Extraction and characterization of anti-virus anthraquinones from Nicotiana tabacum-derived Aspergillus oryzae YNCA1220.

In this study, seven novel anthraquinones (1-7) and four described anthraquinones (8-11) were purified from Nicotiana tabacum-derived Aspergillus oryzae YNCA1220. It is worth noting that only analogs of 4 and 5 have been reported as natural products to date, while the nuclei of compounds 1-3, 6 and 7 were isolated for the first time in nature. Among them, compounds 1-3 bear an unusual anthra[2,3-b]furan-9,10-dione nucleus, 4 and 5 possess a rare 3-methyl-1H-pyrrol-2-yl substituent, and 6 and 7 are new framework anthraquinones bearing a 6-methyl-1,7-dihydro-2H-azepin-2-one ring. Interestingly, the in vivo assays indicated that 1, 4 and 5 had inactivation effects against tobacco mosaic virus (TMV) with inhibition rates of 41.6%, 55.4% and 38.6%, respectively, at a concentration of 50 µg/mL, which were better than that of the positive control agent, ningnanmycin (33.8%). Compounds 1, 4 and 5 also had protective effects with inhibition rates of 48.7%, 60.2% and 43.5% at the same concentration, while 4 had a better curative effect than ningnanmycin at a concentration of 100 µg/mL. In addition, mechanistic studies also revealed that a potent direct effect on TMV, the induction of SAR in tobacco plants, and the effective regulation of defense enzymes, defense genes, and defense hormones may be the reasons for the significant effects of 4 against TMV. At the same time, downregulation of the expression of total NtHsp70 protein by inhibiting the related Hsp70 genes may also be involved in tobacco resistance to TMV. To evaluate whether compounds have broader antiviral activities, the antirotavirus activities of new isolates were also evaluated and found to be highly effective with a therapeutic index (TI) value ranging from 11.6 to 17.7. This study suggests that the above anthraquinone compounds, particularly 4, have broad spectrum antiviral activities. The successful isolation and structure identification of the above anthraquinones provide new materials for the screening of anti-TMV agents and contribute to the improved utilization of N. tabacum-derived fungi.

Forest characteristics, population structure and growth trends of threatened relict Pseudotsuga forrestii in China.

Pseudotsuga forrestii is a relict evergreen coniferous tree species in Pinaceae endemic to China. P. forrestii tree numbers have greatly decreased due to deforestation, over-utilization and habitat degradation. Here we clarify P. forrestii community types, structure, species diversity, seedling recruitments and growth trends. We identified four P. forrestii community types: (1) Pseudotsuga forrestii - Quercus guyavifolia - Acer davidii evergreen coniferous and broad-leaved mixed forest; (2) Pseudotsuga forrestii - Pinus yunnanensis - Quercus guyavifolia evergreen coniferous and broad-leaved mixed forest; (3) Pseudotsuga forrestii evergreen coniferous forest; (4) Pseudotsuga forrestii - Abies georgei var. smithii evergreen coniferous forest. P. forrestii forests are characterized by both warm temperate and temperate affinities. Simpson diversity, Pielou evenness, Shannon-Wiener diversity indices ranged from 0.75 to 0.76, 0.74-0.81, and 1.62-1.93, respectively, with no significant differences among the four forest types. The forest stratification was multilayered. The canopy layer was generally 10-25 m tall, with the emergent layer reached 25-42 m. DBH and age structures of P. forrestii showed multimodal distributions. Its maximum age P. forrestii was 570 years with a DBH of 143 cm. The growth of annual ring width of P. forrestii was slow, and generally decreased with age, whereas the basal area at the breast height increased with age. Established seedlings/saplings were mainly found in unstable micro-habitats. Regeneration of P. forrestii depends on moderate natural disturbances. Finally, we provide recommendations for P. forrestii conservation.

Bloodletting Acupuncture at Jing-Well Points on Hand Induced Autophagy to Alleviate Brain Injury in Acute Altitude Hypoxic Rats by Activating PINK1/Parkin Pathway.

OBJECTIVE: To explore the protective effect of bloodletting acupuncture at twelve Jing-well points on hand (BAJP) on acute hypobaric hypoxia (AHH)-induced brain injury in rats and its possible mechanisms. METHODS: Seventy-five Sprague Dawley rats were divided into 5 groups by a random number table (n=15), including control, model, BAJP, BAJP+3-methyladenine (3-MA), and bloodletting acupuncture at non-acupoint (BANA, tail tip blooding) groups. After 7-day pre-treatment, AHH models were established using hypobaric oxygen chambers. The levels of S100B, glial fibrillary acidic protein (GFAP), superoxide dismutase (SOD), and malondialdehyde (MDA) in serum were measured by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method were used to assess hippocampal histopathology and apoptosis. Transmission electron microscopy assay was used to observe mitochondrial damage and autophagosomes in hippocampal tissues. Flow cytometry was used to detect mitochondrial membrane potential (MMP). The mitochondrial respiratory chain complexes I, III and IV activities and ATPase in hippocampal tissue were evaluated, respectively. Western blot analysis was used to detect the protein expressions of Beclin1, autophagy protein 5 (ATG5), microtubule-associated protein 1 light chain 3 beta (LC3B), phosphatase and tensin homolog induced kinase 1 (PINK1), and Parkin in hippocampal tissues. The mRNA expressions of Beclin1, ATG5 and LC3-II were analyzed by quantitative real-time polymerase chain reaction. RESULTS: BAJP treatment reduced hippocampal tissue injury and inhibited hippocampal cell apoptosis in AHH rats. BAJP reduced oxidative stress by decreasing S100B, GFAP and MDA levels and increasing SOD level in the serum of AHH rats (P<0.05 or P<0.01). Then, BAJP increased MMP, the mitochondrial respiratory chain complexes I, III and IV activities, and the mitochondrial ATPase activity in AHH rats (all P<0.01). BAJP improved mitochondrial swelling and increased the autophagosome number in hippocampal tissue of AHH rats. Moreover, BAJP treatment increased the protein and mRNA expressions of Beclin1 and ATG5 and LC3-II/LC3-I ratio in AHH rats (all P<0.01) and activated the PINK1/Parkin pathway (P<0.01). Finally, 3-MA attenuated the therapeutic effect of BAJP on AHH rats (P<0.05 or P<0.01). CONCLUSION: BAJP was an effective treatment for AHH-induced brain injury, and the mechanism might be through reducing hippocampal tissue injury via increasing the PINK1/Parkin pathway and enhancement of mitochondrial autophagy.

Dispersal Behavior Characters of Spodoptera frugiperda Larvae.

The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a major pest of corn worldwide. FAW larval dispersal is an important life strategy that influences FAW population distribution in corn fields and subsequent plant damage. We studied FAW larval dispersal in the laboratory with sticky plates placed around the test plant and a unidirectional airflow source. Crawling and ballooning were the main dispersal means of FAW larvae both within and between corn plants. All larval instars (1st-6th) could disperse by crawling, with crawling being the only dispersal mechanism for 4th-6th instars. By crawling, FAW larvae could reach all aboveground parts of a corn plant as well as adjacent corn plants where leaves overlapped. Ballooning was used primarily by 1st-3rd instar larvae, and the proportion of these larvae that used ballooning decreased with age. Ballooning was largely governed by the larva's interaction with airflow. Airflow influenced the direction and distance of larval ballooning. With an airflow speed of about 0.05 m/s, 1st instars could travel up to 196 cm from the test plant, indicating that long-distance FAW larval dispersal depends on ballooning. These results increase our understanding of FAW larval dispersal and provide scientific information for the development of FAW monitoring and control strategies.

Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress.

OBJECTIVE: To explore the protective effect and possible mechanisms of bloodletting acupuncture at Jing-well points (BAJP) pre-treatment on acute hypobaric hypoxia (AHH)-induced myocardium injury rat. METHODS: Seventy-five rats were randomly divided into 5 groups by a random number table: a control group (n=15), a model group (n=15), a BAJP group (n=15), a BAJP+3-methyladenine (3-MA) group (n=15), and a BANA (bloodletting at nonacupoint; tail bleeding, n=15) group. Except for the control group, the AHH rat model was established in the other groups, and the corresponding treatment methods were adopted. Enzyme-linked immunosorbent assay (ELISA) was used to detect creatine kinase isoenzyme MB (CK-MB) and cardiac troponins I (CTnI) levels in serum and superoxide dismutase (SOD) and malondialdehyde (MDA) levels in myocardial tissue. Hematoxylin-eosin (HE) staining was used to observe myocardial injury, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to observe cell apoptosis. Transmission electron microscopy detection was used to observe mitochondrial damage and autophagosomes in the myocardium. The mitochondrial membrane potential of the myocardium was analyzed with the fluorescent dye JC-1. Mitochondrial respiratory chain complex (complex I, III, and IV) activities and ATPase in the myocardium were detected by mitochondrial respiratory chain complex assay kits. Western blot analysis was used to detect the autophagy index and hypoxia inducible factor-1α (HIF-1α)/Bcl-2 and adenovirus E1B 19k Da-interacting protein 3 (BNIP3) signaling. RESULTS: BAJP reduced myocardial injury and inhibited myocardial cell apoptosis in AHH rats. BAJP pretreatment decreased MDA levels and increased SOD levels in AHH rats (all P<0.01). Moreover, BAJP pretreatment increased the mitochondrial membrane potential (P<0.01), mitochondrial respiratory chain complex (complexes I, III, and IV) activities (P<0.01), and mitochondrial ATPase activity in AHH rats (P<0.05). The results from electron microscopy demonstrated that BAJP pretreatment improved mitochondrial swelling and increased the autophagosome number in the myocardium of AHH rats. In addition, BAJP pretreatment activated the HIF-1α/BNIP3 pathway and autophagy. Finally, the results of using 3-MA to inhibit autophagy in BAJP-treated AHH rats showed that suppression of autophagy attenuated the treatment effects of BAJP in AHH rats, further proving that autophagy constitutes a potential target for BAJP treatment of AHH. CONCLUSION: BAJP is an effective treatment for AHH-induced myocardial injury, and the mechanism might involve increasing HIF-1α/BNIP3 signaling-mediated autophagy and decreasing oxidative stress.

[Effect of blood-letting puncture of "Well-points" on hippocampal mitophagy-related protein expression in rats with acute brain injury due to hypobaric hypoxia].

OBJECTIVE: To study the effect of blood-letting puncture at "Well-points" of the twelve meridians on hippocampal mitophagy of hypobaric hypoxia-induced brain injury (HHIBI) rats, so as to explore its biological mechanisms underlying improvement of high altitude hypoxia-induced brain injury. METHODS: Male SD rats were randomly divided into normal control group （n=9）， and model and blood-letting groups which were further divided into 6, 12, 24, 48 and 72 h subgroups (n=9 in each subgroup). The HHIBI model was established by putting the rats into a hypobaric hypoxia chamber (equivalent to 5 000 m above sea level).The blood-letting groups were given blood-letting therapy at "Shaoshang"(LU11), "Shangyang"(LI1), "Zhongchong"(PC9), "Guanchong"(SJ1), "Shaochong"(HT9), "Shaoze"(SI9), once a day for 7 days. H.E. staining was used to observe the histopatholo-gical changes of hippocampus tissue. Serum hypoxia inducible factor(HIF)-1α and vascular endothelial growth factor(VEGF) contents were assayed using ELISA, and the expression levels of hippocampal Beclin-1 and LC3-Ⅱ proteins detected using Western blot. RESULTS: Compared with the normal control group, the levels of serum HIF-1α and VEGF at each time point, and the expressions of hippocampal Beclin-1 at 12 and 24 h, LC3-Ⅱat each time point were significantly increased in the model group (P<0.05, P<0.01); while in comparison with the model group, the levels of serum HIF-1α and VEGF contents, and the expressions of Beclin-1 at 12 h, LC3-Ⅱ at 24, 48 and 72 h were further significantly up-regulated in the blood-letting group (P<0.01, P<0.05). H.E. staining revealed that the pyramidal cells in the hippocampal CA1 region had a disordered arrangement, and some of them presented swelling with loose and pale cytoplasm or vacuolation at 6, 12 and 24 h, and showed indistinct nucleolus, irregular shape, pyknosis and deep staining and an obvious edema at 48 and 72 h, which was relatively milder in the blood-letting group. CONCLUSION: Blood-letting of "Well-points" can up-regulate serum HIF-1α and VEGF contents and hippocampal Beclin-1 and LC3-Ⅱ (mitophagy related proteins) expressions in HHIBI rats, which may contribute to its effect in reducing hypoxic brain injury.

Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions.

Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression levels in luffa plants exposed to various simulated abiotic stresses [i.e., cold, drought, heat, salt, H2O2, and abscisic acid (ABA) treatments] were analyzed by qRT-PCR. The stability of the reference gene expression levels was validated using the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. The results indicated that EF-1α was the most stably expressed and suitable reference gene overall and for the heat, cold, and ABA treatments. Additionally, UBQ expression was stable following the salt treatment, whereas TUB was identified as a suitable reference gene for H2O2 and drought treatments. The reliability of the selected reference genes was verified by analyzing the expression of copper/zinc superoxide dismutase (Cu/Zn-SOD) gene in luffa. When the most unstable reference genes were used for data normalizations, the resulting expression patterns had obvious biases when compared with the expression patterns for the most ideal reference genes used alone or combined. These results will be conducive to more accurate quantification of gene expression levels in luffa.

The Energetic Origin of Different Catalytic Activities in Temperature-Adapted Trypsins.

Psychrophilic enzymes were always observed to have higher catalytic activity (k cat) than their mesophilic homologs at room temperature, while the origin of this phenomenon remains obscure. Here, we used two different temperature-adapted trypsins, the psychrophilic Atlantic cod trypsin (ACT) and the mesophilic bovine trypsin (BT), as a model system to explore the energetic origin of their different catalytic activities using computational methods. The results reproduce the characteristic changing trends in the activation free energy, activation enthalpy, and activation entropy between the psychrophilic and mesophilic enzymes, where, in particular, the slightly decreased activation free energy of ACT is determined by its considerably reduced activation enthalpy rather than by its more negative activation entropy compared to BT. The calculated electrostatic contributions to the solvation free energies in the reactant state/ground sate (RS/GS) and transition state (TS) show that, going from BT to ACT, the TS stabilization has a predominant effect over the RS stabilization on lowering the activation enthalpy of ACT. Comparison between the solvation energy components reveals a more optimized electrostatic preorganization to the TS in ACT, which provides a larger stabilization to the TS through reducing the reorganization energy, thus resulting in the lower activation enthalpy and hence lower activation free energy of ACT. Thus, it can be concluded that it is the difference in the protein electrostatic environment, and hence its different stabilizing effects on the TS, that brings about the different catalytic activities of different temperature-adapted trypsins.

Spatial Distribution of Pollinating Butterflies in Yunnan Province, Southwest China with Resource Conservation Implications.

Pollinating butterflies are an important asset to agriculture, which still depends on wild resources. Yunnan Province in Southwest China is a region with typical montane agriculture, but this resource is poorly investigated. From literature reference and specimen examination, the present study identified 554 species of pollinating butterflies (50.8% of the total butterflies) from Yunnan, with family Nymphalidae possessing the least number of pollinators (80 species, 16.0%), while the remaining four families are pollinator-rich (>73%). Tropical lowlands and mountain-valley areas possess higher species richness than those with plain terrains. The species richness of pollinating butterflies in Yunnan does not simply decline with the increase of latitude, nor is significantly different between West and East Yunnan. Zonation of pollinating butterflies using the parsimony analysis of endemicity (PAE) identified nine distribution zones and ten subzones. Most areas of endemism (AOE) are found in lowlands or mountain-valley areas, complexity of terrains, climates, and vegetation types are believed to be the main causes of such endemicity. The potential pollinating service of these butterflies could be great to montane agriculture with expanding areas of cash crops and fruit horticulture. Conservation strategies for pollinating butterflies may consist of preserving habitats and establishing butterfly-friendly agriculture based on local traditions.

Facile Preparation of Highly Stretchable TPU/Ag Nanowire Strain Sensor with Spring-Like Configuration.

Stretchable nano-fibers have attracted dramatic attention for the utility in wearable and flexible electronics. In the present case, Ag nanowires (AgNWs)-intertwined thermoplastic polyurethanes (TPU) unwoven nano-membrane is fabricated by an electrospinning method and dip coating technique. Then a strain sensor with a spring-like configuration is fabricated by a twisted method. The sensor exhibits superior electrical conductivity up to 3990 S cm-1 due to the high weight percentage of the Ag nanowires. Additionally, thanks to the free-standing spring-like configuration that consists of uniform neat loops, the strain sensor can detect a superior strain up to 900% at the point the sensor ruptures. On the other hand, the configuration can mostly protect the AgNWs from falling off. Furthermore, major human motion detection, like movement of a human forefinger, and minor human motion detection, such as a wrist pulse, show the possible application of the sensor in the field of flexible electronics.

An efficient TILLING platform for cultivated tobacco.

Targeting-induced local lesions in genomes (TILLING) is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants. Although TILLING has been developed for many diploid plants, the technology has been used in very few polyploid species due to their genomic complexity. Here, we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco (Nicotiana tabacum L.) using an ethyl methanesulfonate (EMS)-mutagenized population of 1,536 individuals. We optimized the procedures for endonuclease preparation, leaf tissue sampling, DNA extraction, normalization, pooling, PCR amplification, heteroduplex formation, and capillary electrophoresis. In a test screen using seven target genes with eight PCR fragments, we obtained 118 mutants. The mutation density was estimated to be approximately one mutation per 106 kb on average. Phenotypic analyses showed that mutations in two heavy metal transporter genes, HMA2S and HMA4T, led to reduced accumulation of cadmium and zinc, which was confirmed independently using CRISPR/Cas9 to generate knockout mutants. Our results demonstrate that this powerful TILLING platform (available at http://www.croptilling.org) can be used in tobacco to facilitate functional genomics applications.

Probing primordial gravitational waves: Ali CMB Polarization Telescope.

In this paper, we will give a general introduction to the Ali CMB Polarization Telescope (AliCPT) project, which is a Sino-US joint project led by the Institute of High Energy Physics and involves many different institutes in China. It is the first ground-based Cosmic Microwave Background (CMB) polarization experiment in China and an integral part of China's Gravitational-wave Program. The main scientific goal of the AliCPT project is to probe the primordial gravitational waves (PGWs) originating from the very early Universe. The AliCPT project includes two stages. The first stage, referred to as AliCPT-1, is to build a telescope in the Ali region of Tibet at an altitude of 5250 meters. Once completed, it will be the highest ground-based CMB observatory in the world and will open a new window for probing PGWs in the northern hemisphere. The AliCPT-1 telescope is designed to have about 7000 transition-edge sensor detectors at 95 GHz and 150 GHz. The second stage is to have a more sensitive telescope (AliCPT-2) with more than 20 000 detectors. Our simulations show that AliCPT will improve the current constraint on the tensor-to-scalar ratio r by one order of magnitude with three years' observation. Besides the PGWs, AliCPT will also enable a precise measurement of the CMB rotation angle and provide a precise test of the CPT symmetry. We show that three years' observation will improve the current limit by two orders of magnitude.

Identification of the Genetic Variation and Gene Exchange between Citrus Trifoliata and Citrus Clementina.

To identify the genetic variation between Citrus trifoliata and Citrus clementina, we performed genome resequencing on the two citrus species. Compared with the citrus reference genome, a total of 9,449,204 single-nucleotide polymorphisms (SNPs) and 846,615 insertion/deletion polymorphisms (InDels) were identified in the two citrus species, while 1,868,115 (19.77%) of the SNPs and 190,199 (22.47%) of the InDels from the two citrus species were located in the genic regions. Meanwhile, a total of 8,091,407 specific SNPs and 692,654 specific InDels were identified in the two citrus genotypes, yielding an average of 27.32 SNPs/kb and 2.34 InDels/kb. We identified and characterized the patterns of gene exchanges in the grafted citrus plants by using specific genetic variation from genome resequencing. A total of 4396 transporting genes across graft junctions was identified. Some specific genetic variation and mobile genes was also confirmed by Sanger sequencing. Furthermore, these mobile genes could move directionally or bidirectionally between the scions and the rootstocks. In addition, a total of 1581 and 2577 differentially expressed genes were found in the scions and the rootstocks after grafting compared with the control, respectively. These genetic variations provide fundamental information on the genetic basis of important traits between C. trifoliata and C. clementina, as the transport of genes would be applicable to horticulture crops.

[Effect of Acupuncture on Reproductive Endocrine and p-Akt Protein Expression in Polycystic Ovary Syndrome Rats with Insulin Resistance].

OBJECTIVE: To observe the effect of manual acupuncture intervention on the reproductive endocrine and p-Akt protein expression in polycystic ovary syndrome with insulin resistance (PCOS-IR) rats, so as to reveal its mechanism in relieving PCOS-IR. METHODS: Female Wistar rats were randomly divided into control, model, medication (Metformin) and acupuncture groups (n＝8 in each). The PCOS-IR model was established by subcutaneous injection of Dehydroepiandrosterone (DHEA, 6 mg/100 g), once daily for 20 days. Rats of the medication group were treated by gavage of Metformin solution (100 mg/kg), once daily for 28 days. For rats of the acupuncture group, manual acupuncture stimulation (lifting-thrusting and rotating of needles) was applied to the bilateral "Shenshu"(BL 23), "Yishu"(EX-B 3), "Zigong" (EX-CA 1), "Sanyinjiao"(SP 6), "Fenglong"(ST 40) and "Guanyuan"(CV 4) for 5 min, once a day for 28 days. H．E. staining was used to observe the histopathologic changes of the ovary tissue. Serum levels of follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH), testosterone (T) and fasting insulin (FINS) were measured by ELISA. Fasting plasma glucose (FPG) was measured by blood glucose meter, and homeostasis model assessment-insulin resistance index (HOMA-IR) was calculated. Western blot was used to detect the expression of Akt and p-Akt proteins in ovarian tissues. RESULTS: Following modeling, the ovaries showed polycystic changes and an increase of atresia follicles with a larger diameter and no corpus luteum in the model group. After the intervention, there were various types of follicles and a small amount of corpus luteum at different stages of development in both acupuncture and medication groups. In comparison with the control group, serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression were considerably decreased (P<0.05, P<0.01), and serum LH, T and FINS contents and HOMA-IR were significantly increased in the model group (P<0.01). Following the intervention, the decreased serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression, and the increased serum LH, T and FINS contents and HOMA-IR were remarkably reversed in both acupuncture and medication groups (P<0.05，P<0.01), without significant differences between the two groups in the levels of those serum and ovary indexes (P>0.05)．. CONCLUSION: Acupuncture intervention can regulate the levels of reproductive endocrine hormones and reduce insulin resistance in PCOS-IR rats, which may be related to its effect in up- regulating the expression of p-Akt protein in the ovaries.

Expression of multidrug-resistance associated proteins in human retinoblastoma treated by primary enucleation.

AIM: To reveal the expression of multidrug-resistance associated proteins: glutathione-S-transferase π (GSTπ), P-glycoprotein (P-gp) and vault protein lung resistance protein (LRP) in retinoblastoma (RB) without any conservative treatment before primary enucleation and to correlate this expression with histopathological tumor features. METHODS: A total of 42 specimens of RB undergone primary enucleation were selected for the research. Sections from the formalin-fixed, paraffin-embedded specimens were stained with HE and immunohistochemistry to detect the expression of GSTπ, P-gp and LRP. RESULTS: GSTπ was expressed in 39/42 (92.86%) RBs and in 9/9 (100%) well-differentiated RBs. P-gp/GSTπ was found in 30 (71.42%) of 42 RBs. Totally 9 (21.43%) tumors were well differentiated and 33 (78.57%) were poorly differentiated. Totally 15 (35.71%) eyes had optic nerve (ON) tumor invasion, 36 (85.71%) had choroidal tumor invasion, and 14 (33.33%) had simultaneous choroidal and ON invasion. There was no statistically significant relationship between P-gp, GSTπ, LRP positivity and the degree of ocular layer tumor invasion and ON tumor invasion (P>0.05). CONCLUSION: RB intrinsically expresses GSTπ, P-gp and LRP. GSTπ expression is positive in 100% well-differentiation ones, so in which way it is correlated with differentiation. But the other two proteins expressions are not related to tumor differentiation and to the degree of tumor invasion. GSTπ may be a new target of chemotherapy in RB.

Genome-wide genetic variation and comparison of fruit-associated traits between kumquat (Citrus japonica) and Clementine mandarin (Citrus clementina).

KEY MESSAGE: The comprehensive genetic variation of two citrus species were analyzed at genome and transcriptome level. A total of 1090 differentially expressed genes were found during fruit development by RNA-sequencing. Fruit size (fruit equatorial diameter) and weight (fresh weight) are the two most important components determining yield and consumer acceptability for many horticultural crops. However, little is known about the genetic control of these traits. Here, we performed whole-genome resequencing to reveal the comprehensive genetic variation of the fruit development between kumquat (Citrus japonica) and Clementine mandarin (Citrus clementina). In total, 5,865,235 single-nucleotide polymorphisms (SNPs) and 414,447 insertions/deletions (InDels) were identified in the two citrus species. Based on integrative analysis of genome and transcriptome of fruit, 640,801 SNPs and 20,733 InDels were identified. The features, genomic distribution, functional effect, and other characteristics of these genetic variations were explored. RNA-sequencing identified 1090 differentially expressed genes (DEGs) during fruit development of kumquat and Clementine mandarin. Gene Ontology revealed that these genes were involved in various molecular functional and biological processes. In addition, the genetic variation of 939 DEGs and 74 multiple fruit development pathway genes from previous reports were also identified. A global survey identified 24,237 specific alternative splicing events in the two citrus species and showed that intron retention is the most prevalent pattern of alternative splicing. These genome variation data provide a foundation for further exploration of citrus diversity and gene-phenotype relationships and for future research on molecular breeding to improve kumquat, Clementine mandarin and related species.

The cubital tunnel syndrome caused by the intraneural or extraneural ganglion cysts: Case report and review of the literature.

Cubital tunnel syndrome is the second most common nerve compression syndrome in peripheral nerve compression disease. Although potential ulnar nerve entrapment can occur at multiple points along its course, such as the arcade of struthers, the medial intermuscular septum, the medial epicondyle, the cubital tunnel, and the deep flexor pronator aponeurosis, the most common site of entrapment is the cubital tunnel. However, cubital tunnel syndrome could also be caused by the occupying masses along the course of ulnar nerve, such as intraneural or extraneural ganglia. The cubital tunnel syndrome caused by intraneural or extraneural ganglion cysts has been rarely reported. In our hospital, there were 184 patients with cubital tunnel syndrome who underwent surgical treatment from January 2010 to January 2014. Of these patients, 16 had extraneural cysts and 3 had intraneural ganglion cysts. The incidence rate of cysts in the cubital tunnel was 10.33%. Electromyography was used as routine examination. Ultrasound was used only in some patients in whom elbow mass was suspected. In the surgery of the cubital tunnel syndrome combined with cyst, if any other cysts were found, we should be remove completely the cyts and decompress the ulnar nerve thoroughly with the ulnar nerve being anterior transposition. These cysts were confirmed by histopathological examination. Finally, we compared the clinical features of patients who had a medial elbow ganglion with those of patients who had only cubital tunnel syndrome. B ultrasound can significantly improve the diagnosis. All patients were followed up for 4 months to 2 years, and the curative effect was good.

Case report of ocular Kaposi's sarcoma.

BACKGROUND: Kaposi's sarcoma (KS) is generally considered a neoplastic disorder of vascular origin and occurs in patients with acquired immunodeficiency syndrome (AIDS) or who have received immunosuppressive treatments after an organ transplant (Soulier et al., Blood 86(4):1276-80, 1995; Viejo-Borbolla and Schulz, AIDS Rev 5(4):222-9, 2003; Schulz, J Antimicrob Chemother 45(Suppl T3):15-27, 2000; Aversa et al. Crit Rev Oncol Hematol 53(3):253-65, 2005; Mbulaiteye and Engels, Int J Cancer 119(11):2685-91, 2006; Tessari et al., Eur J Dermatol 16(5):553-7, 2006). Several Kaposi's sarcoma case reports involving eyelids and conjunctiva have been published (Bavishi et al., Int J STD AIDS 23(3):221-2, 2012; Baumann et al., Ger J Ophthalmol 4(4):239-45, 1995). CASE PRESENTATION: we report a 13 years old asian male patient rare case of ocular KS that was initiated from the sclera and progressed into the cornea and conjunctiva without an human Immunodeificiency Virus (HIV) or HHV-8 infection after a peripheral blood stem cells transplantation. In this case, anti- vascular endothelial growth factor (VEGF) therapy was attempted to stop the advance of ocular lesions and failed. Eventually, the KS was cured by a limbo-corneal lamellar graft, an amniotic membrane and scleral allograft transplantation plus intraoperative mitomycin C(MMC) after the complete excision of the tumors. CONCLUSION: A compete surgical excision combined with the intraoperative application of MMC, as well as grafts to repair the scleral, conjunctival, and corneal surfaces, could prevent a recurrence of KS.

The Role of miRNAs in Cartilage Homeostasis.

Osteoarthritis (OA) is an age-related disease with poorly understood pathogenesis. Recent studies have demonstrated that miRNA might play a key role in OA initiation and development. We reviewed recent publications and elucidated the connection between miRNA and OA cartilage anabolic and catabolic signals, including four signaling pathways: TGF-ß/Smads and BMPs signaling, associated with cartilage anabolism; and MAPK and NF-KB signaling, associated with cartilage catabolism. We also explored the relationships with MMP, ADAMTS and NOS (NitricOxide Synthases) families, as well as with the catabolic cytokines IL-1 and TNF-α. The potential role of miRNAs in biological processes such as cartilage degeneration, chondrocyte proliferation, and differentiation is discussed. Collective evidence indicates that miRNAs play a critical role in cartilage degeneration. These findings will aid in understanding the molecular network that governs articular cartilage homeostasis and in to elucidate the role of miRNA in the pathogenesis of OA.

Prognostic value of p53 alterations in human osteosarcoma: a meta analysis.

Tumor suppressor gene p53 functions as the guardian of the human genome and mutations in p53 contribute to cancer development. However, studies that investigated the potential of p53 as a prognostic marker in osteosarcoma patients have yielded inconclusive results. Based on recommendation of the Cochrane Collaboration, this meta-analysis was conducted using data from the 17 published studies to evaluate the association of p53 alterations with clinical outcome of osteosarcoma patients. Different databases, including MEDLINE, PsycINFO, Scopus, EMBASE, and the Cochrane Central Register of Controlled Trials (CENTRAL) were searched. Prognostic value of p53 alterations was determined by risk ratio (RR). The data showed that p53-positive immunostaining tended to associate with decreased 2-year survival rates (RR, 1.94; 95% CI, 1.43 to 2.64; p < 0.0001, I(2) = 10%). However, the prediction value of RR was smaller with p53 expression than with p53 mutations. Moreover, patients who received neoadjuvant chemotherapy and surgery tended to have a stronger association between p53-positive staining and 2-year mortality compared to the patients treated with surgery only. However, p53-positive staining was not associated with 3-year (RR, 1.64; 95% CI, 0.84 to 3.20; P = 0.15; I(2) = 56%) and 5-year survival (RR, 1.25; 95% CI, 0.78 to 2.01; P = 0.36; I(2) = 70%). The data from the current study suggest that p53-positive osteosarcoma only predicted a decreased short-term survival rate, but not 3- or 5-year survival.