Background: Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans. Objectives: In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood.The antifungal mechanism of KTZ needs to be studied in detail. Methods: In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis.The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto. Results: At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ. Conclusions The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.

Objective:To study the independent risk factors of tumor recurrence after radiofrequency ablation (RFA) in patients with hepatocellular carcinoma (HCC), and to establish a preoperative prediction score.Methods:A retrospective study was conducted on 168 HCC patients treated with RFA at Zhongshan Hospital affiliated to Sun Yat-sen University from June 2016 to September 2019. The X-tile software was used to determine the optimal cut-off value of preoperative circulating tumor cells (CTC) which was then used to analyze the relationship between different CTCs values with various clinical factors. The Cox regression model was used to analyze independent risk factors of recurrence after RFA, and each independent risk factor was assigned a score of 1 to compose the prediction score. The patients were divided into the low-risk group (0-2 scores), intermediate-risk group (3 scores) and high-risk group (4-5 scores). The Kaplan-Meier method was used to draw cumulative recurrence curves in calculating the cumulative recurrence rates of the 3 different groups.Results:Of 168 patients, there were 151 males and 17 females. Their age (Mean±SD) was 58.33±9.53 years. CTC≥1/3.2 ml was detected in 131 patients (77.98%) (range 0-20/3.2 ml). The X-tile software determined the preoperative CTC cut-off value of HCC patients to be 2/3.2ml which separated a CTC-negative group with 93 patients, and a positive group of 75 patients. On analyses, the relationship between preoperative CTC and various preoperative clinical parameters were related to number of tumor nodules, tumor maximum diameter and alpha-fetoprotein (AFP) levels ( P<0.05). Multivariate analysis showed that CTC positivity[ HR(95% CI): 1.990(1.332-2.974)], AFP>20 ng/ml[ HR(95% CI): 1.659(1.111-2.477)], PIVKA-II>40 mAU/ml[ HR(95% CI): 1.580 (1.022-2.443)], number of tumor nodules ≥2[ HR(95% CI): 1.568 (1.057-2.326)], and tumor diameter>30 mm[ HR (95% CI): 1.544 (1.007-2.369)] were independent risk factors of recurrence ( P<0.05) after RFA in HCC patients. The cumulative recurrence rates of patients at 6 months, 12 months, and 18 months were 14.9%, 35.6%, and 56.4% in the low-risk group, 38.9%, 70.5%, and 85.0% in the intermediate-risk group, and 64.5%, 84.5% and 100% in the high-risk group. The differences were significant ( P<0.05). Conclusion:Preoperative CTC positivity, AFP>20 ng/ml, PIVKA-II>40 mAU/ml, tumor nodules ≥2, and tumor diameter>30 mm were independent risk factors of recurrence after RFA in HCC patients. This preoperative predictive score could be used to guide clinical treatment strategies.

Objective:To analyze the risk factors of hepatocellular carcinoma microvascular invasion (MVI) and to construct a preoperative prediction clinical scoring system.Methods:A retrospective analysis was made on 113 patients with hepatocellular carcinoma undergoing hepatectomy at Zhongshan Hospital from March 2018 to Jun 2019.Postoperative pathology confirmed 35 cases with microvascular invasion.Results:The multivariate logistic regression model showed that the maximum tumor diameter( OR: 1.028, 95% CI: 1.001-1.005), the smoothness of the capsule edge( OR: 0.208, 95% CI: 0.062-0.699), the positive circulating tumor cells (CTC)( OR: 3.728, 95% CI: 1.029-13.501) and abnormal prothrombin(PIVKA-Ⅱ)( OR: 1.001, 95% CI: 1.000-1.002) were risk factors for MVI. The area, sensitivity and specificity of the clinical score constructed by assigning 1 point to each risk factor were 0.906, 74.29% and 92.31%, respectively. Clinical scores of 0, 1, 2, 3, and 4 predict MVI positive rates of 0 (0/26), 9.09% (3/33), 28.57% (6/21), 77.78% (14/ 18), 85.71% (12/14). Conclusions:Tumor maximum diameter>62 mm, PIVKA-Ⅱ>115 mAU/ml, unsmooth tumor capsule and CTC in peripheral blood are independent high risk factors in patients with MVI.

Objective:To explore the application value of detecting circulating tumor cells (CTC) before liver transplantation for predicting the recurrence and survival of hepatocellular carcinoma (HCC).Methods:From October 2015 to October 2019, 62 HCC patients at Affiliated Zhongshan Hospital were collected and analyzed by Cyttel method before liver transplantation. CTC was determined by X-tile software and Kaplan-Meier method for determining the optimal cutoff value of CTC before liver transplantation and the relationship between CTC and clinical factors was analyzed. Univariate and multivariate COX regression analyses were performed for determining the independent risk factors affecting the prognosis. Kaplan Meier method was employed for describing the survival curve of tumor-free survival and overall survival after transplantation.Results:The optimal preoperative critical value of CTC was 3.2 ml. CTC ≥3/3.2 mL was set as CTC positive group while CTC <3/3.2 mL CTC negative group. The positive/negative CTC before transplantation was significantly correlated with preoperative Alpha-fetoprotein(AFP) level, maximal tumor diameter, lymph node metastasis, liver transplantation criteria and degree of differentiation ( P<0.05). Univariate and multivariate COX regression models indicated that the number of preoperative CTC (HR: 1.262, 95%CI: 1.069-1.489, P=0.006) and microvascular invasion (HR: 2.657, 95%CI: 1.120-6.305, P=0.027) were independent risk factors for tumor-free survival after transplantation while microvascular invasion (HR: 3.738, 95%CI: 1.219-11.459, P=0.027) was the sole independent risk factor affecting the overall survival of HCC after transplantation. Statistically significant difference existed between preoperative CTC positive/negative and tumor recurrence or metastasis (no recurrence, intrahepatic recurrence, and distant metastasis)( χ2=7.790, P=0.020). The disease-free survival rates of 1/2/3-year CTC-negative/positive patients were 82.90%, 68.70%, 58.90% and 49.00%, 29.40%, 22.10%; the 1/2/3-year overall survival rates of preoperative CTC-negative/positive patients were 85.50%, 77.10%, 69.79% and 64.90%, 47.20%, 40.50% respectively. The disease-free survival curve of CTC-negative patients was significantly higher than that of CTC-positive counterparts ( P<0.001) and the overall survival curve of CTC-negative patients was significantly higher than that of CTC-positive counterparts ( P<0.005). Conclusions:Preoperative CTC detection has certain application value in evaluating the prognosis of liver cancer after liver transplantation, which has important clinical significance and application prospects.

Objective:To investigate the correlation between preoperative circulating tumor cells (CTC) and microvascular invasion (MVI) in patients with hepatocellular carcinoma.Methods:The data of 227 patients who underwent hepatocellular carcinoma resection in Zhongshan Hospital Affiliated to Sun Yat-sen University from January 2018 to March 2020 were retrospectively analyzed. The peripheral blood CTC was detected by Cyttel detection before operation. The relationship between preoperative peripheral blood CTC and clinical characteristics of patients was analyzed; the multivariate logistic regression model was used to analyze the independent risk factors for MVI; the receiver operating characteristic (ROC) curve was used to compare the efficacy of each independent risk factor in predicting the occurrence of MVI, and the relationship between CTC and MVI was clarified.Results:According to the ROC curve, the cut-off values for predicting MVI of CTC, alpha-fetoprotein (AFP), protein induced by vitamin K absence or antagonist Ⅱ (PIVKA-Ⅱ), and tumor long-axis diameter were 3 CTC/3.2 ml, 158 μg/L, 178 AU/L and 59 mm. CTC-positive group had ≥3 CTC/3.2 ml in peripheral blood, and CTC-negative group had <3 CTC/3.2 ml, and there were 117 and 110 cases in the two groups. The median AFP levels of preoperative CTC-positive group and CTC-negative group were 123.0 μg/L (0-20 000.0 μg/L) and 9.6 μg/L (0-18 676.0 μg/L), and the median tumor long-axis diameter was 50.0 mm (5.0-200.0 mm) and 36.0 mm (2.0-150.0 mm), the differences between the two groups were statistically significant (both P < 0.05). Before operation, AFP≥158 μg/L ( OR = 3.551, 95% CI 1.426-8.843, P = 0.006), PIVKA-Ⅱ≥178 AU/L ( OR = 12.250, 95% CI 4.384-34.231, P < 0.01), peripheral blood CTC ≥ 3 CTC/3.2 ml ( OR = 8.913, 95% CI 3.561-22.306, P < 0.01) and tumor long-axis diameter ≥59 mm ( OR = 3.250, 95% CI 1.339-7.885, P = 0.009) were independent risk factors for the occurrence of MVI; the area under the ROC curve (AUC) of these factors for predicting MVI was 0.752, 0.777, 0.857 and 0.743. CTC was more effective in predicting MVI than AFP and tumor long-axis diameter, and the differences were statistically significant (both P < 0.05). The efficacy of CTC in predicting MVI was slightly better than that of PIVKA-Ⅱ, but the difference was not statistically significant ( P > 0.05). Conclusion:CTC may be one of the important indicators of hepatocellular carcinoma MVI in clinical practice.

Objective:To study the value hyperthermic intraperitoneal chemoperfusion after ruptured and hemorrhage of hepatocellular carcinoma.Methods:A retrospective study was conducted on 53 patients with ruptured hepatocellular carcinoma treated in Zhongshan Hospital Affiliated to Sun Yat-sen University from January 1, 2009 to January 1, 2019.Patients who underwent surgical resection combined with hyperthermic intraperitoneal chemoperfusion were included into the experimental group. Those who underwent surgical treatment only were included into the control group. The clinical data, postoperative hospital stay, complications, long-term tumor-free survival and overall survival were analyzed. Independent risk factors affecting prognosis were also determined.Results:Of the 33 patients in the experimental group, there were 27 males and 6 females, with a mean age ± s. d. being 50.49±11.59 years. There were 20 patients in the control group, including 17 males and 3 females, with a mean ± s. d. of 53.70±13.89 years. There were no significant differences in the length of postoperative hospital stay and complication rates between the two groups ( P>0.05). The tumor-free survival rate of the experimental group was significantly higher than that of the control group ( P<0.05). There was no significant difference in overall survival rates between the two groups ( P>0.05). Cox multivariate analysis showed that histological classification [ HR(95% CI): 27.700(1.695-452.794); 42.754(2.091-874.034)] and hyperthermic intraperitoneal chemoperfusion [ HR(95% CI): 0.238(0.086-0.661); 0.205(0.069-0.611)] were independent risk factors affecting tumor-free survival and overall survival (all P<0.05). Conclusion:Hyperthermic intraperitoneal chemoperfusion after surgical resection for ruptured hepatocellular carcinoma is a safe and effective treatment.

Objective:To explore the effect of family doctor system on the risk stratification of community hypertensive population by application of Markov modelMethods:Retrospective investigation was conducted on hypertensive patients under continuing management from 13 community health service centers in Shanghai Xuhui District from January 2014 to December 2016. Among 98 996 subjects, 50 920 (51.45%) were contracted to family doctors (contracted group) and 48 046 (48.55%) did not contracted to family doctors (non-contracted group). According to the risk stratification of hypertension, the four-state Markov model (low-risk, medium-risk, high-risk, and extremely high-risk) was established. The prediction effect of the model was validated, and the changes in the risk stratification status of hypertension in the study subjects from 2017 to 2020 was predicted by using the Markov model.Results:Among all subjects the number of medium-risk and extremely high-risk accounted for the majority(>80%). Compared with 2014, in 2016 the number of low-risk patients with hypertension was decreased from 9 042 cases (17.76%) to 6 851 cases (13.45%) in contracted group; and from 9 971 cases (20.75%) to 7 906 cases (16.46%) in non-contracted group; the number of people at extremely high risk of hypertension was increased from 15 609 cases (30.65%) to 17 639 cases (34.64%) in the contracted group; from 13 847 cases (28.82%) to 15 641 cases (32.55%) in the non-contracted group. According to the Markov model one year after the risk stratification, the risk status of most subjects remained in the original one. There was not transform from extremely high-risk to low-risk state (0%), but there was transform from low-risk to extremely high-risk state in some extend, and the degree of transform in non-contracted group [2.06%(205/9 971)] was higher than the contracted group [1.85%(167/9 042)]. Predicted by the Markov model, between 2017 and 2020 the number and proportion of the medium-risk>extremely high-risk>low-risk>high-risk in both contracted group and non-contracted group. With the extension of time, low-risk proportion is gradually reduces, and the rate of reduction of the contracted group was lower than that of the non-contracted group, while proportion of medium-risk, high-risk and extremely high-risk is gradually increased.Conclusions:The constructed Markov model is accruable and stable, which can be used in the study of hypertension prognosis. The study indicates that the contracted services of family doctor have positive effects on the management of community hypertensive patients.

Objective: To investigate the expression of LC3B, p-AMPKα and p27 in cortical tuberous sclerosis complex (TSC). Methods: Nineteen specimens of surgically resected TSC cortical tubers were collected at Xuanwu Hospital, Capital Medical University, from 2014 to 2017. The expression of the three proteins in the lesions and the adjacent relatively normal regions was detected by immunohistochemical staining (EnVision two-step method). Results: LC3B was mainly expressed in the dysmorphic neuron and giant cell in TSC cortical tubers and in the adjacent relatively normal neurons, and the expression was diffuse or perinuclear cytoplasmic. There was no significant difference in the average optical density between abnormal cells and neurons adjacent to the lesions (0.343±0.195 vs. 0.419±0.088, P>0.05). p-AMPKα was localized in the cytoplasm of dysmorphic neurons and giant cell in TSC cortical tubers. The average optical density of abnormal cells in the lesions was significantly higher than that of neurons adjacent to the lesions (0.306±0.123 vs. 0.233±0.654, P<0.05). P27 showed nuclear positivity, mainly expressed in the neurons and glial cells close to TSC cortical tubers, while the positive rate in the abnormal cells in TSC cortical tubers was low (15/19 vs. 7/19, P<0.05). Conclusion There is no significant decrease in the level of autophagy in dysmorphic neurons and giant cells in TSC cortical tubers, which may be related to the compensatory mechanism of AMPK signaling pathway, but without activation of downstream p27.

investigate the expression of LC3B, p?AMPKα and p27 in cortical tuberous sclerosis complex (TSC). Methods Nineteen specimens of surgically resected TSC cortical tubers were collected at Xuanwu Hospital, Capital Medical University, from 2014 to 2017. The expression of the three proteins in the lesions and the adjacent relatively normal regions was detected by immunohistochemical staining (EnVision two?step method). Results LC3B was mainly expressed in the dysmorphic neuron and giant cell in TSC cortical tubers and in the adjacent relatively normal neurons, and the expression was diffuse or perinuclear cytoplasmic. There was no significant difference in the average optical density between abnormal cells and neurons adjacent to the lesions (0.343±0.195 vs. 0.419±0.088, P>0.05). p?AMPKα was localized in the cytoplasm of dysmorphic neurons and giant cell in TSC cortical tubers. The average optical density of abnormal cells in the lesions was significantly higher than that of neurons adjacent to the lesions (0.306 ± 0.123 vs. 0.233 ± 0.654, P<0.05). P27 showed nuclear positivity, mainly expressed in the neurons and glial cells close to TSC cortical tubers, while the positive rate in the abnormal cells in TSC cortical tubers was low (15/19 vs. 7/19, P<0.05). Conclusion There is no significant decrease in the level of autophagy in dysmorphic neurons and giant cells in TSC cortical tubers, which may be related to the compensatory mechanism of AMPK signaling pathway, but without activation of downstream p27.

Objective To analyze the gait kinematics parameters of patients with unilateral leg amputation after wearing prosthesis using the computer assisted rehabilitation environment (CAREN) gait assessment system,and the reasons for the differences.Methods Nine patients with unilateral mid-leg amputation were selected as the prosthesis group,and 11 healthy subjects were selected as the standard group.The gait kinematics parameters of the two groups were collected,processed and analyzed by using the CAREN gait evaluation system.Results The gait phase index of the prosthetic limb group was (0.88±0.04).Significant differences were observed between the prosthetic and healthy limbs in terms of step length,stance phase percentage,maximum hip extension angle and maximum knee flexion angle,maximum dorsiflexion and plantar flexion angle of the ankle joint during the stance phase,as well as the dorsiflexion angle of ankle joint during heel strike to the ground (P＜0.05).Moreover,there were significant differences between the affected limbs of the prosthetic limb group and limbs of the standard group in terms of the walking speed,gait cycle,stride length,percentage of stance phase,hip flexion angle,knee flexion and ankle dorsiflexion during heel strike,maximum hip extension and flexion angle,maximum dorsi-and plantar-flexion of ankle joint during stance phase (P＜0.05).Conclusion The relative symmetry of the gait of the unilateral leg amputee is (0.88±0.04),with their kinematics parameters of the prosthetic limb significantly weaker than those of the contralateral side and the healthy controls.