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 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 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 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 investigate the characteristics and risk factors of infection after liver transplantation from donor liver of donation after citizen's death. Methods Clinical data of 68 recipients after liver transplantation from donor liver of donation after citizen's death were analyzed retrospectively. The recipients were divided into infection group (33 cases) and non-infection group (35 cases) according to the presence of infection after operation. Major infection characteristics of the 68 recipients after liver transplantation were summarized. Univariate analysis was conducted on the possible risk factors of infection after liver transplantation, and multivariate analysis was further conducted on the risk factors with statistical significance, so as to find out the independent risk factors. In addition, accuracy of predicting infection after liver transplantation was analyzed using receiver operating characteristic (ROC) curves. Results Thirty-three recipients were infected after liver transplantation, accounting for 49% of the total recipients with bacterial infection and fungal infection mainly. These recipients mainly presented pulmonary infection and abdominal cavity infection. Univariate analysis results showed that a total of 8 factors contributed to infection after liver transplantation from donor liver of organ donation, including the donors' open injury, recipients' preoperative hemoglobin level, platelet count, Child-Pugh classification of liver function, model for end-stage liver disease (MELD) score, intraoperative erythrocyte infusion, gamma-glutamyl transpeptidase (GGT) on day 1 after operation and postoperative stay time of intensive care unit (ICU) (all P<0.05). Multivariate Logistic regression results analysis showed that preoperative hemoglobin level <120 g/L and postoperative stay time of ICU >96 h were the independent risk factors of infection after liver transplantation from donor liver of organ donation (both P<0.05). Analysis results of ROC curves showed that preoperative hemoglobin level<114 g/L and postoperative stay time of ICU >102 h resulted in higher accuracy for predicting postoperative infection. Conclusions Infection after liver transplantation from donation after citizen's death presents high incidence, dominated by bacterial infection and fungal infection in lung and abdominal cavity. Low preoperative hemoglobin level and long postoperative stay time of ICU of recipients can increase the risk of infection after liver transplantation.

Background::Links between alterations in gut microbiota composition and amyotrophic lateral sclerosis (ALS) have previously been reported. This study aimed to examine the microbiota in the nasal cavity of ALS.Methods::Sixty-six ALS patients and 40 healthy caregivers who live in close proximity with patients were enrolled. High throughput metagenomic sequencing of the 16S ribosomal deoxyribonucleic acid (rDNA) gene V3–V4 region of nasal microbiota was used to characterize the alpha and beta diversity and relative abundance of bacterial taxa, predict function, and conduct correlation analysis between specific taxa and clinical features.Results::The nasal microbiome of ALS patients showed lower alpha diversity than that of corresponding healthy family members. Genera Gaiella, Sphingomonas, Polaribacter_1, Lachnospiraceae_NK4A136_group, Klebsiella, and Alistipes were differentially enriched in ALS patients compared to controls. Nasal microbiota composition in ALS patients significantly differed from that in healthy subjects (unweighted UniFrac P = 0.001), while Linear discriminant analysis Effect Size (LEfSe) analysis indicated that Bacteroidetes and Firmicutes dominated healthy nasal communities at the phylum level, whereas Actinobacteria was the predominant phylum and Thermoleophilia was the predominant class in ALS patients. Genus Faecalibacterium and Alistipes were positively correlated with ALS functional rating scale revised (ALSFRS-R; rs = 0.349, P = 0.020 and rs = 0.393, P = 0.008), while Prevotella-9 and Bacteroides operational taxonomic units (OTUs) were positively associated with lung function (FVC) in ALS patients ( rs = 0.304, P = 0.045, and rs = 0.300, P = 0.048, respectively). Prevotella-1 was positively correlated with white blood cell counts (WBC, rs = 0.347, P = 0.021), neutrophil percentage (Neu%, rs = 0.428, P = 0.004), and neutrophil-to-lymphocyte ratio (NLR, rs = 0.411, P = 0.006), but negatively correlated with lymphocyte percentage (Lym%, rs = -0.408, P = 0.006). In contrast, Streptococcus was negatively associated with Neu% ( rs = -0.445, P = 0.003) and NLR ( rs = -0.436, P = 0.003), while positively associated with Lym% ( rs = 0.437, P = 0.003). No significant differences in nasal microbiota richness and evenness were detected among the severe and mild ALS patients. Conclusions::ALS is accompanied by altered nasal microbial community composition and diversity. The findings presented here highlight the need to understand how dysbiosis of nasal microbiota may contribute to the development of ALS.

Objective To summarize the experience of prevention of biliary tract complications after liver transplantation from organ donation after citizen's death. Methods Clinical data of 88 cases undergoing liver transplantation from organ donation after citizen's death in the Affiliated Zhongshan Hospital of Sun Yat-sen University from October 2008 to December 2016 were retrospectively analyzed. Results Eighty-eight cases were eligible for the standards for organ donation after brain death plus cardiac death according to the Ⅲ national system for organ donation in China. According to the standard procedures, donor livers were successfully harvested and transplanted in 88 recipients. The biliary tract was reconstructed using the bile duct end-to-end anastomosis. The length of bile duct in the donors was shortened as possible. Slight tension should be maintained during anastomosis. Neither primary liver graft nonfunction nor rejection reaction occurred. One recipient suffered from bile leakage and recovered after drainage for 3 weeks. Two patients presented with biliary tract stenosis and mitigated after the placement of biliary tract stent. Conclusions The harvesting of donor liver should be in accordance with the standard procedures. The advantages of extracorporeal membrane oxygenation (ECMO) should be fully utilized to shorten warm and cold ischemia time as possible. Much attention should be diverted to the reconstruction of biliary tract, which contributes to decreasing the risk of biliary tract complications. Favorable clinical efficacy can be achieved in liver transplantation from organ donation after citizen's death.

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.