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 compare the clinical efficacy of arthroscopy-assisted and open reduction and internal fixation in the treatment of Schatzker type I-III tibial plateau fractures.Methods:The data of patients with Schatzker type I-III tibial plateau fractures who were treated from August 2017 to July 2019 were retrospectively analyzed. According to the treatment, the patients were divided into the arthroscopic-assisted minimally invasive reduction and internal fixation group (arthroscopy group) and the conventional open reduction and internal fixation group (incision group). In the arthroscopy group, there were 30 patients, 19 males and 11 females were included; the age was 45.13±7.12 years old (range, 29-60 years). Among them, 13 cases were Schatzker type I fractures, 14 cases were Schatzker type II fractures, and 3 cases were Schatzker type III fractures. In the incision group, there were 30 patients, 17 males and 13 females were included; the age was 43.53±7.79 years old (range, 31-58 years). Among them, 11 cases were Schatzker type I fractures, 15 cases were Schatzker type II fractures, and 4 cases were Schatzker type III fractures. The operation time, intraoperative blood loss, postoperative ambulation time, postoperative complete weight-bearing time and postoperative complications were recorded. The degree of knee joint swelling, knee flexion and extension range of motion and the American knee society knee score (AKS score) were compared between the arthroscopy group and the incision group.Results:Both groups were followed up. The follow-up time of the arthroscopy group were 10-18 months, with an average of 14 months; the follow-up time in the incision group were 12-18 months, with an average of 15 months. In the arthroscopy group, the operation time (87.60±9.20 min vs. 94.33±10.65 min), intraoperative blood loss (57.16±9.63 ml vs. 71.93±11.15 ml), postoperative ambulation time (5.13±1.28 d vs. 6.17±1.53 d) and postoperative complete weight-bearing time (12.83±1.68 weeks vs. 14.23±1.77 weeks) were superior to the incision group, and the differences were statistically significant ( t=2.62, 5.49, 2.83, 3.94; all P<0.05). The healing time was 13.33±1.37 weeks in the arthroscopy group and 14.86±1.63 weeks in the incision group, and the difference was statistically significant ( t=3.94, P<0.001). At 1 year after surgery, the range of flexion and extension of knee joint in the arthroscopy group was 116.77°±12.46°, which was better than that in the incision group, which was 109.13°±9.89°, and the difference was statistically significant ( t=2.63, P=0.011). The AKS score in the arthroscopy group was 164.57±11.16 points, and the score in the incision group was 149.53±14.77 points, and the difference was statistically significant ( t=4.45, P<0.001). There were no malunion or compartment syndrome in the arthroscopy group and the incision group. The total incidence of complications in the arthroscopy group was 13% (4/30), including 2 cases of poor wound healing, 2 of poor knee range of motion after operation. The total incidence of complications in the incision group was 23% (7/30), including 4 cases of poor wound healing, 1 of wound infection, 2 of poor knee range of motion after operation. And the difference between the two groups in complication incidence was not statistically significant (χ 2=1.00, P=0.317). Conclusion:Arthroscopic-assisted reduction and internal fixation in the treatment of Schatzker I-III tibial plateau fractures has the advantages of less trauma, less bleeding, early mobility, fewer complications, and better knee joint function, and there are no obvious arthroscopic-related complications, which is a safe and reliable treatment method.

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 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 morphological characteristics, treatment strategies and clinical results of complex hyperextension tibial plateau fractures.Methods:From October 2017 to January 2019, data of 27 patients with complex hyperextension tibial plateau fractures were retrospectively analyzed. There were 19 males and 8 females with an average age of 43.4 years (range, 23-68 years). According to Schatzker classification of tibial plateau fractures: there are 8 cases of type IV, 5 of type V, and 14 of type VI; according to the three-column theory classification: there are 8 cases of two-column fracture and 19 cases of three-column fracture. Bicondylar fractures were treated with medial Tomofix locking plate and anterolateral L-shaped locking plate through medial and anterolateral approach; tibialmedial condylar fractures was treated with T-shaped plate and posteromedial locking plate through extended medial approach. Patients with anterior tibial fractures were treated with horizontal strip plate through modified anterior median approach. Combined soft tissue or bone injury was repaired. The fracture healing and reduction were evaluated by X-ray and CT scan. The reduction of tibial plateau fracture was evaluated by Rasmussen radiology standard, and the knee joint function was evaluated 12 months after the operation by the score of American hospital for special surgery (HSS).Results:All the 27 surgeries were performedsuccessfully. The operation time was 130-350 minutes, with an average time of 165 minutes. Twenty-seven cases were followed up for 12-24 months, with an average period of 15.8 months. All fractures were healed. The average clinical healing time was 13.5 weeks (range, 10-18 weeks). Twelve months after operation, Rasmussen's radiology score was 13-18, with an average of 16.7 points, among them there were 19 excellent and 8 good. Twelve months after the operation, the score of HSS knee joint was 82-98, with an average score of 93.2 points, and there were 22 cases excellent, 4 cases good and 1 case fair. The excellent and good rate was 96.2% (26/27).Conclusion:Complex hyperextension tibial plateau fractures often combined with tibial bicondylar, medial tibial condyle or anterior tibial fractures. According to the morphological characteristics of complex hyperextension tibial plateau fractures, using appropriate surgical approach and internal fixation, repairing ligament soft tissue structure and reconstructing knee joint stability can achieve satisfactory results.

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 discuss how to make the surgical strategy for tibial tubercle fracture associated with bicondylar tibial plateau fracture.Methods:Data of thirty-five patients of tibial tubercle fractures associated with bicondylar tibial plateau fractures who were treated from October 2014 to May 2018 were retrospectively analyzed. There were 26 males and 9 females with an average age of 37.6 years (range, 21-68 years). According to Schatzker classification in tibial plateau fracture, 16 cases were type V and 19 cases were type VI. According to the integrity of tibial tubercle fracture and cortical bone of the proximal tibia in bicondylar tibial plateau fracture, they were divided into four types: type A, tibial tubercle fracture fragment and cortical bone of the proximal tibia are both complete; type B, tibial tubercle fracture fragment is complete but cortical bone of the proximal tibia is comminuted; type C, tibial tubercle fracture fragment is comminuted but cortical bone of the proximal tibia is complete; type D, both of them are comminuted. The surgical approaches and fixation methods of all the tibial tubercle fractures were according to the four different types. There were 22 cases with type A and B that were treated via an anterolateral and a medial incision, 13 cases with type C and D were treated via an anterior midline and a medial incision. There were 4 cases belonging to type A fixed with lag screws singly, 18 cases with type B fixed with 1/4 tubular plates, 7 cases with type C and 6 cases with type D fixed by 1/4 tubular plates combined with lag screws.Results:Thirty-five patients were followed up for 16.8 months (range, 12-24 months). All fractures healed with an average time of 4.7 months (range, 3-6 months). Loss of reduction didn’t occur in 34 cases except one. According to Rasmussen radiographic evaluation, the average score was 14.1 (range, 10-18) and clinical outcomes were rated with excellent in 11 cases, good in 19, fair in 5. The excellent and good rate was 85.7% (30/35) . The mean Hospital for Special Surgery (HSS) scores of all cases were 86.8 (range, 64-98) and the functional scores were excellent in 22 cases, good in 10 cases and fair in 3 cases with the excellent and good rate of 91.4% (32/35) . Surgical complications included fat liquefaction in 2 cases, superficial wound infection in 1, loosening of implant in 1and traumatic arthritis in 1.Conclusion:This kind of tibial tubercle fracture associated with bicondylar tibial plateau fracture is rare and special. Therefore, the preoperative plan should be made by considering the morphological features of the tibial tubercle fragments and the cortical bone of the proximal tibia. The middle longitude approach is the best way to expose tibial tubercle fragments which should be fixed with 1/4 tubularplate and/or lag screws.

Objective:To summarize the clinical characteristics and laboratory diagnostic methods of infant botulism caused by Clostridium botulinum type B. Methods:Clinical data of 3 infants with type B botulism who were admitted to Children′s Hospital Affiliated to Capital Institute of Pediatrics from May to November 2018 were retrospectively analyzed. Botulinum toxin was detected in fecal samples or fecal enrichment solution of the patients, and Clostridium botulinum was cultured and isolated from fecal samples. Results:The age of onset of the patients (two boys and one girl) was 3, 3 and 8 months old, respectively. Two cases had the onset in May and one case had the onset in November. There were two cases with mixed feeding and one case with breast feeding. One case′s family members engaged in meat processing. All of them were previously healthy. All the children presented with acute flaccid paralysis, cranial nerve involvement and difficult defecation. Two cases had secondary urinary tract infection. Electromyograms of two cases showed that action potential amplitude of the motor nerve were lower than those of their peers. After treatments including intravenous human immunoglobulin, respiratory tract management, urethral catheterization, nasal feeding, etc., three cases recovered completely 2 to 4 months later. Type B botulinum toxin was detected in the fecal diluent of one patient, and the TPGYT enrichment solution and cooked meet medium of the feces of 3 patients, respectively. Clostridium botulinum B was identified from the feces of 3 infants after culture, isolation and purification. Conclusions:Combined with typical clinical manifestations including acute flaccid paralysis, cranial nerve involvement symptoms and difficult defecation examination, infant botulism can be clinically diagnosed. The detection of fecal botulinum toxin and the culture and isolation of Clostridium botulinum are helpful for the diagnosis.

Objective: To explore the clinical features and treatment strategies of tibial plateau fractures sustained with hyperextension varus. Methods: Data of 11 patients of tibial plateau fractures with hyperextension varus treated from January 2008 to November 2017 were retrospectively analyzed. There were 7 males and 4 females with an average age of 41.2 years old (range, 25-67 years). Injuries were caused by falling down in 7 cases, traffic accident in 3 cases, and falling from height in 1 case, respectively. On the basis of Luo's three columns classification in tibial plateau, there were 9 cases of medial column fracture and 2 cases of medial combined with posterior column fracture. Six cases were concomitant with fibular head fracture and 2 cases with the injury of common peroneal nerve. Preoperative magnetic resonance imaging showed that there were anterior cruciate ligament injury in 3 cases, posterior cruciate ligament injury in 4 cases, medial meniscus injury in 5 cases, lateral meniscus injury in 3 cases, medial collateral ligament injury in 6 cases, iliotibial band injury in 2 cases and posterolateral complex injury of the knee joint in 9 cases, respectively. All tibial plateau fractures were treated firstly by open reduction and internal fixation via medial approach of the knee. The medial meniscuses and collateral ligaments were explored, in which of them there were 2 medial meniscuses with the marginal tear been sutured simultaneously. Then the knee joints which were still unstable after the examination of stable tests in 6 cases with posterolateral complex injuries were repaired surgically via lateral approach. Fibular head fractures were fixed with anchor nails or cannulated screws in 6 cases. The ruptured posterior cruciate ligaments in 2 cases were reconstructed with autologous tendon transplantation under endoscopy. Results: All the patients were followed up for an average period of 16.2 months (range, 12-22 months). All fractures were healed in 10-20 weeks with an average time of 16.5 weeks. The range of extension of the affected knee joint in all patients was 0° and the average flexion was 135° (range, 120°-145°) one year after surgery. The average flexion of affected knee in 4 cases which were only treated with the tibial plateau fracture without the mild ligament injuries was 137° (range, 132°-145°) and the average flexion of affected knee in 7 cases who were treated with tibial plateau fracture and severe posterolateral complex included posterior cruciate ligaments completely broken with reconstruction was 132° (range, 120°-140°). According to Rasmussen radiographic evaluation, the average score of all patients was 16.3 (range, 14 to 18) and clinical outcomes were rated with excellent in 10 cases and good in 1. The excellent and good rate was 100% (11/11). The mean of the hospital for special surgery (HSS) score was 86.7 (range, 79-96) and the functional scores were excellent in 9 cases, good in 2 cases thus the excellent and good rate was 100% (11/11). Both varus stress test 30° and dial test were positive in one case considered for the ligament laxity postoperatively who didn’t accept further treatment and the stabilization tests were negative in the other 10 cases. There were no intraoperative complications in all patients such as neurovascular injury. No incision infection, failure of the implants and fracture nonunion occurred postoperatively. Traumatic arthritis of the affected knee occurred one year after surgery in 1 case who had no obvious pain after treated with oral medicine. Conclusion The hyperextension varus injuries of the knee are rare clinically. The posterolateral complex should be evaluated thoroughly for this injury pattern. If it's necessary, the posterolateral structures must be repaired surgically after the tibial plateau fractures are fixed.