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Soluble sugars, including glucose, fructose and sucrose, are the most important determinants that affect the flavor and quality of red pitaya (Hylocereus polyrhizus) fruit. Vacuolar invertase (VIN), which catalyzes sucrose hydrolysis into glucose and fructose, is a key type of enzyme responsible for soluble sugar metabolism in plant growth and development. Herein, we conducted genome-wide identification, gene expression analysis, subcellular localization and an enzymatic properties assay for the VIN-encoding genes from red pitaya. During red pitaya fruit development towards ripening, the enzymatic activities of VIN showed an up-regulated trend towards ripening. In total, four isoforms (HpVIN1–4) of the VIN-encoding gene were identified from the pitaya genome. Sequence alignment results revealed that the HpVIN1, HpVIN3 and HpVIN4 proteins contained essential motifs for targeting the vacuole and conserved motifs or residues responsible for sucrose binding and hydrolysis. Gene expression pattern analyses revealed that the level of HpVIN4 was obviously increasing during fruit development and acted as the most abundant VIN isoform towards ripening. Subcellular localization detection via transient expression in Arabidopsis thaliana mesophyll protoplasts revealed that the HpVIN4 protein was localized in the vacuole. Growth complementation tests of heterologous expression in the invertase-deficient baker's yeast strain suggested that the HpVIN4 protein had a sucrose hydrolysis activity and could restore the yeast growth in vivo. The identification of enzymatic properties in vitro demonstrated that the HpVIN4 protein could degrade sucrose into glucose and fructose with an optimum pH of 4.0. Specifically, the HpVIN4 protein had an estimated Km value of 5.15 ± 1.03 mmol·L−1 for sucrose hydrolysis. Ultimately, this study provides a comprehensive understanding of the potential roles of VINs during fruit development and towards ripening and provides functional gene resources for regulating soluble sugar accumulation in red pitaya fruit. [ABSTRACT FROM AUTHOR]

From the opening of the silo to the end of its feeding, the silage face undergoes materials and energy flow with the external environment. Oxygen is the cause of the deterioration of silage quality. It promotes a series of aerobic microorganisms to eat high-nutrient substances (such as water-soluble sugars and lactic acid), which leads to a decrease in the dry matter and energy content of the silage, and may even result in the production of mycotoxins. The silage face is susceptible to oxidation, but effective management of the silage face can help preserve more nutrients for ruminants. The paper introduces the structural characteristics of silage, gas exchange, volatilization of fermentation ends, and microbial activities that occur on the silage face. It also describes the conditions that lead to aerobic deterioration on the silage face. Additionally, the paper reviews the role of silage face management, such as management principles, daily removal rate, silage remover, and feeding principles, in ensuring personnel safety, preserving nutrients and reducing dry matter losses, to provide a reference for implementing effective feed-out practices. [ABSTRACT FROM AUTHOR]

The article presents information on a study related to the establishment and application of fluorescent reverse transcriptase polymerase chain reaction (RT-PCR) for detection of Newcastle disease virus. It informs that in the study pair of primers and a probe were designed based on the relatively conservative sequence. It further informs that the development of fluorescent RT-PCR method provided a rapid and effective method for Newcastle disease virus detection.

Fire is an important ecological factor in boreal coniferous forest, which directly affects plant taxonomic and functional diversity and consequently forest succession. We analyzed the changes of soil nutrient contents, leaf functional traits, taxonomic and functional diversity of a Larix gmelinii community under different fire intensity levels (low, medium, heavy). The forest stand had been naturally recovered for 12 years in Yakeshi area. The results showed that fire significantly reduced soil total nitrogen content, but did not affect soil total phosphorus content. Low and medium fire intensities could maintain higher species diversity and functional diversity of the community. Medium fire disturbance significantly increased species diversity of the community, with maximum species richness index, Shannon index, Simpson index and Pielou evenness index. Fire disturbance reduced functional richness and functional divergence of the community, which were the largest in medium and low fire intensity respectively (except unburned). Functional evenness and Rao qua-dratic entropy index increased after forest fire disturbance, and the largest was in the low intensity burned area. With the increases of fire intensity, leaf dry matter content, leaf tissue density and leaf total phosphorus content showed a significantly increased trend, while specific leaf area, leaf water content, leaf total nitrogen content and leaf N:P showed the trend unburned ＞ medium fire ＞ low fire ＞ heavy fire, leaf thickness increased first and then decreased. Fire intensity had significant effects on leaf functional traits and functional diversity of forest community. Moderate fire distur-bance could promote forest restoration.

Aim: To evaluate the application of bipolar coagulation (BIP) in hepatectomy by comparing the efficacy of BIP alone, cavitron ultrasonic surgical aspirator (CUSA) + BIP and conventional clamp crushing (CLAMP)., Methods: Based on our database of patient records, a total of 380 consecutive patients who underwent hepatectomy at our hospital were retrospectively studied for the efficacy of BIP alone, CUSA + BIP and CLAMP. Of all the patients, 75 received saline-coupled BIP (Group A), 53 received CUSA + BIP (Group B), and 252 received CLAMP (Group C). The pre-, mid-, and postoperative clinical manifestations were compared, and the effects of those maneuvers were evaluated., Results: There was no obvious difference among the preoperative indexes between the different groups. The operative time was longer in Groups A and B than in Group C (P < 0.001 for both). The amount of bleeding and the rate of transfusion during the operation were significantly higher in Group C than in Groups A and B (P < 0.001 for all). The incidence of postoperative complications in Group C (46.43%) was higher than that in Groups A (30.67%, P = 0.015) and B (28.30%, P = 0.016). The patients' liver function recovery and postoperative hospital stay were not significantly different. BIP could decrease intraoperative hemorrhage and postoperative complications compared to CLAMP., Conclusion: Simple saline-coupled BIP should be considered a safe and reliable technique for liver resection to decrease intraoperative hemorrhage and postoperative complications.

Objective: To investigate the change of aberrant methylation of p16, CDH, RASSF1A and TIMP3 in cervical carcinoma and their significance in cervical carcinoma., Methods: Using the bisulfite-modification technique and methylation-specific PCR (MSP), we examined the aberrant promoter hypermethylation patterns of 4 tumor suppressor genes (p16, CDH1, RASSF1A, TIMP3) in 140 samples of cervical intraepithelial neoplasia (CINI, n = 40), CINII-III (n = 40), cervical carcinomas (CC, n = 40), and normal cervical tissue as a control group (n = 20)., Results: (1) Methylation was completely absent in control tissues. (2) Significant differences between CINII-III group and CINI group were detected for p16 and CDH1 (22% vs 2%, P < 0.05; 35% vs 5%, P < 0.05), while there were no significant differences between the two groups for RASSF1A and TIMP3 (12% vs 2%, P > 0.05; 15% vs 2%, P > 0.05). (3) The presence of methylation of p16 (40%), CDH1 (58%), RASSF1A (20%) and TIMP3 (35%) in CC were higher than the corresponding CINII-III group, but with no significant differences (P > 0.05). (4) Significant differences between CC and CINIfor p16, CDH1, RASSF1A and TIMP3 genes (P < 0.05) were observed. (5) Methylation for at least one gene was a frequent event. These figures in CC 90% (36/40) were significantly different from CINII-III 55% (22/40; P < 0.05). In comparison between CINI8% (3/40) and CC and CINII-III, these figures were significantly different (P < 0.05)., Conclusions: Among the four genes, p16, CDH, RASSF1A and TIMP3, there is a significant trend for increased methylation with increasing degree of histopathological change. It suggests that the aberrant methylation of tumor suppressor genes plays a role during cervical cancer development. This may help identify women at increased risk for or cancer development and progression.

Objective: To study the inhibitory effect of T-2 toxin on the expression of aggrecan and collagen II in chondrocytes and the protection of selenium against this effect., Methods: Human chondrocytes cultured in vitro were treated with T-2 toxin at different concentrations for varied time periods (1-5 days), and the cell viability was measured by MTT assay. Aggrecan expression was detected by toluidine blue staining and collagen II expression by immunostaining using monoclonal antibody of collagen. Aggrecan and collagen II mRNA expressions were measured by semiquantitative RT-PCR., Results: T-2 toxin dose- and time-dependently affected chondrocyte viability within the concentration range of 0.001-2 mg/L, the prolonged treatment time further enhanced the dose dependence of the inhibitory effect. T-2 toxin lowered aggrecan and collagen II synthesis in the chondrocytes and reduced their mRNA expressions. Selenium could partly attenuate the inhibitory effects of T-2 toxin on aggrecan mRNA expression, but showed no such effect against T-2-induced collagen II expression., Conclusion: T-2 toxin can obviously inhibit aggrecan and collagen II synthesis in human chondrocytes, and selenium can partly antagonize the inhibitory effects of T-2 toxin on aggrecan.

Objective: To investigate the role of matrix metalloproteinase-2,9 (MMP-2, MMP-9) and tissue inhibitor of matrix metalloproteinases-2,1 (TIMP-2, TIMP-1) in human amniochorionic membrane., Methods: Amniochorionic membranes were collected from the following groups of women: (1) women with spontaneous premature rupture of membrane (PROM) before onset of labor (PROM group, n = 8), (2) Women with term labor after vaginal delivery (vaginal delivery group, n = 8), (3) Women undergoing elective cesarean section (C-section group, n = 8). Messenger ribonucleic acid expression for MMP-2, MMP-9, and their specific inhibitors TIMP-2 and TIMP-1 were studied with reverse transcriptase polymerase chain reaction., Results: (1) MMP-2 level in PROM group was 0.849 +/- 0.037; in vaginal delivery group 0.327 +/- 0.023; in C-section group 0.307 +/- 0.028. Expression in PROM group was highest, with significant difference compared with the other groups (P < 0.05). Its expression in vaginal delivery group and C-section group had no significant difference (P > 0.05). (2) MMP-9 level in PROM group was 0.026 +/- 0.004; in vaginal delivery group 0.008 +/- 0.001, with significant difference between the two groups (P < 0.05), while the expression was absent in C-section group. (3) TIMP-2 level in PROM group was 0.420 +/- 0.122; in vaginal delivery group 0.730 +/- 0.148; in C-section group 0.885 +/- 0.065. The expression in PROM group was significantly lower than in vaginal delivery group and C-section group (P < 0.05). And it was significantly lower in vaginal delivery group than in C-section group (P < 0.05). (4)TIMP-1 level in PROM group was 0.442 +/- 0.020; in vaginal delivery group 0.431 +/- 0.016; in C-section group 0.427 +/- 0.011. The expression in three groups had no significant difference (P > 0.05)., Conclusions: In PROM group, the expression of MMP-2, MMP-9 and their inhibitors TIMP-2, TIMP-1 is imbalanced, leading to increased extracellular matrix degradation, and weakening of the fetal membranes, and eventually premature rupture of the membranes.