Abstract To determine whether ozone can regulate fruit ripening and delay fruit aging, Kiwifruit (Actinidia deliciosa 'Guichang') was fumigated with different ozone concentrations (100 μL/L, 200 μL/L, 300 μL/L) for 3 h, stored at 1 °C and 85 ± 5% RH for three months, and then matured at 20 °C and 85% RH for 12 days. Compared with controls, the optimal concentration of ozone delayed kiwifruit ripening during cold storage by delaying the respiratory burst that precedes fruit decay. Consequently, fruit firmness, nutrient content, and enzyme activity remained stable; the expression of AdACS1 and AdACO1 as well as the activity of ACC synthetase (ACS) and ACC oxidase (ACO) was downregulated, whereby ACC and MACC were accumulated. In addition, ozone-treated fruit showed significantly reduced infection with grey mold. The effect of ozone on kiwifruit at 200 μL/L was the best suited for delaying fruit senescence, maintaining high quality during storage and fruit ripening compared with other fruits. Overall, our study indicated that ozone may play a major residual role in the physiological process of fruit ripening; furthermore, ozone can affect ethylene biosynthesis and fruit changes associated with endogenous ethylene production.

Abstract We investigated the effects of melatonin (MT) treatment on the physiological quality and cell wall metabolism of kiwifruit during a 12-day shelf life period (20 ± 1 °C, 90 ± 5% RH), after 90 days of cold storage (0 ± 0.5 °C, 90 ± 5% RH). Our results showed that MT treatment delayed kiwifruit softening, as evidence by the enhanced endurance of a healthy appearance, quality, color difference L* value, and firmness of the fruit flesh. MT treatment reduced fruit decay rate, respiration rate, and malondialdehyde content; furthermore, MT-treated kiwifruit maintained high levels of total soluble solids, titratable acid, and ascorbic acid. Additionally, MT inhibited the decrease in fruit pectin, cellulose, and hemicellulose contents; it delayed the increase in water-soluble pectin content and kept pectin methyl esterase, polygalacturonase, cellulase, and β-galactosidase activities low. These results indicated that melatonin may have reduced the speed of softening of kiwifruit and preserved fruit quality by inhibiting enzymatic degradation of the cell wall, and by delaying pro-pectin, cellulose, and hemicellulose degradation, as well as the increase of water-soluble pectin content. These findings indicate that MT treatment may be an effective measure to delay kiwifruit postharvest cell-wall metabolism and preserve fruit quality.

Abstract The effects of 1-methylcyclopropene (1-MCP) treatment and controlled-atmosphere (CA) storage and their combination on quality and antioxidant capacity of postharvest blueberries were evaluated here. To investigate the most appropriate approach for maintaining fruit quality, blueberries were subjected to four different treatments, a control (CK), 1-MCP, 3% O2 + 10% CO2 (CA), and 1-MCP combined with 3% O2+10% CO2 (1-MCP + CA), then stored for 80 days at 0 °C. It was found that the applications of 1-MCP, CA and 1-MCP + CA were effective at delaying the increase in weight loss and inhibiting the decrease in firmness, total soluble solids (TSS) contents and titratable acidity (TA) contents. In addition, 1-MCP, CA and 1-MCP + CA treatment could significantly suppress MDA accumulation and relative electrolytic leakage promotion. Moreover, the production of reactive oxygen species (ROS), such as O2- and H2O2, was reduced by all the treatment, which was associated with increased activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). Taken together, our findings indicate 1-MCP, CA and 1-MCP + CA have potential applications in maintaining the quality and improving the antioxidant capacity of harvested blueberries, and 1-MCP + CA is the most effective treatment.

Abstract Pitaya fruits have high senescence rates throughout their postharvest storage period. Additionally, studies have confirmed that melatonin plays a regulatory role in plant senescence. However, the involvement of melatonin in the postharvest senescence of fruits remains unclear. In this study, two cultivars of pitaya fruit, ‘Zihonglong’ and ‘Jinghonglong’, were treated with melatonin and then evaluated for characteristics of senescence while in storage for 10 days. The results showed that melatonin treatment delayed fruit senescence in both pitaya cultivars, as indicated by the inhibition of weight loss, decay incidence, relative membrane permeability, and malondialdehyde (MDA) content, as well as the maintenance of the total soluble solids and ascorbic acid contents and the reduced respiration intensity. In addition, melatonin treatment reduced the O2·- production rates, H2O2 contents, and lipoxygenase activities but enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) in both pitaya cultivars. These results indicate that melatonin may contribute to delaying senescence in pitaya fruits. This study shows the potential of the use of melatonin in the postharvest storage of pitaya fruits, as well as other horticultural fruits.

Abstract Melatonin is known to influence various physiological processes, including plant ripening; however, the effect of melatonin on postharvest blueberry fruit softening is unknown. Herein, the effects of melatonin at different concentrations (100, 300, and 500 μmol L-1) on the quality parameters, cell wall substances, cell-wall-degrading enzymes, and cell wall ultrastructure of blueberry fruits were investigated during low-temperature storage. Melatonin treatment was found to effectively delay the decline in fruit firmness, slow the loss of soluble solids and titratable acids, and decrease the respiration rate and decay rate of blueberry fruits. The blueberry fruits treated with 300 μmol L-1 melatonin exhibited the best quality. Melatonin treatment promoted anthocyanin accumulation, delayed ascorbic acid degradation, maintained a low relative membrane permeability in pulp, inhibited increases in lipoxygenase (LOX) activity, and reduced membrane lipid peroxidation. Moreover, melatonin controlled the formation of soluble pectin by inhibiting the activities of pectin methylesterase (PME), polygalacturonase (PG), cellulase (Cx), and β-glucosidase (β-Glu) while maintaining high contents of protopectin and cellulose. These results indicate that melatonin can effectively improve blueberry fruit quality and antioxidant capacity as well as and delay fruit softening by inhibiting the degradation of cell wall substances.

Abstract The study was designed to investigate the effects of cryopreservation on bovine, ovine, and goat sperm motility, acrosome structure, enzyme activity, and fertilization ability. Percentage of sperm with hyaluronidase enzyme (HYD) activity was detected by a modified sodium hyaluronate-gelatin membrane. The N-α-benzoyl-DL-arginine-p-nitroanilide (BNPNA) method was used to assess the sperm acrosome enzyme (ACE). The mean percentage of sperm acrosome integrity dropped significantly (P < 0.01) after cryopreservation. The ACE activity of bovine sperm (100.48) was higher (P < 0.01) than that of ovine (57.88) or goat sperm (50.30), while the percentage of sperm with HYD activity of bovine (71.10%) and ovine (67.60%) sperm was higher than that of goat sperm (58.52%) after cryopreservation (P < 0.01). Sperm motility was positively correlated with the activity of the two acrosome enzymes before and after cryopreservation (P < 0.01). Cryopreservation had a negative effect on acrosomal morphology, motility, and acrosomal enzyme activity in their sperm. The fertilization ability of ovine and goat sperm decreased significantly after cryopreservation, but that of frozen bovine sperm did not differ significantly when compared with fresh sperm. There was no significant difference between ovine and goat sperm indices, except for percentage of sperm with HYD activity.

Abstract Purpose Patients with diabetes are vulnerable to myocardial I/R (ischaemia/reperfusion) injury, but are not responsive to IPO (ischaemic post-conditioning). We hypothesized that decreased cardiac Adiponectin (APN) is responsible for the loss of diabetic heart sensitivity to IPO cardioprotecton. Methods Diabetic rats were subjected to I/R injury (30 min of LAD occlusion followed by 120 min of reperfusion). Myocardial infarct area was determined by TTC staining. Cardiac function was monitored by a microcatheter. ANP, 15-F2t-isoprostane, nitrotyrosine and MDA were measured by assay kits. Levels of p-Akt, total-Akt and GAPDH were determined by Western Blot. Results Diabetic rats subjected to myocardial IR exhibited severe myocardial infarction and oxidative stress injury, lower APN in the plasma and cardiac p-Akt expression ( P <0.05). IPO significantly attenuated myocardial injury and up-regulated plasma APN content and cardiac p-Akt expression in non-diabetic rats but not in diabetic rats. Linear correlation analysis showed that the expression of adiponectin was positively correlated with p-Akt and negatively correlated with myocardial infarction area ( P <0.01). Conclusion Protective effect of IPO was tightly correlated with the expression of adiponectin, exacerbation of I/R injury and ineffectiveness of IPO was partially due to the decline of adiponectin and inactivation of Akt in diabetes mellitus.

Abstract This study aimed to develop a simple and rapid method to detect KRAS gene mutations for conventional clinical applications under laboratory conditions. The genotype of mutation sites was determined based on the occurrence of target bands in the corresponding lanes of the reaction tubes through polymerization-conjunction of the probes, probe purification and amplification, and agarose gel electrophoresis. Circulating DNA samples were obtained from the plasma of 72 patients with lung cancer, which were identified based on six mutation sites (G12S, G12R, G12C, G12D, G12A, and G12V) of codon 12 of the KRAS gene. The detection results were compared with direct sequencing data. The proposed detection method is characterized by simple operation, high specificity, and high sensitivity (2%). This method can detect the mutations of three samples at G12S, G12R, and G12A. In the direct sequencing spectra of these samples, the genotype could not be determined due to the lack of evident sequencing peaks that correspond to the basic group of mutations. In conclusion, a simple and rapid method was established based on probe polymerization-conjunction-agarose gel electrophoresis for detecting KRAS gene mutations. This method can be applied to the conventional mutation detection of inhomogeneous samples.

Abstract Purpose: To investigate the specific molecular mechanisms and effects of curcumin derivative J147 on diabetic peripheral neuropathy (DPN). Methods: We constructed streptozotocin (STZ)-induced DPN rat models to detected mechanical withdrawal threshold (MWT) in vivo using Von Frey filaments. In vitro, we measured cell viability and apoptosis, adenosine 5‘-monophosphate-activated protein kinase (AMPK) and transient receptor potential A1 (TRPA1) expression using MTT, flow cytometry, qRT-PCR and western blot. Then, TRPA1 expression level and calcium reaction level were assessed in agonist AICAR treated RSC96cells. Results: The results showed that J147reduced MWT in vivo, increased the mRNA and protein level of AMPK, reduced TRPA1 expression and calcium reaction level in AITCR treated RSC96 cells, and had no obvious effect on cell viability and apoptosis. Besides, AMPK negative regulated TRPA1 expression in RSC96 cells. Conclusions: J147 could ameliorate DPN via negative regulation AMPK on TRPA1 in vivo and in vitro. A curcumin derivative J147might be a new therapeutic potential for the treatment of DPN.

Abstract In this work, the effects of 1-methylcyclopropene (1-MCP) on texture properties, titratable acidity (TA) and total soluble solids (TSS) in blueberry were examined during long term storage and simulated transportation. Blueberry fruits were stored at 0 ± 0.5 °C after being treated with 0.1, 0.2 and 0.3μL L-11-MCP. After 40 and 80 days of storage, quickly put 8 punnets into a foam box with commercial cold pack to simulate the commercial transportation for 2 days at 25 °C. There were considerable differences in the texture properties during long term storage and simulated transportation with 1-MCP. The 0.3 μL L-1 1-MCP can effectively decreased the decline of gumminess and chewiness, retarded the reduction of TA, and delayed blueberry fruit softening. There was significant positive correlation among firmness, springiness, cohesiveness, gumminess, chewiness, resilience, TSS, and TA. These results demonstrate that the 0.3 μL L-1 1-MCP combines with commercial polyethylene bags were successful effect in maintaining the texture properties of blueberry.

Abstract Large quantities of kitchen waste are produced in modern society and its disposal poses serious environmental and social problems. The aim of this study was to isolate degradative strains from kitchen waste and to develop a novel and effective microbial agent. One hundred and four strains were isolated from kitchen waste and the 84 dominant strains were used to inoculate protein-, starch-, fat- and cellulose-containing media for detecting their degradability. Twelve dominant strains of various species with high degradability (eight bacteria, one actinomycetes and three fungi) were selected to develop a compound microbial agent "YH" and five strains of these species including H7 (Brevibacterium epidermidis), A3 (Paenibacillus polymyxa), E3 (Aspergillus japonicus), F9 (Aspergillus versicolor) and A5 (Penicillium digitatum), were new for kitchen waste degradation. YH was compared with three commercial microbial agents-"Tiangeng" (TG), "Yilezai" (YLZ) and Effective Microorganisms (EM), by their effects on reduction, maturity and deodorization. The results showed that YH exerted the greatest efficacy on mass loss which decreased about 65.87% after 14 days. The agent inhibited NH3 and H2S emissions significantly during composting process. The concentration of NH3 decreased from 7.1 to 3.2 ppm and that of H2S reduced from 0.7 to 0.2 ppm. Moreover, E4/E6 (Extinction value460nm/Extinction value665nm) of YH decreased from 2.51 to 1.31, which meant YH had an obvious maturity effect. These results highlighted the potential application of YH in composting kitchen waste.

The effects of Dangguibuxue Tang (DBT) on growth performance and immunity response in immunosuppressed broiler chicks were investigated in this study. 240 one-d-old broiler chicks (DaHeng S01) were randomly divided into 4 groups, 2.0% DBT-treatment (A), 0.5% DBT-treatment (B), cyclophosphamide-control (C), and control group (D). From 4 d to 7 d of age, chicks in group A, B and C were given cyclophosphamide (CY) at a dosage of 100mg/kg body weight (BW) daily by intraperitoneal injection to induce immunosuppression. Chicks in group D were given an equal volume of physiological saline daily by intraperitoneal injection and considered normal chicks. Groups A and B were supplemented with 2.0% or 0.5% of DBT in the drinking water from 8 d to 42 d of age. Groups C and D did not receive any additional medication. The results revealed that chicks from group B had lower feed:gain rate (FGR), lower total mortality, higher immunity organ indexes, higher levels of Newcastle disease (ND) antibody and infectious bursal disease (IBD) antibody, higher interleukin-2 and interleukin-6 levels, and greater lymphocyte proliferative responses to concanavalin A (ConA) during the experiment than those from group C. However, no significant difference in the immunity status in the two levels of DBT-treatment was observed. These results indicate that supplementation of 0.5% of DBT can improve both cellular immunity and humoral immunity in immunosuppressed broiler chicks.

OBJETIVO: Comparar prospectivamente o efeito terapêutico da preparação do túnel femoral através do túnel tibial (TT) ou da porta ântero-medial(AM) na reconstrução do ligamento cruzado anterior(LCA) em feixe único. MÉTODOS: Entre junho de 2008 e outubro de 2010, 76 pacientes foram submetidos à reconstrução do LCA em feixe único pelo enxerto autógeno de tendão semitendíneo egrácil.Todos os casos foram divididos aleatoriamente em dois grupos de acordo como método de preparação do túnel femoral: grupo transtibial (TT) (n=38) e grupo ântero-medial (AM) (n=38). Foi usado o escore Lysholm para joelho. O relaxamento anterior do joelho a 30° sob força tênsil de 133,32N foi determinado com o medidor KT-1000 no pré e no pós-operatório nos dois grupos. RESULTADOS: Sessenta e cinco pacientes (grupo TT, 34; grupo AM,31)foram acompanhados por mais de 12 meses, com uma taxa de follow-up de 86%.A pontuação do Lysholm para joelho e do relaxamento anterior medido pelo KT-1000 aos 12 meses de pós-operatório foi significativamente melhor do que antes da reconstrução.As pontuações de Lysholme do relaxamento KT-1000 não foram significativamente diferentes comparando os grupos TT e AM após a operação. CONCLUSÃO: A preparação do túnel femoral através do túnel tibial ou da porta ântero-medial na reconstrução do ligamento cruzado anterior em feixe único mostrou os mesmos efeitos terapêuticos.

PURPOSE: To prospectively compare therapeutic effect of femoral tunnel preparation through the tibial tunnel and the anteromedial (AM) portal in single-bundle anterior cruciate ligament (ACL) reconstruction. METHODS: Between June 2008 and October 2010, 76 patients underwent single-bundle ACL reconstruction by autogenous grafting of semitendinosus and gracilis tendon. All cases were randomly divided into two groups according to the method of femoral tunnel preparation: transtibial (TT) group (n=38) and anteromedial (AM) group (n=38). Lysholm knee score and the KT-1000 anterior laxity at 30° of pre-and post-operation were assessed for two groups. RESULTS: Sixty-five patients (TT group, 34; AM group, 31) were followed up for more than 12 months, with a follow-up rate of 86%. The Lysholm knee score and the KT-1000 anterior laxity 12 months after operation were significantly better than before reconstruction. The Lysholm knee score and the KT-1000 anterior laxity were not significantly different between the TT and AM groups after operation. CONCLUSION: Femoral tunnel preparation through tibial tunnel or the anteromedial portal in single-bundle anterior cruciate ligament reconstruction shows same therapeutic effects.