Differences of bioelectrical impedance in the development and healing phase of pressure ulcers and erythema in mouse model.

Pressure ulcers (PUs) are economically burdensome medical conditions. Early changes in pressure ulcers are associated with erythema. In this study, bioelectrical impedance was used to measure the differences between PUs and blanchable erythema. We divided 21 ICR mice into three groups: control, 1000 mmHg-1h, and 1000 mmHg-6h. Healthy skin, blanchable erythema, and PUs were induced on the dorsal skin. The results indicated an immediate increase in impedance, resistance, and reactance values in the pressure group after release, followed by a subsequent decrease until two days after release. Compared with the control group, impedance and reactance significantly increased by 30.9% (p < 0.05) and 30.1% (p < 0.01), respectively, in the 6 h-loading group immediately after release. One and two days after release, the 1 h-loading and 6 h-loading groups exhibited significantly different degrees of decline. One day after release, impedance and resistance decreased by 30.2% (p < 0.05) and 19.8% (p < 0.05), respectively, in the 1 h-loading group; while impedance, resistance, and reactance decreased by 39.2% (p < 0.01), 26.8% (p < 0.01), and 45.7% (p < 0.05), respectively, in the 6 h-loading group. Two days after release, in the 1 h-loading group, impedance and resistance decreased by 28.3% (p < 0.05) and 21.7% (p < 0.05), respectively; while in the 6 h-loading group, impedance, resistance, and reactance decreased by 49.8% (p < 0.001), 34.2% (p < 0.001), and 59.8% (p < 0.01), respectively. One and two days after release the pressure group reductions were significantly greater than those in the control group. Additionally, we monitored changes during wound healing. Distinguishing early PUs from blanchable erythema by noninvasive bioelectrical impedance technology may have applications value in early assessment of PUs.

[Establishment and application of animal models of pulmonary alveolar proteinosis].

Pulmonary alveolar proteinosis (PAP) is a rare respiratory disease, but this disease has slow research progress. Animal model is an effective tool for basic research. Current PAP animal models are based on the main pathogenesis of granulocyte-macrophage colony stimulation factor (GM-CSF) signal disorder and environmental homeostasis imbalance in the alveoli. Application researches focus on the treatment strategies of PAP. The existing PAP animal models cannot fully reflect to the development of human PAP, which should be further developed and improved to provide the basis for clinical practice.

[Clinical analysis of liver chemistries in children and adolescent with 2019-nCoV infection].

Objective: To describe the clinical features of liver involvement in children and adolescent with 2019-nCoV infection. Methods: The clinical data of 77 hospitalized cases admitted to the Children's Hospital of Fudan University were collected from January 19 to November 28, 2020. The characteristics and risk factors of abnormal liver chemistries in children with laboratory-confirmed 2019-nCoV infection were analyzed. Results: Of the 77 cases, 44 were male (57.1%) and 33 were female (42.9%), with a median age of 10 years. 27(35.1%) were asymptomatic, 28(36.4%) had mild illness, 22(28.6%)had non-severe pneumonia. Hydroxychloroquine was used in 7 cases. Of the 75 children without underlying diseases, alanine aminotransferase was elevated in 1 case (1.5%, during hydroxychloroquine therapy), aspartate aminotransferase was elevated in 7 cases (10.3%), alkaline phosphatase was elevated in 7 cases (28%), and total bilirubin, direct bilirubin, albumin, international normalized ratio were in normal range. There was no statistical difference between the pneumonia group and the non-pneumonia group in term of liver chemistries (P > 0.05), same as between the elevated erythrocyte sedimentation rate group and the normal group. There was no aggravation of liver injury in the child with biliary atresia. The child with epilepsy showed no abnormal liver chemistries after infection. Conclusion: Children with 2019-nCoV infection had mild clinical symptoms with few cases of liver injury. The abnormal liver chemistries in children with COVID-19 infection may be related to the underlying disease and the use of antiviral drugs.

Characterization of a novel allelic variant in HLA-B*40 lineage, HLA-B*40:298:02, by cloning and sequencing.

A novel allelic variant in HLA-B*40 lineage, HLA-B*40:298:02, has been identified in an individual of Han ethnicity afflicted with nasopharyngeal carcinoma in Hunan province, southern China. Following polymerase chain reaction-Sanger sequence-based typing (PCR-SBT), this new variant was further confirmed by two distinct strategies of cloning and sequencing. HLA-B*40:298:02 differs from HLA-B*40:298:01 by a single synonymous cytosine substitution at nucleotide position 26 (T→C) in exon 3, which corresponds to codon 99 of the mature HLA-B mRNA molecule. This new allele has an estimated frequency of 0.0002, in about 2,500 sequence-based typed subjects from the same population.

Characterization of a novel HLA-B*39:01:01-related allele, HLA-B*39:130, by cloning and phasing.

A novel HLA-B*39:01:01-related variant, HLA-B*39:130, has been identified in a normal individual of Han ethnicity in Hunan province, southern China. Following Sanger polymerase chain reaction-sequence-based typing (PCR-SBT), this new allele was further confirmed by cloning, phasing and sequencing. Aligned with HLA-B*39:01:01, HLA-B*39:130 has a nonsynonymous thymine substitution at nucleotide position 94 in exon 4, resulting in amino acid change from threonine to isoleucine at codon 214 (ACA→ATA) of the mature HLA-BmRNA molecule.

Characterization of a novel HLA-B*40 allele, HLA-B*40:186:02, by cloning and sequencing.

A novel HLA-B*40 variant, HLA-B*40:186:02, has been identified by cloning and sequencing in a southern Chinese Han population. Aligned with HLA-B*40:01:01, HLA-B*40:186:02 has a nonsynonymous cytosine mutation at nucleotide position 165 in exon 2, leading to amino acid change from glycine to arginine at codon 56. It differs from HLA-B*40:186:01 by a synonymous change (adenine to cytosine) at position 165 in exon 2.

Sequence-based typing of HLA-A gene in 930 patients with nasopharyngeal carcinoma in Hunan province, southern China.

In this study, we typed 930 cases of nasopharyngeal carcinoma (NPC) and 1134 normal controls recruited from Hunan province, southern China for human leukocyte antigen-A (HLA-A) locus by sequencing exons 2-4. Very significant associations between HLA-A*02:07, HLA-A*11:01 and NPC were established [25.7% vs 16.18%; odds ratio, OR (95% confidence interval, CI) = 1.79 (1.54-2.09), P < 0.0001 and 21.1% vs 30.42%, OR (95% CI) = 0.61 (0.53-0.70), P<0.0001, respectively]. Further analysis of the molecular basis underlying these associations suggests that cysteine (C) at codon 99 of α2-helix of HLA-A protein is probably deleterious and confers risk to NPC. Convincing evidence was uncovered for negative association of a rare allele in southern Chinese populations, HLA-A*31:01, with NPC [0.22% vs 2.12%, OR (95% CI) = 0.1 (0.04-0.28), P < 0.0001]. rs1059449-A, which encodes arginine (R) at codon 56 of α1-helix of HLA-A protein, was postulated to be crucial for such a pattern of negative association with NPC. A subset of NPC cases (N = 632) and normal controls (N=712) were tested for anti-virus capsid antigen (anti-VCA) immunoglobulin A (IgA), very significant difference in seropositivity for anti-VCA IgA was observed between the two groups [67.56% vs 6.46%, OR (95% CI) = 30.16 (21.42-42.46), P < 0.0001]. However, seropositivity for anti-VCA IgA did not correlate with HLA-A allelic typing in both groups.

Component spectra extraction from terahertz measurements of unknown mixtures.

The aim of this work is to extract component spectra from unknown mixtures in the terahertz region. To that end, a method, hard modeling factor analysis (HMFA), was applied to resolve terahertz spectral matrices collected from the unknown mixtures. This method does not require any expertise of the user and allows the consideration of nonlinear effects such as peak variations or peak shifts. It describes the spectra using a peak-based nonlinear mathematic model and builds the component spectra automatically by recombination of the resolved peaks through correlation analysis. Meanwhile, modifications on the method were made to take the features of terahertz spectra into account and to deal with the artificial baseline problem that troubles the extraction process of some terahertz spectra. In order to validate the proposed method, simulated wideband terahertz spectra of binary and ternary systems and experimental terahertz absorption spectra of amino acids mixtures were tested. In each test, not only the number of pure components could be correctly predicted but also the identified pure spectra had a good similarity with the true spectra. Moreover, the proposed method associated the molecular motions with the component extraction, making the identification process more physically meaningful and interpretable compared to other methods. The results indicate that the HMFA method with the modifications can be a practical tool for identifying component terahertz spectra in completely unknown mixtures. This work reports the solution to this kind of problem in the terahertz region for the first time, to the best of the authors' knowledge, and represents a significant advance toward exploring physical or chemical mechanisms of unknown complex systems by terahertz spectroscopy.

Induction function of siRNA-mediated survivin gene silencing on nasopharyngeal carcinoma cell apoptosis.

We examined the function of survivin gene expression in patients with nasopharyngeal carcinoma (NPC), as well as small interfering RNA (siRNA) on controlling CNE-2 NPC proliferation and apoptosis. Immunohistological methods, in situ hybridization, and reverse transcription-polymerase chain reaction technique were used to detect survivin protein and mRNA expression. We designed an siRNA sequence to inhibit survivin gene expression. The MTT method was used to examine the function of siRNA on controlling cell growth and proliferation. Induction of cell apoptosis by siRNA was examined by flow cytometry; electron microscopy was used to observe ultrastructure changes in CNE-2 cells. Western blotting was used to detect survivin gene expression. The survivin protein was expressed in 71.9% of cells, while its mRNA was expressed in 65.6% of cells. Relative mRNA expression was 4.16 x 10(-2); these data for the control groups were 23.3, 33.3, and 4.42 x 10(-4), respectively. Following transfection with 3 different siRNA sequences, survivin mRNA expression in CNE-2 cells was decreased. Inhibition of cell proliferation and rate of apoptosis increased with increasing siRNA concentration. Western blotting revealed decreased survivin expression and electron microscopy revealed ultrastructural changes in cancer cells. Survivin gene expression in NPC generally increased. In vitro transcription of siRNA decreased CNE-2 survivin gene expression, and different sequences of siRNA decrease gene expression in CNE-2 cells to varying degrees. Transfected siRNA3 can effectively inhibit CNE-2 cell proliferation and induce apoptosis; gene silencing using siRNA may represent a new treatment for NPC.

Association of cyclin D1 and survivin expression with sensitivity to radiotherapy in patients with nasopharyngeal carcinoma.

The association between cyclin D1 and survivin protein expressions with radiotherapy sensitivity in patients with nasopharyngeal carcinoma was investigated. Biopsy specimens of 72 patients with nasopharyngeal carcinoma were collected before the initiation of radiotherapy (49 cases were in the radiation-sensitive group and 23 cases were in the radiation-insensitive group). Conventional hematoxylin and eosin staining was used for tissue typing. The immunohistochemical SP method was used to detect cyclin D1 and survivin protein expression levels. The IBM SPSS Statistics 20 statistical software was applied for conducting the chi-squared test and the Spearman correlation analysis. In the 72 cases, the high expression rates of cyclin D1 were 28.6% (14/49) and 69.6% (16/23) in the radiotherapy-sensitive group and in the radiotherapy-insensitive group, respectively, and the differences between groups were statistically significant (P<0.05). The high expression rates of survivin were 34.7% (17/49) and 73.9% (17/23) in the radiotherapy-sensitive group and in the radiotherapy-insensitive group, respectively, which differed significantly (P<0.05). The protein expressions of cyclin D1 and survivin were positively correlated (Spearman's r=0.353, P<0.05). Cyclin D1 and survivin expression levels were negatively correlated with the radiosensitivity of nasopharyngeal carcinoma. Cyclin D1 and survivin may be used as molecular markers to predict the sensitivity of radiotherapy.

Screening of the residual normal ovarian tissue adjacent to orthotopic epithelial ovarian carcinomas in nude mice.

The objective of this study was to explore the feasibility and methods of screening the residual normal ovarian tissue adjacent to orthotopic ovarian carcinomas in nude mice. Human epithelial ovarian cancer cells (OVCAR3) were subcutaneously implanted for a tumor source and ovarian orthotopic transplantation. The cancer tissue, proximal paraneoplastic tissue, middle paraneoplastic tissue, remote paraneoplastic tissue, and normal ovarian tissue were removed. CK-7, CA125, p53, survivin, MMP-2, and TIMP-2 expression was detected by reverse transcription polymerase chain reaction. We obtained 35 paraneoplastic residual ovarian tissues with normal biopsies from 40 cases of an orthotopic epithelial ovarian carcinoma model (87.5%). CK-7, CA125, p53, survivin, MMP-2, and TIMP-2 expression was lower in proximal paraneoplastic tissue than in cancer tissue (P < 0.05) and higher than in middle and remote paraneoplastic tissue (P < 0.01). There was no statistically significant difference between the expression of these genes in middle and proximal paraneoplastic tissue as well as among residual normal ovarian tissues with different severity (P > 0.05). In ovarian tissues of 20 normal nude mice, the expression of CK- 7, CA125, p53, survivin, MMP-2, and TIMP-2 was negative. Overall, the expression levels of CK-7, CA125, p53, survivin, MMP-2, TIMP-2, and other molecular markers showed a decreasing trend in the non-cancer tissue direction. The expression levels can be used as standards to screen residual normal ovarian tissue. We can obtain relatively safe normal ovarian tissues adjacent to epithelial ovarian cancer.

Molecular characterization of a novel monopartite begomovirus isolated from Pouzolzia zeylanica in China.

The complete genome sequence of a monopartite begomovirus isolate TY01 was obtained from diseased Pouzolzia zeylanica plants exhibiting golden mosaic symptoms in Baise, Guangxi Province, China. It consisted of 2723 nucleotides (nt) and encoded two ORFs (CP and AV2) in the virion-sense DNA and five ORFs (AC1-AC5) in the complementary-sense DNA. Compared with the DNA-A sequences of other begomoviruses, it has the highest (78.5 %) nucleotide sequence identity with ageratum yellow vein virus (AYVV) isolate AFSP6D from Thailand, which is less than the 89 % identity in the complete genome that has been defined as the threshold value for demarcation of species in the genus Begomovirus, family Geminiviridae. Phylogenetic analysis showed that TY01 was grouped in a separate clade from the other 28 begomovirus isolates. These results indicate that isolate TY01 is a member of a novel Begomovirus species, for which the name "Pouzolzia golden mosaic virus" (PGMV) is proposed.

First Report of Ageratum yellow vein China virus Infecting Zinnia elegans in Vietnam.

Whitefly-transmitted geminiviruses belong to the Geminiviridae family, which includes a group of plant DNA viruses that occurs in tropical and subtropical regions worldwide and caused severe leaf curl diseases on many crops. In July 2011, symptoms of mild yellowing and curling of leaves were observed on Zinnia elegans in Hue City, Vietnam. To identify possible begomoviruses, two samples with typical symptoms (Z1 and Z2) were collected and total DNA was extracted using PlantGen DNA Kit (CoWin Biotech Co., China). Using begomovirus-specific degenerate primer pair AV494 and AC1048 to amply geminivirus capsid protein gene (3), a 586-bp DNA fragment was amplified from both collected samples. The PCR products from Z1 and Z2 were cloned and sequenced. The partial sequences from two isolates had 96.8% similarity to each other and most closely related to Ageratum yellow vein China virus (AYVCNV) (98% identity to GenBank Accession Nos. AM940137.1 and AJ849916.1). Using two degenerate primer pairs bego1 (5'-ACMGGCCYATGWAYMGGAAG-3')/bego6 (5'-ACNGGDAARACRATGTGGGC-3') and bego2 (5'-CTTDATRTTBTCRTCCATCC-3')/bego3 (5'-CCWGGATTGCANARSAMGAT-3') to amplify full-length viral genome from Z1 (1). The complete DNA-A sequence from Z1 were determined to be 2,741 nucleotides (JX911332), showed high levels of sequence identity (89% [JN809812] to 98% [AJ849916.1 and AJ558120]) with AYVCNV and shared 98% nucleotide sequence identity with AYVCNV-G68 (AJ849916.1) and AYVCNV-G13 (AJ558120). No DNA-B component nor DNA-ß could be detected using primers described previously (4). Zinnia leaf curl virus, Ageratum enation virus, Tobacco leaf curl virus, and Alternanthera yellow vein virus have been reported to be associated with Zinnia leaf curl disease (2). To our knowledge, this is the first report of AYVCNV infecting Z. elegans. AYVCNV was first reported in A. conyzoides on Hainan Island, China, and could infect L. esculentum (Solanaceae), A. conyzoides (Asteraceae), or tobacco (Solanaceae). Our results further corroborate that AYVCNV can infect Asteraceae plants. Therefore, this virus may pose a serious threat to many plants. References: (1) L. Lin et al. Plant Prot. 37:44, 2011. (2) P. Naveen et al. J. Biotechnol. Bioinform. 2:6, 2012. (3) S. D. Wyatt et al. Phytopathology 86:1288, 1996. (4) X. P. Zhou et al. J. Gen. Virol. 84:237, 2003.

First Report of a Tospovirus in Mulberry.

Mulberry (Morus alba L.) is an economically important crop grown widely throughout Asia. Various virus-like symptoms including mosaics, vein banding, and chlorotic ringspots have been observed and reported on mulberry trees in China and Japan for decades. However, the etiology of mulberry viral diseases is generally understudied, although two mulberry-infecting viruses, Mulberry latent virus (genus Carlavirus) (2) and Mulberry ringspot virus (genus Nepovirus) (3), have been partially characterized. In a recent (2010 to 2011) field survey in Guangxi Province, China, supported by the local government, the incidence of virus-like diseases of mulberry ranged between 40 and 80%. To identify the viruses infecting mulberry, deep sequencing of small RNAs (4) was conducted using an Illumina Genome Analyzer. Small RNAs were isolated from five samples of mulberry leaves showing various virus-like symptoms and sequenced. Among the contigs assembled, a 445-bp contig (GenBank Accession No. JX268597) was found to share 76.6% nucleotide identity and 83.0% amino acid identity to Groundnut bud necrosis virus (genus Tospovirus, family Bunyaviridae; Accession Nos. U42555 and AAC55521). To obtain a longer cDNA fragment of this virus, a reverse transcription (RT)-PCR was done with primers MV-N-F (5'-AAGCCATCAATGTGCCTCCGGA-3') and MV-N-R (5'-AACACCATGTCTACCGTCCGTC-3') that align to the S-RNA sequence encompassing the nucleocapsid (N) gene and a portion of the intergenic region (IGR) of the Tospovirus. PCR products of about 1,000 bp were successfully amplified from the total RNA of the three mulberry samples (sl-1, xcsy-1, and xcsy-4) showing vein banding symptoms, but not from asymptomatic mulberry (jk-1). These PCR products were cloned and sequenced. The lengths of the amplicons were 1,027 bp (isolate sl-1, JX173786), 987 bp (isolate xcsy-1, JX173787), and 979 bp (isolate xcsy-4, JX173788) and the partial IGRs of the sl-1, xcsy-1, and xcsy-4 isolates were 187 bp, 147 bp, and 139 bp, respectively. The coding regions for the N protein were 831 bp and the deduced proteins of 277 amino acid residues were 100% identical for all three isolates. Since the N protein of this virus shared up to only 74.4% identity to other tospoviruses (74.4% to Capsicum chlorosis virus, ABB83818; and 71.5% to Watermelon bud necrosis virus, ABY79095), it may represent a new member of the Tospovirus genus, temporarily named Mulberry vein banding virus (MuVBV), according to the species demarcation criteria for the Bunyaviridae (1). To the best of our knowledge, this is the first report of a Tospovirus infecting M. alba. In an RT-PCR screening of 48 randomly selected mulberry samples suspected to be virus-infected, 32 were MuVBV-positive. Giving the high incidence and the high yield loss associated with Tospovirus and the presence of thrips, suspected vectors for the virus, MuVBV may represent a substantial threat to the silkworm industry in China. References: (1) M. Q. K. Andrew et al. Virus Taxonomy: 9th Report of the ICTV. Elsevier Academic Press, San Diego, 2012. (2) T. Tsuchizaki. Annu. Phytopath. Soc. Japan 42:304, 1976. (3) T. Tsuchizaki et al. Annu. Phytopath. Soc. Japan 37:266, 1971. (4) Q. Wu et al. PNAS. 107:1606, 2010.

Early strategy vs. late initiation of renal replacement therapy in adult patients with acute kidney injury: an updated systematic review and meta-analysis of randomized controlled trials.

OBJECTIVE: The optimal time to start renal replacement therapy (RRT) for acute kidney injury (AKI) remains controversial. We aim to compare the effects of early vs. delayed RRT initiation on clinical outcomes in adult patients with AKI. MATERIALS AND METHODS: PubMed, Embase, Cochrane Library, Web of Science, Chinese Biomedical Literature Database, ClinicalTrials.gov, and the International Clinical Trial registry platform were systematically searched from inception to 7 August 2022. The review included randomized clinical trials (RCTs) comparing early and delayed initiation of RRT in AKI patients. The selected primary outcomes were short-term and long-term mortality. Secondary outcomes included RRT dependency, intensive care unit (ICU) length of stay, hospital length of stay, mechanical ventilator-free days, vasoactive agents-free days, RRT-free days, and adverse events. RESULTS: Overall, 15 RCTs, including 5,625 patients, were analyzed. Early RRT showed no survival benefit when compared to the delayed therapy (28-or 30-day mortality: RR, 1.01, 95% CI: 0.94-1.08, p = 0.87; 60-day mortality: RR, 0.87, 95% CI: 0.71-1.06, p = 0.16; 90-day mortality: RR, 1.00, 95% CI: 0.88-1.13, p = 0.97; in-hospital mortality: RR, 1.05, 95% CI: 0.88-1.24, p = 0.58; ICU mortality: RR, 1.00, 95% CI: 0.91-1.10, p = 0.98). The delayed RRT did not lead to a higher risk of RRT dependency, ICU, or hospital length of stay than the early RRT. Similarly, early initiation of RRT did not lead to longer ventilator-free, vasoactive agent-free, and RRT-free days. However, early RRT initiation was associated with more adverse events. CONCLUSIONS: Our study suggested that early RRT initiation was not associated with survival benefits or better clinical outcomes and increased the risk of RRT-associated adverse events. Current evidence does not support the use of early RRT for AKI patients without urgent indications.

[Clinicopathological characteristics and differential diagnosis of 6 cases of congenital granular cell tumor].

To provide references for the diagnosis and treatment of congenital granular cell tumor (CGCT), by comprehensive analysis of the clinical data, histopathological and immunohistochemical results. Patients with CGCT were involede, from March 2015 to November 2020, at the Department of Oral and Maxillofacial Surgery of the First Affiliated Hospital of Zhengzhou University. A total of 6 children, aged 3-16 days, 1 male and 5 female, 5 maxillary and 1 mandibular, with maximum tumor diameter of 6-70 mm, were included. The lesions of CGCT were single and connected to the alveolar ridge by a pedicle. The surface of the tumor was covered with a vascular network, and two cases had ulcers on the surface of the tumor. All 6 cases had the tumor removed surgically and there was no recurrence or metastasis in the follow-up visit. Although CGCT is rare, it is a benign tumor and generally does not recur or metastasize after surgery, and has a good prognosis. The prenatal imaging, clinical manifestations after delivery, pathological characteristics and immunohistochemical analyses may provide reference for early diagnosis and treatment of CGCT.

Preparation and characterization of iridium dioxide-carbon nanotube nanocomposites for supercapacitors.

A thin film of novel hierarchical structure, suitable for supercapacitor applications, has been developed through combining conductive multi-wall carbon nanotubes (MWCNTs) and square IrO(2) nanotubes (IrO(2)NT) of nanometer size. Synthesis of this hierarchical structure with open porosity is performed by depositing IrO(2) short tubes densely along the long wires of carbon nanotube on a substrate of stainless steel. A IrO(2) tube of rutile structure grows in the [001] direction, with an opening at its top, surrounded by very thin walls. The IrO(2) addition on the MWCNT template increases the capacitance of the CNT thin film effectively, because of pseudocapacitance of the IrO(2) surface. For this particular composite, featured with two tubular nanostructures, the specific capacitance increases from 15 F g(-1) (MWCNT) to 69 F g(-1) (IrO(2)NT/MWCNT), measured using the galvanostatic discharge experiment. Its property of fast retrieval of the stored charge is assured in the impedance measurement, showing that the internal resistance of the IrO(2)NT/MWCNT nanocomposite electrode is lower than that of the bare MWCNTs.

Molecular Identification and Characterization of Tomato zonate spot virus in Tobacco in Guangxi, China.

In Guangxi Province of southwest China, diseases caused by Tospoviruses (family Bunyaviridae) pose a serious threat to tobacco (Nicotiana tobacum) production. During surveys conducted annually at Xinrong Village in Jingxi County from 2008 to 2010, more than 130 ha of fields were found to have 10 to 50% of plants exhibiting symptoms similar to spotted wilt caused by Tomato spotted wilt virus (TSWV). During this period, disease symptoms at similar prevalence and incidence were also found at Fushan, Debao County in most of the cultivars produced in these areas, including Yunyan 85, 87, 92, 97, and K326. Symptoms on tobacco varied but commonly included dwarfing, midrib browning, distorted apical buds, and concentric ringspots that coalesced to form large areas of dead leaf tissue. Mechanical inoculation from diseased tobacco leaves with concentric ringspots back to tobacco cv. Yunyan 85 or 87, resulted in 12 of 16 plants with symptoms similar to those observed in the field. No symptoms on plants developed following inoculation with buffer only. Symptoms found in the field resembled those caused by TSWV. However, testing using TSWV-specific antiserum was shown to be negative by double-antibody sandwich-ELISA (Agdia, Elkhart, IN). Total RNA was extracted from 27 diseased tobacco plants collected from different regions in Guangxi using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. RNA extracts were amplified by reverse transcription (RT)-PCR using the degenerate primers T2740 (ATGGGDATNTTTGATTTCATG) and T3920c (TCATGCTCATSAGRTAAATYTCTCT) designed to target the partial RNA-dependent RNA polymerase (RdRp) sequence of members in the genus Tospovirus (3). Amplification was performed at 42°C for 60 min, followed by 35 cycles of PCR (30 s denaturation at 94°C, 45 s annealing at 55°C, and 30 s extension at 72°C) and a 7-min final extension at 72°C. A PCR product of approximately 1.2 kb was amplified from 21 diseased plants. RT-PCR amplicons were cloned into the pUC19-T Simple Vector (TaKaRa, Dalian, China) and sequenced in both directions. Sequences were assembled and analyzed by DNAStar 5.01 (DNASTAR, Madison, WI). Sequences of representative isolates were deposited in GenBank (Accession Nos. JN020022 to JN020027). The 1.2-kb partial RdRp sequences of these isolates were shown to have 94.4 to 95.3% nucleotide identity and 96.5 to 97.5% amino acids identity to Tomato zonate spot virus (TZSV) (GenBank Accession No. NC_010491) (1). Among these TZSV isolates from Guangxi, the partial RdRp sequences have 98.0 to 99.4% nucleotide identity and 98.8 to 100% amino acids identity with each other. The presence of TZSV was further confirmed in diseased tobacco plants by indirect ELISA using antiserum of TZSV (provided by Prof. Zhongkai Zhang, Agricultural Academy of Yunnan, China). TZSV has been characterized as a novel tospovirus on various hosts including tobacco in Yunnan province (1,2). To our knowledge, this is the first report of TZSV-associated disease on tobacco in Guangxi Province, southwest China. Further work is necessary to study the epidemiology and management of the disease. References: (1) J. Dong et al. Arch. Virol. 153:855, 2008. (2) J. Dong et al. J. Insect Sci. 10:166, 2010. (3) Y. Lin. Master Thesis. National Chung Hsing University, Taichung, Taiwan, Republic of China, 2007.

[Assessment of the quality of life of mandibular ameloblastoma patients after reconstruction with double-barrel fibula flap].

Objective: To analyse the quality of life of patients receiving repair of bone defect with folded fibula flap after removal of mandibular ameloblastoma. Methods: The case data of 39 patients with ameloblastoma admitted to the First Affiliated Hospital of Zhengzhou University from August 2013 to April 2016 were retrospectively analysed, including 21 males and 18 females, from 18 to 58 years old. 3D printing and digital technology were used in flap preparation before surgery in all patients. The folded fibular flaps were used to repair mandibular defects and the implants were placed between 6-9 months after surgery. The short form-36 health survey questionnaire (SF-36) and the university of Washington quality of life questionnaire (UW-QOL) were applied to evaluate the quality of life of patients before surgery and at 6 months and 24 months after surgery. The higher the score, the better the condition. SPSS 20.0 was adopted for statistical analysis. Results: The SF-36 survey showed that the mean score of body role before surgery (72.4±11.7) was significantly higher than that at 6 months after surgery (39.6±11.1, t=23.580, P<0.05) or that at 24 months after surgery (59.8±6.4, t=8.358, P<0.001). Compared with the preoperative mean scores of Physical Pain (73.0±11.0), General Health (73.4±10.4) and Health Changes (79.2±3.9) before surgery, the mean scores Physical Pain (53.1±7.7), General Health (53.5±7.5) and Health Changes (63.9±11.7) at 6 months after surgery were decreased significantly respectively (t=13.068, 13.756 and 10.880, respectively, all P<0.05), but the mean scores Physical Pain (78.8±14.0), General Health (80.9±12.6) and Health Changes (84.4±4.6) at 24 months after surgery were increased significantly respectively (t=-2.904, -4.027 and -7.586, respectively, all P<0.05), with significant differences in the mean scores of Physical Pain, General Health and Health Changes between 6 and 24 months after surgery (t=-14.241, -16.490, -14.294, respectively, all P<0.001). The UW-QOL survey showed that the mean scores of chewing, language and taste functions decreased at 6 months after surgery (53.1±6.7, 53.0±7.7 and 62.2±9.9, respectively), but improved at 24 months after surgery (67.9±3.9, 63.9±2.9 and 68.4±11.1, respectively), with statistically significant difference (t=-16.765, -11.675 and 2.498, respectively, all P<0.001). Conclusion: The application of folded fibula flaps to repair bone defects after sugery of mandibular ameloblastoma can better meet the needs of language and chewing functions and improve the quality of life of patients.

[Application of the posteromedial thigh flap for oral and maxillofacial defect reconstruction].

To explore the feasibility of using the posteromedial thigh flap as an alternative source for oral and maxillofacial reconstruction. During January 2019 to January 2020, twenty-three patients underwent oral and maxillofacial tumor ablation and defect reconstruction with 23 posteromedial thigh flaps were enrolled in the Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University. Thirteen of the patients were male and ten were female, with age of (54.5±9.7) years (33-72 years). The numbers and types of perforators, the dimension of flap and the vascular pedicle length were measured. The outcomes of flaps and donor-site complication were recorded. The patients' satisfaction with donor-site cosmesis were evaluated by the visual analogue scale (VAS). More than one sizable perforators was found in each case and the median number of perforators was 2 (range, 1 to 4), and all of the perforators were musculocutaneous. The pedicle length was (9.8±1.5) cm (range, 7.3 to 13.4 cm). The diameters of artery and the larger vein were 2.0 mm (range, 1.5 to 2.5 mm) and 2.0 mm (range, 1.5 to 3.0 mm), respectively. The dimension of the flaps ranged from 8 cm×6 cm to 12 cm×8 cm, and the donor sites were all closed primarily. All of the flaps were clinically survived, only one patient experienced partial wound dehiscence of donor site 14 days postoperatively and no donor site infection or permanent muscular weakness was reported. The VAS scores of the patients' satisfaction with donor-site cosmesis were all more than 8. The perforators of the posteromedial thigh flap is consistent and the donor-site scar is well concealed, which make the posteromedial thigh flap an excellent option for oral and maxillofacial reconstruction.