Visceral adiposity index and cervical arterial atherosclerosis in northeast China: a population based cross-sectional survey.

BACKGROUND AND PURPOSE: The burden of cervical arterial atherosclerosis, which is an important risk factor of stroke, is huge in China. The visceral adiposity index (VAI) is a convenient index that is related to a series of metabolic abnormalities and a variety of diseases, including cerebrovascular diseases. This study aimed to explore the relationship between VAI and cervical arterial atherosclerosis. METHODS: This cross-sectional study used the multistage stratified random cluster sampling method to acquire a representative sample of residents aged ≥40 in the general population of northeast China. All participants completed a detailed questionnaire and provided blood samples. Cervical vascular ultrasound was performed by experienced ultrasound doctors. VAI was calculated by a well-established formula and analyzed by quartiles (Q1-Q4). RESULTS: A total of 3688 people aged ≥40 years who underwent cervical arterial ultrasound were finally included, amongst whom 3.2% had increased intima-media thickness (IMT), 48.6% had carotid plaque and 4.0% had cervical artery stenosis (CAS). After adjusting for several parameters, little statistical significance was found between VAI and increased IMT; compared with the VAI-Q1 group, the VAI-Q2, VAI-Q3 and VAI-Q4 groups were more likely to have CAS [odds ratio (OR) 2.01, 95% confidence interval (CI) 1.17-3.43; OR 1.82, 95% CI 1.05-3.15; OR 1.79, 95% CI 1.03-3.12] and the VAI-Q3 and VAI-Q4 groups were at greater risk of carotid plaque (OR 1.30, 95% CI 1.05-1.62; OR 1.34, 95% CI 1.07-1.67). CONCLUSIONS: The VAI was related to carotid plaque and CAS in the general population that is older than 40 years.

Investigating the Potential Mechanism of Pydiflumetofen Resistance in Sclerotinia sclerotiorum.

The necrotrophic pathogen Sclerotinia sclerotiorum is one of the most damaging and economically important plant pathogens. Pydiflumetofen, which was developed by Syngenta Crop Protection, has already been registered in China for the management of Sclerotinia stem rot, which was caused by S. sclerotiorum in oilseed rape. In an attempt to preempt and forestall the development of resistance to this useful fungicide, the current study was initiated to investigate the potential mechanism of resistance in laboratory mutants. Five pydiflumetofen-resistant S. sclerotiorum mutants were successfully generated by repeated exposure to the fungicide under laboratory conditions. Although the mutants had greatly reduced sensitivity to pydiflumetofen, they were also found to have significantly (P < 0.05) reduced fitness, exhibiting reduced mycelial growth and sclerotia formation on potato dextrose agar medium. However, three of the four mutants had significantly (P < 0.05) increased pathogenicity on detached soybean leaves compared with their respective parental isolates, indicating a moderate to high level of fungicide resistance risk according to the criteria of the Fungicide Resistance Action Committee. Sequence analysis of four succinate dehydrogenase (Sdh) target genes identified several nucleotide changes in the sequences of the pydiflumetofen-resistant mutants, most of which were synonymous and caused no changes to the predicted amino acid sequences. However, all of the pydiflumetofen-resistant mutants had two amino acid point mutations (A11V and V162A) in their predicted SsSdhB sequence. No similar changes were found in the SsSdhA, SsSdhC, and SsSdhD genes of any of the mutants tested. In addition, there was a positive cross-resistance between pydiflumetofen and boscalid, and no cross-resistance between pydiflumetofen and other commonly used fungicides, including tebuconazole, fludioxonil, cyprodinil, dimethachlone, prochloraz, pyraclostrobin, fluazinam, procymidone, and carbendazim. These results indicate that pydiflumetofen has great potential as an alternative fungicide for the control of S. sclerotiorum, especially where resistance to other fungicides has already emerged. Mixing or alternate application with fludioxonil, prochloraz, and fluazinam could be used to limit the risk of resistance to pydiflumetofen.

Exploring the Biological and Molecular Characteristics of Resistance to Fludioxonil in Sclerotinia sclerotiorum From Soybean in China.

Sclerotinia sclerotiorum is one of the most damaging and economically important necrotrophic plant pathogens, infecting more than 400 plant species globally. Although the phenylpyrrole fungicide fludioxonil has high activity against S. sclerotiorum, reports indicate that there is also substantial potential for the development of fungicide resistance. However, the current study investigating five fludioxonil-resistant laboratory mutants found a significant fitness cost associated with fludioxonil resistance resulting in significantly (P < 0.05) reduced mycelial growth and sclerotia formation on potato dextrose agar as well as significantly (P < 0.05) lower pathogenicity on detached tomato leaves, with one mutant, LK-1R, completely losing the capacity to cause infection. In addition, all of the fludioxonil-resistant mutants had significantly (P < 0.05) increased sensitivity to osmotic stress (0.5 M of potassium chloride and 1.0 M of glucose), which is consistent with the proposed fludioxonil target sites within the high osmolarity glycerol stress response mitogen-activated protein kinase (HOG1-MAPK) signaling transduction pathway. Sequence analysis of six genes from this two-component pathway, including SsHk, SsYpd, SsSk1, SsSk2, SsPbs, and SsHog, revealed several mutations that may be associated with fludioxonil resistance. For example, six separate point mutations were found in SsHk that led to changes in the predicted amino acid sequence, including A136G, F249V, G353A, E560K, M610K, and K727R. Similarly, SsPbs had three mutations (D34G, S46L, and L337E), SsSk1 and SsYpd had two (S53G and A795V for SsSk1, and E67G and Y141H for SsYpd), and SsHog and SsSk2 had one each (V220A and S763P, respectively). To our knowledge, these constitute the first reports of amino acid changes in proteins of the HOG1-MAPK pathway being associated with fludioxonil resistance in S. sclerotiorum. This study also showed a positive cross-resistance between fludioxonil and dimethachlone and procymidone, but none with tebuconazole or carbendazim, indicating that the inclusion of tebuconazole within an integrated pest management program could reduce the risk of fludioxonil resistance developing in field populations of S. sclerotiorum and ensure the sustainable production of soybeans in China into the future.

[Short-term effect of sodium zirconium cyclosilicate on potassium lowering in chronic kidney disease patients with hyperkalemia].

Objective: To investigate the efficacy of sodium zirconium cyclosilicate on emergency correction of hyperkalemia in chronic kidney disease patients. Methods: Patients with chronic kidney disease who were admitted to the Department of Nephrology of the First Affiliated Hospital of Zhengzhou University from May to June 2020 were selected. Those who had hyperkalemia and took sodium zirconium cyclosilicate powder were finally included. The patient's clinical data and laboratory results were collected. Results: A total of 24 results were included from 21 patients. The age of patients was (48.9±13.5) years old. Fourteen patients were male, and 7 patients were female. After 2 hours of administration, the venous potassium level decreased from (5.85±0.52) mmol/L to (5.15±0.43) mmol/L (P<0.001, n=21), with an average decline of (0.71±0.43) mmol/L. Meanwhile, the arterial potassium level decreased from (5.50±0.40) mmol/L to (4.88±0.33) mmol/L (P<0.001, n=10), with an average decline of (0.62±0.29) mmol/L. Based on the initial venous potassium level, the patients were further divided into three groups. The average potassium decrease in<5.5 mmol/L group (4 patients), 5.5-<6.0 mmol/L group (11 patients) and ≥6.0 mmol/L group (6 patients) was (0.46±0.26) mmol/L, (0.62±0.38) mmol/L and (1.04±0.45) mmol/L, respectively. There was statistically significant difference of potassium reduction among the three groups (P=0.045). Moreover, the extent of potassium reduction was positively correlated with baseline venous potassium level (r=0.603, P=0.004, n=21). The study did not reveal any treatment-related adverse event. Conclusion: Sodium zirconium cyclosilicate powder can rapidly and effectively reduce the serum potassium level in chronic kidney disease patients with hyperkalemia.

[Prediction of clinical outcomes of patients with surplus cleavage-stage embryos for blastocyst culture after D3 embryo transfer].

Objective: To find the best strategy of embryo transfer, so as to provide theoretical basis for improving the clinical outcomes of in vitro fertilization-Embryo transfer (IVF-ET), we investigate the blastocyst culture of surplus cleavage-stage embryos after D3 embryo transfer and the prediction of clinical outcomes with or without blastocyst formation. Methods: A retrospective study was conducted on 3 568 patients who underwent IVF-ET in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2016 to May 2018, whotransplanted two embryos in D3 with blastocyst culture of surplus cleavage-stage embryos, according to their age, they were divided into three groups: <35 years old group, 35-38 years old group, and>38 years old group.And according to the presence or absence of blastocyst formation, they were also divided into two subgroups: blastocyst formation group and non-blastocyst formation group. The embryo development and clinical outcomes in each group were compared. Results: (1) Comparisons of the embryo development in the three age groups with the first cycle. The total fertilization rate, cleavage rate and high quality embryo rate of the blastocyst formation group in the three groups were higher than those in the non-blastocyst formation group, P<0.05; In<35 years old group, the embryo utilization rate (75.0% vs 70.6%), pregnancy rate (74.9% vs 70.3%), planting rate (53.6% vs 48.6%), delivery rate (66.7% vs 61.1%) and live birth rate (66.5% vs 61.0%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. (2) Comparisons of embryo development in the three age groups with multiple cycles (≥2 cycles). In<35 years old group, the total fertilization rate (75.0% vs 70.6%),delivery rate (62.7% vs 43.8%) and live birth rate (62.7% vs 43.8%) of the blastocyst formation group were significantly higher than those in the non-blastocyst formation group, P<0.05; In>38 years old group, the pregnancy rate (56.3% vs 25.8%), implantation rate (34.4% vs 14.5%), delivery rate (43.8% vs 11.3%), live birth rate (43.8% vs 11.3%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. Conclusions: The results of blastocyst culture in different groups can predict the outcomes of embryo transfer in D3. For patients<35 years old with the first cycle, the clinical outcomes of the blastocyst formation group after D3 embryo transfer is better than that of the non-blastocyst formation group. For Patients with multiple cycles (≥2 cycles),the clinical outcomes of the embryo formation group is superior to that of the non-blastocyst formation group<35 years old or>38 years old.

[Downregulated PI3K-Akt-eNOS expression is related to increased atrial fibrillation inducibility in diabetic rats].

Objective: To explore the impact of PI3K-Akt-eNOS signaling on atrial fibrillation inducibility in diabetic rats. Method: Eight-week-old male diabetic rats were randomized assigned into GK group, IGF group and L-NAME group (n=8 each) which respectively received normal saline (NS), insulin like growth factor (IGF-1) or L-NAME+IGF-1 through tail vein daily for 4 weeks. Eight 8-week-old male homologous Wister-Kyoto(WKY) rats treated with intravenous NS served as control group (WKY group). Blood glucose was measured once per week. The left atrial diameter (LAD) was measured by echocardiography, the atrial electrical parameters, including the P-wave duration, the atrial effective refractory period (AERP) and its dispersion (AERP-d), the incidence and the duration of atrial fibrillation induced by atrial burst pacing, were evaluated by electro-physiological instrument at 4 weeks post various treatments. Rats were then sacrificed, left atrial (LA) cell morphology was determined on HE stained sections, LA interstitial collagen was determined on Masson stained sections. The protein expression of phosphatidylinositol 3-kinase (PI3K) and phosphate endothelial nitric oxide synthase (p-eNOS) were detected by Western blot. Results: (1) At the beginning of the study, the random blood glucose (GLU) level was significantly higher and LAD was large in GK, IGF and L-NAME groups than in WKY group;after 4 weeks, GLU level and LAD dimension of IGF group were lower than GK and L-NAME groups (P<0.01 or 0.05). (2) One rat in L-NAME group died during operation. Four weeks later, the incidence of atrial fibrillation in GK group, IGF group, L-NAME group and WKY group was 7/8, 2/8, 6/7 and 3/8. The median duration of atrial fibrillation in GK group, IGF group, L-NAME group and WKY group was 11.9(9.3, 13.1), 0(0, 1.8), 11.5(4.4, 12.0), and 0(0, 3.0) s. Compare with WKY group, the P-wave duration and PR interval were significantly longer, AERP-d, incidence, and duration of atrial fibrillation were significantly higher in GK group (P<0.01), these changed were significantly reversed in IGF group compared to GK and L-NAME groups (all P<0.01). Heart rate and AERP were similar among the 4 groups on (P>0.05). (3) Four weeks later, the CSA and CVF of LA were significantly larger in GK group than in WKY group (P<0.01), which were significantly reversed in IGF group (P<0.01 vs. GK group), and the beneficial effects of IGF disappeared by co-treatment with L-NAME (P<0.01 vs. IGF group). (4) Four weeks later, compare with WKY group, the protein expression of PI3K (P<0.01) and p-eNOS (P<0.05) of LA were significantly downregulated in GK group, which could be significantly upregulated by IGF (P<0.01 and 0.05 vs. GK group), these effects diminished by co-treatment with L-NAME (P<0.01 or 0.05 vs. IGF group). Conclusion: Increased atrial fibrillation susceptibility in diabetic rat is linked with structural and electrical remodeling in LA, possibly mediated through downregulated PI3K-Akt-eNOS signaling.

[Chinese multicenter randomized trial of customized chemotherapy based on BRCA1 (breast cancer susceptibility gene 1)-RAP80 (receptor-associated protein 80) mRNA expression in advanced non-small cell lung cancer (NSCLC) patients].

Objective: BRCA1 (breast cancer susceptibility gene 1) and RAP80 (receptor-associated protein 80) play key roles in predicting chemosensitivity of platinum and taxanes. A randomized trial was carried out to compare non-selected cisplatin-based chemotherapy with therapy customized according to BRCA1 and RAP80 expression. Methods: Advanced stage NSCLC patients whose tumor specimen was sufficient for molecular analysis were randomized (1∶3) to the control or experimental arm. Patients in the control arm received docetaxel/cisplatin; in the experimental arm, patients with low RAP80 expression received gemcitabine/cisplatin (Arm 1), those with intermediate/high RAP80 expression and low/intermediate BRCA1expression received docetaxel/cisplatin (Arm 2), and those with intermediate/high RAP80 expression and high BRCA1 expression received docetaxel alone (Arm 3). The primary end point was progression-free survival (PFS). Results: 226 patients were screened and 124 were randomized in this trial. ORR in the four subgroups was 22.6%, 48.4%, 30.3% and 19.2%, respectively (P=0.08); PFS was 4.74, 5.59, 3.78 and 2.73 months, respectively (P=0.55); and OS was 10.82, 14.44, 10.86 and 10.86 months, respectively (P=0.84). The common adverse effects included neutropenia, nausea, anemia and fatigue. Conclusions: No statistically significant difference of ORR, PFS or OS is observed in the experimental arms compared with the control arm. Patients with low RAP80 mRNA levels have a trend of better survival and higher response rate to gemcitabine/cisplatin chemotherapy.

[Evaluation and analysis of the tibial coronal alignment after total knee replacement with the extramedullary tibial cutting guided by the tibial tubercle and anterior tibial tendon in Chinese patients].

OBJECTIVE: To evaluate the efficiency of the tibial coronal alignment after total knee arthroplasty with the extramedullary cutting system guided by the tibial tubercle and anterior tibial tendon, and to discuss the potential risk factors for the postoperative malalignment. METHODS: A retrospective study of 212 primary unilateral TKA surgeries (in 188 patients) in our hospital between June to December in 2014 had been analysed. All the cases were performed by one surgical group. An extramedullary tibial cutting guide system had been used, with a landmark of one third inner-medial tibial tubercle as a proximal anatomy reference, and anterior tibial tendon as a distal marker. The mechanical axis of lower extremity in full-length X-ray was measured before surgery, and the tibial prosthetic coronal alignment was checked two weeks postoperation, evaluating the accuracy of this extramedullary cutting system guided by our method. RESULTS: Since good alignment was defined as an angle within 3 degrees of the perpendicular to the mechanical axis, the tibial component positions were acceptable in 191 knees (90.1%), five knees were in valgus postoperative, and sixteen knees were in varus. There were seventeen(8.7%) in 179 knees with a preoperative varus presented malalignment after surgery, and four in 12 preoperative valgus kneess howed malalignment also, no statistical difference was found by Chi-square test(χ(2)=2.778,P=0.096), which cannot define the relationship between the varus or valgus deformity preoperation and the malalignmentposition in tibial prosthesis after surgery. Twenty-two knees suffered a severe preoperative deformity as a varus or valgus angle larger than 20 degrees with absolute value of mechanical axis before surgery and tibial prosthetic coronal alignment were 21.95 °(20.00°,26.90°)and 1.85°(0.10°, 7.10°), showed a significant difference (Z=2.11,P=0.035) compared with the data [10.65°(0.50°,19.80°)in preoperative mechanical axis and 1.10°(0.00°,4.60°)in the tibial prosthetic coronal alignment]of 190 knees who presented a mild deformity before surgery(less than 20 degrees), the result indicated the severe preoperative deformity might be a potential mal-alignment risk factor within this cutting system in TKA surgery. CONCLUSION: The tibial coronal alignment after total knee arthroplasty could achieve good results with extramedullary cutting guide, by using one third inner-medial tibial tubercle and anterior tibial tendon as a proximal and distal anatomy marker. Postoperative varus might occur in this system, and tibial prosthetic malalignment appeared more often in the patients who suffered a severe deformity before surgery.

[Ideal current of injury and R-wave sensingvalues for identifying optimized placement of right ventricular active-fixation pacing leads].

OBJECTIVE: To define the ideal COI and R-wave sensing values for optimized placement of right ventricular active-fixation pacing leads. METHODS: Three hundred and six patients underwent right ventricular active-fixation lead implantation in our department from January 2013 to December 2014 were enrolled in this prospective pilot cohort study. According to the times needed for successful implantation, patients were divided into group SP (success poisoning after single procedure, n=229) and group MP (success poisoning after multiple procedures, n=77). Current of injury (COI) was characterized as the magnitude of ST-segment elevation. Pacing parameters were measured up to 10 minutes after lead fixation. RESULTS: (1) COI at 1 minute (COI1, (9.34±2.44)mV vs. (3.19±1.36)mV, P<0.001) and 10 minutes (COI10, (4.99±2.04)mV vs. (1.91±0.62)mV, P<0.001) after lead fixation was significantly higher in SP group compared to MP group. R-wave sensing measured at 10 minutes after lead fixation (R10 min) was also significantly higher in SP group than in MP group ((14.2±4.5)mV vs. (5.4±0.7)mV, P<0.001). Pacing threshold measured at 10 minutes after lead fixation was significantly lower ((0.65±0.18)V vs. (1.90±0.40)V, P<0.001) in SP group than in MP group. (2) The optimized placement of pacing lead was related with COI1(OR=0.247, 95%CI 0.098-0.623, P=0.003) and R10 min(OR=0.081, 95%CI 0.015-0.428, P=0.003). (3) The cut-off value of COI1 was 4.77 mV(sensibility 0.97, specificity 0.95), the cut-off value of R10 min was 7.25 mV (sensibility 0.96, specificity 1.00) for optimized lead fixation. After 3 months follow up, pacing threshold was (0.68±0.19)V, R-wave sensing was (12.1±3.6)mV, and pacing impedance was (543±107)Ω for right ventricular active-fixation pacing leads. CONCLUSIONS: COI1>4.77 mV and R10 min>7.25 mV are the ideal parameters for identifying optimized placement of right ventricular active-fixation pacing leads.

The effects of germacrone on lipopolysaccharide-induced acute lung injury in neonatal rats.

Germacrone is one of the main bioactive components in the traditional Chinese medicine Rhizoma curcuma and has been shown to possess an anti-inflammatory activity. Our present study aimed to investigate the protective effects of germacrone on lipopolysaccharide (LPS)-induced acute lung injury in neonatal rats. Results showed that germacrone treatment significantly decreased the expression of pro-inflammatory cytokines IL-6 and TNF-α. Meanwhile, the expression of anti-inflammatory mediators TGF-ß1 and IL-10 was obviously increased following germacrone administration. The LPS-induced pathological changes in neonatal rats were also attenuated by germacrone treatment. In vitro, MTT and EdU incorporation assay indicated that germacrone administration significantly increased the A549 cell viabilities in a dose-dependent manner. Besides, flow cytometry and TUNEL analysis showed that the cell apoptosis rate was significantly reduced in a concentration-dependent manner after germacrone injection. At the molecular level, we found that germacrone treatment promoted the expression of claudin-4 both in vivo and in vitro as shown by real time PCR and western blot. Collectively, our study demonstrated that germacrone protected neonatal rats against LPS-induced ALI partially by modulation of claudin-4.

First Report of Powdery Mildew Caused by Erysiphe betae on Swiss Chard in China.

Swiss chard (Beta vulgaris L. subsp. cicla) is a widely planted vegetable in China. From May to June 2013, an outbreak of powdery mildew on Swiss chard cultivar Fangzheng was observed in the commercial fields in Zhoukou city of Henan Province, located in central China. More than 80% of the plants exhibited symptoms of the disease. At the beginning of infection, circular, white, dust-like colonies of powdery mildew occurred mainly on adaxial surfaces of leaves. As the disease progressed, white mycelia covered the foliar parts of plant. No cleistothecia were found on or in collected samples. Upon microscopic evaluation, conidiophores were unbranched with the length of 63 to 126 and width of 7 to 10 µm (n = 50), produced conidia singly, and composed of a cylindrical foot cell followed by one to three short cells. Conidia were colorless, hyaline, ovoid, measured 29 to 40 × 12 to 18 µm (n = 100), lacked fibrosin bodies, and produced germ tubes on the ends of the conidia. The fungus was identified as Erysiphe betae according to the morphological features (1). To verify the identity, the internal transcribed spacer (ITS) region was amplified with the universal primers ITS1 and ITS4 (2) and sequenced. The ITS sequence obtained was assigned as Accession No. KF268348 in GenBank, which showed 100% homogeneity with two ITS sequences of E. betae isolates from UK (DQ164432 and DQ164436). Koch's postulates were conducted by inoculating 15 healthy 5-week-old plants (cv. Fangzheng) with detached infected leaves, which grew in a growth chamber under 22/16°C (day/night), 50% relative humidity, 120 µmol/m2/s light and a 16-h photoperiod. Fifteen non-inoculated plants grew in another growth chamber with the same conditions as control. Symptoms consistent with the infected field plants were observed on the inoculated plants, while no symptoms were found on the control plants. Microscopic observation revealed that the pathogen growing on the inoculated plants was consistent with the morphology of the original fungus. To our knowledge, this is the first report of E. betae infection on Swiss chard in China (3). References: (1) S. Francis. Mol. Plant Pathol. 3:119, 2002. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990. (3) R. Y. Zheng et al. Page 63 in: Flora Fungorum Sinicorum, Vol. 1, Erysiphales. Science Press, Beijing, 1987.

First Report of Powdery Mildew Caused by Erysiphe heraclei on Carrot in Central China.

Carrot (Daucus carota) is an important root vegetable crop in China, which accounted for 46% of global production in 2011. Carrot was grown in Henan Province on >20,000 ha/year, which ranks first in China for area of carrots harvested. In October 2012, a powdery mildew outbreak was observed in 16 investigated carrot production fields in Zhoukou, Henan Province, in central China. White colonies typical of powdery mildew were seen on leaves of affected plants. The colonies enlarged and finally coalesced. Small, scattered fruiting bodies found on the adaxial and abaxial leaf surfaces were determined microscopically to be chasmothecia. Examining the pathogen morphologically revealed that appressoria were lobed, conidiophores were straight and bore single conidia, and cylindrical foot cells were followed by one to three shorter cells in the conidiophores. Conidiophores were subhyaline and 54.1 to 66.1 × 6.1 to 8.1 µm. Conidia were barrel-cylindrical and 28.8 to 38.6 × 11.4 to 14.8 µm. Chasmothecia were subspherical, dark brown to black, formed hyphoid appendages, and 110 to 122 µm in diameter. Appendages typically had one to five branches, which were nearly dichotomous or irregular, flexuous or almost straight, and 30 to 165 µm long. Each chasmothecium contained multiple asci that were saccate, multiguttulate, short-stipitate or not, 62.5 to 63.8 × 43.2 to 45.9 µm, and each contained two to six ascospores. Ascospores were subhyaline, ovoid to ellipsoid, and 16.5 to 17.7 × 11.2 to 12.7 µm. Based on characteristics of the anamorphic and teleomorphic stages, the fungus was identified as Erysiphe heraclei (2,4). To verify the identity, the internal transcribed spacer (ITS) region of ribosomal DNA was amplified with universal primers ITS1 and ITS4, and sequenced. The ITS sequence was assigned GenBank Accession No. KC480605, and showed 100% similarity to ITS sequences of E. heraclei on carrot in GenBank (EU371725 and GU252368). Koch's postulates were completed by using detached infected leaves from 10-week-old carrot plants growing in a field to inoculate 10 healthy, 5-week-old plants of the carrot cultivar Dinghong, growing in a growth chamber under 22/16°C (day/night) cycle at 50% relative humidity with 120 µmol/m2/s light and a 14-h photoperiod. Ten non-inoculated plants served as replicates of a control treatment. Symptoms consistent with those in the field were observed on inoculated plants 20 days post-inoculation. No symptoms were observed on the control plants. Microscopic observation of the pathogen growing on the inoculated plants revealed that it was the same as the original fungus. Powdery mildew on carrot has been observed in many countries including Australia (1), Mexico (3), and the United States (2). To our knowledge, this is the first report of E. heraclei infection on carrot in central China, a major region of carrot production, although the disease has previously been observed in northwestern China (4). Further research should help to reduce losses in carrot crops caused by E. heraclei in central China. References: (1) J. H. Cunnington et al. Australas. Plant Dis. Notes 3:38, 2008. (2) D. A. Glawe et al. Plant Health Progress doi: 10.1094/PHP-2005-0114-01-HN, 2005. (3) G. Rodríguez-Alvarado et al. Plant Dis. 94:483, 2010. (4) R. Zheng and G. Chen. Pp. 97-99 in: Flora Fungorum Sinicorum Vol. 1. Erysiphales. R. Zheng et al., eds. Science Press, Beijing, 1987.

First Report of Powdery Mildew Caused by Blumeria graminis on Festuca arundinacea in China.

Tall fescue (Festuca arundinacea Schreb), a predominant cool-season perennial grass, is widely used as forage and turf grasses in China. In July 2013, powdery mildew was observed on 10 F. arundinacea lawns (about 0.5 ha in total) in Urumchi, Xinjiang Province, China, with 20 to 30% of the area being infected. Signs of the disease initially appeared as irregular white mycelial colonies on the adaxial surface of infected leaves. As the disease progressed, the colonies covered the whole adaxial surface and white patches appeared on the abaxial surface of infected leaves. Conidiophores were unbranched and cylindrical with swollen bases, measuring 13.3 to 15 × 16.7 to 20 µm, and borne vertically on hyphae. Each conidiophore produced 10 to 18 conidia in a chain. The conidia were oval, one-celled, and colorless, measuring 8.1 to 9.8 × 26 to 29.7 µm. Cleistothecia were black, spherical, and 164.3 to 207.3 µm in diameter, each of which contained 9 to 26 asci. Asci were oblong or ovate, measuring 32.1 to 40 × 85.7 to 96.4 µm. Asci were petiolate, containing eight ascospores. Ascospores were round to oval, colorless, one-celled, measuring 19.1 to 22.5 × 11.7 to 13.6 µm. Based on morphological characteristics of the anamorph and the teleomorph, the fungus was identified as Blumeria graminis (DC.) Speer. Additionally, the internal transcribed spacer (ITS) of 563 bp was amplified from DNA of conidia using ITS1 and ITS4 primers (4). The ITS sequence was deposited in GenBank (Accession No. KF545644). The ITS sequence showed 100% homogeneity with those of B. graminis on Poa pratensis in Swizerland (AB273540) and on P. bulbosa in Iran (AB273551) (1), which further confirmed the identification. Ten 3-week-old healthy plants were inoculated by spraying a spore suspension (1 × 105 conidia ml-1) made from conidia brushed from infected plants, and 10 plants sprayed with sterile distilled water were served as controls. All the plants were placed in the same growth chamber at 20°C, 80% humidity, and 16-h photoperiod. Twenty days after inoculation, typical signs and symptoms of powdery mildew were observed on all the inoculated plants, whereas no symptoms were observed on the controls. Microscopic and ITS analysis showed that the fungus on the inoculated plants is identical to that on diseased field plants. B. graminis on F. arundinacea has been observed in a few European countries (1), Israel (3), and the United States (2). To our knowledge, this is the first report of powdery mildew caused by B. graminis on F. arundinacea in China, which will increase the difficulty to prevent powdery mildew on grasses including cereals. References: (1) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustav Fischer Verlag, Jena-Stuttgart-New York, 1995. (2) F. M. Dugan and G. Newcombe. Pacific Northwest Fungi. 2:1-5, 2007. (3) S. O. Voytyuk et al. Biodiversity of the Powdery Mildew Fungi (Erysiphales, Ascomycota) of Israel Vol. 7. Biodiversity of Cyanoprocaryotes, Algae and Fungi of Israel. Koeltz Scientific Books, 2009. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

Loop-tail phenotype in heterozygous mice and neural tube defects in homozygous mice result from a nonsense mutation in the Vangl2 gene.

N-ethyl-N-nitrosourea (ENU) is a powerful point mutagen that can generate random mutations. It has been used to generate mouse mutations to produce phenotypic models of human disease. Neural tube defects (NTD) are common birth defects in which the brain and/or spinal cord can be exposed; however, the mechanisms of these defects are poorly understood. Craniorachischisis is one type of NTD that bears a close resemblance to the phenotype of the loop-tail (Lp) mouse. Here we describe a C57BL/6J Lp mouse generated by ENU-induced mutagenesis. The mutation was mapped to the Vangl2 gene on chromosome 1, near markers D1Mit113 and D1Mit149. Sequence analysis of Vangl2 heterozygotes (Vangl2(m1Yzcm)/+) revealed a C/T transition mutation that resulted in substitution of a glutamine codon for a stop (nonsense) codon at position 449. The Vangl2 protein is involved in epithelium planar cell polarity. The predicted truncated protein would lack the PDZ-domain binding motif involved in protein-protein interaction; therefore, Vangl2(m1Yzcm) may be a loss-of-function mutant. Morphological and histological examination of homozygous mouse embryos revealed a neural tube closure defect that leads to craniorachischisis. This Vangl2(m1Yzcm) mouse represents a valuable model for the study of NTDs in humans.

The effects of the Chinese medicine ZiBu PiYin recipe on the hippocampus in a rat model of diabetes-associated cognitive decline: a proteomic analysis.

AIMS/HYPOTHESIS: Increasing evidence suggests that diabetes is associated with an enhanced risk of cognitive decline. The precise mechanisms underlying diabetes-associated cognitive decline (DACD) remain unclear. Here we investigated the molecular changes associated with DACD using a comparative proteomics study of hippocampus in a rat model of type 2 diabetes. In addition, we tested the effects of the Chinese medicine ZiBu PiYin recipe (ZBPYR) on DACD. METHODS: The hippocampus was dissected from control, diabetic and diabetic rats treated with ZBPYR (DM/ZBPYR). Soluble proteins were separated using fluorescence-based difference gel electrophoresis. Protein spots were visualised with fluorescent dyes and spot density was compared between each pair of groups. Proteins of interest were identified using mass spectrometry. Proteins of specific interest were also tested by western blot and real-time PCR analysis. RESULTS: We found 13 spots that were altered between control and diabetes groups, and 12 spots that were changed between diabetes and DM/ZBPYR groups. The identities of nine proteins were determined by mass spectrometry. The identified proteins were largely involved in energy metabolism, cytoskeleton regulation and oxidative stress. The protein alterations observed in the diabetes group were ameliorated to varying degrees following ZBPYR treatment. CONCLUSIONS/INTERPRETATION: The protein changes identified in hippocampus from a rat model of type 2 diabetes suggest that specific cellular alterations contribute to DACD. The Chinese medicine ZBPYR was found to affect multiple targets and partially repaired the original cellular balance. This study may provide important insights into the molecular events underlying DACD and allow the identification of novel therapeutic targets.

Celastrol enhances AAV1-mediated gene expression in mice adipose tissues.

The transduction of adeno-associated virus (AAV) in adipose tissues was not well characterized and appeared to be insufficient as compared with other targeted tissues in gene therapy. We have found that celastrol, a chemical from a traditional Chinese herb known to inhibit the proteasome activity, was able to enhance the transgene expression mediated by AAV1 in 3T3-L1 preadipocytes both before and after induced differentiation. A synergism of celastrol and nonionic surfactant pluronic F68 cotreatment on AAV1 transduction was observed in the experiments with rat primary preadipocyte cultures and in adipose tissues in vivo. By fluorescent microscopy using Alexa Fluor 647-labeled AAV and quantitative PCR assays, we found that celastrol treatments increased the nuclear distribution of AAV genomic DNAs, but not the total amount of viral cellular uptake in preadipocytes, which was different from the effect of pluronic F68 treatment to significantly promote the AAV internalization. Our data suggested that bioactive monomeric compounds extracted from herbal medicines might be used to facilitate AAV-mediated gene transfer applications.

LncRNA HOTAIR inhibited osteogenic differentiation of BMSCs by regulating Wnt/ß-catenin pathway.

OBJECTIVE: This study aims to investigate whether HOX transcript antisense RNA (HOTAIR) can participate in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the Wnt/ß-catenin pathway, thereby participating in the pathogenesis of osteoporosis. PATIENTS AND METHODS: We detected the expression level of HOTAIR in 60 osteoporosis patients and 60 normal controls by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Meanwhile, BMSCs derived from human or rats were subjected to determination of HOTAIR level. Subsequently, the effects of HOTAIR on osteogenic differentiation were evaluated by the activity of Alkaline Phosphatase (ALP), Alizarin Red S (ARS) staining, ALP staining and osteogenic-specific gene expression. The expression level of proteins related to the Wnt/ß-catenin was determined by Western blot, and ALP activity was detected by ALP activity determination kit and alizarin red staining after knockdown or overexpression of HOTAIR, as well as the treatment of DKK1 or the Wnt pathway antagonist. Finally, osteoporosis model in rats was established by ovariectomy (OVX). We examined protein levels of HOTAIR, ß-catenin, CyclinD, C-myc, and Runx2 in rat bone tissues. Bone morphology was observed in each group as well. RESULTS: The serum and BMSCs levels of HOTAIR in patients with osteoporosis were remarkably higher than that in normal people. Inhibition of HOTAIR induced increased ALP activity increased osteogenic marker genes and enhanced number of calcified nodules in BMSCs. However, the overexpression of HOTAIR exhibited the opposite effects. HOTAIR inhibited the expression level of Wnt/ß-catenin pathway-related protein. Also, Wnt pathway antagonist DKK1 partially reversed the regulatory effects of HOTAIR on Wnt/ß-catenin. DKK1 treatment markedly reduced the promotive effect of HOTAIR knockdown on ALP activity, ALP content and calcification ability of BMSCs. DKK1 administration in rats undergoing OVX showed worse bone morphology relative to controls. Protein levels of HOTAIR, ß-catenin, CyclinD, C-myc and Runx2 remarkably downregulated in OVX rats administrated with DKK1. CONCLUSIONS: HOTAIR inhibits osteoblast differentiation of rat BMSCs. The underlying mechanism of which may be related to the mediation of Wnt/ß-catenin pathway.