[Clinical features of renal cell carcinoma associated with Xp11.2 translocation/TFE3 gene fusions-single-center study].

Objective: To analyze the clinical characteristics, treatment methods and prognosis of renal cell carcinoma associated with Xp11.2 translocation/TFE3 gene fusions (Xp11.2 tRCC). Methods: From January 2007 to February 2018, 48 patients were diagnosed with Xp11.2 tRCC at Nanjing Drum Tower Hospital. The epidemiological features, treatment methods and long-term follow-up results were retrospectively reviewed. Results: Of the 48 patients, 20 cases were female and 28 cases were male, aged from 2 to 72 years. Gross hematuria and flank pain were the most frequent symptoms, which occurred on 14 cases and 8 cases respectively. The mean tumor size of 48 cases was (5.3±2.5)cm. Among the 34 cases who were classified as stageⅠ/Ⅱ, 14 cases received laparoscopic nephron-sparing surgery(NSS)and 20 cases received radical nephrectomy(RN). The other 14 cases who were classified as stage Ⅲ/Ⅳ received RN but one case received target therapy. On univariate analysis, tumor diameter, adjuvant treatment, AJCC stage, lymph node metastasis and vein tumor thrombosis showed association with progression-free survival (PFS) and overall survival (OS) (P<0.05). Multivariate analysis indicated that AJCC stage (P=0.023, 95% CI: 0.048-0.081)and vein tumor thrombosis (P=0.046, 95% CI: 1.004-1.590)were independent prognostic factors of PFS. Conclusions: Xp11.2 tRCC mainly occurs in females. RN was the major method for Xp11.2 tRCC. However, NSS can also receive satisficed results for stage T1a case. High AJCC stage and the occurrence of vein tumor thrombosis indicated poor prognosis.

Discovery of a second species of Hyalocylindrophora and the phylogenetic position of the genus in Bionectriaceae.

A new holomorphic species, Hyalocylindrophora bispora, is described and illustrated based on a collection on rotten branches from Guangdong Province, China. The fungus is characterized by fleshy perithecia that become deeply cupulate when dry, covered with long and stiff hairs on the surface, and not change color in KOH or lactic acid. Asci are two-spored and evanescent at maturity. Ascospores are ellipsoidal to elongate-ellipsoidal, unicellular, and warted. Conidiogenous cells are phialidic and cylindrical. Conidia are thick-walled, unicellular, ellipsoidal to somewhat lemon-shaped. This is the first report of sexual state for Hyalocylindrophora. The phylogenetic position of the genus in Bionectriaceae is confirmed by sequence analyses of the combined nuc rDNA 28S, α-actin, and DNA-directed RNA polymerase II subunit 1 regions. Distinctions between the new taxon and the only known species of the genus are compared.

Revision of the genus Thyronectria (Hypocreales) from China.

Recent collections and herbarium specimens of Thyronectria from different regions in China were examined. Using combined analyses of morphology and molecular data, we recognized eight species. Among them, Thyronectria atrobrunnea, T. orientalis, and T. sinensis are described and illustrated as new species. Thyronectria atrobrunnea is characterized by blackish brown perithecia that become cupulate when dry, and 8-spored asci containing ellipsoidal to broadly fusiform or subcylindrical ascospores that bud to form bacillar to subellipsoidal ascoconidia within the asci. Thyronectria orientalis can be easily recognized by stromata that are erumpent through the epidermis of the host, immersed or semi-immersed perithecia covered with yellowish green scurf, and ellipsoidal to subfusiform, muriform ascospores. Thyronectria sinensis on Pinus features solitary ascomata that are rarely aggregated, and 8-spored asci giving rise to subcylindrical to vermiform, multiseptate ascospores that form bacillar to allantoid ascoconidia that fill the asci. The new species and their close relatives are compared and differences between them are discussed. Thyronectria strobi is reported for the first time in China. Name changes for the previously recorded species are noted. Phylogenetic analyses inferred from 28S, ITS, RPB1, TEF1, and TUB2 hint that phenotypic characters, viz. stromata, ascospores, appendage of perithecial wall, and host specificity may carry phylogenetic information as previous papers discussed.

Botrytis pyriformis sp. nov., a novel and likely saprophytic species of Botrytis.

A novel species of Botrytis from Sedum sarmentosum was described based on morphology and analyses of DNA sequences of nuc rDNA ITS regions and three nuclear genes (G3PDH, HSP60, RPB2). Meanwhile pathogenicity in 32 plant species, response to temperature for growth and conidial germination for the species were determined. The Botrytis species was named Botrytis pyriformis sp. nov. It was characterized by formation of grayish mycelia, brownish conidia and melanized sclerotia on PDA. The conidia are pear-shaped, melanized and covered with abundant villiform appendages on the conidial surface. Comparison of the ITS sequences confirmed its placement in the genus Botrytis Phylogenetic analysis based on DNA sequences of G3PDH, HSP60 and RPB2 genes indicated that B. pyriformis and other 30 Botrytis species form a monophyletic clade, which was further divided into three subclades. Subclade I comprised B. pyriformis alone, whereas subclades II and III comprised six and 24 Botrytis species, respectively. Botrytis pyriformis could not infect 32 plant species including S. sarmentosum, possibly due to deficiency in formation of infection cushions. This study presents a formal description and illustrations for B. pyriformis and provides experimental evidence, indicating that B. pyriformis might be a saprophytic species.

Two new hyaline-ascospored species of Trichoderma and their phylogenetic positions.

Collections of hypocrealean fungi found on decaying wood in subtropical regions of China were examined. Two new species, Trichoderma confluens and T. hubeiense, were discovered and are described. Trichoderma confluens is characterized by its widely effuse to rarely pulvinate, yellow stromata with densely disposed yellowish brown ostioles, simple acremonium- to verticillium-like conidiophores, hyaline conidia and multiform chlamydospores. Trichoderma hubeiense has pulvinate, grayish yellow stromata with brownish ostioles, trichoderma- to verticillium-like conidiophores and hyaline conidia. The phylogenetic positions of the two fungi were investigated based on sequence analyses of RNA polymerase II subunit b and translation elongation factor 1-α genes. The results indicate that T. confluens belongs to the Hypocreanum clade and is associated with but clearly separated from T. applanatum and T. decipiens. Trichoderma hubeiense belongs to the Polysporum clade and related to T. bavaricum but obviously differs from other members of the clade in sequence data. Morphological distinctions between the new species and their close relatives are noted and discussed.

Trichoderma (Hypocrea) species with green ascospores from China.

Stromata of Trichoderma species having green ascospores were collected in various regions of China. Based on morphology of the sexual and asexual morph, culture characteristics, and sequence analyses of rpb2 and tef1 genes, 17 species with green ascospores were identified. Among them, Trichoderma rosulatum, T. rufobrunneum and T. stipitatum are described as new species, and seven other species are reported for the first time from China. Trichoderma rosulatum produces small bright yellow or pale greenish stromata with dense dark green ostioles and gliocladium-like conidiophores, shows a close relationship to T. thelephoricola, and belongs to the Chlorospora clade. Trichoderma rufobrunneum, which typically forms reddish brown stromata, is recognised as a member of the Harzianum clade. Trichoderma stipitatum is characterised by turbinate, pale yellow to nearly orange stromata and verticillium-like conidiophores; it forms a distinct, independent lineage with strong bootstrap support in the phylogenetic trees. The distinctions between the new species and their close relatives are discussed, and their phylogenetic positions are explored.

Three new species of Trichoderma with hyaline ascospores from China.

Collections of Trichoderma having hyaline ascospores from different areas of China were examined. Using combined analyses of morphological data, culture characters and phylogenetic information based on rDNA sequences of partial nuc translation elongation factor 1-α encoding gene (TEF1-α) and the gene encoding the second largest nuc RNA polymerase subunit (RPB2), three new species, Trichoderma applanatum, T. oligosporum and T. sinoluteum, were discovered and are described. Trichoderma applanatum produces continuous flat to pulvinate, white to cream stromata with dense orange or pale brown ostioles, and simple acremonium-like to verticillium-like conidiophores, belongs to the Hypocreanum clade and is closely related to T. decipiens. Trichoderma oligosporum forms reddish brown stromata with a downy surface, hyaline conidia and gliocladium-like conidiophores, and is closely related to but distinct from T. crystalligenum in the Psychrophila clade. Trichoderma sinoluteum, as a member of the Polysporum clade, is characterized by pale yellow stromata, white pustulate conidiomata, pachybasium-like conidiophores, and hyaline conidia. Differences between the new species and their close relatives are discussed.

Morphological and phylogenetic identification of Botrytis sinoviticola, a novel cryptic species causing gray mold disease of table grapes (Vitis vinifera) in China.

Seventy-five isolates of Botrytis collected from table grapes (Vitis vinifera) with gray mold symptoms in China were identified based on morpho-cultural characteristics on potato dextrose agar (20 C) and/or phylogenetic analysis using the sequences of three nuclear genes (G3PDH, HSP60, RPB2). Isolates of different species of Botrytis were compared with fenhexamid sensitivity, Bc-hch gene-RFLP haplotyping and pathogenicity to V. vinifera. The 75 isolates comprise two species, B. cinerea (63 isolates) and an undescribed Botrytis sp. (12 isolates) described here as Botrytis sinoviticola Zhang et al. sp., nov. Both B. sinoviticola (Bs) and B. cinerea (Bc) were found to have 20 C optimum for mycelial growth and 25 C for conidial germination. Sensitivity to fenhexamid was significantly greater (P < 0.05) for Bc (EC50 = 0.04 ± 0.01 µg mL(-1)) than for Bs (EC50 = 0.08 ± 0.02 µg mL(-1)). Digestion of the PCR amplicons of the Bc-hch gene with Hha I generated two haplotypes, Group I haplotype for Bs and Group II haplotype for Bc. Bs infected table grapes (leaves, berries) only through wounds, whereas Bc infected both injured and non-injured tissues of table grapes. This study suggests that Bs is a cryptic species sympatric with Bc on table grapes in China.

Caulobacter FliQ and FliR membrane proteins, required for flagellar biogenesis and cell division, belong to a family of virulence factor export proteins.

The Caulobacter crescentus fliQ and fliR genes encode membrane proteins that have a role in an early step of flagellar biogenesis and belong to a family of proteins implicated in the export of virulence factors. These include the MopD and MopE proteins from Erwinia carotovora, the Spa9 and Spa29 proteins from Shigella flexneri, and the YscS protein from Yersinia pestis. Inclusion in this family of proteins suggests that FliQ and FliR may participate in an export pathway required for flagellum assembly. In addition, mutations in either fliQ or fliR exhibit defects in cell division and thus may participate directly or indirectly in the division process. fliQ and fliR are class II flagellar genes residing near the top of the regulatory hierarchy that determines the order of flagellar gene transcription. The promoter sequence of the fliQR operon differs from most known bacterial promoter sequences but is similar to other Caulobacter class II flagellar gene promoter sequences. The conserved nucleotides in the promoter region are clustered in the -10, -20 to -30, and -35 regions. The importance of the conserved bases for promoter activity was demonstrated by mutational analysis. Transcription of the fliQR operon is initiated at a specific time in the cell cycle, and deletion analysis revealed that the minimal sequence required for transcriptional activation resides within 59 bp of the start site.

Anti-arrhythmic effects of captopril on coronary occlusion and reperfusion periods in pigs.

Anti-arrhythmic effects of captopril (Cap) were studied in the anesthetized pigs using a reversible balloon catheter. Results showed that Cap did not exert any influence on the weight percentage of ischemic area to the whole left ventricle, on the levels of serum creatine kinase (CK) and creatine kinase isozyme (CK-MB), nor on the incidence and duration of transient and persistent tachycardia, but reduced the incidence of ventricular fibrillation (2/12, 1/12 in high-dose group pigs treated with Cap 6 mg.kg-1 in the first 10 min, 25 micrograms.kg-1.min-1 in the later 90 min and 12/21, 11/21 in control group treated with normal saline through the occlusion and reperfusion periods, respectively, P < 0.05). It was suggested that Cap did not exhibit direct (or non-specific, if any) effects on anti-arrhythmias.

Expression of an early gene in the flagellar regulatory hierarchy is sensitive to an interruption in DNA replication.

Genes involved in the biogenesis of the flagellum in Caulobacter crescentus are expressed in a temporal order and are controlled by a trans-acting regulatory hierarchy. Strains with mutations in one of these genes, flaS, cannot transcribe flagellar structural genes and divide abnormally. This gene was cloned, and it was found that its transcription is initiated early in the cell cycle. Subclones that restored motility to FlaS mutants also restored normal cell division. Although transcription of flaS was not dependent on any other known gene in the flagellar hierarchy, it was autoregulated and subject to mild negative control by other genes at the same level of the hierarchy. An additional level of control was revealed when it was found that an interruption of DNA replication caused the inhibition of flaS transcription. The flaS transcript initiation site was identified, and an apparently unique promoter sequence was found to be highly conserved among the genes at the same level of the hierarchy. The flagellar genes with this conserved 5' region all initiate transcription early in the cell cycle and are all sensitive to a disruption in DNA replication. Mutations in these genes also cause an aberrant cell division phenotype. Therefore, flagellar genes at or near the top of the hierarchy may be controlled, in part, by a unique transcription factor and may be responsive to the same DNA replication cues that mediate other cell cycle events, such as cell division.

Molecular cloning and restriction mapping of human lymphotoxin gene.

In order to clone the human lymphotoxin (HuLT) gene, we practiced a concise and time-saving method: homologous recombination in vivo (1). By using the mouse lymphotoxin (MuLT) cDNA (1.3 kb) as a probe, we isolated the HuLT gene from a human genomic library which was constructed with cosmid pcos2EMBL as a vector. After linearization, the recombinant cosmid was partially digested with BamHI, EcoRI, PstI, and PvuII respectively, and either cos end was labelled by hybridization with radioactive oligos complementary to the cohesive end sequence (2). The physical map of HuLT gene was made by this method.