic muscle fibers.

MicroRNA-27a promotes porcine myoblast proliferation by downregulating myostatin expression.

MicroRNAs are endogenous ~22nt RNAs that negatively regulate gene expression at the posttranscriptional level via binding to the 3'-untranslated region (3'UTR) of target mRNAs. The microRNA miR-27a was reported to depress the expression of myostatin, a critical inhibitor of skeletal myogenesis, by binding to its 3'UTR in mouse. In this study, we cloned the full-length 3'UTR of porcine myostatin by rapid amplification of 3'-cDNA ends (3'-RACE) and demonstrated that the 3'UTR of porcine myostatin is targeted by miR-27a. The phenomenon that the level of myostatin inversely correlated with miR-27a was observed in fat and heart of pigs and also in proliferating porcine myoblasts. Besides, overexpression of miR-27a in porcine myoblasts promoted cell proliferation by reducing the expression of myostatin. Our data suggest that miR-27a positive regulates porcine myoblast proliferation via targeting myostatin.

Improved fluorimetric determination of dissolved aluminium by micelle-enhanced lumogallion complex in natural waters.

A modification of the aluminium-lumogallion fluorescence measurement in the presence of the non-ionic surfactant Triton X-100 is presented. The detection limit for dissolved Al is 0.7 nM, with a relative standard deviation of 3.6% at an Al level of 5.0 nM. Compared with previously reported methods in the literature, the method described here is free from matrix effects and can be used for the determination of aluminium in fresh, estuarine and saline waters. The interferences from iron and fluoride were minimized by the addition of o-phenanthroline and Be2+, respectively. The analysis of NIST SRM 1643C and PRC standard 2430101 by the proposed method provides results consistent with the certified values. A successful inter-laboratory calibration exercise also demonstrates the merit of the proposed method for the determination of Al in environmental and marine sciences.

Mitogenic phospholipase D activity is restricted to caveolin-enriched membrane microdomains.

Phospholipase D (PLD) activity is elevated in response to the oncogenic stimulus of several signaling oncogenes. PLD activity is also elevated in response to peptide growth factors, indicating that PLD likely plays an important role in mitogenic signaling. Many proteins that mediate mitogenic signaling are localized in caveolin-enriched membrane microdomains (CEMMs). We report here that the elevated PLD activity in NIH 3T3 cells transformed by activated oncogenic forms of Src, Ras, and Raf is largely restricted to the CEMMs. Likewise, the PLD activity stimulated by epidermal growth factor is also restricted to the CEMMs. Although both PLD1 and PLD2 were found in CEMMs, neither was particularly enriched in the CEMMs of the transformed relative to the parental cells, indicating that it is the specific activity of PLD that is increased in the CEMMs. An apparent PLD substrate specificity in transformed cells for phosphatidylcholine lacking arachidonate acyl groups is also explained by the localization of activity in the CEMMs where [(3)H]arachidonate-labeled PC was excluded. These data indicate that mitogenic signals through PLD are initiated in CEMMs where many signaling molecules colocalize.

[Discriminant function based on serum analyte for diagnosis of primary hepatic cancer].

OBJECTIVE: The aim of this study was to diagnose primary hepatic cancer(PHC). METHODS: Seven serum indexes of 92 serum samples consisting of 48 controls and 44 PHC were detected. Four discriminant functions based on seven serum indexes were obtained with computer SPSS statistic package. RESULTS: The rate of coincidence, sensitivity, and specificity of the general discriminant functions were 86.96%, 77.27%, and 95.83% respectively; and that of the stepwise discriminant functions were 84.78%, 79.55%, and 89.59% respectively. CONCLUSION: Usually PHC can be diagnosed by analyzing patients serum indexes with the discriminant functions.

SPECT brain perfusion findings in mild or moderate traumatic brain injury.

BACKGROUND: The purpose of this manuscript is to present the findings in the largest series of SPECT brain perfusion imaging reported to date for mild or moderate traumatic brain injury. PATIENTS AND METHODS: This is a retrospective evaluation of 228 SPECT brain perfusion-imaging studies of patients who suffered mild or moderate traumatic brain injury with or without loss of consciousness (LOC). All patients had no past medical history of previous brain trauma, neurological, or psychiatric diseases, HIV, alcohol or drug abuse. The patient population included 135 males and 93 females. The ages ranged from 11-88 years (mean 40.8). The most common complaints were characteristic of the postconcussion syndrome: headaches 139/228 (61%); dizziness 61/228 (27%); and memory problems 63/228 (28%). LOC status was reported to be positive in 121/228 (53%), negative in 41/228 (18%), and unknown for 63/228 (28%). RESULTS: Normal studies accounted for 52/228 (23%). For abnormal studies (176/228 or 77%) the findings were as follows: basal ganglia hypoperfusion 338 lesions (55.2%); frontal lobe hypoperfusion 146 (23.8%); temporal lobes hypoperfusion 80 (13%); parietal lobes hypoperfusion 20 (3.7%); insular and or occipital lobes hypoperfusion 28 (4.6%). Patients' symptoms correlated with the SPECT brain perfusion findings. The SPECT BPI studies in 122/228 (54%) were done early within 3 months of the date of the accident, and for the remainder, 106/228 (46%) over 3 months and less than 3 years from the date of the injury. In early imaging, 382 lesions were detected; in 92 patients (average 4.2 lesions per study) imaging after 3 months detected 230 lesions: in 84 patients (average 2.7 lesions per study). CONCLUSIONS: Basal ganglia hypoperfusion is the most common abnormality following mild or moderate traumatic brain injury (p = 0.006), and is more common in patients complaining of memory problem (p = 0.0005) and dizziness (p = 0.003). Early imaging can detect more lesions than delayed imaging (p = 0.0011). SPECT brain perfusion abnormalities can occur in the absence of LOC.

Attenuation-Correction Induced Artifact in F-18 FDG PET Imaging Following Total Knee Replacement.

Purpose: The clinical use of PET FDG in the work-up of patients with bone and soft tissue malignant tumors is rapidly increasing. The recognition of any source of artifact, therefore, is important to avoid interpretation pitfalls.Procedures: Two patients with complete knee joint replacement by metallic prosthesis in the course of their treatment for malignant bone and soft tissue sarcoma were evaluated by PET F-18 FDG imaging using a dual head coincidence gamma camera.Results: Both studies demonstrated in the attenuation-corrected images intense increase activity at the joint space between the metallic prosthetic surfaces at the level of the knee joint. No uptake, however, was noted in the same location on the non-attenuation-corrected images. Subsequent bone and thallium-201 scans confirmed the absence of tumor recurrence in the first patient. The second patient had multiple follow up F-18 FDG scans over a period of 16 months that show no changes from the baseline study.Conclusion: In the F-18 FDG PET images of patients with total knee metallic prosthesis, an intense activity tends to be seen in the joint space, only in the attenuation-corrected images. Such pattern of uptake is considered artifactual and should always be verified in the non-attenuated images.

Functional association between Arf and RalA in active phospholipase D complex.

Activation of phospholipase D1 (PLD1) by Arf has been implicated in vesicle transport and membrane trafficking. PLD1 has also been shown to be associated with the small GTPase RalA, which functions downstream from Ras in a Ras-RalA GTPase cascade that facilitates intracellular signal transduction. Although PLD1 associates directly with RalA, RalA has no effect upon the activity of PLD1. However, PLD1 precipitated from cell lysates with immobilized glutathione S-transferase-RalA fusion protein is active. This suggests the presence of an additional activating factor in the active RalA-PLD1 complexes. Because Arf stimulates PLD1, we looked for the presence of Arf in the active RalA-PLD1 complexes isolated from v-Src- and v-Ras-transformed cell lysates. Low levels of Arf protein were detected in RalA-PLD1 complexes; however, if guanosine 5'-[gamma-thio]triphosphate was added to activate Arf and stimulate translocation to the membrane, high levels of Arf were precipitated by RalA from cell lysates. Interestingly, deletion of 11 amino-terminal amino acids unique to Ral GTPases, which abolished the ability of RalA to precipitate PLD activity, prevented the association between RalA and Arf. Brefeldin A, which inhibits Arf GDP-GTP exchange, inhibited PLD activity in v-Src- and v-Ras-transformed cells but not in the nontransformed cells, suggesting that the association of Arf with RalA is required for the increased PLD activity induced by v-Src and v-Ras. These data implicate Arf in the transduction of intracellular signals activated by v-Src and mediated by the Ras-RalA GTPase cascade. Because both Arf and PLD1 stimulate vesicle formation in the Golgi, these data raise the possibility that vesicle formation and trafficking may play a role in the transduction of intracellular signals.