Objective:To identify and characterize one Spiroplasma strain (designated as DGKH1) isolated from the blood of a patient with sepsis. Methods:The traditional bacterial culture, staining, morphological observation, physiological and biochemical identification, 16S rRNA gene sequencing, phylogenetic analysis, genome sequencing, and the genome-related index analysis were performed to accurately determine the taxonomic status of the strain DGKH1. Antibiotic susceptibility testing was performed using a specific kit for culturing and testing Ureaplasma urealyticum/ Metamycoplasma hominis. Results:The strain DGKH1 could weakly grow on Columbia blood agar, chocolate agar, and Haemophilus chocolate 2 agar. However, it did not grow in liquid culture medium containing tetracycline (4 μg/ml), doxycycline (1 μg/ml), minocycline (1 μg/ml), josamycin (2 μg/ml), roxithromycin (1 μg/ml), clarithromycin (1 μg/ml), or telithromycin (1 μg/ml). DGKH1 resembling Metamycoplasma hominis formed "fried egg-like colonies" on Mycoplasma solid culture medium. DGKH1 could not be stained by Gram staining. When observed under transmission electron microscopy (TEM) using phosphate buffer as the matrix, the bacteria were spiral-shaped. Results of 16S rRNA gene sequence alignment showed that DGKH1 was highly similar (99.85%) to Spiroplasma eriocheiris CCTCC M 207170 T. However, the urea decomposition test was positive, which was different from all of the known Spiroplasma species. The phylogenetic analysis based on whole genome showed that DGKH1 was clustered in a small branch along with Spiroplasma eriocheiris CCTCC M 207170 T. However, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the two strains were 94.14% and 56.00%, respectively, both below the threshold for prokaryotic species identification. Conclusions:DGKH1 represented a potential new species of genus Spiroplasma, closely related to Spiroplasma eriocheiris. Some microbiological characteristics of DGKH1 were similar to Mycoplasmas. However, the natural host and epidemiological data of DGKH1 need to be further studied.

Objective:To analyze the biological characteristics, phylogenic features and clinical significance of SQ219 and SQ220 isolated from clinical sputum and midstream urine specimens.Methods:The culture and biochemical characteristics of the two strains were observed. VITEK2 System, drug sensitivity testing and MALDI-TOF mass spectrometry were used for bacterial identification. Phylogenetic analysis based on 16S rRNA and core genome was performed. The average nucleotide identity (ANI) based on whole genome sequences was calculated.Results:SQ219 and SQ220 were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and non-motile bacteria. Their optimum growth was observed in NaCl-free medium at 30℃ and pH7. Flexirubin-type pigments were produced by SQ220 on Colombia blood agar, but not by SQ219. Both SQ219 and SQ220 were resistant to aztreonam, amikacin, tobramycin and colistin, which was consistent with the drug resistance phenotype of genus Chryseobacterium. The genome sequences of SQ219 and SQ220 were 5.08 Mb and 4.80 Mb in length, and the G+ C contents were 36.72% and 36.36%, respectively. Both strains carried β-lactam resistance gene ( blaCGA). 16S rRNA phylogenetic analysis showed that SQ219 and SQ220 were closely related to Chryseobacterium gambrini DSM18014 T with the similarities of 98.93% and 98.36%, respectively. Core genome phylogenetic analysis revealed that SQ219 and SQ220 were highly homologous to Chryseobacterium gambrini DSM18014 T. However, the ANI values between the two strains and Chryseobacterium gambrini DSM18014 T were 92.49% and 93.27%, respectively, below the threshold for prokaryotic species identification. Conclusions:Based on the phenotypic and phylogenetic data, SQ219 and SQ220 represent a novel species of the genus Chryseobacterium. This study would help promote the understanding of the evolution of Chrysobacterium and provide reference for the identification of new species of Chrysobacterium.

Objective: To investigate the oxidative stress injury of nano zinc oxide nanoparticles(ZnO NPs) on human myocardial cells (AC16) ，and to analyze the mechanism of ZnO NPs from the transcriptome level． Methods: Dynamic light scattering (DLS) was used to characterize and detect ZnO NPs．After AC16 cells were exposed to ZnO NPs at different doses and at different times，the cell survival rate was determined by CCK-8 method．AC16 cells were divided into control group，ZnO NPs (50，100，200 μmol /L) ，after 6 h treatment，the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were measured．AC16 cells were divided into control group，50 μmol /L ZnO NPs group and 200 μmol /L ZnO NPs group．After 6 h exposure，total RNA was extracted by TRIzol for transcriptome analysis ，and the differentially expressed genes were enriched by gene body ( GO) ，Kyoto Encyclopedia of Genes and Genomes (KEGG) . Results : The results of DLS showed that the hydrodynamic diameter was ( 192. 2 ± 1. 63 ) nm and the Zeta potential was ( －23. 26 ± 1. 05 ) mV． CCK-8 results showed that the survival rate of AC16 cells decreased with the increase of dose and time of exposure to ZnO NPs. Fluorescence quantification showed that with the increase of ZnO NPs exposure dose，MMP significantly decreased at 100 μmol /L ZnO NPs(P＜0. 05) ，and ROS significantly increased at 50 μmol /L ZnO NPs(P＜0. 05) ．Using the multifunctional microplate reader，it was observed that MMP and ROS were statistically significant at 100 and 50 μmol /L ZnO NPs，respectively，showing a decrease in MMP and an increase in ROS．Transcriptome analysis showed that 1 071 genes were enriched in the 50 μmol /L ZnO NPs group compared with the control group，including 561 up-regulated genes and 510 down-regulated genes．Compared with the control group，7 164 genes were enriched in 200 μmol /L ZnO NPs group，including 4 098 up-regulated genes and 3 066 down-regulated genes．GO and KEGG analysis showed that the differential genes were mainly concentrated in ROS，antioxidant activity，mitochondrial cytochrome C release，apoptosis and other signaling pathways． Conclusion ZnO NPs can decrease the survival rate of AC16 cells and induce mitochondrial damage and oxidative stress，among which ROS-mediated oxi- dative stress and mitochondrial function changes are important toxic mechanisms of ZnO NPs induced AC16 cytotoxicity.

Objective:To investigate the value of the vesical imaging reporting and data system score (VI-RADS) in the prognostic assessment of patients with bladder cancer.Methods:The data of 294 patients with pathologically confirmed bladder cancer in our department from February 2012 to September 2019 were retrospectively analyzed. Divide the patients into two groups based on the surgical method. In the transurethral resection of bladder tumor (TURBT) group, there were 121 cases, 102 males and 19 females; The average age of the patients was (66.7±12.3) years old, 52 cases <65 years old, 69 cases ≥65 years old, with VI-RADS <3 in 84 cases, VI-RADS ≥3 in 37 cases. In the radical cystectomy (RC) group, there were 173 cases, including 154 males and 19 females; The average age of the patients was (65.7±10.8) years, 77 cases <65 years old, 96 cases ≥65 years old, with VI-RADS <3 in 51 cases and VI-RADS ≥3 in 122 cases. The prognostic factors of TURBT group and RC group were analyzed, and the predictive value of VI-RADS score on overall survival (OS) and progression free survival (PFS) of bladder cancer patients after surgery was analyzed.Results:In this study, there were 294 cases with postoperative pathological diagnosis of urothelial carcinoma. The pathological staging was Ta stage in 104 cases (35.4%), T 1stage in 82 cases (27.9%), T 2 stage in 58 cases (19.7%), T 3 stage in 34 cases (11.6%), and T 4stage in 16 cases (5.4%). Pathological grading: 11 cases (3.7%) were low malignant potential, 77 cases (26.2%) were low grade, and 206 cases (70.1%) were high grade. There were 186 cases (63.3%) in the NMIBC group and 108 cases (36.7%) in the MIBC group. In the TURBT group, there were 114 cases (94.2%) in the NMIBC group and 7 cases (5.8%) in the MIBC group; In the RC group, there were 72 cases (41.6%) in the NMIBC group and 101 cases (58.4%) in the MIBC group. In the NMIBC group, the VI-RADS<3 and ≥3 were 128 cases (68.8%) and 58 cases (31.2%), respectively ( P<0.01); In the MIBC group, 101 cases (93.5%) had a VI-RADS <3 and 7 cases (6.5%) had a VI-RADS ≥ 3, respectively ( P<0.01). In the high grade of postoperative pathological group, 62 cases (30.1%) had a VI-RADS <3 and 144 cases (69.9%) had a VI-RADS ≥ 3, respectively ( P<0.01); In the non-high grade of postoperative pathological group, the VI-RADS <3 and ≥ 3 were 73 cases (83.0%) and 15 cases (17.0%), respectively ( P<0.01). The median OS survival for all patients in this study was 27.4 (16.6, 38.1)months and the median PFS survival was 24.7(14.0, 36.8) months. The results of univariate analysis showed that age ≥ 65 years old (OS: HR=6.09, P=0.001; PFS: HR=1.71, P=0.035), postoperative pathological diagnosis of tumor muscle infiltration (OS: HR=4.66, P<0.01; PFS: HR=2.24, P=0.001), postoperative high-grade tumor (OS: HR=4.26, P=0.008; PFS: HR=1.92, P=0.023), and VI-RADS score ≥ 3 (OS: HR=4.24, P=0.001; PFS: HR=2.21, P=0.002) were associated with poorer OS and PFS in patients. Multifactorial Cox model analysis revealed that a score of VI-RADS ≥3 was an independent risk factor for OS ( HR=3.41, P=0.012) and PFS ( HR=2.23, P=0.016). In the TURBT group, univariate analysis found that VI-RADS ≥3 ( HR=2.05, P=0.053) and high grade of postoperative pathology ( HR=2.77, P=0.005) were associated with poor PFS in patients, multifactorial Cox model analysis found only high grade of postoperative pathology ( HR=2.54, P=0.013) to be an independent risk factor for PFS. In the RC group, VI-RADS ≥3 ( HR=3.29, P=0.032) and age ≥65 years ( HR=5.37, P=0.001) were found to be independent risk factors for OS. The survival curve showed that the 5-year OS rates for groups with a VI-RADS ≥ 3 and <3 were 93.9% and 73.1%, respectively ( P<0.01), and the 5-year PFS rates for groups with a VI-RADS ≥ 3 and <3 were 76.5% and 53.0%, respectively ( P<0.01), with statistically significant differences. Conclusions:This study showed that VI-RADS ≥3 was an independent risk factor for prognosis in patients with bladder cancer and was more significant in patients receiving RC, but was not a significant predictor of prognosis in patients receiving TURBT.

Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-

Objective:To investigate the short-term outcomes of laparoscopic simultaneous resection of primary colorectal cancer and liver metastases in patients with resectable synchronous colorectal liver metastases (SCRLM).Methods:A descriptive case series study was performed. Clinicopathological data of patients with SCRLM who underwent laparoscopic simultaneous resection of colorectal cancer and liver metastases in Zhongshan Hospital between December 2015 and September 2018 were retrieved from a prospective colorectal cancer database. Perioperative presentations and short-term outcomes were analyzed.Results:A total of 53 patients were enrolled with average age of(61.7±11.3) years. Among them, 32 were male (60.4%) and 21 were female (39.6%). Twenty-five patients (47.2%) were American Society of Anesthesiologists (ASA) grade I and 28 (52.8%) were grade II. All the patients completed laparoscopic simultaneous resection without conversion. The average operation time was (320.2±114.5) min. The estimated blood loss was 150.0 (45.0-2000.0) ml, and only 2 cases (3.8%) received intraoperative transfusion. Postoperative pathologic results revealed that the average primary tumor size was (5.4±1.9) cm; 4 cases (7.5%) were T1-2 stage and 48 cases (90.6%) were T3-4 stage; 40 patients (75.5%) had lymph node metastasis; 19 (35.8%) had vascular involvement; 24 (45.3%) had neural invasion. The median number of liver metastases was 1.0 (1-8), and the average size of largest liver metastases was (3.0±1.9) cm. The median margin of liver metastases was 1.0 (0.1-3.5) cm, and only 1 case was R1 resection. The average time to the first postoperative flatus was (67.9±28.9) h, and the average time to the liquid diet was (107.0±33.8) h. The average postoperative indwelling catheterization time was (85.6±56.4) h. The average postoperative hospital stay was (9.2±4.4) d, and the average cost was (82±26) thousand RMB. No death within postoperative 30-day was found. The morbidity of postoperative complication was 32.1% (17/53) and 3 patients developed grade III to IV complications which were improved by conservative treatment. The median follow-up period was 23.2 months. During follow-up, 19 patients (35.8%) developed recurrence or metastasis, and 4 (7.5%) died. The 1- and 2-year disease-free survival (DFS) rates were 68% and 47% respectively, and the 1- and 2-year overall survival rates were 95% and 86% respectively.Conclusions:Laparoscopic simultaneous resection of primary colorectal cancer and liver metastases is safe and feasible in selected patients with SCRLM. Postoperative intestinal function recovery is enhanced, and morbidity and oncological outcomes are acceptable.

Objective:To investigate the short-term outcomes of laparoscopic simultaneous resection of primary colorectal cancer and liver metastases in patients with resectable synchronous colorectal liver metastases (SCRLM).Methods:A descriptive case series study was performed. Clinicopathological data of patients with SCRLM who underwent laparoscopic simultaneous resection of colorectal cancer and liver metastases in Zhongshan Hospital between December 2015 and September 2018 were retrieved from a prospective colorectal cancer database. Perioperative presentations and short-term outcomes were analyzed.Results:A total of 53 patients were enrolled with average age of(61.7±11.3) years. Among them, 32 were male (60.4%) and 21 were female (39.6%). Twenty-five patients (47.2%) were American Society of Anesthesiologists (ASA) grade I and 28 (52.8%) were grade II. All the patients completed laparoscopic simultaneous resection without conversion. The average operation time was (320.2±114.5) min. The estimated blood loss was 150.0 (45.0-2000.0) ml, and only 2 cases (3.8%) received intraoperative transfusion. Postoperative pathologic results revealed that the average primary tumor size was (5.4±1.9) cm; 4 cases (7.5%) were T1-2 stage and 48 cases (90.6%) were T3-4 stage; 40 patients (75.5%) had lymph node metastasis; 19 (35.8%) had vascular involvement; 24 (45.3%) had neural invasion. The median number of liver metastases was 1.0 (1-8), and the average size of largest liver metastases was (3.0±1.9) cm. The median margin of liver metastases was 1.0 (0.1-3.5) cm, and only 1 case was R1 resection. The average time to the first postoperative flatus was (67.9±28.9) h, and the average time to the liquid diet was (107.0±33.8) h. The average postoperative indwelling catheterization time was (85.6±56.4) h. The average postoperative hospital stay was (9.2±4.4) d, and the average cost was (82±26) thousand RMB. No death within postoperative 30-day was found. The morbidity of postoperative complication was 32.1% (17/53) and 3 patients developed grade III to IV complications which were improved by conservative treatment. The median follow-up period was 23.2 months. During follow-up, 19 patients (35.8%) developed recurrence or metastasis, and 4 (7.5%) died. The 1- and 2-year disease-free survival (DFS) rates were 68% and 47% respectively, and the 1- and 2-year overall survival rates were 95% and 86% respectively.Conclusions:Laparoscopic simultaneous resection of primary colorectal cancer and liver metastases is safe and feasible in selected patients with SCRLM. Postoperative intestinal function recovery is enhanced, and morbidity and oncological outcomes are acceptable.

Objective:To evaluate the safety and efficacy of eltrombopag combined with immunosuppressive therapy in patients with aplastic anemia (AA) in China.Methods:We investigated and analyzed the clinical data of AA patients from 14 hematological treatment centers who were treated with oral eltrombopag for at least 3 mon.Results:We enrolled 56 AA patients, including 19 treatment-na?ve patients and 37 IST-refractory patients. The median administration period for eltrombopag was 7 (3-31) months, and the median maximum stable dosage was 75 mg/d (50-150 mg/d) . The 3-month hematological response (HR) rate was 60%, and the complete response (CR) rate was 30% in 10 SAA patients who were treated with first-line eltrombopag and standard IST (ATG+CsA) . Eight of 9 eltrombopag and CsA ± androgen first-line treated SAA patients responded (8/9, 89%) and 4 (44%) gave CR. The overall HR and CR rates were 79% and 52.6%, respectively, among these 19 patients by the end of the follow-up period. Of the 19 AA patients who were refractory to CsA ± androgen, 11 achieved HR (57.9%) at 3 mon, and the best HR rate was 44% in standard IST (ATG+CsA) refractory 18 patients after eltrombopag treatment. Fifty-one percent of the patients experienced mild or moderate adverse events, and gastrointestinal discomfort was the most common adverse effect reported by the study subjects.Conclusion:Adding Eltrombopag in first-line IST can accelerate the acquisition and improve the quality of hematological responses in AA patients. AA with relatively more residual hematopoietic cells may be well treated with eltrombopag and non-ATG IST. Eltrombopag can be used as salvage therapy for CsA±androgen refractory patients. Eltrombopag was generally safe and well tolerated by AA patients in China.

Objective To study the use of preoperative indocyanine green retention test at 15 minutes (ICG R15) in the prediction of liver functional reserve in patients with hilar cholangiocarcinoma (HCCA).Methods The clinical data of 62 patients with HCCA treated in our department from March 2016 to March 2018 was reviewed.The relationship between preoperative ICG R15 and postoperative hepatic insufficiency was analyzed.The relationship between preoperative ICG R15 and Child-Pugh scoring was also studied.Univariate analysis was used to evaluate the risk factors of postoperative liver dysfunction.Logistic regression was used to assess the independent risk factors of postoperative liver dysfunction.The regression equation between independent risk factors and postoperative liver dysfunction was established.Results Among the 62 patients,ICG R15 was less than 10.0％ in 26 patients,between 10.0％ and 19.0％ in 17 patients,between 20.0％ and 29.0％ in 9 patients,between 30.0％ and 39.0％ in 5 patients,and over 40.0％ in 5 patients.There were 29 patients with a Child-Pugh A grading and 33 patients with a Child-Pugh B grading in the preoperative evaluation of liver function.The Wilcoxon W rank sum test was used to compare the preoperative ICG R15 in patients with Child-Pugh grading A and B separately.The ICG R15 in Child-Pugh grading A patients was significantly lower than those in Child-Pugh B grading patients (P ＜0.05).There were no significant differences in age,gender,history of previous liver diseases,duration of operation,and intraoperative blood loss (P ＞ 0.05) between the normal liver function group and the liver dysfunction group.However,there was a significant difference in the preoperative ICG R15 and preoperative bilirubin levels (P ＜ 0.05) between the two groups.The preoperative ICG R15 and preoperative bilirubin levels were significant risk factors of postoperative hepatic insufficiency.Regression analysis suggested that preoperative ICG R15 level was an independent risk factor of postoperative hepatic insufficiency (P ＜ 0.05).A regression equation:logit(P) =0.185 × preoperative ICG R15-3.152 could be constructed.Conclusions ICG R15 is an ideal clinical indicator for evaluation of preoperative liver functional reserve in patients with HCCA.It predicted the recovery of postoperative liver function.

Objective To investigate dermoscopic features of onychomycosis and nail psoriasis.Methods From March 2017 to March 2018,128 patients with onychomycosis and 128 with nail psoriasis were enrolled from Department of Dermatology,Xiangyang Central Hospital.Dermoscopic images of the lesional nails were compared.Enumeration data were analyzed by chi-square test and Fisher's exact test.Results There were 7 kinds of dermoscopic patterns in onychomycosis,including jagged edge (69/128,53.91％),longitudinal striae (72/128,56.25％),cone-shaped keratosis (30/128,23.44％),nail plate thickening (21/128,16.41％),black dots (5/128,3.91％),onycholysis (26/128,20.31％),and nail discoloration (23/128,17.97％).The jagged edge and longitudinal striae occurred more frequently in distal/ lateral subungual onychomycosis (DLSO) than in the other 3 clinical subtypes of onychomycosis (all P ＜ 0.001),and cone-shaped keratosis occurred more frequently in total dystrophic onychomycosis (TDO) than in the other 3 clinical subtypes of onychomycosis (x2 =42.020,P ＜ 0.001).There were no significant differences in the prevalance of nail plate thickening,black dots,onycholysis and nail discoloration among the 4 clinical subtypes of onychomycosis (all P ＞ 0.05).There were 7 kinds of dermoscopic patterns in nail psoriasis,including pitting corrosion (61/128,47.66％),oil drop sign (41/128,32.03％),splinter hemorrhage (41/128,32.03％),dilated hyponychial capillaries (23/128,17.97％),nail plate thickening (20/128,15.63％),onycholysis (20/128,15.63％),and nail discoloration (13/128,10.16％).The oil drop sign occurred more frequently in psoriasis vulgaris and psoriasis arthropathica than in the other 2 clinical subtypes of psoriasis (P =0.019).The dilated hyponychial capillaries occurred more frequently in pustular psoriasis than in the other 3 clinical subtypes of psoriasis (P =0.047).There were no significant differences in the prevalance of pitting corrosion,splinter haemorrhage,nail plate thickening and nail discoloration among the 4 clinical subtypes of psoriasis (all P ＞ 0.05).Conclusions The dermoscopic features of onychomycosis include jagged edge,longitudinal striae,cone-shaped keratosis and black dots,and the dermoscopic features of nail psoriasis include pitting corrosion,oil drop sign,splinter haemorrhage and dilated hyponychial capillaries.Their common dermoscopic features include nail plate thickening,onycholysis and nail discoloration.