A New Source of Diterpene Lactones From Andrographis paniculata (Burm. f.) Nees-Two Endophytic Fungi of Colletotrichum sp. With Antibacterial and Antioxidant Activities.

Endophytic fungi of medicinal plants are abundant, and their metabolites often have antioxidant, antibacterial, and antitumor effects and can produce secondary metabolites identical or similar to those of their hosts, which can mitigate the problem of insufficient supply of medicinal plants. In this study, we screened endophytic fungi for strains that produce the same diterpene lactones as Andrographis paniculata based on their biological activity. Firstly, the dominant group of endophytic fungi of Andrographis paniculata was screened and pathogenicity was studied using Koch's rule. Secondly, DPPH, ABTS, OH, PTIO radical scavenging, and FRAP assays were used to detect the antioxidant activity of the extracellular extracts of the strains, and total phenol and total flavonoid contents of the strains with high antioxidant capacity were determined. S. aureus, B. subtilis, E. coli, and P. aeruginosa were used to determine the antibacterial activity of the mycelial extracts of the strains. Finally, the secondary metabolites of the mycelial extracts of the strains were examined by high-performance liquid chromatography. The results showed that 32 strains of Andrographis paniculata were relatively isolated > 70% and non-pathogenic. Extracellular extracts of strains AP-1 and AP-4 showed vigorous antioxidant activity, and AP-4, AP-12, AP-47, and AP-48 showed antibacterial activity against four strains of bacteria. The HPLC results indicated that the mycelial extracts of AP-4 and AP-12 contained diterpene lactones. The two endophytic fungi were recognized as Colletotrichum sp. The study successfully obtained diterpene lactones from the endophytic fungus of Andrographis paniculata and confirmed the feasibility of using endophytic fungal strains to produce active substances consistent with the host. It was also useful for exploring endophytic fungi and medicinal plants. The relationship provides theoretical guidance.

Predictive risk scales for development of pressure ulcers in pediatric patients admitted to general ward and intensive care unit.

BACKGROUND: More than ten special scales are available to predict the risk of pressure ulcers in children. However, the performances of those scales have not yet been compared in China. AIM: To compare the Waterlow, Braden Q, and Glamorgan scales, and identify more suitable pressure ulcer evaluation scale for the pediatric intensive care unit (PICU). METHODS: Trained nurses used the Waterlow, Braden Q, and Glamorgan scales to assess pediatric patients at Sun Yat-sen Memorial Hospital (China) within 24 h of admission from May 2017 to December 2020 in two stages. Skin examination was carried out to identify pressure ulcers every 3 d for 3 wk. RESULTS: The incidence of pressure ulcers was 3/28 (10.7%) in the PICU and 5/314 (1.6%) in the general pediatric ward. For children in the general ward, the Waterlow, Braden Q, and Glamorgan scales had comparable area under the operating characteristic curve (AUC) of 0.870, 0.924, and 0.923, respectively, and optimal cut-off values of 14, 14, and 29 points. For PICU, the Waterlow, Braden Q, and Glamorgan scales had slightly lower AUC of 0.833, 0.733, and 0.800, respectively, and optimal cut-off values of 13, 16, and 27 points. Braden Q demonstrated a satisfactory specificity, and during the second stage of the study for PICU patients, the AUC of the Braden Q scale was 0.810, with an optimal cut-off value of 18.35 points. CONCLUSION: The Waterlow, Braden Q, and Glamorgan scales have comparable performance, while the Braden Q scale demonstrates a better specificity and can be successfully used by pediatric nurses to identify patients at high risk of pressure ulcers in PICU.

Progress in the physiological and genetic mechanisms underlying the high prolificacy of the Erhualian pig.

The Erhualian pig, originally distributed in the Taihu area, is well known for its universally high fertility. Previous studies have found that high ovulation numbers, low embryo mortality and high uterine volumes are important physiological characteristics underlying the high prolificacy of the Erhualian pig. Although candidate genes such as follicle-stimulating hormone ß (FSHß) and several quantitative trait loci (QTLs) on chromosome 2, 6, 7, 8, 12, 13 and 15 have been reported to affect the litter size in the Erhualian pig, the key genes related to high prolificacy remain poorly understood. In this review, we summarize the recent research progress in the physiological and genetic mechanisms underlying the high prolificacy of the Erhualian pig. First we review the role of high ovulation numbers, low embryo mortality rates and high uterine volumes in the formation of the high litter size in the Erhualian pig. Then we summarize candidate genes and QTLs for the high litter size detected by classical strategies, as well as by genomic strategies. Moreover, we describe the methods to investigate the causative genes of the high prolificacy through integrative analysis of multi-omics data including genomics, transcriptomics, proteomics and functional genomics. This review will provide insights to understand the molecular basis of the high prolificacy in the Erhualian pig.

Isolation, Culture and Identification of Porcine Skeletal Muscle Satellite Cells.

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

Expression of bone morphogenetic protein 2, 4, and related components of the BMP signaling pathway in the mouse uterus during the estrous cycle.

The objective was to investigate the expression of bone morphogenetic protein (BMP) family members in the mouse uterus during the estrous cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry. Uterine samples from Swiss ICR mice were collected and dissected free of surrounding tissue. One uterine horn was snap frozen in liquid nitrogen immediately after collection and stored at -80 °C for RNA extraction, and the other was fixed in 40 mg/ml paraformaldehyde at room temperature for immunolocalization of BMP2 protein. Real-time PCR analysis showed that the expression level of Bmp2 was significantly higher at proestrus than at estrus and metestrus (P<0.05). The relative abundance of Bmp4 exhibited significant fluctuations, but there were no statistically significant differences between the expression levels of Bmp2 and Bmp4 (P>0.05). The expression levels of Bmpr1a and Bmpr2 remained unchanged during estrous cycles. However, the level of Bmpr1b mRNA decreased significantly at estrus (P<0.05), increasing subsequently at metestrus. Furthermore, the level of Bmpr1b mRNA was significantly lower than those of Bmpr1a and Bmpr2 mRNA at the corresponding stages (P<0.05). All three receptor-regulated Smads (R-Smads) detected were differentially expressed in the mouse uterus and the expression levels of Smad1 and Smad5 were significantly higher than that of Smad8 (P<0.05). In addition, the expression level of Smad4 did not change substantially throughout the estrous cycle. Immunohistochemical experiments revealed that BMP2 protein was differentially expressed and localized mainly in the uterine luminal and glandular epithelial cells throughout the estrous cycle. In conclusion, our results provide information about the variation in the mRNA levels of Bmp2 and Bmp4 and related components of the BMP signaling pathway. The data provide quantitative and useful information about the roles of endometrial BMP proposed and demonstrated by others, such as the degradation and remodeling of the endometrium.

Cell-specific expression and immunolocalization of nitric oxide synthase isoforms and soluble guanylyl cyclase α and ß subunits in postnatal porcine uteri.

The aim of the present study was to investigate the cellular expression and immunolocalization of nitric oxide synthase (NOS) isoforms and soluble guanylyl cyclase (sGC) subunits in postnatal porcine uteri. Immunohistochemical experiments showed that three isoforms of NOS were mainly localized in the uterine luminal and glandular epithelium and myometrium, and the intensity of immunostaining for iNOS and eNOS was increased gradually with temporal development of the postnatal uterus. In addition, sGC subunits, sGCα1 and ß, were present in the uterine luminal and glandular epithelium, myometrium and stromal cells. The uterine NOS activity data showed that the total NOS and iNOS activities were significantly increased at postnatal days 21 and 35. Although constitutive NOS activity was increased at postnatal day 21, it decreased subsequently at postnatal day 35. Immunoblot analysis revealed that iNOS protein expression was significantly increased at postnatal days 21 and 35. Furthermore, sGCα1 protein expression was not significantly changed throughout days 7 to 35. Collectively, our findings suggest that NO/cGMP signaling is involved in the process of postnatal porcine uterine development.

[Infrared spectrum denoising with combination of lifting wavelet domain thresholding and median filtering].

Infrared spectra are often corrupted by noise, which may greatly influence the accuracy and precision of the analytical result. To improve the analytical precision, the authors need to denoise the spectrum data first. In the present paper, a spectrum denoising method by the second generation wavelet transform domain thresholding combined with the median filtering is introduced. The spectrum of a certain kind of wheat was used to test the performance of the proposed denoising method. In the experiment,noise with signal to noise ratio 21.17 dB was first added to the spectrum, and then removed by the proposed denoising method. The signal to noise ratio (SNR), the root mean square error (RMSE), the average relative error of the peak value (AREPV) and the average error of the peak position (AEPP) were used to evaluate the performance of the proposed denoising method. Experimental results show that the proposed method can remove the spectrum noise and keep the useful information more effective than Donoho's soft and hard threshold method. At the same time, it can achieve a higher PSNR, a lower RMSE, a lower AREPV and a lower AEPP than the other two denoising methods.

Age-dependent expression of forkhead box O proteins in the duodenum of rats.

The O subfamily of forkhead box (FoxO) proteins is the downstream effector of the insulin-like growth factor-1/phosphoinositide 3-kinase/protein kinase B (IGF-1/PI3K/PKB) signal pathway. The objective of the present study was to examine the expressions of three members of FoxO proteins, FoxO1, FoxO3a, and FoxO4 in the duodenum of Sprague-Dawley rats at different ages. The result demonstrated that the expression of FoxO4 in rat duodenum showed an age-dependent manner. At Day 21, there were no detectable localization and expression of FoxO4 in the duodenum, while, at Months 2 and 6, localization and expression of FoxO4 were distinct. In addition, FoxO4 staining was primarily concentrated in the cell nuclei of the lamina propria around the intestinal gland of the duodenum in 2-month-old rats, but was not detectable in the same area in 6-month-old rats. Our results showed also that although FoxO3a existed in the cytoplasm of the lamina propria at a low level at the 2- and 6-month marks, it was still not detectable at Day 21. Besides, FoxO1 was not detectable in all parts and stages. Taken together, our findings suggested that the cell-specific and age-dependent expressional patterns of FoxO4 and FoxO3a proteins in the duodenum play some roles in the development and growth performance of the rat duodenum.

[The genetic effects of IGF2 gene intron3 variance in pigs].

A mispairing PCR-RFLP technique was applied in this study to determine the Insulin-like Growth Factor 2(IGF2) gene intron3 G3072A mutation in an outbred Landrace and Large White, and the gelded boars from Landrace x Large White cross. The difference of corresponding traits and the genetic effects of the boars inherited from parental A allele and inherited from parental G allele were analyzed. The results indicated that comparing with the boars inherited from parental G allele, the boars inherited from parental A allele increased significantly in the circumference 3.06% (P< 0.05) and index of body 3.01% (P< 0.05), respectively. The boars inherited from parental A allele had a significantly less average buttock fat thickness (15.31%, P< 0.01), thorax-waist fat thickness (23.74%, P< 0.01), skin thickness 9.38% (P< 0.01), fiber density (20.03%, P< 0.01) and had more less 6th-7th rib fat thickness (20.27%, P< 0.05), tendernce (17.32%, P< 0.05), and had more thick shoulder fat thickness (7.97%, P< 0.05), and had bigger the loin eye area (22.58%, P< 0.01) and fiber cross-sectional area (32.70%, P< 0.01) and fiber diameter (15.38%, P< 0.01) and lean meat (2.18%, P< 0.01) than the boars inherited from parental G allele. The results were suggested that the parental A allele has highly significant genetic effects in improving pig body development and carcass lean percent by increasing fiber diameter and the loin eye area, and decreasing the skin thickness and fat percent.

FoxO4 is the main forkhead transcriptional factor localized in the gastrointestinal tracts of pigs.

Forkhead box (Fox) proteins play critical roles in the regulation of differentiation, proliferation, immunity and aging of cells. Most studies on Fox proteins are limited to cultured cells and rodent. The aim of the current study is to detect by immunohistrochemistry whether FoxO1, FoxO3a and FoxO4 proteins are localized in the stomach and intestine of the pig. The results showed that FoxO4 exists in the mucosa in all parts of the stomach and intestine; FoxO3a exists mainly in the lamina propria and muscularis of some parts. However, FoxO1 is not detectable in all parts of the stomach and intestine. Collectively, the results of the present study indicate that there exists a distinct expression pattern of Fox proteins, and that FoxO4 is a primary forkhead transcriptional factor localized in the gastrointestinal tracts of the pig.

The correlation of reproduction-related gene expression with germ cell number in DM and PLL gilts.

In this study, the ovarian germ cell number was counted in 3-week-old Duroc x Meishan (DM, n=30) and PIC x (Landrace x Large White) (PLL, n=53) gilts, and the mRNA expression levels of four reproduction-related genes were investigated by quantitative RT-PCR. Correlation of germ cell number with the expression level of these genes was analyzed. Results showed that the germ cell number of DM was significantly higher than that of PLL gilts (P<0.01), although there was no significant difference between the ovarian weight of DM and PLL gilts (P=0.269). No significant correlation existed between germ cell number and ovarian weight in the two gilt groups (R=0.335, P=0.07; R=0.119, P=0.398, respectively). A significant correlation was found between the germ cell number and expression level of ESR and IGF1R mRNA in DM gilts (R=0.648, P<0.05; R=0.757, P<0.01, respectively), but the correlation between the germ cell number and expression level of FSHR and INHBA mRNA did not reach statistical significance. Significant correlation was found between the germ cell number and the expression level of ESR, FSHR, and IGF1R mRNA in PLL gilts (R=0.435, P<0.01; R=0.438, P<0.01; R=0.292, P<0.05, respectively), but not with INHBA mRNA in PLL gilts.

[Effects of microsatellite DNA markers on meat quality traits in pig chromosome 13].

In this reseanch, 7 microsatellite DNA loci linked with PPAR gene were selected from the published genetic map of chromosome 13 in pig,and polymorphisms of these microsatellites in 100 samples from Sutai pigs (Duroc x Erhualian) populations were detected. Results revealed that the number of alleles were 6-9, heterozygosity 0.59 - 0.81, polymorphism information content 0.51 - 0.76. Effects of S0021, SW1937, SW482, S0222, S0293, S0281 and SWR2054 on meat quality traits were analyzed with PROC GLM of SAS. Results showed that the effects of S0021 on pH value and SW937 on water-holding capacity reached a significant level at P < 0.01 respectively. The effect of S0293 on tenderness and SW482 on BFT were also significant (P < 0.05). S0222, S0281 and SWR2054 had no significant effect on the 7 selected meat qualitytraits (P>0.05).

[Analysis of gene expression information in the fat and muscle tissues of pig].

An in silico study was developed to detect the gene expression profile of pig fat and muscle tissues by using the porcine EST resources and human gene sequences,and thus provided candidate information for basic genetic analysis in meat quality improvement of pig. In this study,a BLAST search was performed to identify homologies between the cDNA sequences of human genes and the ESTs sequences of pig,and the high homologous records were screened out. Four Java programs were developed to retrieve and collect sequences, and analyze the BLAST alignment results. By statistical analysis, it was found that there were at least 2002 genes expressed in the fat and muscle tissues of pig, and 1087 in the fat tissue, 1205 in the muscle respectively (290 genes co-expressed in the two tissues). The top-ranking records were screened out,and meantime, 114 basic active genes (BAGs) were found to express in the two tissues,80 in the fat tissue and 34 in the muscle tissue respectively. The top 10 records were described in the paper. This study was summarized in relation to current meat quality improvement of pig at molecular level.

[Study on water extraction process of Herba epimedii with microwave technology].

OBJECTIVE: To study, the high polar media-water extraction process of Herba epimedii with the microwave technology. METHOD: The single parameters of extraction process was investigated with dry extract yield and extraction efficiency of icariin as content index. RESULT: Through studying the single parameter, the optimal conditions are as follows: 70% ethanol as wetting solvent, volume of solvent to weight of material equal to 6:1 in wetting, 30 min wetting time, 2.5 min microwave irradiation time, volume of solvent to weight of material equal to 50:1 in extraction process, 10 min microwave-assisted extraction time. CONCLUSION: Compared with conventional methods, microwave-assisted extraction (MAE) after Microwave Pretreament (MP) of raw material was higher extraction efficiency and time-saving.