["Double Grooves-Double Rings" technique of transurethral Thulium laser enucleation of the prostate: learning curve of single surgeon].

Objective: To evaluate the learning curve of the "Double Grooves-Double Rings" (DGDR) technique of transurethral Thulium laser enucleation of the prostate (ThuLEP) for benign prostatic hyperplasia (BPH) by a single surgeon. From June 2021 to July 2022, 84 patients mean age (69.0±8.0) years,preoperative prostate volume (90.9±40.3)ml with BPH underwent ThuLEP in the Department of Urology, Peking University First Hospital.Performed by a single surgeon who had no experience of transurethral resection of prostate (TURP) and any laser surgeries. The case scatter plots with the best fitting line were drawn to analyze the learning curve. According to the date of the surgeries, the patients were equally divided into three learning stages (28 patients for each group). The T-PSA,prostate volume,operative time,enucleation time, enucleation efficiency,catheter indwelling time, hemoglobin drop and perioperative complications (including re-TURP, blood transfusion, stress incontinence≥3 months and urethral stricture) were compared among the groups. The learning curve was divided into three stages, and the cutting point was shown on the 14th case. Except the prostate volume [stage1 (75.7±30.7) ml, stage2 (93.40±39.6)ml, stage3 (103.5±46.2) ml, P<0.05], there was no significant difference of the baseline data between three groups (P>0.05). Compared with those of stage 1(100.6±24.7) min,(0.55±0.22) g/min, a statistically significant improvement was observed in both of the operative time and the enucleation efficiency among stage 2[(84.5±36.6) min, (0.87±0.33) g/min and stage 3 (71.2±26.3) min, (1.27±0.45) g/min, P<0.05]. The learning curve of the DGDR technique for ThuLEP can be divided into three stages. A ThuLEP beginner can preliminarily master this technique after completing 14 cases.

Analysis of risk factors for stone remnants and recurrence after lateral decubitus percutaneous nephrolithotomy.

OBJECTIVE: This study aims to explore the risk factors for stone remnants and recurrence after lateral decubitus percutaneous nephrolithotomy (PCNL), providing insights to enhance the stone-free rate and reduce the stone recurrence rate. PATIENTS AND METHODS: A retrospective analysis was conducted on 356 patients with renal or upper ureteral stones who underwent lateral decubitus PCNL from January 2015 to August 2022. Among them, 271 patients had complete clinical and follow-up data. General clinical information, perioperative data, and follow-up data were collected. Univariate and multivariate logistic regression analyses were performed to identify risk factors for stone remnants and recurrence after lateral decubitus PCNL. RESULTS: The stone-free rate after lateral decubitus PCNL was 88.6% (195/271), and the stone recurrence rate within three years was 28.1% (76/271). Stone size (p<0.001) and stone co-infection (p=0.047) were identified as independent risk factors for stone remnants after lateral decubitus PCNL. Multiple stones (p=0.003) were an independent risk factor for stone recurrence after lateral decubitus PCNL. CONCLUSIONS: Stone size and stone co-infection are independent risk factors for stone remnants after lateral decubitus PCNL. Multiple stones are an independent risk factor for stone recurrence after lateral decubitus PCNL.

Updating species diversity of Colletotrichum, with a phylogenomic overview.

The genus Colletotrichum includes important plant pathogens, endophytes, saprobes and human pathogens. Even though the polyphasic approach has facilitated Colletotrichum species identification, knowledge of the overall species diversity and host distribution is largely incomplete. To address this, we examined 952 Colletotrichum strains isolated from plants representing 322 species from 248 genera, or air and soil samples, from 87 locations in China, as well as 56 strains from Saudi Arabia, Thailand, Turkey, and the UK. Based on morphological characteristics and multi-locus phylogenetic analyses, the strains were assigned to 107 species, including 30 new species described in this paper and 18 new records for China. The currently most comprehensive backbone tree of Colletotrichum, comprising 16 species complexes (including a newly introduced C. bambusicola species complex) and 15 singleton species, is provided. Based on these analyses, 280 species with available molecular data are accepted in this genus, of which 139 have been reported in China, accounting for 49.6 % of the species. Colletotrichum siamense, C. karsti, C. fructicola, C. truncatum, C. fioriniae, and C. gloeosporioides were the most commonly detected species in China, as well as the species with the broadest host range. By contrast, 76 species were currently found to be associated with a single plant species or genus in China. To date, 33 Colletotrichum species have been exclusively reported as endophytes. Furthermore, we generated and assembled whole-genome sequences of the 30 new and a further 18 known species. The most comprehensive genome tree comprising 94 Colletotrichum species based on 1 893 single-copy orthologous genes was hence generated, with all nodes, except four, supported by 100 % bootstrap values. Collectively, this study represents the most comprehensive investigation of Colletotrichum diversity and host occurrence to date, and greatly enhances our understanding of the diversity and phylogenetic relationships in this genus. Taxonomic novelties: New species: Colletotrichum arecacearum F. Liu, Z.Y. Ma & L. Cai, Colletotrichum bicoloratum F. Liu, W.P. Wu & L. Cai, Colletotrichum bromeliacearum F. Liu & L. Cai, Colletotrichum buxi F. Liu, W.P. Wu & L. Cai, Colletotrichum chamaedoreae F. Liu, W.P. Wu & L. Cai, Colletotrichum crousii F. Liu, Z.Y. Ma & L. Cai, Colletotrichum danxiashanense F. Liu, W.P. Wu & L. Cai, Colletotrichum diversisporum F. Liu, W.P. Wu & L. Cai, Colletotrichum diversum F. Liu & L. Cai, Colletotrichum dolichoconidiophori F. Liu, W.P. Wu & L. Cai, Colletotrichum iris F. Liu & L. Cai, Colletotrichum monsterae F. Liu, W.P. Wu & L. Cai, Colletotrichum multiseptatum F. Liu, W.P. Wu & L. Cai, Colletotrichum nageiae F. Liu, W.P. Wu & L. Cai, Colletotrichum obovoides F. Liu & L. Cai, Colletotrichum parabambusicola F. Liu, W.P. Wu & L. Cai, Colletotrichum paraendophytum F. Liu, W.P. Wu & L. Cai, Colletotrichum reniforme F. Liu, Z.Y. Ma & L. Cai, Colletotrichum schimae F. Liu, W.P. Wu & L. Cai, Colletotrichum shivasii F. Liu & L. Cai, Colletotrichum sinuatum F. Liu, W.P. Wu & L. Cai, Colletotrichum subacidae F. Liu, Z.Y. Ma & L. Cai, Colletotrichum subsalicis F. Liu & L. Cai, Colletotrichum subvariabile F. Liu, W.P. Wu & L. Cai, Colletotrichum syngoniicola F. Liu, Z.Y. Ma & L. Cai, Colletotrichum telosmae F. Liu, W.P. Wu & L. Cai, Colletotrichum tibetense F. Liu & L. Cai, Colletotrichum variabile F. Liu, W.P. Wu & L. Cai, Colletotrichum zhaoqingense F. Liu & L. Cai, Colletotrichum zhejiangense F. Liu, W.P. Wu & L. Cai. Citation: Liu F, Ma ZY, Hou LW, Diao YZ, Wu WP, Damm U, Song S, Cai L (2022). Updating species diversity of Colletotrichum, with a phylogenomic overview. Studies in Mycology 101: 1-56. doi: 10.3114/sim.2022.101.01.

[The incidence and causes of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia].

Objective: To observe the incidence of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia and to explore the risk factors for its occurrence. Methods: The data of patients suffered from Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia from June 2018 to May 2021 were retrospectively reviewed. The type of procedure, surgeon, age and gender were selected as matching factors, 4 patients without Tapia syndrome were selected as control group for each case. The radiological parameters including mandibular-vertebral distance, thyroid-vertebral distance, thyroid cartilage-vertebral distance, and C2-C7 lordotic Cobb angle were measured on lateral radiographs of the cervical spine. The above parameters were measured on neutral, over-flexion and over-extension radiographs. The difference between the Tapia group and the control group were analyzed. Results: There were 9 patients (0.37%) suffered from Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia in 2 431 patients, and it happened in 0.67 days (0-2 days) after the operation. There were 3 males and 6 females with a mean age of (61±5) years. The clinical manifestations was tongue extension deviation in 8 cases (88.9%), dysarthria in 6 cases (66.7%), dysphagia in 3 cases (33.3%), tongue stiffness in 3 cases (33.3%), hoarseness in voice and pharyngeal discomfort in 1 case (11.1%). All of the symptoms were relieved in all patients at 3 months postoperative follow-up. In neutral position, the mandibular-vertebral distance was (7.19±3.96) mm in the control group and it was (3.98±3.01) mm in Tapia group (P<0.05). From neutral position to hyperflexion position, the distance between mandible and vertebral body was reduced from 3.98 mm to 1.95 mm in the Tapia group and decreased for 51.0%, and it decreased from 7.19 mm for 31.8% to 4.90 mm in the control group. Conclusions: The incidence of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia is low. A smaller mandibular-vertebral distance on pre-operative cervical spine lateral view radiograph maybe a risk factor for Tapia syndrome after posterior cervical surgery under oral tracheal intubation general anesthesia.

[The correlation between the severity of uncovertebral joints degeneration and heterotopic ossification after single-level artificial cervical disc replacement].

Objective: To investigate the correlation between the severity of uncovertebral joints degeneration and heterotopic ossification (HO) after single-level artificial cervical disc replacement (ACDR). Methods: From January 2005 to January 2016, 70 patients who had undergone single-level ACDR in Peking University Third Hospital and had at least 5 years follow-up were included in this study. There were 35 males and 35 females with an average age of (42±8) years (range, 25-62 years). Cervical spine A-P X-rays were taken to assess the degeneration of uncovertebral joints and lateral X-rays were taken to assess the degeneration of intervertebral space. Cervical spine lateral and the flexion-extension X-rays at 5 years follow up were taken to assess HO. Degeneration of uncovertebral joints were evaluated by the classification system set-up in Peking University Third Hospital. Kellgren&Lawrence grading system was used to evaluate the degeneration of intervertebral space. HO was evaluated by the McAfee grading standards. The data were collected before surgery and at 5-years follow-up, then the correlation between degeneration of uncovertebral joints, degeneration of intervertebral space and HO was analyzed with Spearman non-parametric test. Results: The average follow-up time of 70 patients was (62.7±4.8) years (range, 52-74 months). There was a significant positive correlation between preoperative uncovertebral joints degeneration and HO after ACDR (r=0.585, P<0.01). There was a significant positive correlation between preoperative intervertebral space degeneration and HO (r=0.557, P<0.01). There was a significant positive correlation between preoperative intervertebral space degeneration and preoperative uncovertebral joints degeneration (r=0.727, P<0.01). Conclusion: There is a significant positive correlation between preoperative uncovertebral joints degeneration and HO after ACDR.

[Surgical treatment of severe congenital cervical kyphosis].

Objective: To investigate the surgical treatment of severe congenital cervical kyphosis. Methods: The clinical data of patients with severe congenital cervical kyphosis (Cobb>40°) treated in Peking University Third Hospital from March 2004 to March 2018 were retrospectively summarized. In this series, 8 cases were enrolled, included 4 males and 4 females; the patients were 5-45 years old. According to the etiology, 4 patients were diagnosed with vertebral body underdevelopment, 2 with vertebral insufficiency, 1 with cervical spine congenital fusion and 1 with C(2) spinous process mecism. Five cases were treated with traction before final surgical correction. The surgical strategy was anterior correction or posterior correction or combined procedure in regards to different situation. The curvature of cervical angle was measured by two-line Cobb method, and the cervical kyphosis angle was measured on lateral radiographs in the neutral and extended position at the pre-operation and post-operation in each patient. The correction rate and evaluated Japanese Orthopedic Association (JOA) scoring for the function of spinal cord were also measured. The data before and after the operation were compared with t test. Results: In this series, the average kyphotic Cobb angle was 67°±18° and 8°±8° before and after surgical correction, respectively (t=8.471,P<0.05).The final correction rate was 87%±13%.The JOA score improved from 11.1±2.7 to 14.0±1.5 (t=-2.656, P<0.05) at the end of follow up. Conclusions: The pre-correction by cervical spine traction and final surgical correction by anterior, posterior or combined approaches of internal fixation and fusion can achieve good results and reduce risk and difficulty in operation for severe congenital cervical spine kyphosis with vertebral body underdevelopment without vertebral insufficiency and cervical spine congenital fusion. The final surgical correction by one-stage anterior, posterior or combined approaches of internal fixation and fusion can achieve good results for severe congenital cervical spine kyphosis with vertebral insufficiency and/or cervical spine congenital fusion.

Fusarium incarnatum-equiseti complex from China.

The Fusarium incarnatum-equiseti species complex (FIESC) is shown to encompass 33 phylogenetic species, across a wide range of habitats/hosts around the world. Here, 77 pathogenic and endophytic FIESC strains collected from China were studied to investigate the phylogenetic relationships within FIESC, based on a polyphasic approach combining morphological characters, multi-locus phylogeny and distribution patterns. The importance of standardised cultural methods to the identification and classification of taxa in the FIESC is highlighted. Morphological features of macroconidia, including the shape, size and septum number, were considered as diagnostic characters within the FIESC. A multi-locus dataset encompassing the 5.8S nuclear ribosomal gene with the two flanking internal transcribed spacers (ITS), translation elongation factor (EF-1α), calmodulin (CAM), partial RNA polymerase largest subunit (RPB1) and partial RNA polymerase second largest subunit (RPB2), was generated to distinguish species within the FIESC. Nine novel species were identified and described. The RPB2 locus is demonstrated to be a primary barcode with high success rate in amplification, and to have the best species delimitation compared to the other four tested loci.

[Midterm clinical outcomes and radiological results of surgical treatment for Hirayama disease].

OBJECTIVE: To assess the midterm clinical and radiological outcomes of internal fixation and fusion for the treatment of Hirayama disease and to evaluate the clinical significance and value of this procedure. METHODS: In the study, 36 patients were treated with anterior cervical internal fixation and fusion. The clinical outcomes including muscle strength and atrophy were recorded. The radiological outcomes including range of motion of cervical spine and the cross-sectional area of spinal cord at each level on MRI scan were measured before and at 3 month, 1 year and 2 years follow-up time points after surgery. RESULTS: (1) Clinical outcomes: all the patients showed no further progression of symptoms except one patient with mild progression of muscular weakness and atrophy. As the time passed by, the ratio of the patients with muscle strength and atrophy improvement increased. There were 26.5% of patients in 3 months, 36.0% in 1 year and 85.7% in 2 years who experienced muscle strength improvement. 8.8% of patients in 3 months, 24.0% in 1 year and 35.8% in 2 years felt muscle atrophy improvement. And 12 of the 14 patients showed improved muscle strength and atrophy at the end of 2 years period follow-up. (2) Radiological outcomes: the range of motion (ROM) of C2-C7 was significantly decreased after the operation. The ROM of preoperation was 62.25°±2.10° and that of 2 years postoperation was 13.67°±7.51°(P<0.01). The spinal cord was of no compression on flexion MRI. The cross-section area of spinal cord on MRI was significantly increased only at C6 level (P<0.05) at the end of three months follow-up. The level of increased cross-section area rose to C4-C5-C6 levels (P<0.01) in 1 year and to C4-C5-C6-C7 levels at the end of 2 years follow-up (P<0.05). The cross-section area increased 15.60% at C4, 19.08% at C5, 21.60% at C6 and 23.91% at C7 with significant difference (P<0.05) 2 years after the operation. CONCLUSION: Anterior cervical internal fixation and fusion is an effective surgical treatment for Hirayama disease and may provide preferable midterm clinical and radiological outcomes. This procedure has clinical significance and value in terms of control of the progression and outcome of this disease.

Colletotrichum species causing anthracnose disease of chili in China.

Anthracnose caused by Colletotrichum species is a serious disease of more than 30 plant genera. Several Colletotrichum species have been reported to infect chili in different countries. Although China is the largest chili-producing country, little is known about the species that have been infecting chili locally. Therefore, we collected samples of diseased chili from 29 provinces of China, from which 1285 strains were isolated. The morphological characters of all strains were observed and compared, and multi-locus phylogenetic analyses (ITS, ACT, CAL, CHS-1, GAPDH, TUB2, and HIS3) were performed on selected representative strains. Fifteen Colletotrichum species were identified, with C. fioriniae, C. fructicola, C. gloeosporioides, C. scovillei, and C. truncatum being prevalent. Three new species, C. conoides, C. grossum, and C. liaoningense, were recognised and described in this paper. Colletotrichum aenigma, C. cliviae, C. endophytica, C. hymenocallidis, C. incanum, C. karstii, and C. viniferum were reported for the first time from chili. Pathogenicity of all species isolated from chili was confirmed, except for C. endophytica. The current study improves the understanding of species causing anthracnose on chili and provides useful information for the effective control of the disease in China.

First Report of Colletotrichum truncatum Causing Anthracnose of Tomato in China.

Colletotrichum truncatum (syn. C. capsici) is an important plant pathogen that has a wide host range including pepper, eggplant, muskmelon, chickpea, grapes, and many other species of plants (1,2). Anthracnose fruit rot incited by C. coccodes is a prevalent disease in some major tomato (Solanum esculentum) producing regions in China. In October 2012, anthracnose symptoms (circular, sunken lesions with spore masses produced in black acervuli) were observed on ripe tomato fruit in Fuzhou City of Fujian Province, China. Three single-spore isolates (FZ-1, FZ-2, and FZ-3) were derived from fungal cultures isolated from different infected fruits from the same farm. A mycelial plug (5 mm in diameter) from the growing edge of an active colony of each isolate was transferred onto potato dextrose agar (PDA) and incubated at 28°C. Colonies grown on PDA changed from grayish to dark grey with an average colony diameter of 71.8 mm after 7 days. Conidia were falcate and 17.6 to 21.6 × 2.57 to 3.31 µm. Growth rate measured by colony diameter was greater at 25 to 30°C than at other temperatures tested (12, 18, 20, and 37°C). Based upon these morphological and cultural characteristics, the causal agent was identified as C. truncatum (3). To test the three isolates for pathogenicity, detached ripe tomato fruits were each inoculated with 1 µl of a conidial suspension (106 conidia/ml) using either injection or applying a droplet of the spore suspension on the surface of each fruit; the control treatment consisted of fruit that was treated with 1 µl of sterilized water using the two methods of inoculation mentioned above. Five replicate ripe fruits were inoculated with each of the three isolates using each method described above, and incubated in a moist chamber at 25°C. Five ripe fruits were used the negative control treatment for each inoculation method. After 7 days, typical anthracnose symptoms had developed on the inoculated fruit but not on control fruit. To confirm identity of the three isolates, the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, a partial sequence of the actin (ACT) gene, and partial histone 3 (HIS3) gene were amplified with ITS1 and ITS4 universal primers, GDF1 and GDR1 primers, ACT-512F and ACT-783R primers, and CYLH3F and CYLH3R primers, respectively (1). The ITS consensus sequence (Accession No. KC460308) of these isolates shared 99% homology with the ITS sequence of C. truncatum in GenBank (AJ301945), and the three other consensus sequences were all 100% homologous with the corresponding sequences of C. truncatum in Genbank (GU228254, GU227960, and GU228058, respectively). The pathogen was re-isolated from the inoculated fruit but not from the control fruit, and the identity of the re-isolates was confirmed to be C. truncatum by morphological features and based on the ITS, GAPDH, ACT, and HIS3 sequences as described above. To our knowledge, this is the first report of C. truncatum causing anthracnose on tomato in China. References: (1) U. Damm et al. Fungal Divers. 39:45, 2009. (2) K. K. Pandey. J. Mycol. Plant Pathol. 36:104, 2006. (3) I. S. Sawant et al. New Dis. Rep. 25:2, 2012.

First Report of Colletotrichum boninense Causing Anthracnose on Pepper in China.

Anthracnose, caused by Colletotrichum spp., is a severe disease and results in large losses in pepper (Capsicum frutescens) production in China (4). Colletotrichum boninense is one of the Colletotrichum species in pepper in China. In August 2011, anthracnose symptoms (circular, sunken lesions with orange to black spore masses) were observed on pepper fruits in De-Yang, Sichuan Province, China. Three single-spore isolates (SC-6-1, SC-6-2, SC-6-3) were obtained from the infected fruits. A 5-mm diameter plug was transferred to potato dextrose agar (PDA); the isolates formed colonies with white margins and circular, dull orange centers. The conidia were cylindrical, obtuse at both ends, and 10.5 to 12.6 × 4.1 to 5.0 µm. The colonies grew rapidly at 25 to 28°C, and the average colony diameter was 51 to 52 mm after 5 days on PDA at 25°C. Based upon these characters, the causal agent was identified as C. boninense. To confirm the identity of the isolates, the internal transcribed spacer (ITS) regions were amplified with the ITS1/ITS4 universal primers (1). The internal transcribed spacer (ITS) sequences (Accession No. JQ926743) of the causal fungus shared 99 to 100% homology with ITS sequences of C. boninense in GenBank (Accession Nos. FN566865 and EU822801). The identity of the causal agent as C. boninense was also confirmed by species-specific primers (Col1/ITS4) (2). In a pathogenicity test, five detached ripe pepper fruits were inoculated with 1 µl of a conidial suspension (106 conidia/mL) or five fruits with 1 µl of sterile water were kept as control. After 7 days in a moist chamber at 25°C, typical anthracnose symptoms had developed on the five inoculated fruits but not on control fruits. C. boninense was reisolated from the lesions, and which was confirmed by morphology and molecular methods as before. There have reports of C. boninense infecting many species of plants, including pepper (3). To our knowledge, this is the first report of C. boninense causing anthracnose on pepper in China. References: (1) A. K. Lucia et al. Phytopathology 93:581, 2002. (2) S. A. Pileggi et al. Can. J. Microbiol. 55:1081, 2009. (3) H. J. Tozze et al. Plant Dis. 93:106, 2009. (4) M. L. Zhang. J. Anhui Agri. Sci. 2:21, 2000.