[Relationship of symptoms with sleep-stage abnormalities in obstructive sleep apnea-hypopnea syndrome].

Objective:To study the relationship between symptoms and sleep staging in OSAHS patients.Method:A cross-sectional study. Adult subjects who attended a sleep laboratory for diagnostic polysomnography for a period of 1 month were recruited consecutively.OSAHS was diagnosed using American Academy of Sleep Medicine criteria.Subjects filled a questionnaire for symptoms prior to polysomnography.Result:Thirty subjects, of whom 83.3% were obese, met diagnostic criteria, with males constituting 46.7% and females constituting 53%.Mean age was (53.40±11.60) years.Sleep architecture comprised N1 (19.50±19.00)%, N2 (53.93±13.39)%,N3 (3.90±19.50)%, and rapid eye movement (8.92±6.21)%.Excessive fatigue or sleepiness, waking up tired, falling asleep during the day, trouble paying attention, snoring and insomnia were significantly related to decreased N3 sleep.Conclusion:Most of the symptoms in OSAHS in adults are related to decreased stage N3 sleep.If confirmed by larger controlled studies, correcting N3 sleep deficiency by pharmacotherapy may become an important adjunct to CPAP/BIPAP therapy to alleviate symptoms.

Identification of Pseudomonas syringae pv. actinidiae strains causing bacterial canker of kiwifruit in the Anhui Province of China, and determination of their streptomycin sensitivities.

Bacterial canker, caused by Pseudomonas syringae pv. actinidiae, is one of the most severe diseases of kiwifruit. It has become an international pandemic and threatens the sustainable development of kiwifruit production in all main kiwi-growing regions worldwide. Streptomycin has been the major bactericide for the control of kiwifruit canker, especially in Anhui Province, one of the main kiwifruit production regions in China. However, until now, no studies on the baseline sensitivity to streptomycin of field isolates of P. syringae pv. actinidiae from China have been available. During 2012-2013, a total of 102 single-colony P. syringae pv. actinidiae strains were isolated: 36, 12, 13, 26, and 15 strains from Yuexi, Jinzhai, Huoshan, Qianshan, and Taihu counties, respectively. All strains were confirmed by production of a 280-bp fragment using the specific primers PsaF1/R2 upon polymerase chain reaction amplification, followed by an assay for confirmation of pathogenicity to fulfill Koch's postulates. In this study, the streptomycin sensitivity of the 102 isolated strains was determined. The half-maximal effective concentration values for inhibition of growth by streptomycin were 0.03-0.42 µg/mL (average 0.12 ± 0.06 µg/mL). The baseline sensitivity curve was unimodal, representing range-of-variation factors of 14.0. No resistant subpopulation was identified among the strains used in the study. Thus, these sensitivity data could be used as a baseline for monitoring the shift in sensitivity of P. syringae pv. actinidiae populations to streptomycin in Anhui Province. Continuous resistance monitoring should be carried out, as streptomycin is an at-risk bactericide agent.

Detection of a mutation at codon 43 of the rpsL gene in Xanthomonas oryzae pv. oryzicola and X. oryzae pv. oryzae by PCR-RFLP.

The aim of this study was to develop a method to detect a point mutation in the ribosomal S12 protein (rpsL) gene in streptomycin-resistant strains of Xanthomonas oryzae pv. oryzicola and X. oryzae pv. oryzae. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was developed to detect a point mutation in codon 43 of the rpsL gene in X. oryzae pv. oryzicola and X. oryzae pv. oryzae. The 304-bp PCR product from the rpsL gene was digested by MboII to form two fragments (201 and 103 bp) if there was a mutation at codon 43, or three fragments (146, 103, and 55 bp) if there was no mutation. Compared with the results from nucleotide sequencing, the PCR-RFLP method was accurate in detecting the point mutation at codon 43 of the rpsL gene in streptomycin-resistant strains of X. oryzae pv. oryzicola and X. oryzae pv. oryzae. These results indicate that the PCR-RFLP is a simple, rapid and reliable method for detecting the point mutation at codon 43 of the rpsL gene.

Sodium hyaluronate-CNTF gelatinous particles promote axonal growth, neurogenesis and functional recovery after spinal cord injury.

OBJECTIVES: Currently, effective therapeutic strategy for spinal cord injury (SCI) is not clinically available. To establish a better method that may help repair the injured spinal cord, sodium hyaluronate-ciliary neurotrophic factor (CNTF) gelatinous particles were generated. METHODS: A segment of spinal cord tissue was excised to form a 2.5-mm-long cavity at thoracic level in an adult rat, and sodium hyaluronate-CNTF gelatinous particles were implanted into the lesion cavity. The recovery of the injured spinal cord was evaluated by immunohistochemistry, nerve tracing, electrophysiological test and Basso-Beattie-Bresnahan locomotor rating scale. RESULTS: Open-field locomotion of the sodium hyaluronate-CNTF rats was significantly enhanced up to 12 weeks postoperation. Together with the evidence of enhanced cortical motor evoked potentials and cortical somatosensory evoked potentials in the sodium hyaluronate-CNTF group, these findings suggested a powerful functional recovery component. Immunohistochemical analyses suggested that the functional recovery might be attributable partly to an increase in axonal regrowth as well as in replenishment of ß-tubulin-III-positive neuron-like cells. CONCLUSION: Sodium hyaluronate-CNTF gelatinous particles may provide an effective method for treating SCI.

First Report of a Fruit Rot and Twig Blight on Pomegranate (Punica granatum) Caused by Pilidiella granati in Anhui Province of China.

Pomegranate, Punica granatum Linn., is widely planted as an ornamental and a fruit crop in Huaiyuan County, Anhui Province, which is the primary pomegranate production area in China. In the early summer of 2012, twig dieback and fruit rot were observed on about 10% and 30% of the pomegranate trees, respectively, in several villages of Huaiyuan County. Necrosis was observed in the twigs, resulting in death of the branches. On fruit, dry rot started at the sepals, covered the entire surface in severely infected fruit, and eventually resulted in shriveling of the fruit. Abundant, black, and solitary pycnidia were observed on diseased twigs and fruit. Pieces of tissue (3 mm in size) from diseased twigs and sepals were surface-disinfected in 75% ethanol for 1 min, washed in sterile water three times, plated on potato dextrose agar (PDA) acidified with 2.5 ml 85% lactic acid per liter, and incubated at 25°C. Resulting fungal cultures produced pale green or white aerial mycelia and sporulated after 5 to 7 days. Pycnidia, ~80 to 130 µm in diameter, were globose and black with thin and membranous walls and contained hyaline, one-celled, and ellipsoid to fusiform conidia, averaging 10.8 to 17.2 × 2.9 to 4.7 µm in size. These morphological features were consistent with Pilidiella granati Sacc. (= Coniella granati Sacc.) (2). Genomic DNA from each of the 10 isolates was extracted and purified using a DNA Gel Extraction Kit (AxyPrep, Hangzhou, China), and PCR was conducted using a DNA Engine System PTC-200 (BIO-RAD, Watertown, MA) with ITS1 and ITS4 internal transcribed spacer universal primers. A single 616-bp fragment was amplified from all 10 isolates and sequenced. Sequence analysis revealed that the ITS from these isolates were identical (100% similarity, GeneBank Accession No. KF560320) to each other and showed >99.5% similarity with those of the P. granti isolates deposited in GenBank (AY339342.1). To evaluate pathogenicity, mycelial plugs (5 mm diameter) from 7-day-old PDA cultures were transferred onto the non-wounded sepals of pomegranate fruit (one plug per fruit, six fruits per isolate), and then all inoculated fruit were placed in plastic bags and maintained at 25°C for 14 days. In addition, twigs on pomegranate plant growing in the field were inoculated by placing mycelial plugs of the fungus on young bark and covered with cotton saturated with sterile water (one plug per twig, six twigs per isolate). Sterile PDA plugs were used as controls in both tests. All 10 isolates colonized the fruit after 5 to 8 days; this was followed by the appearance of dry rot and formation of abundant pycnidia after 10 to 12 days. No decay was observed on the control fruit. Isolates were also pathogenic on twigs, resulting in brown lesions after 2 months that were 2 to 5 cm long. No lesions were observed on the control twigs. Furthermore, the pathogen was isolated from all inoculated fruit and twig tissues and identified to be P. granati as described above, fulfilling Koch's postulates. This pathogen has been reported in Spain (3), Greece (4), and Iran (1), causing crown rot on pomegranate in addition to infecting fruit, but has not been reported previously in Anhui Province of China. This disease is an emerging problem in Anhui Province and will necessitate the development of new disease management practices to sustain commercial production in this region. References: (1) M. Mirabolfathy et al. Plant Dis. 96:461, 2012. (2) Niekerk et al. Mycol. Res. 108:283, 2004. (3) L. Palou et al. New Dis. Rep. 22:21, 2010. (4) T. Thomidis et al. Plant Dis. 95:79, 2011.

First Report of Pestalotiopsis theae on Loquat (Eriobotrya japonica) in Anhui Province of China.

Loquat, Eriobotrya japonica (Thunb.) Lindl., is an important fruit that is widely planted and used as an ornamental in Jingxian, Anhui Province, China. Loquat branches with severely spotted leaves and fruits were observed in this region in 2012. Symptoms on leaves consisted of small (0.5 to 1.2 cm in diameter), circular to oblong, greenish-brown lesions that coalesced to form isolated or confluent, dark brown spots. On fruit, the disease appeared as circular to elongated, sunken spots. Expanding lesions spread over the surface resulting in death of the fruit. Acervuli were observed within lesions. Isolations from symptomatic tissue onto potato dextrose agar (PDA) medium consistently yielded white fungal colonies of sparse aerial mycelium with acervuli containing black, slimy spore masses on the surface. The colony reached 8.0 cm diameter after 7-day culture on PDA at 24°C. Conidia produced in the culture were five-celled, narrow fusiform, straight or slightly curved, with a tapering base and 2 to 4 hyaline appentages (apical appentages measured 15 to 34 µm long and a single basal appentage was 5 to 9 µm long). Conidia were 24 to 32 × 5 to 8 µm with median cells 15 to 20 µm and two hyaline, cylindrical to conical apical cells typical of Pestalotiopsis spp. (3). A total of 12 isolates were obtained by isolation from the diseased fruit or leaves. Genomic DNA from the fungal isolates was purified using a DNA Gel Extraction Kit (AxyPrep, Hangzhou, China), and applied to a DNA Engine System PTC-200 (BIO-RAD, Watertown, MA) with ITS1 and ITS4 internal transcribed spacer (ITS) universal primers. The amplified sequences (533 bp) were analyzed together with other Pestalotiopsis sequences (1). ITS from all 12 of the fungal isolates were identical (99.5% similarity) to each other and to isolates of Pestalotiopsis theae, which infects tea trees in China (2). To demonstrate pathogenicity, suspensions (prepared in distilled water) of 106 conidia ml-1 of each isolate were sprayed on the loquat leaves in vivo and mature fruits in vitro. Distilled water was used as the control. More than 20 leaves and 10 mature fruits were sprayed for the treated and control plants, respectively, and the inoculation tests were repeated twice. The inoculated plants and fruit were kept in a humidity chamber for 7 days. Approximately 50% of the inoculated leaves and fruits developed blight symptoms similar to natural infections. P. theae was reisolated from diseased plants to complete Koch's postulates. Control plants sprayed with distilled water remained symptomless. There is a previous study reporting that P. guepini infected loquat in Argentina (4); however, to our knowledge, this is the first report of P. theae causing leaf and fruit spots on loquat in China. References: (1) R. Jeewon et al. Mol. Phylogenet. Evol. 25:378, 2002. (2) J. Y. Lu. Diagnosis of plant diseases. Page 194 in: Pestalotiopsis. J. Y. Lu, Z. G. Xu, Y. X. Chen, D. R. Shen, X. B. Zheng, and Y. Q. Cao, eds. China Agriculture Press, Beijing, 1995. (3) T. R. Nag Raj. Coelomycetous Anamorphs with Appendage-Bearing Conidia. Mycologue Publications, Waterloo, Canada, 1993. (4) A. E. Perelló and S. Larran. Plant Dis. 83:695, 1999.