Discovery of Fusarium proliferatum f. sp. malus domestica Causing Apple Replant Disease in China.

Apple replant disease (ARD) is the most serious threat facing the apple industry globally. ARD is mainly manifested as decreased plant growth, serious root rot disease, and considerable yield loss. Microbial factors are the dominant factors leading to the occurrence of ARD. Research on soil-borne pathogenic fungi leading to the occurrence of ARD in China is limited. In the present study, we selected 16 replanting orchards from the Northwest Loess region and around the Bohai Gulf. Diseased roots and rhizosphere soil from healthy apple trees and trees showing ARD symptoms were sampled at random. High-throughput sequencing was used to study the fungal communities in the rhizosphere soil, which showed that the composition of the rhizosphere soil fungal community of ARD-symptomatic and healthy apple trees was different. Nectriaceae at the family level and Fusarium at the genus level dominated the rhizosphere soil fungal community in the two regions, while for healthy apple trees, the relative abundance of Mortierella, Minimedusa, Tetracladium, and Chaetomium was higher. Tissue separation and serial dilution were used to separate fungi, and a total of 89 genera and 219 species were obtained, most of which were Fusarium. Fusarium was further confirmed to be the most abundant pathogen species leading to the occurrence of ARD in China through pathogenicity assays. A pathogenicity assay was carried out by the dip-and-cut technique using different host plants. It was found that Fusarium MR5 showed strong aggressiveness to apple rootstocks. Diseased seedlings specifically exhibited chlorosis of the leaves, browning from the edge of the leaf, followed by rolling and yellowing of the leaves, resulting in wilting and eventually death. Strain MR5 was preliminarily identified as F. proliferatum according to the morphological and cultural characteristics. A maximum likelihood analysis of identities based on six gene sequence (ITS, TUB2, IGS, mtSSU, RPB2, and the TEF gene) alignments between the MR5 strain and other strains showed 99 to 100% homology with F. proliferatum. Based on our test results, strain MR5 was identified as F. proliferatum f. sp. malus domestica, which is of great significance for finding new measures to control ARD in China.

[Nonalcoholic fatty liver disease and liver transplantation].

The rising prevalence of nonalcoholic fatty liver disease (NAFLD) is now the second largest indication for liver transplantation in Western countries, but viral hepatitis B and end-stage alcohol-related liver disease are still the main indications in China. With the improvement of people's living standards, the prevalence of metabolic syndrome, and the number of NAFLD patients has also gradually increased. At the same time, with the hepatitis B vaccination popularization and the nucleos(t)ide analogues and other drugs uses, it is predicted that NAFLD-related end-stage liver disease may become one of the main indications for liver transplantation in our country in the future. This article reviews the research progress of NAFLD and liver transplantation.

[The clinical effect of ultrasonic bone curette-assisted "zoning" style laminectomy for the treatment of severe ossification of thoracic ligamentum flavum].

Objective: To investigate the safety and clinical efficacy of "zoning" style laminectomy by ultrasonic bone curette in patients with severe thoracic ossification of the ligamentum flavum(TOLF). Methods: The clinical data of 36 patients with severe TOLF treated by "zoning" style laminectomy at Department of Spinal Surgery,Zhengzhou Orthopaedic Hospital from October 2015 to October 2018 were respectively analyzed.There were 17 males and 19 females,aged(57.3±10.2)years(range:43 to 80 years).According to the anatomical characteristics of the thoracic ligamentum flavum and the pathological process of ossionization,each decompression segment was divided into the upper 1/3 area of the lamina,the bilateral area of the ossionum flavum,the transitional area,and the area of close contact between the ossionum flavum and the spinal cord.Different surgical strategies were used for decompression in turn.The modified Japanese Orthopedic Association (mJOA) was used to evaluate the neurological function status before and after surgery,to evaluate the surgical effect of patients,and to observe the surgical complications.Paired sample T test was used for data analysis. Results: All 36 patients successfully completed the operation,the operation time was (88.6±24.6) minutes(range:60 to 150 minutes).The intraoperative blood loss was (426.7±167.4) ml(range:250 to 800 ml).Follow-up time was (27.2±7.7) months(range:12 to 48 months).The mJOA score at the last follow-up was 9.0±1.5,which was statistically significant compared with the preoperative score 5.4±1.8 (t=13.59,P<0.01).The improvement rate of mJOA score was (65.7±22.1) %,of which 17 cases were excellent (47.2%),13 cases were good (36.1%),4 cases were normal (11.1%),2 cases were ineffective (5.6%).Ten patients had cerebrospinal fluid leakage during the separation or removal of dural ossification and were cured after a series of comprehensive conservative treatment.Two patients showed transient neurological deterioration,and the neurological function gradually recovered to the preoperative state after comprehensive treatment such as increasing the mean arterial pressure and using neurotrophic drugs.During the follow-up,no aggravation of neurological dysfunction and segmental kyphosis were found. Conclusions: The ultrasonic bone curette-assisted "zoning" style laminectomy for the treatment of severe TOLF can directly observed the position relationship between ossification of the ligamentum flavum and the spinal canal structure during the operation,and accurately guide the surgical decompression.It has the advantages of safe operation and complete decompression,which provides an important reference for the selection of clinical surgery.

[Initial exploration of transfusion-free liver transplantation].

Objective: To evaluate the effect of transfusion-free techniques on the prognosis of liver transplant patients. Methods: The recipients of adult liver transplantation at Tianjin First Central Hospital from August to December 2019 were included in the clinical observation. Liver transplantation without allogeneic blood transfusion was performed through anesthesia management techniques such as acute hemodilution or phlebotomy without volume replacement,maintaining decreased baseline central venous pressure and cell saver. According to the actual results,the patients were divided into two groups: transfusion-free group(n=21) and allogeneic transfusion group(n=28). There were 13 males and 8 females aged of (56.3±11.6) years in the transfusion-free group;and there were 16 males and 12 females aged (54.3±14.2)years in the allogeneic transfusion group. The transplant recipients who had not adopted transfusion management strategy from January to July 2019 were included as control group(27 males and 13 females,aged of (58.9±14.1)years). The clinical data of patients in perioperative period were collected to compare whether there were differences in the recovery of liver function and early complications among the three groups, one-way ANOVA test, rank-sum test, and χ2 test were used for data analysis. Results: The amount of intraoperative blood loss in both the transfusion-free group and the transfusion group was less than that in the control group((454.2±271.3)ml vs.(673.6±333.4)ml vs.(890.3±346.7)ml;q=-6.342,-5.286,both P<0.05).The duration of stay in ICU of the transfusion-free group was less than that of the transfusion group and control group((36.4±9.1)hours vs.(44.3±14.9)hours vs.(58.2±21.1)hours;q=-4.432,-3.824,both P<0.05).The mean ALT level at 7 days after operation was significantly lower in the transfusion-free group than in the control group((56.8±32.1)U/L vs.(89.6±45.6)U/L;q=-3.358,P<0.05). Conclusions: The improvement of multi-disciplinary transfusion management technology aimed at transfusion-free liver transplantation can effectively reduce intraoperative hemorrhage and help to avoid surgical transfusion. Transfusion-free liver transplantation is beneficial to the early postoperative recovery,and its long-term clinical significance is worthy of further clinical research.

[Impact of psoas muscle index on early postoperative mortality and complications after liver transplantation].

Objective: To investigate the relationship between psoas muscle index (PMI) and early postoperative survival rate and the incidence of complications after liver transplantation in adults. Methods: The clinical data of 225 patients (male, n=184; female, n=41) underwent liver transplantation at the Organ Transplantation Department of First Central Clinic Institute of Tianjin Medical University from January 2014 to December 2016 were analyzed, retrospectively.Original disease: hepatitis B liver cirrhosis(44 cases), hepatitis C cirrhosis(10 cases), autoimmune liver cirrhosis(29 cases), other benign liver diseases(24 cases), liver cirrhosis with liver cancer(116 cases), hilar cholangiocarcinoma(1 case) and hepatic vascular sarcoma(1 case). The area of bilateral psoas muscle on the lower edge level of the third lumbar vertebral body was measured through preoperative CT image.The PMI was calculated using this formula: bilateral psoas muscle area (mm(2))/the square of the body height (m(2)). According to the receiver operating characteristic curve and cut-off values, the male and female patients were divided into low PMI group and high PMI group respectively.The χ(2) test, Fisher exact test and t-test was used to compare the differences in perioperative data, survival rate and postoperative complications between the two groups, respectively. Results: There were 44 patients in the low PMI group, and 181 patients in the high PMI group. ICU time was longer (82.5(62.0-128.0) hours vs.69.1(56.0-104.0) hours; P=0.006) and preoperative blood urea nitrogen level (5.86(4.35-15.52) mmol/L vs. 4.94(4.05-7.06) mmol/L; P=0.012) was higher in the low PMI group than those in the high PMI group. Incidence rates of grade 5 complication (18.2%) and grade 4a complication (18.2%) were higher in the low PMI group, and 120-day cumulative survival rate was lower than that in high PMI group(81.8% vs. 95.6%, P=0.001). On the other hand, there were no significant differences in preoperative white blood cell count level, serum creatinine level, operative time, anhepatic period time, intraoperative blood loss, and incidence of postoperative grade 3 complications between the two groups(all P>0.05). Conclusions: There is a significant correlation between PMI and early postoperative survival rate and incidence of complications.Patients with lower PMI has poor prognosis after liver transplantation.

Liver injury possibly related to drug interaction after liver transplant: a case report.

WHAT IS KNOWN AND OBJECTIVE: Drug-induced hepatotoxicity is potentially lethal. Liver transplant patients receive a large number of medications and adverse drug reactions, and drug-drug interactions must be closely monitored. CASE SUMMARY: We report a case of a 29-year-old liver transplant patient who suffered liver injury most likely induced by drug interaction between capecitabine and warfarin. Vitamin K1 caused skin rash possibly because of the distribution and metabolism characteristic of the drug in this patient. WHAT IS NEW AND CONCLUSION: Close monitoring and prompt discontinuation of the drugs with high volume of distribution and metabolized through the liver are necessary to avoid drug-drug interaction in liver transplant patients.

Effect of CDH1 gene methylation on transforming growth factor (TGF-ß)-induced epithelial-mesenchymal transition in alveolar epithelial cell line A549.

The aim of this study was to investigate the specific molecular mechanism of the transforming growth factor ß (TGF-ß)-induced epithelial-mesenchymal transition in a lung cancer cell line, and to provide new ideas for targeting therapy of lung cancer. A549 cells were treated with different concentrations of TGF-ß and 5-aza-deoxycytidine (5-aza-dC). The morphological changes after the intervention were observed. The change in the expression of the epithelial marker E-cadherin (E-cad) was detected by Western blot. The proliferation of A549 cells was measured using the MTT assay. Cell movement and invasion capacity was evaluated with the cell scratch test and invasion test. TGF-ß induced A549 cells to transform to a mesenchymal cell morphology and downregulated the expression of E-cad, and 5-aza-dC inhibited this phenomenon. Compared with the control group, the number of transmembrane cells was higher and cell migration was markedly increased in the experimental group with continued culture in the presence of 10 ng/mL TGF-ß, showing significant differences (P < 0.05). CDH1 gene methylation is involved in TGF-ß-induced epithelial-mesenchymal transition in the alveolar epithelial cell line A549.

First Report of Powdery Mildew Caused by Erysiphe macleayae on Torenia fournieri in China.

Torenia fournieri (Linderniaceae) is a common ornamental plant in China. It is also an important Chinese medicinal herb for its heart clearing and toxin removal properties. In October 2013, severe powdery mildew infections were observed on T. fournieri in Baihuayuan Garden (125.35°E, 43.88°N), China. Voucher specimens were deposited in the Herbarium of Mycology of Jilin Agricultural University under the accession number HMJAU02176. Whitish colonies covered the surface of leaves, petioles, sepals, and stems. The infected leaves became yellow and necrotic by advanced stages of the infection. Chasmothecia with a diameter between 63.5 and 95 µm were present singly or in groups, and bore dark brown mycelioid. The appendages were 0.5 to 4 times as long as the chasmothecial diameter, brown at the base and paler toward the apex. Asci were 2 to 6 per chasmothecium, short-stalked or sessile, 50 to 62 × 30 to 50.5 µm, and 2- to 4-spored. Ascospores were pale brown, oval to ellipsoid, 27 to 43 × 13 to 17 µm. Hyphae were flexuous to straight, branched, and septate. Appressoria were well-developed, lobed, and solitary or in opposite pairs. Conidiophores were unbranched, cylindrical, and 94 to 185 × 9 to 15 µm. Foot-cells were straight, cylindrical, 19 to 40 µm long, and followed by 1 to 3 cells shorter or nearly equal to the foot-cell. Conidia were singly produced, cylindrical or oval, 34 to 44 × 16 to 20 µm, and without distinct fibrosin bodies. Lobed germ tubes were produced at the tip of conidia. The morphological characteristics of sexual and asexual structures were consistent with Erysiphe macleayae (1,2). To confirm the identification, the complete internal transcribed spacer (ITS) region of rDNA of the pathogen was amplified with the primers ITS1/ITS4 and sequenced (3). The resulting 574-bp sequence (KJ600796) showed 100% similarity with E. macleayae (KF856294) and Oidium sp. isolated from Chelidonium majus (HQ286645 to 46) and one base different from M. microcarpa (JQ681217). Koch's postulate was completed by gently pressing a diseased leaf onto three young excised leaves of asymptomatic seedlings. Three non-inoculated leaves were used as controls. Inoculated leaves were incubated in separate petri dishes in a greenhouse at 20 to 25°C. Symptoms developed 5 days after inoculation, whereas the control leaves remained symptomless (voucher specimens HMJAU02176I). The morphology of the fungus on the inoculated leaves was identical to that observed on the originally diseased leaves. Powdery mildew on T. fournieri has only been reported as Golovinomyces sp. in Hungary (4). To our knowledge, this is the first report of powdery mildew caused by E. macleayae on T. fournieri worldwide. Infection of Torenia by E. macleayae, a common powdery mildew on various hosts of the Papaveraceae, is very unusual and unexpected. It demonstrates a wider host range of this species beyond the limits of the Papaveraceae. References: (1) U. Braun and R. T. A. Cook. Taxonomic manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series 11. CBS, Utrecht, the Netherlands, 2012. (2) M. J. Park et al. Plant Dis. 96:1376, 2012. (3) S. Takamatsu et al. Mycol. Res. 113:117, 2009. (4) P. Vági et al. Eur. J. Plant Pathol. 117:89, 2007.

First Report of Powdery Mildew on Euphorbia pekinensis Caused by Podosphaera euphorbiae-helioscopiae in China.

Euphorbia pekinensis Rupr, the Peking spurge, is a perennial grass that is planted throughout northeast China. Its roots can be used as a Chinese medicine for diuretic or purgative purposes. Some diterpenoids isolated from Euphorbia spp. show strong antitumor, antivirus, skin-irritating, and tumor-promoting activities. Some species contain chemicals that can be used as pesticides. In September and October 2009, spurge plants were seriously infected by powdery mildew fungi in Changchun Agri-Expon, Jilin Province, China. White mildew colonies appeared in early autumn and initial colonies coalesced to cover the entire upper sides of leaves, stems, and petals 3 to 4 weeks after the disease was noticed. Until late October, chasmothecia were observed on leaves and stems and the infected areas also became dark brown. Mycelium was amphigenous, in patches, white, and persistent. Conidia with distinct fibrosin bodies were ellipsoid-cylindrical, 21.6 to 33.8 × 12 to 15.6 µm, and produced in chains. Germ tubes arose near a pole of the conidia and terminated in simple, unlobed apices. Conidiophores were 59.8 to 139.9 µm; foot cells were straight to slightly flexuous, followed by one to three cells, and 24 to 48 × 8.4 to 10.8 µm. Hyphal appressoria were papillate. Chasmothecia were 70 to 95 µm in diameter, averaging 79.4 µm, and were immersed in mycelia. Mycelioid appendages (5 to 12) occurred in the lower part of the chasmothecia and were flexuous, curved, thick walled, and strongly undulate. Appendages were long (349-) 490 to 1,000 (-1,300) µm, averaging 759.4 µm. They were often densely crowded around the chasmothecia. Asci were single, sessile or short stalked, 60 to 80 × 45 to 70 µm, and eight spored. Ascospores were ellipsoid-ovoid and 16.8 to 28.8 × 10.8 to 18 µm. The fungus was identified as Podosphaera euphorbiae-helioscopiae on the basis of morphological characteristics of the anamorphic and teleomorphic stages. An herbarium specimen was deposited in the Herbarium of Mycology of Jilin Agriculture University (CJLCC100). Genomic DNA was extracted from the chasmothecia, and the internal transcribed spacer region (ITS) and the D1/D2 domains of the 28S rDNA were amplified and sequenced (GenBank Nos. JF795490 and JF795491). The ITS sequence has 98% identity with those of Podosphaera lini, P. macularis, and P. spiraeae, and the D1-D2 sequence of 28S rDNA has 99% identity with those of P. negeri and P. spiraeae, which indicated that this fungus belongs to the genus Podosphaera, even though there is no direct sequence evidence that it is P. euphorbiae-helioscopiae. P. euphorbiae-helioscopiae has only been previously reported in Japan (2), occurring on Euphorbia helioscopia. This species is unique because of the long and curved, thick-walled appendages, and there is no comparable species (1). P. euphorbiae-hirtae and P. euphorbiae are reported to infect Euphorbiaceae plants. Both species differ from P. euphorbiae-helioscopiae in their conspicuously shorter appendages (1,3). To our knowledge, this is the first reported occurrence of powdery mildew on E. pekinensis, and P. euphorbiae-helioscopiae is new to China. This is also the first description of the anamorph of the fungus since there is no record in the Erysiphales Database. References: (1) U. Braun. Nova Hedwigia 89:1, 1987. (2) Y. Nomura. Taxonomical Study of Erysiphaceae of Japan. Yokendo LTD, Tokyo, 1997. (3) R. Y. Zheng and Y. N. Yu. Flora Fungorum Sinicorum. Vol. 1. Erysiphales. Science Press, Beijing, 1987.

Milan criteria, University of California, San Francisco, criteria, and model for end-stage liver disease score as predictors of salvage liver transplantation.

The selection criteria for salvage liver transplantation (SLT) candidates have not been previously established. A global analysis for the association between the criteria and prognosis is required. All of the adult patients who underwent liver transplantation with a diagnosis of hepatocellular carcinoma (HCC) from January 1, 2000, to December 31, 2011, were retrospectively analyzed. A total of 1,554 cases were involved, including 1,392 primary liver transplantation (PLT) and 162 SLT cases. All the cases were classified into 3 groups according to the Milan criteria combined with the University of California, San Francisco (UCSF), criteria, and significant differences were found between the 2 groups. The overall graft survival rate was lower in all cases of SLT than in PLT (P = .030). Within the Milan criteria, no significant difference in the graft survival rate was found between PLT and SLT. In a Cox regression analysis, the Model for End-Stage Liver Disease (MELD) score and tumor levels graded according to the Milan/UCSF criteria were found to be independent risk factors for the graft survival rate. Receiver operating characteristic (ROC) curves were generated by the fatality risk values calculated by means of the Cox model and the 1-year graft survival rates of all the patients and of the SLT patients. The areas under the ROC curves were 0.922 and 0.935, respectively. Compared with PLT, the global graft survival rate of SLT was compromised. The MELD score and Milan/UCSF criteria were effective in predicting the prognosis of PLT and SLT. Therefore, when the recurrent lesions of HCC are within the Milan criteria, SLT can be performed with a good prognosis.

[The development of a residual voltage tester].

The paper introduces the key points of the residual voetage tester's designing and its circuit frame diagram.

Inverted Duplication and Deletion of 10q25q26 in a Patient without Any Obvious Skeletal Anomalies.

We report on a girl with inverted duplication and deletion of 10q25q26 revealed by array-CGH and FISH analysis. Array-CGH analysis demonstrated a ∼13.1-Mb duplication encompassing 10q25.3q26.2 and a ∼5-Mb deletion at 10q26.2q26.3. No single-copy region was detected between the deleted and duplicated segments. FISH analysis found the arrangement duplicated in an inverted position. FISH analysis using the same probes did not show any abnormality in both parents, which indicates a de novo occurrence. The frequently reported features of distal 10q duplication include developmental delay, blepharophimosis, hypotonia, skeletal anomalies and some facial dysmorphisms. The girl presented with many features of distal 10q duplication with the exception of skeletal anomalies. To our knowledge, this is the fourth patient reported in the literature with inv dup del 10q. 10q duplication seems to account for most of the phenotypes for our patient. Although no obvious skeletal feature was found in our patient at present, follow-up assessment of skeletal development should be planned with the increase of age.