First Survey of the Pathogenic Fungus Batrachochytrium dendrobatidis in Wild Populations of the Yunnan Caecilian (Ichthyophis bannanicus) in Guangxi, China.

Batrachochytrium dendrobatidis (Bd), which causes chytridiomycosis, mainly infects Anura and Caudata but is poorly known in Gymnophiona. We conducted a survey of Bd in the Yunnan caecilian (Ichthyophis bannanicus) and found that 6 of 71 samples (8.4%) tested positive for Bd. To our knowledge, this is the first detection of Bd in wild I. bannanicus.

Discovery of novel inhibitors of RNA silencing suppressor P19 based on virtual screening.

To control plant viruses, viral RNA silencing suppressors are important drug targets due to their key roles in interfering antiviral RNA silencing. Here we have presented a strategy, combining virtual and experimental screening, to discover the inhibitors of viral suppressor. By docking 157 026 compounds from a natural product library into P19 model, eighteen candidate compounds were selected. Candidates VS2, VS12, VS14 and VS15 displayed strong binding ability to P19 in the surface plasmon resonance imaging assay with K D values of 136.2, 111.6, 81.2 and 124.5 nM, respectively. Then the inhibition activities of these inhibitors on the association between P19 and siRNA were also affirmed by electrophoretic mobility shift assay. Moreover, the antiviral effects on plants showed that compounds VS14 and VS15 both exhibited antiviral activities against Tomato bushy stunt virus (TBSV) in vivo with inhibition rates of 32.35% and 16.61% in 11 dpi, respectively. This strategy would be a powerful tool for the discovery of novel antiviral agents and provide new insights into the control of plant viruses.

Rapid Detection of Prunus Necrotic Ringspot Virus by Reverse Transcription-cross-priming Amplification Coupled with Nucleic Acid Test Strip Cassette.

Prunus necrotic ringspot virus (PNRSV) is one of the most devastating viruses to Prunus spp. In this study, we developed a diagnostic system RT-CPA-NATSC, wherein reverse transcription-cross-priming amplification (RT-CPA) is coupled with nucleic acid test strip cassette (NATSC), a vertical flow (VF) visualization, for PNRSV detection. The RT-CPA-NATSC assay targets the encoding gene of the PNRSV coat protein with a limit of detection of 72 copies per reaction and no cross-reaction with the known Prunus pathogenic viruses and viroids, demonstrating high sensitivity and specificity. The reaction is performed on 60 °C and can be completed less than 90 min with the prepared template RNA. Field sample test confirmed the reliability of RT-CPA-NATSC, indicating the potential application of this simple and rapid detection method in routine test of PNRSV.

Efficacy of calcitriol in treating glucocorticoidinduced osteoporosis in patients with nephrotic syndrome: an open-label, randomized controlled study.

OBJECTIVE: To evaluate the efficacy and safety of calcitriol in the prevention and treatment of glucocorticoid-induced osteoporosis. METHODS: 66 patients treated with glucocorticoids (GC) for primary nephrotic syndrome (NS) were randomly assigned to 3 groups. Groups were designated as follows: calcitriol alone (n = 22), calcitriol plus calcium carbonate (n = 23), or calcium carbonate alone (n = 21). Serum markers of bone metabolism and bone mineral density (BMD) were tested at 3 different time points: the initiation of GC treatment (baseline), 12 weeks, and 24 weeks after the initiation of treatment. RESULTS: Levels of serum 25-hydroxy vitamin D, serum osteocalcin, and total serum collagen type N-terminal extension of the peptide were significantly decreased following GC therapy (p < 0.05). ß-collagen serum-specific sequences were significantly increased following GC therapy. The above-mentioned changes were less dramatic in patients treated with calcitriol, although the differences were significant (p < 0.05). Changes in serum levels of calcium, phosphorus, alkaline phosphatase, and parathyroid hormone (PTH) were not significant. 24 weeks after the initiation of treatment, BMD of the lumbar spine and femoral bone significantly decreased in all of 3 groups. However, patients who received calcitriol had significantly higher BMD of the lumbar spine than patients who received calcium carbonate alone (calcitriol plus calcium carbonate vs. calcium carbonate alone: 0.82 ± 0.19 g/cm2 vs. 0.62 ± 0.23 g/cm2 p < 0.05; calcitriol vs. calcium carbonate alone 0.805 ± 0.203 g/cm2 vs. 0.615 ± 0.225 g/cm2 p < 0.05), respectively. No serious adverse events were observed. CONCLUSION: Calcitriol may be more effective than calcium carbonate in preventing and treating GC-induced osteoporosis in patients with NS.

[Clinical efficacy and safety of sequential treatment with alprostadil and beraprost sodium for chronic renal failure induced by chronic glomerulonephritis].

OBJECTIVE: To evaluate the clinical efficacy and safety of sequential treatment with alprostadil and beraprost sodium for chronic renal failure caused by chronic glomerulonephritis. METHODS: Sixty-three patients with chronic renal failure due to chronic glomerulonephritis, after receiving a 2-week-long conventional treatment, were randomly divided into alprostadil group (n=20, with alprostadil injection at 10 µg/d for 2 weeks), sequential treatment group (n=21, with alprostadil injection at 10 µg/d for 2 weeks and oral beraprost sodium at 20 µg three times a day for 12 weeks), and strengthened sequential treatment group (n=22, with alprostadil injection at 20 µg/d for 2 weeks and a double dose of oral beraprost sodium for 12 weeks). Urinary albumin excretion rate (UAER), cystatin C (Cys C), blood urea nitrogen, creatinine, fibrinogen, D-dimer, prothrombin time (PT), and platelets were tested before and after the treatment, and the changes in urinary albumin discharge rate, serum creatinine, and glomerular filtration rate were determined. RESULTS: The patients in strengthened sequential treatment group showed a significantly decreased change rate of urinary albumin discharge rate (P<0.01) than those in the other two groups. In the two sequential treatment groups, especially the strengthened treatment group, the change rate of glomerular filtration rate increased significantly compared with that in alprostadil group (P<0.01). Strengthened sequential treatment resulted also in significantly decreased increment of serum creatinine compared that in the other 2 groups (P<0.01). After 14 weeks of treatment, fibrinogen and D-dimer were decreased in all the 3 groups (P<0.05) to a comparable level between the 3 groups (P>0.05), and prothrombin time (PT) or platelet showed no significant changes (P>0.05). CONCLUSION: Sequential treatment with alprostadil and beraprost sodium can improve the glomerular filtration rate and decrease urine albumin excretion rate, serum creatinine increase rate, and lower blood fibrinogen and D-dimer levels, thus delaying the progression of chronic renal failure caused by chronic glomerulonephritis. This therapy shows a dose-related effect with good clinical safety.

Transcriptome analysis of Nicotiana tabacum infected by Cucumber mosaic virus during systemic symptom development.

Virus infection of plants may induce a variety of disease symptoms. However, little is known about the molecular mechanism of systemic symptom development in infected plants. Here we performed the first next-generation sequencing study to identify gene expression changes associated with disease development in tobacco plants (Nicotiana tabacum cv. Xanthi nc) induced by infection with the M strain of Cucumber mosaic virus (M-CMV). Analysis of the tobacco transcriptome by RNA-Seq identified 95,916 unigenes, 34,408 of which were new transcripts by database searches. Deep sequencing was subsequently used to compare the digital gene expression (DGE) profiles of the healthy plants with the infected plants at six sequential disease development stages, including vein clearing, mosaic, severe chlorosis, partial and complete recovery, and secondary mosaic. Thousands of differentially expressed genes were identified, and KEGG pathway analysis of these genes suggested that many biological processes, such as photosynthesis, pigment metabolism and plant-pathogen interaction, were involved in systemic symptom development. Our systematic analysis provides comprehensive transcriptomic information regarding systemic symptom development in virus-infected plants. This information will help further our understanding of the detailed mechanisms of plant responses to viral infection.

[Expressions and clinical significance of urokinase-type plasminogen activator and its receptor in vulvar tissues].

OBJECTIVE: To explore the expressions of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in vulvar tissues and elucidate the patterns. METHODS: Thirty cases of vulvar neoplasms, 30 vulvar intraepithelial neoplasia (VIN) and 30 cases of normal epithelial tissue were harvested. And the expressions of uPA and uPAR in paraffin-embedded tissues were detected by immunohistochemistry. And the differential expressions of various vulvar tissues were observed. And the differences were compared in clinical stage and lymph node metastasis. RESULTS: (1) The expressions of uPA and uPAR in normal tissues (0.422 ± 0.022, 0.431 ± 0.027), vulvar intraepithelial neoplasia (VIN) (0.462 ± 0.018, 0.467 ± 0.015) and vulvar neoplasms (0.508 ± 0.020, 0.510 ± 0.020) were significantly different (P < 0.01). (2) The expressions of uPA and uPAR in vulvar tissue significantly increased with a rising stage-level (P < 0.05). (3) Compared with those without lymph node metastasis, the patients with lymph node metastasis had significantly higher expressions of uPA and uPAR in vulvar tissue (P < 0.05). CONCLUSION: Thus uPA and uPAR play an important role in the development process of vulvar neoplasms with an elevated expression in normal vulvar tissue, VIN and vulvar neoplasms. The invasiveness and prognosis of vulvar neoplasms may be evaluated through detecting the tissue expressions of uPA and uPAR.

[A preliminary approach of covering corrugator with rectangle periosteous flap in rhytidectomy].

OBJECTIVE: To evaluate the aesthetic outcome after application of rectangle periosteous flap to cover corrugator. METHODS: On the basis of regular rhytidectomy, a inter-eyebrow periosteous flap is applied to cover the fascia flap of corrugator by means of turnover. This manipulation helps to separate the skin from the muscle and prevent the re-adherence between the skin and muscle. RESULTS: The approach was applied on 15 cases. The follow-up ranged from 6 months to 1 year. The results were satisfactory. CONCLUSIONS: The method is effective in elimination of inter-eyebrow crease.

[A new method of pulmonary artery cannulation with minimally invasive decannulation for right ventricular assistance:an animal experiment].

OBJECTIVE: To create a new method of pulmonary artery cannulation during operation for right ventricular assistance, for which minimally invasive decannulation can be done after operation. METHODS: A specially-made disassemblable composite graft consisting of a woven dacron tube and a bovine pericardial tube (composite graft) was anastomosed to the pulmonary artery as a pathway in seven dogs by median sternotomy. The end of the woven dacron tube and a link-thread for connecting the two tubes in the composite graft were drawn outside the body through an intercostal tunnel. A predisposed drawing-tieing thread in the composite graft was drawn outside through another intercostal tunnel and then tightened. An inflow cannula was placed into the left femoral vein or jugular vein via an incision of skin so as to establish a right ventricular assist system. By means of the centrifugal pump, the right ventricular bypass was established. After stopping the pumping, the 2 sets of predisposed prolene thread in the composite graft were tied by catheter and knotting tool via the small skin incision, 8 knots for each set, to close the bovine pericardial tube, and then the drawing-tieing thread, link-thread, and the dacron tube in the composite graft were removed. Median sternotomy was performed again to observe the anastomotic stoma, the closure of the stump of bovine pericardial tube, and whether hemorrhage occurred. RESULTS: The mean maximum output of right ventricular assist system from left femoral or jugular vein to pulmonary artery was (64.4 +/- 12.3) ml.min(-1).kg(-1). The second sternotomy showed satisfactory knotting on the bovine pericardial tube except in one case, whose second set of drawing-tieing thread was tied in the seventh knot and failed to be drawn out, thus partially remained in the body. No bleeding was found in all cases. CONCLUSION: The new cannulating methods with minimally invasive decannulation after operation created in this study is simple, easy and effective.