Viscoelastic properties of decellularized and freeze-dried human dermis between i909c and 40cC.

BACKGROUND: Human donor skin is processed to make the acellular dermis matrix (ADM) for tissue repair and regeneration. There is no data on the viscoelastic properties of ADM at room and subzero temperatures. OBJECTIVE: The work evaluated the temperature dependence of viscoelastic properties of freeze-dried ADM. MATERIALS AND METHODS: Donor skin was de-epidermized, de-cellularized and freeze-dried with trehalose as the lyo-protectant. Glass transition of freeze-dried ADM was measured by differential scanning calorimeter (DSC), and viscoelastic properties were examined by dynamic mechanical analyzer (DMA). RESULTS: At the low moisture range (1.4 +/- 0.5%), the glass transition temperature (Tg) of freeze-dried ADM was 90 degree C to 100 degree C. As the moisture content increased, the Tg decreased steadily. At the high moisture range (10.8 +/- 2.9%), the Tg was 40 degree C to 60 degree C. There were large donor-to-donor variations in viscoelastic properties of freeze-dried ADM as demonstrated by the changes in storage modulus (G'), loss modulus (G") and damping factor tan delta (G"/G'). However, the trends of the temperature dependence for G', G" and tan delta were similar among all 8 donors. For each donor, changes in G' and G" were relatively small between -90 degree C and 40 degree C, and G' was at least one order of magnitude greater than G". Two viscoelastic relaxations were observed in freeze-dried ADM, one at -20 degree C and the other at -60 degree C respectively. CONCLUSION: Freeze-dried ADM was protected in the glassy carbohydrate matrix. DMA observed two viscoelastic relaxations (i.e., alpha process at -20 degree C and beta process at -60 degree C). Overall changes in G' and G'' of freeze-dried ADM were relatively small within one order of magnitude between -90 degree C and 40 degree C. https://doi.org/10.54680/fr24110110612.

[Status of occupational exposure limits of air toxic substances in the GESTIS Substances Database].

Objective: In order to understand the current situation of air toxic substances without occupational exposure limits (OELs) in the workplace in the Germany GESTIS Substance Database, and to provide an effective reference for formulating OELs of corresponding toxic substances and improving health standards. Methods: From March 2022 to May 2023, based on the standard of GBZ 2.1-2019 Occupational Exposure Limits for Hazardous Agents in the Workplace-Part 1: Chemical Hazardous Agents, air toxic substances without OELs in the standard of GBZ/T 300.1-2017 Determination of Toxic Substances in Workplace Air-Part 1: General Principles were screened out, then corresponding OELs in other countrie/regions were queried through the Germany GESTIS Substance Database. Results: Among the 333 kinds (classes) of air toxic substances in 160 parts of GBZ/T 300.1-2017 standard, 48 kinds (classes) of air toxic substances were screened out and had not yet been formulated OELs in GBZ 2.1-2019 standard. By querying the Germany GESTIS Substance Database, it was found that among the 48 kinds (classes) of air toxic substances, 35 kinds (classes) of air toxic substances had both 8-hour occupational exposure limit and short-term occupational exposure limit, 4 kinds (classes) of air toxic substances had 8-hour occupational exposure limit but no short-term occupational exposure limit, 9 kinds (classes) of air toxic substances hadn't been retrieved any OELs. In addition, standard test methods of 7 kinds of air toxic substances hadn't been published in the present, including trimethylchlorosilane, trimethylbenzenes, cumene, chloroethane, chloropropane, dibromoethane and acetophenone. Conclusion: In the process of formulating or revising the standards of GBZ 2.1-2019 and GBZ/T 300, the latest published OELs in the Germany GESTIS Substance Database could be used as a reference basis.

[Efficacy comparison of zero-profile intervertebral fusion and stand-alone interbody cage combined with cage-titanium plate construct in treatment of two-segment skip cervical spondylosis].

Objective: To investigate the difference in clinical efficacy between zero-profile interbody fusion (ROI-C) and stand-alone interbody cage combined with cage-titanium plate construct for patient with two-segment skipped cervical spondylosis who received the anterior cervical discectomy and fusion (ACDF) surgical strategies. Methods: The clinical data of 62 patients with two-segment skipped cervical spondylosis who underwent surgical treatment in the First Affiliated Hospital of Zhengzhou University from June 2017 to June 2020 were retrospectively analyzed, included 38 males and 24 females, aged (53.3±8.5) years. Thirty-three cases were treated with ROI-C (ROI-C group), and 29 cases with stand-alone interbody cage combined with cage-titanium plate construct (cage+titanium plate group). The following parameters, including operation time, intraoperative blood loss, dysphagia Bazaz grade, Japanese Orthopaedic Association (JOA) score, visual analogue scale (VAS) of pain, neck disability index (NDI), average intervertebral height of operated segments, C2-7 Cobb angle, and related complications, were compared between the two groups. Results: The patients were followed up for (28±5) months (16-34 months). The operation time of ROI-C group was (127.6±34.2) min, which was shorter than that in cage+titanium plate group [(157.1±43.9) min, P=0.004]. The scores of JOA and VAS in both ROI-C group and cage+titanium plate group were significantly improved 3 months after operation when compared with those before operation, and there was no significant difference between the two groups (all P>0.05). The average intervertebral space height of fusion segment in ROI-C group was (6.02±1.03) mm before operation, it was improved to (8.38±1.47) mm at 3 months after operation, (8.16±1.40) mm at 12 months after operation, and it was (6.24±1.05) mm, (8.58±1.18) mm and (7.87±0.73) mm in cage+titanium plate group, respectively, and there was no significant difference between the two groups at each time point (all P>0.05). The Cobb angle of cervical vertebrae in ROI-C group was 10.5°±6.8° before operation, improved to 19.2°±9.0° at 3 months after operation, 18.2°±5.8° at 12 months after operation, and it was 10.9°±4.6°, 18.5°±7.8°, 17.1°±5.2° in cage+titanium plate group, respectively, and there was no significant difference between the two groups at each time point (all P>0.05). The incidence of postoperative dysphagia was 9.1%(3/33) in the ROI-C group and 37.9%(11/29) in the cage+titanium plate group, and the difference was statistically significant (P=0.007). Conclusions: Both ROI-C and stand-alone interbody cage combined with cage-titanium plate construct can achieve good results for two-segment skipped cervical spondylosis. However, ROI-C is more advantageous in shortening the operation time and reducing early postoperative dysphagia.

[Characteristics and regulatory mechanisms of lipid metabolism remodeling after malignant transformation of glioma-associated macrophages].

Objective: To observe the lipid metabolism characteristics of tumor-associated macrophages (TAM) after malignant transformation in the glioma micro-environment, and analyze the biological phenotype changes and regulatory mechanisms after inhibiting the lipid metabolism remodeling. Methods: Twelve male Balb/c mice of 6-8 weeks were used in the study. Macrophages (Mφ) were derived from mouse bone marrow, and malignantly transformed macrophages (tMφ1 and tMφ2) were cloned from the model of glioma stem cell (GSC) through interaction with Mφ in vivo and in vitro. Intracellular lipid droplet formation and cellular cholesterol content were measured respectively in Mφ, tMφ1 and tMφ2. qRT-PCR was performed to detect the genes expression level related with lipid metabolism, including sterol regulatory element binding protein (SREBP), fatty acid synthase (FASN), and 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase (HMG-CoA). Simvastatin (SIM) was used to analyze the proliferation, immigration and invasiveness ability in tMφ1 and tMφ2 after inhibition of the lipid metabolism. Differential expression profiles of miRNAs after SIM treatment were constructed in t-Mφ1 and bio-informatics analysis was screened and verified for miR449a and its target gene sorting micro-tubule connectin 17 (SNX17) associated with lipid metabolism remodeling. The effect on SNX17 by up-regulated miR-449a were analyzed by qRT-PCR and Western blot, meanwhile, the biological phenotype and cholesterol content were observed after up-regulation of miR449a. Low-density lipoprotein receptor (LDLR) protein levels after SNX17 knockdown and intracellular cholesterol content after LDLR knockdown were detected respectively. Results: The numbers of intracellular lipid droplet formation in tMφ1 and tMφ2 were more than that in Mφ (P<0.001). Likewise, the relative contents of cholesterol (3.89±0.68 and 3.56±0.53), SREBP (4.78±0.60 and 2.84±0.41), FASN (4.65±0.70 and 3.01±0.45), and HMG-CoA (5.74±0.55 and 2.97±0.34) were significantly higher in tMφ1 and tMφ2 than those of Mφ (1.01 wel, 1.02 wel and 0.99 wel, respectively) (all P<0.001). The proliferation rates of tMφ1 and tMφ2 decreased from (47.06±5.88) % and (45.29±5.64)% to (23.53±4.70)% and (18.74±5.76)%, respectively after treatment with SIM (both P<0.05). The numbers of migrated cells decreased from 1 025±138 and 350±47 to 205±63 and 99±25, respectively (both P<0.001). And the numbers of invasiveness cells decreased from 919±45 and 527±34 to 220±23 and 114±21, respectively (both P<0.001). While the relative intracellular cholesterol content decreased to 0.52±0.08 and 0.58±0.07 (both P<0.05), respectively. MiR-449a was screened from tMφ1 by SIM, and the target gene was analyzed and verified to be SNX17. SNX17 expression was down-regulated, and the proliferation rate, the number of migration and invasiveness was significantly decreased after miR-449a over-expression (all P<0.05). Low-density lipoprotein receptor (LDLR) expression was down-regulated after knock-down of SNX17, while the cholesterol content was decreased after knock-down of LDLR in tMφ1 and tMφ2 (all P<0.05). Conclusions: Malignantly transformed TAMs undergo lipid metabolism remodeling characterized with enhanced lipid metabolism. MiR-449a regulates the LDLR by targeting SNX17, thereby affecting the lipid metabolism of malignantly transformed macrophages, and subsequently inhibiting its proliferation, migration, and invasion ability. Precise intervention with miR-449a/SNX17/LDLR axis could provide an experimental basis for reversing its tumor-promoting micro-environment remodeled by GSC through metabolic intervention.

Incidence and mortality of nasopharyngeal carcinoma: interim analysis of a cluster randomized controlled screening trial (PRO-NPC-001) in southern China.

BACKGROUND: Previous mass screening studies have shown that IgA antibodies against Epstein-Barr Virus (EBV) can facilitate early detection of nasopharyngeal carcinoma (NPC), but the impact of EBV-antibody screening for NPC-specific mortality remains unknown. PATIENTS AND METHODS: A prospective, cluster randomized, controlled trial for NPC screening (PRO-NPC-001) was conducted in 3 selected towns of Zhongshan City and 13 selected towns of Sihui City in southern China beginning in 2008. Serum samples of the screening group were tested for two previously selected anti-EBV antibodies. Subjects with serological medium risk were subsequently retested annually for 3 years, and those with serological high risk were referred to otorhinolaryngologists for diagnostic check-up. An interim analysis was carried out to evaluate the primary end points of the NPC-specific mortality and the early diagnostic rate, and the secondary end point of the NPC incidence, through linkage with the database of Zhongshan City. RESULTS: Among 70 296 total subjects, 29 413 screened participants (41.8% of the total subjects) in the screening group and 50 636 in the control group, 153 (43.3 per 100 000 person-year), 62 (55.3 per 100 000 person-year) and 99 (33.1 per 100 000 person-year) NPC cases were identified. The early diagnostic rates of NPC were significantly higher in the participants (79.0%, P < 0.0001) and the screening group (45.9%, P < 0.0001) compared with the control group (20.6%). Although no differences were found between NPC-specific mortality of the screening group and the control group [relative risk (RR)= 0.82, 95% confidence interval (CI) 0.37-1.79], lower NPC-specific mortality was noticed among participants from the screening group versus the control group (RR = 0.22, 95% CI 0.09-0.49). CONCLUSION: IgA antibodies against EBV can identify high-risk population and was effective in screening for early asymptomatic NPC. Although the mortality reduction was not significant in the primary end point, we noted encouraging evidence of a mortality reduction in screening participants in this interim analysis. CLINICAL TRIAL NUMBER: NCT00941538.

Construction of a recombinant baculovirus expressing swine hepatitis E Virus ORF2 and preliminary research on its immune effect.

In the swine hepatitis E virus (HEV), open reading frame 2 (ORF2) is rich in antigenic determinants and neutralizing epitopes that could induce immune protection. We chose the Bac-to-Bac® Baculovirus Expression System to express fragments containing the critical neutralizing antigenic sites within the HEV ORF2 protein of pigs to obtain a recombinant baculovirus. The fragment of swine HEV ORF2 region (1198-1881bp) was cloned into vector pFastBacTM. A recombinant baculovirus, rBacmid-ORF2, was obtained after transposition and transfection. The molecular mass of the recombinant protein was 26 kDa. Mice were immunized by the intraperitoneal and oral routes with cell lysates of recombinant baculovirus rBacmid-ORF2. Serum and feces of the mice were collected separately at 0, 14, 28, and 42 d after immunization and the antibody levels of IgG and secretory IgA against swine HEV were determined using an enzyme-linked immunosorbent assay. The results suggested that rBacmid-ORF2 induced antibodies of the humoral and mucosal immune responses in mice and that the oral route was significantly superior to the intraperitoneal route. This is the first study to demonstrate that that recombinant baculovirus swine HEV ORF2 could induce humoral and mucosal immune responses in mice.

[Analysis on characteristics of hearing loss in occupational noise-exposed workers in automotive manufacturing industry].

Objective: To investigate the characteristics of hearing loss in occupational noise-exposed workers in automotive manufacturing industry. Methods: According to Technical Specifications for Occupational Health Surveillance (GBZ188-2014) , 530 noise-exposed workers were examined for health and pure tone audiometry during January to June, 2017, and the data were collected for analysis. Poisson regression analysis was used to analysis the relationship between work age and hearing loss. Results: After correction of age and gender, pure-tone threshold of 500, 1 000, 2 000, 3 000, 4 000, 6 000 of the noise-exposed workers in the <3 year-exposure group were 20.15±3.60, 18.80±3.43, 18.54±3.38, 19.10±4.03, 20.32±6.79, 23.48±8.92 dB (A) ; the values in the 3-10 year-exposure group were 20.96±5.38, 19.25±4.76, 19.22±5.29, 20.17±7.50, 21.36±8.88, 25.39±11.60 dB (A) ; the values in the >10 year-exposure group were 20.18±3.67, 18.34±3.60, 18.36±4.25, 18.49±4.41, 20.32±9.24, 22.50±9.57 dB (A) . Poisson regression analysis showed that the relative risk (RR) of hearing loss of the noise-exposed workers compared with the noise-free workers was 4.38 (P<0.05) . Conclusion: The noise-exposed workers in automotive manufacturing industry revealed hearing loss, which was associated with their actual working age of noise exposure.

[Role of ORAI1 in the balloon injury-induced rat carotid artery neointimal formation and the relationship between ORAI1 and sodium and calcium exchange 1].

Objective: To investigate the role of ORAI1 in the balloon injury-induced rat carotid artery neointimal formation and the relationship between ORAI1 and sodium and calcium exchanger(NCX)1. Methods: According to condition of carotid artery balloon injury and observation time, Sprague Dawley rats were divided into 3 groups including sham group, 7 days injury group, and 14 day injury group(n=3 each). According to virus of transfection, Sprague Dawley rats were divided into 2 groups including negative control group (negative lentiviruses particles transfection after establishment of carotid artery balloon injury model, n=3) and siORAI1 group(siORAI1 transfection after operation, n=3). Carotid artery neointimal formation was analyzed after HE staining. The ORAI mRNA expression level was detected using real-time PCR. The proliferating cell nuclear antigen (PCNA), ORAI and NCX1 protein expression level were measured by Western blot. Results: (1) Carotid artery intima was significantly thicker in 7 days and 14 days injury group than in sham group((0.54±0.11)µm2 and (0.89±0.12)µm2 vs.(0.11±0.08)µm2, both P<0.05). Relative expression level of PCNA protein was significantly higher in 7 days and 14 days injury group than in sham group(1.43±0.16 and 1.95±0.16 vs. 1, both P<0.05). Relative mRNA expression level of ORAI1 (1.39±0.14 and 1.78±0.21 vs. 0.56±0.09, both P<0.05) and protein expression level (1.42±0.19 and 1.78±0.22 vs. 1, both P<0.05) were significantly higher in 7 days and 14 days injury group than in sham group.(2) After 14 days, relative expression level of ORAI1 protein was significantly lower in siORAI1 group than in negative control group (0.21±0.161, P<0.05). Carotid artery intima thickness was significantly reduced in siORAI1 group compared to negative control group((0.19±0.14)µm2 vs.(0.91±0.23)µm2,P<0.05). Relative expression level of NCX1 protein was also significantly lower in siORAI1 group than in negative control group(0.53±0.131, P<0.05). Conclusions: ORAI1 may play a key role in the balloon injury-induced neointimal formation and vascular smooth muscle cells proliferation in rat carotid artery.ORAI1 knockdown could down regulate the NCX1 expression and attenuate the balloon injury-induced neointimal formation and vascular smooth muscle cells proliferation in rat carotid artery.

[The value of urinary neutrophil gelatinase-associated lipocalin for early diagnosis of contrast-induced nephropathy].

Objective: To explore the value of urinary neutrophil gelatinase-associated lipocalin (NGAL) for early diagnosis of contrast-induced nephropathy (CIN) in patients who underwent percutaneous coronary angiography (CAG) or percutaneous coronary intervention (PCI). Methods: From May 2015 to January 2016, 506 consecutive patients who underwent CAG or PCI in our hospital were enrolled in this prospective study. Patients were divided into CIN group (n=47) and non-CIN group (n=459). Clinical and interventional data were compared between the two groups. Spearman ranking correlation coefficient was used to define the relation between NGAL and CIN, and multivariable logistic regression analysis was performed to identif independent predictors of CIN. Receiver-operator characteristic (ROC) curve was generated, and area under the curve (AUC) was calculated and sensitivity and specificity for CIN diagnosis were analyzed. Results: (1) Basic clinical and interventional data including age, incidence of diabetes, hypertension and chronic heart failure, level of systolic blood pressure, serum creatine before procedure, use of isotonic contrast agent, contrast volume, Mehran score, operation time, treatment number of coronary artery, hydration and medication were all similar between two groups (all P>0.05). (2) Urinary NAGL level at 24 and 48 h after procedure, serum creatinine (Scr) level at 48 h after procedure, and ΔNGAL24-0 h were significantly higher in CIN group than in non-CIN group (all P<0.01), but the differences of urinary NGAL level before procedure, Scr level 24 h after procedure, and ΔNGAL48-24 h were similar between two groups (all P>0.05). (3)The positive correlation was found by Spearman ranking correlation between ΔNGAL24-0 h and CIN (r=0.478, P<0.001). (4) Multivariable logistic regression analysis showed that estimated glomerular filtration rate(OR=1.020, 95%CI 1.005-1.035, P=0.007) and ΔNGAL24-0 h (OR=1.020, 95%CI 1.014-1.027, P<0.001) were the independent predictors of CIN.(5) ROC curve showed that the AUC of NGAL with the cutoff value 4.65 was 0.899(0.854, 0.944)for diagnosis of CIN (sensitivity 93.6%, and specificity 0.944). Conclusion: The rise of urinary NGAL level at 24 and 48 hours after CAG or PCI is suggestive of CIN and could be used as a reliable parameter for the early diagnose of CIN.

Allogeneic hematopoietic stem cell transplantation in children with aplastic anemia.

The aim of this study was to prospectively investigate the efficacy and safety of fully matched allogeneic hematopoietic stem cell transplants in children with severe aplastic anemia in China. A total of twenty patients with severe aplastic anemia were enrolled in our study. Thirteen cases underwent transplantation with fully human leukocyte antigen (HLA)-matched, granulocyte-colony stimulating factor (G-CSF)-primed bone marrow and peripheral blood stem cells (PBSCs) from matching sibling donors. One patient received fully HLA-matched bone marrow from an unrelated donor. Six patients received fully HLA-matched G-CSF-primed PBSCs from unrelated donors. The conditioning regimen included fludarabine, cyclophosphamide, and rabbit anti-thymocyte globulin. Graft-versus-host disease prophylaxis was conducted with cyclosporin A and short-course methotrexate. The median follow-up duration was 3.08 years (range, 0.83-8.41years). The median time of neutrophil recovery (>0.5 x 10(9)/L) was 14 days (range, 10-20 days), and the median time of platelet recovery (>20 x 10(9)/L) was 19 days (range, 14-31 days). The survival rate at the cutoff point of follow-up was 95.0% (19/20). Initial engraftment rate was 95% (19/20). Late graft failure (graft failures occurring 1 year or longer after transplantation) was observed in one patient. Only one patient developed Grade I acute graft-versus-host disease. Two cases suffered from Epstein- Barr virus (EBV)-associated post-transplant lymphoproliferative disorder and remitted after treatment with rituximab. One patient was diagnosed with hyperthyroidism 2.5 years after transplantation. Our study indicated that allogeneic hematopoietic stem cell transplantation is an effective and safe treatment for children with severe aplastic anemia in China.

First Report of Corn Kernel Brown Spot Disease Caused by Mucor irregularis in China.

In October 2012, a brown spot disease was found on corn kernels during a field survey in Nanyang city (33°01' N, 112°29' E), China. The incidences of affected ears and kernels were 2 to 10% (n = 600) and 0.08 to 0.4% (n = 25,000), respectively. Symptoms first appeared as circular or irregular brown spots on the endosperm. These spots subsequently enlarged or coalesced, resulting in the formation of a large light-brown or light-yellow irregular speckle commonly surrounded by a dark-brown edge. Pure fungal cultures with similar morphological characteristics were obtained from surface-disinfected symptomatic kernels using a conventional method for isolation of culturable microbes. The isolated fungal cultures were purified by single-spore isolation (3). A representative isolate F1 was randomly selected, used for pathogenicity tests, and identified using morphological and molecular methods. Colonies on PDA were circular with abundant villiform aerial mycelia. The color of colonies was white-gray at first and turned to light yellow or became ochraceous after 3 days of incubation at 28°C. Hyphae were hyaline and less septate, with rectangular branches. Sporangiophores were erect and unbranched or branched, with globose sporangia formed on their tips. Sporangiospores were elliptical to round, 3.6 to 7.3 × 1.6 to 3.7 µm (n = 100) in size. Two gene regions were amplified for multilocus sequence typing. The D1/D2 region of the nuclear large subunit ribosomal RNA gene (nucLSU) was amplified with primers NL1 and NL4 and the rDNA internal transcribed spacer (ITS) with primers ITS1 and ITS4. PCR products were purified using an Axygen nucleic acid purification kit for sequencing. Both rDNA D1/D2 and rDNA-ITS sequences were submitted to GenBank with accession numbers KM093834 and KM203872, respectively. The isolate F1 showed 98% identity with two isolates of Mucor irregularis (KC524427 and KC461926) in rDNA-ITS sequences and 99% identity with multiple isolates (JX976221, JX976203, and JX976219) of M. irregularis in rDNA D1/D2 sequences. Pathogenicity tests of isolate F1 were conducted based on Koch's postulates. Thirty kernels of fresh ears (milk stage) were pricked by sterilized toothpicks and separately inoculated with a sporangiospore suspension (1 × 106 spores/ml) and 5-day-old mycelial plugs (5 × 5 mm) of isolate F1. Kernels on ears that were inoculated with sterilized water and pure PDA plugs were separately used as controls. After 7 days of incubation, brown spot symptoms developed on the F1-inoculated kernels, which were similar to those observed on the naturally infected ears from the field samples. The control ears remained symptomless during the inoculation tests. Fungal cultures showing the same morphological characteristics as those of isolate F1 were consistently recovered from the diseased cobs inoculated by isolate F1, indicating that M. irregularis was responsible for corn kernel brown spot disease. M. irregularis was reported as a pathogen causing human skin diseases in China (5), America (1), and India (2) and as a phytopathogen causing fruit rot on durian (4). This is the first report of M. irregularis causing corn kernel brown spot disease in China. References: (1) M. M. Abuali et al. J. Clin. Microbiol. 47:4176, 2009. (2) B. M. Hemashettar et al. J. Clin. Microbiol. 49:2372, 2011. (3) S. L. Huang and K. Kohmoto. Bull. Fac. Agric., Tottori Univ. 44:1, 1991. (4) W. F. Wang et al. Plant Quarant. l23:60, 2009. (5) Y. Zhao et al. Mycopathologia 168:243, 2009.

First Report of Fusarium Head Blight of Wheat Caused by Fusarium sacchari in China.

Fusarium head blight (FHB), or scab, caused by Fusarium species, is an economically devastating disease of wheat and other cereal crops worldwide. FHB epidemics in wheat occur frequently in China, especially along the middle and lower reaches of the Yangtze River, including Jiangsu and Shanghai. In 2013, wheat spikes showing clear FHB symptoms were collected from fields in Jiangsu and Shanghai. Symptomatic seeds were surface-sterilized for 1 min with a 5% sodium hypochlorite solution and dipping in 70% ethanol for 30 s, then rinsed three times in sterile distilled water and dried. They were placed onto potato dextrose agar (PDA) and incubated for 3 to 5 days at 28°C in the dark. Fungal colonies displaying morphological characteristics of Fusarium spp. (1,2) were purified by the single-spore technique and characterized at the species level by morphological observations (1,2) and translation elongation factor 1-α (TEF) gene sequencing. The results indicated that members of the Fusarium graminearum clade were predominant on wheat, while the morphological characteristics of 16 isolates were found to be identical to those of F. sacchari (1,2). Colonies on PDA were densely cottony, initially pale but becoming violet with age. The average growth rate was 6 to 8 mm per day at 25°C in the dark. Reverse pigmentation was brownish red to violet-brown. Microconidia, abundant in the aerial mycelium and formed in false heads, were oval to ellipsoidal in shape, primarily zero-septate, measuring 5.7 to 18.8 (average 10.6) µm in length. Macroconidia were slender, three- to five-septate, with a curved apical cell and a poorly developed basal cell, 26.3 to 68.9 (average 44.0) µm in length. No chlamydospores were observed. Two F. sacchari strains (Y37 and S43), isolated from Jiangsu and Shanghai, respectively, were investigated by sequence comparison of their partial TEF gene sequences (Accession Nos. KM233195 and KM233196). BLASTn analysis of the TEF sequences obtained with sequences available in the GenBank database revealed 99.8 and 99.5% sequence identity to F. sacchari (GenBank Accession Nos. JF740708 and JF740709). Pathogenicity tests were conducted by injecting 10 µl of a spore suspension (5 × 105 spores/ml) into wheat florets (20 per isolate of cv. Yangmai16), which were then grown under field conditions in Shanghai. Control plants were inoculated with sterile distilled water. Spikes were harvested and evaluated 14 days post-inoculation. Reddish white mold was observed on inoculated wheat spikes; in addition, spikelets adjacent to the inoculation point and the infected florets were brown. No symptoms were observed on water controls. Koch's postulates were fulfilled by reisolating the pathogen from infected florets and identifying them by TEF gene sequencing. F. sacchari is the cause of an important disease of sugar cane, pokkah boeng (1), and has been reported to produce the mycotoxin beauvericin, which causes toxicosis in human and other animals (3). To our knowledge, this is the first report of F. sacchari causing wheat head blight in China. The report contributes to an improved understanding of the composition of Fusarium species on wheat in the lower reaches of the Yangtze River in China, which will be useful for exploring appropriate disease management strategies in this region. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (2) J. F. Leslie et al. Mycologia 97:718, 2005. (3) A. Moretti et al. Int. J. Food Microbiol. 118:158, 2007.

Cyclooxygenase inhibitors use is associated with reduced risk of esophageal adenocarcinoma in patients with Barrett's esophagus: a meta-analysis.

BACKGROUND: Esophageal adenocarcinoma (EAC) has high mortality and is increasing in incidence. Barrett's esophagus (BE) increases the risk for EAC. Studies have reported inconsistent findings on the association between use of cyclooxygenase (COX) inhibitors and the risk of neoplastic progression in BE patients. Therefore, we performed a meta-analysis to investigate this association. METHODS: A meta-analysis was undertaken among a total of 9 observational studies using fixed- and random-effects models, comprising 5446 participants; 605 had EAC or high-grade dysplasia (HGD). RESULTS: Overall, COX inhibitors use was associated with a reduced risk of EAC/HGD among BE patients (relative risk (RR)=0.64, 95% confidence interval (CI)=0.53-0.77). Aspirin use also reduced the risk of EAC/HGD (RR=0.63, 95% CI=0.43-0.94), as well as non-aspirin COX inhibitors (RR=0.50, 95% CI=0.32-0.78). The chemopreventive effect seemed to be independent of duration response. CONCLUSIONS: Cyclooxygenase inhibitors use is associated with a reduced risk of developing EAC in patients with BE. Both low-dose aspirin and non-aspirin COX inhibitors are associated with a reduced risk of neoplasia. More well-designed randomised controlled trials are needed to increase our understanding of the chemopreventive effect of COX inhibitors.

CT and MRI features of patients with diastematomyelia.

OBJECTIVE: To evaluate computed tomography (CT) and magnetic resonance imaging (MRI) features in patients with diastematomyelia and to investigate clinical characteristics of this lesion. STUDY DESIGN: A retrospectively study. SETTING: The Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University. METHODS: A total of 82 diastematomyelia cases were retrospectively studied. All the patients underwent neurological examinations as well as MRI and CT of the spine. A self-established neurological functional grading system was used, and posterior tibial nerve somatosensory cortical-evoked potential (PTNSCEP) was measured to assess the neurological status of the patients. Imaging features of symmetry of splitting, presence of septum, location of lesion and number of split segments were studied. The neurological functional grading, PTNSCEP, and imaging findings were then analyzed and compared, and the difference was considered to be significant if P-value was lower than 0.05. RESULTS: Neurological functional grading and latency of PTNSCEP were significantly different but related in terms of symmetry of splitting, presence of septum and location of lesion. Although no significant differences were present in the number of split segments, the severity of the neurological functional grading and PTNSCEP impairment were not related to the number of split segments. CONCLUSION: The imaging features in diastematomyelia are characteristic and relate well with the clinical manifestations according to neurological functional grading and PTNSCEP measurement, except the number of split segments.

First Report of Dieback Disease on Cedars Caused by Diplodia seriata in China.

Cedars (Cedrus deodara (Roxb.) G. Don) are well known as evergreen ornamental trees widely used in horticulture in temperate climates. In March 2013, dieback symptoms were found on cedar trees in different locations (including the campus of Nanyang Normal University) in Nanyang (33°01' N, 112°29' E), a southwestern city of Henan Province, China. Characteristic symptoms included needle discoloration and defoliation, canker formation and gummosis on trunks and branches, browning and tissue necrosis under the bark as well as dieback of branches/trunks. Of 873 cedar trees investigated, 139 (16%) were symptomatic. A total of 21 Diplodia sp. isolates were obtained from 102 tissue pieces randomly sampled from the lesion margins of 31 affected trees with a conventional method for isolation of culturable fungal species from plant tissues. Monohyphal cultures were isolated from actively growing edges of colonies to purify the isolates. The purified isolates were grown on 2% water agar with sterilized stems of Foeniculum vulgare to induce their colonies to form pycnidia (4). Unmatured conidia were hyaline, aseptate, and turned light to dark brown with maturity. Mature conidia were aseptate (rarely uniseptate), ovoid with truncated or rounded base and obtuse apex, externally smooth, roughened on the inner surface, and 8 to 11 × 23 to 26 µm (n = 50). These morphological characteristics of the isolates agreed with those of Diplodia seriata (the anamorph of Botryosphaeria obtusa) (5). The rDNA-ITS sequences of two representative isolates (xs-01 and xs-06) were amplified with primers ITS1 and ITS4. PCR products were purified and ligated with PMD-19T vector for sequencing. The rDNA-ITS sequences were submitted to GenBank with accession nos. KJ463386 and KJ549774 for isolates xs-01 and xs-06, respectively, showing 100% identity with multiple isolates of D. seriata (HQ660463, KC461297, and KF535906). Koch's postulates were fulfilled in greenhouse tests on 2-year-old cedar plantlets inoculated by the two isolates. Ten plantlets were used for the inoculation tests for each of the isolates, and their trunks were wounded to a 2 mm depth with a sterilized cork borer (3 mm diameter). The wounds were inoculated by mycelial plugs cut from 7-day-old colonies grown on potato dextrose agar (PDA) plates and wrapped with Parafilm, and those inoculated with pure PDA plugs served as control. Inoculated plantlets were incubated in a greenhouse with alternating cycles of 14 h fluorescent light/10 h darkness under moist conditions for 30 to 60 days at 28°C. Nine of 20 inoculated plantlets developed needle discoloration and shoot blight symptoms similar to those observed on naturally infected cedar trees. The control plantlets remained symptomless during the incubation period. D. seriata cultures were constantly recovered from each diseased plantlet, indicating that the isolated D. seriata isolates were responsible for the disease. D. seriata has been reported as a phytopathogen causing dieback diseases worldwide on multiple woody plant species such as olive (4), mulberry (1), Pinus spp., and Picea glauca (2,3). To our knowledge, this is the first report of D. seriata causing dieback disease on cedars in China. References: (1) M. Arzanlou et al. Arch. Phytopathol. Plant Protect. 46:682, 2013. (2) T. Burgess et al. Appl. Environ. Microbiol. 67:354, 2001. (3) G. Hausner et al. Can. J. Plant Pathol. 21:256, 1999. (4) J. Kaliterna et al. Plant Dis. 96:290, 2012. (5) A. J. L. Phillips et al. Fungal Divers. 25:141, 2007.

Efficacy of Deferasirox for the treatment of iron overload in Chinese thalassaemia major patients: results from a prospective, open-label, multicentre clinical trial.

OBJECTIVE: To assess the efficacy and safety of deferasirox in Chinese thalassaemia major (TM) patients BACKGROUND: EPIC (Evaluation of Patients' Iron Chelation with Exjade(®)) was a large multi-national study and, notably, the first clinical trial of an iron chelator registered with the Chinese State Food and Drug Administration. METHODS: Efficacy and safety of deferasirox were compared in Chinese (n = 117) and non-Chinese (n = 998) TM patients. Deferasirox was initiated at 20 mg kg(-1) day(-1), with titration increments of 5-10 mg kg(-1) day(-1), based on serum ferritin trends and safety parameters. RESULTS: At baseline, Chinese patients were younger than non-Chinese (mean age 6·8 versus 19·5 years), with higher median serum ferritin (4519 vs 3058 ng mL(-1)). Over 1 year, mean actual deferasirox dose was similar for Chinese and non-Chinese patients (24·6 and 24·0 mg kg(-1) day(-1), respectively); median serum ferritin did not change significantly from baseline in Chinese patients (+340 ng mL(-1), P = 0·102) and significantly decreased in non-Chinese patients (-220 ng mL(-1); P < 0·001). In the 1-year extension in Chinese patients, (mean actual deferasirox dose 33·6 mg kg(-1) day(-1)), median serum ferritin decreased (-756 ng mL(-1); P = 0·0397), with a numerically higher reduction in patients aged ≥6 to < 12 than <6 years (-982 vs -457 ng mL(-1), respectively). The safety profile of deferasirox in Chinese patients was similar to the overall population with respect to clinically-relevant findings. CONCLUSION: Age and deferasirox exposure influenced study findings, supporting the need for longer-term treatment and dose escalation to ≥30 mg kg(-1) day(-1) to achieve neutral or negative iron balance in heavily iron overloaded and younger Chinese patients.

Characteristics and management of occult intrasacral extradural cyst in children.

Occult intrasacral extradural cyst is a rare entity. Since little about this lesion has been reported in the literature, this study herein demonstrates by cases some of the clinical features and surgical treatment of occult intrasacral extradural cyst in children. A series of 4 children, 2 boys and 2 girls aged from 4 years and 6 months to 11 years, with occult intrasacral extradural cyst were reviewed. All patients underwent neurological examinations and magnetic resonance imaging. Of these 4 patients two had urinary incontinence in daytime, one frequent micturition, and one numb in saddle area. There were no abnormal findings on physical or laboratory examination. Whole excision of the cyst and ligation of the tract between the cyst and thecal sac were performed for all the patients. No complications such as cerebrospinal fluid leakage and infection were found after operation. All cases made complete recovery and have been asymptomatic at follow-up. The clinical and radiological features of occult intrasacral extradural cyst are characteristic in children. Magnetic resonance imaging is the choice of investigation and surgery is curative.

Arctigenin, a natural compound, activates AMP-activated protein kinase via inhibition of mitochondria complex I and ameliorates metabolic disorders in ob/ob mice.

AIMS/HYPOTHESIS: Arctigenin is a natural compound that had never been previously demonstrated to have a glucose-lowering effect. Here it was found to activate AMP-activated protein kinase (AMPK), and the mechanism by which this occurred, as well as the effects on glucose and lipid metabolism were investigated. METHODS: 2-Deoxyglucose uptake and AMPK phosphorylation were examined in L6 myotubes and isolated skeletal muscle. Gluconeogenesis and lipid synthesis were evaluated in rat primary hepatocytes. The acute and chronic effects of arctigenin on metabolic abnormalities were observed in C57BL/6J and ob/ob mice. Changes in mitochondrial membrane potential were measured using the J-aggregate-forming dye, JC-1. Analysis of respiration of L6 myotubes or isolated mitochondria was conducted in a channel oxygen system. RESULTS: Arctigenin increased AMPK phosphorylation and stimulated glucose uptake in L6 myotubes and isolated skeletal muscles. In primary hepatocytes, it decreased gluconeogenesis and lipid synthesis. The enhancement of glucose uptake and suppression of hepatic gluconeogenesis and lipid synthesis by arctigenin were prevented by blockade of AMPK activation. The respiration of L6 myotubes or isolated mitochondria was inhibited by arctigenin with a specific effect on respiratory complex I. A single oral dose of arctigenin reduced gluconeogenesis in C57BL/6J mice. Chronic oral administration of arctigenin lowered blood glucose and improved lipid metabolism in ob/ob mice. CONCLUSIONS/INTERPRETATION: This study demonstrates a new role for arctigenin as a potent indirect activator of AMPK via inhibition of respiratory complex I, with beneficial effects on metabolic disorders in ob/ob mice. This highlights the potential value of arctigenin as a possible treatment of type 2 diabetes.

Congenital dermal sinus of the cervical spine: clinical characteristics and management.

Congenital dermal sinus (CDS) in the cervical region is extremely rare with only few cases reported in the literature. This study reports a young case of CDS, by which we demonstrate the clinical characteristics and management of cervical CDS, particularly in kids. A 2-year and 7-month-old male child who presented with a one-year history of difficulty in holding objects by hands was diagnosed to have harbored a CDS in the cervical spine since birth. Neurological examination demonstrated mild weakness in both hands and also atrophy in intrinsic hand muscles. Surgery was performed right after diagnosis. Intraoperatively, the sinus tract was totally excised, and untethering was performed. No complications were found after operation. Histopathological examination suggested dorsal dermal sinus. During the follow-up of three years, the patient presented a stable neurologic deficit but did not suffer from an aggravation or improvement of nervous symptoms. At three-year follow-up, the patient was a stable neurologic deficit, and MRI revealed no evidence of the untethered cervical spinal cord and dermal sinus tract. The management strategies of CDS are early surgical treatment to prevent the development of neurological defects.

Moisture retention of glycerin solutions with various concentrations: a comparative study.

Various methods of evaluating a humectant's moisture retention have unique mechanisms. Hence, for designing advanced or efficient ingredients of cosmetic products, a clear understanding of differences among methods is required. The aim of this study was to analyze the moisture-retention capacity of glycerin, a common ingredient in cosmetic products. Specifically, this study applied gravimetric analysis, transepidermal water loss (TEWL) analysis, and differential scanning calorimetry (DSC) to examine the evaporation of glycerin solutions of different concentrations. The results revealed that the moisture-retention capacity of glycerin increased with the glycerin concentration from 0 to 60 wt%, and glycerin at concentration of 60-70 wt% did not exhibit weight change during the evaporation process. When the glycerin concentration exceeded 70 wt%, moisture sorption occurred in the glycerin solution. Furthermore, the results revealed a deviation between the evaporation rates measured using gravimetric analysis and those measured using TEWL analysis. However, normalizing the results of these analyses yielded the relative evaporation rates to water, which were consistent between these two analyses. DSC thermograms further confirmed the consistent results and identified two hydrated water microstructures (nonfreezable water and free water) in the glycerin solutions, which explained why the measured evaporation rate decreased with the glycerin concentration. These findings can be applied to prove the moisture-retention capacity of a humectant in cosmetic products by different measuring methods.