To investigate the antimicrobial resistance of Enterococci isolated from 596 anal swabs,feces and environmental samples were collected from pig,cattle,chicken and quail farms in Chang-sha,Hunan Province,Enterococci were isolated and identified by mass spectrometry.The minimum inhibitory concentrations(MICs)of 10 antimicrobial agents were determined by AGAR diffusion method.Whole genome sequencing(WGS)was used to detect the distribution of multilocus se-quence typing(ST),drug resistance genes and virulence genes.A total of 272 strains of Enterococ-cus were isolated(45.6％).The isolates were resistant to cefoxitin(68.9％)and cefotiofur(58.5％),followed by trimethoprim/sulfamethoxazole(52.2％),vancomycin(4.4％),and linezolid(13.6％).In this study,six linezolid highly resistant Enterococci isolates were analyzed by whole genome sequencing to explore the transmission mechanism of linezolid resistance because linezolid is forbidden to be used in aquaculture.ST403(4/6),ST16(1/6)and ST476(1/6)were the most common ST types,which all originated from the same farm.Three oxazolidinone resistance genes(cfr,poxtA and optrA)were found in all 21 strains.One strain(Ecc60)carried all three oxazo-lidinone resistance genes,but none of them were located on the plasmid.tet(M),aph(3')-Ⅲ,and lsa(A)were found in all six Enterococcus isolates.Interestingly,the present study is the first to i-dentify enterococci carrying the optrA gene in quail fecal samples.The analysis of the genetic envi-ronment of oxazolidinone resistance genes showed that the genetic environment of cfr(D),poxtA and optrA of isolates from the same farm was similar.A total of 19 virulence genes were detected in 6 isolates,of which 12 genes(ElrA,SrtA,ace,agg,cCF10,cOB1,cad,camE,ebpA,ebpC,efaAfs,tpx)were carried by all 6 isolates,and the types of virulence genes in strains from the same farm were extremely similar.The results showed that the drug resistance of Enterococci iso-lated from the fish farms in Changsha was serious,and the resistance rate to linezolid,which was prohibited in the fish farms,was high.The oxazolidinone resistance gene loci were accompanied by other resistance genes,especially the florfenicol resistance gene(FexA),which may be related to the abuse of florfenicol in the fish farms.

To investigate the antimicrobial resistance of Enterococci isolated from 596 anal swabs,feces and environmental samples were collected from pig,cattle,chicken and quail farms in Chang-sha,Hunan Province,Enterococci were isolated and identified by mass spectrometry.The minimum inhibitory concentrations(MICs)of 10 antimicrobial agents were determined by AGAR diffusion method.Whole genome sequencing(WGS)was used to detect the distribution of multilocus se-quence typing(ST),drug resistance genes and virulence genes.A total of 272 strains of Enterococ-cus were isolated(45.6％).The isolates were resistant to cefoxitin(68.9％)and cefotiofur(58.5％),followed by trimethoprim/sulfamethoxazole(52.2％),vancomycin(4.4％),and linezolid(13.6％).In this study,six linezolid highly resistant Enterococci isolates were analyzed by whole genome sequencing to explore the transmission mechanism of linezolid resistance because linezolid is forbidden to be used in aquaculture.ST403(4/6),ST16(1/6)and ST476(1/6)were the most common ST types,which all originated from the same farm.Three oxazolidinone resistance genes(cfr,poxtA and optrA)were found in all 21 strains.One strain(Ecc60)carried all three oxazo-lidinone resistance genes,but none of them were located on the plasmid.tet(M),aph(3')-Ⅲ,and lsa(A)were found in all six Enterococcus isolates.Interestingly,the present study is the first to i-dentify enterococci carrying the optrA gene in quail fecal samples.The analysis of the genetic envi-ronment of oxazolidinone resistance genes showed that the genetic environment of cfr(D),poxtA and optrA of isolates from the same farm was similar.A total of 19 virulence genes were detected in 6 isolates,of which 12 genes(ElrA,SrtA,ace,agg,cCF10,cOB1,cad,camE,ebpA,ebpC,efaAfs,tpx)were carried by all 6 isolates,and the types of virulence genes in strains from the same farm were extremely similar.The results showed that the drug resistance of Enterococci iso-lated from the fish farms in Changsha was serious,and the resistance rate to linezolid,which was prohibited in the fish farms,was high.The oxazolidinone resistance gene loci were accompanied by other resistance genes,especially the florfenicol resistance gene(FexA),which may be related to the abuse of florfenicol in the fish farms.

Objective:To investigate the clinical features and therapeutic efficacy of patients with hereditary leiomyomatosis and renal cell carcinoma(RCC) syndrome-associated RCC (HLRCC-RCC) in China.Methods:The clinical data of 119 HLRCC-RCC patients with fumarate hydratase (FH) germline mutation confirmed by genetic diagnosis from 15 medical centers nationwide from January 2008 to December 2021 were retrospectively analyzed. Among them, 73 were male and 46 were female. The median age was 38(13, 74) years. The median tumor diameter was 6.5 (1.0, 20.5) cm. There were 38 cases (31.9%) in stage Ⅰ-Ⅱand 81 cases (68.1%) in stage Ⅲ-Ⅳ. In this group, only 11 of 119 HLRCC-RCC patients presented with skin smooth muscle tumors, and 44 of 46 female HLRCC-RCC patients had a history of uterine fibroids. The pathological characteristics, treatment methods, prognosis and survival of the patients were summarized.Results:A total of 86 patients underwent surgical treatment, including 70 cases of radical nephrectomy, 5 cases of partial nephrectomy, and 11 cases of reductive nephrectomy. The other 33 patients with newly diagnosed metastasis underwent renal puncture biopsy. The results of genetic testing showed that 94 patients had FH gene point mutation, 18 had FH gene insertion/deletion mutation, 4 had FH gene splicing mutation, 2 had FH gene large fragment deletion and 1 had FH gene copy number mutation. Immunohistochemical staining showed strong 2-succinocysteine (2-SC) positive and FH negative in 113 patients. A total of 102 patients received systematic treatment, including 44 newly diagnosed patients with metastasis and 58 patients with postoperative metastasis. Among them, 33 patients were treated with tyrosine kinase inhibitor (TKI) combined with immune checkpoint inhibitor (ICI), 8 patients were treated with bevacizumab combined with erlotinib, and 61 patients were treated with TKI monotherapy. Survival analysis showed that the median progression-free survival (PFS) of TKI combined with ICI was 18 (5, 38) months, and the median overall survival (OS) was not reached. The median PFS and OS were 12 (5, 14) months and 30 (10, 32) months in the bevacizumab combined with erlotinib treatment group, respectively. The median PFS and OS were 10 (3, 64) months and 44 (10, 74) months in the TKI monotherapy group, respectively. PFS ( P=0.009) and OS ( P=0.006) in TKI combined with ICI group were better than those in bevacizumab combined with erlotinib group. The median PFS ( P=0.003) and median OS ( P=0.028) in TKI combined with ICI group were better than those in TKI monotherapy group. Conclusions:HLRCC-RCC is rare but has a high degree of malignancy, poor prognosis and familial genetic characteristics. Immunohistochemical staining with strong positive 2-SC and negative FH can provide an important basis for clinical diagnosis. Genetic detection of FH gene germ line mutation can confirm the diagnosis. The preliminary study results confirmed that TKI combined with ICI had a good clinical effect, but it needs to be confirmed by the results of a large sample multi-center randomized controlled clinical study.

Objective:To investigate the safety and efficacy of the novel single-port surgical robotic surgical system in extraperitoneal urological surgery.Methods:From February to April 2022, patients was prospectively enrolled who required laparoscopic radical prostatectomy, partial nephrectomy and adrenal tumor resection in urology department. Inclusion criteria were: age ≥ 18 years old; BMI 18.5-30 kg/m 2; American Society of Anesthesiologists (ASA) physical status classification system grades 1 to 3; can cooperate with the completion of the visits and related examinations stipulated in the plan, and participate voluntarily clinical trials, and consent or the guardian agrees to sign the informed consent form; tumor indicators meet one of the following surgical treatment indications: kidney tumor T 1 stage, single, maximum tumor diameter ≤ 4 cm; prostate cancer, stage ≤ T 2b, preoperative PSA ≤ 20 ng /ml; Gleason score ≤ 7; adrenal tumor diameter ≤ 7 cm, for non-functioning adrenal adenoma, tumor diameter ≥ 3 cm. Exclusion criteria were: patients with other malignancies or a history of other malignancies and the investigators believe that they are not suitable for inclusion in this researcher; patients who have received the same type of urological surgery in the past and are not suitable for participating in this study as assessed by the investigators; included Those who have undergone other major surgery within the first 3 months and during the trial period, or who cannot recover from the side effects of any such surgery; syphilis, hepatitis B, HIV infection and carriers; long-term use of anticoagulants or blood system diseases; Unable to use effective contraception during the trial period and other conditions that the investigators deem inappropriate to participate in this trial. All operations were performed by a novel single-port robotic surgical operating system, and all surgical procedures were performed through an extraperitoneal approach. Surgical method: the surgical system is mainly composed of a remote console including a high-definition display, a surgical equipment trolley, a surgical execution system that accommodates a serpentine robotic arm, and a bendable serpentine robotic arm. In this study, the extraperitoneal approach was used. For radical prostatectomy, the patient was placed in a supine position, a longitudinal incision of about 3 cm was made below the umbilicus, the anterior rectus sheath was incised, the extraperitoneal space was separated, and an operating sheath was placed. A 12 mm trocar is placed between the right McBurney point and the umbilicus as an auxiliary hole. For partial nephrectomy and adrenal tumor resection, the patient is placed in the lateral position, and an 3cm incision is made 2 cm above the iliac crest on the midaxillary line as the main operating hole. The skin, subcutaneous tissue, and muscle were incised to the retroperitoneal cavity, and a 12mm trocar was placed at the level of the anterior superior iliac spine on the anterior axillary line as an auxiliary hole. The operation was performed after connecting each robotic arm. After the operation, the specimen was placed in the specimen bag, and a drainage tube is placed in the auxiliary hole, the specimen was taken out, and the incision was closed in turn. Preoperative basic information, operation time, blood loss, incision size, postoperative complications, preoperative and postoperative PSA score, eGFR index, postoperative pathological information and other perioperative information were collected. Results:A total of 17 patients were included in this study, including 6 with prostate cancer, 8 with renal tumor, and 3 with adrenal tumor. There were 9 males and 8 females, with an average age of (56.7±14.6) years and a BMI of (23.3±3.4) kg/m 2. The mean operation time of radical prostatectomy was (244.6±35.1) min, the mean operating time of the chief surgeon was (184.0±39.0) min, and the mean blood loss was (36.6±23.8) ml. Postoperative positive margin was found in 2 cases. The average operation time of partial nephrectomy was (189.6±49.4) minutes, the average operating time of the chief surgeon was (115±39.7) minutes, the average blood loss was (12.7±8.3) ml, and the average warm ischemia time was (23.1±10.8) minutes. There was no significant difference in the eGFR index before and after the operation ( P>0.05). The average operation time of adrenalectomy was (177.6±26.9) min, the average operating time of the chief surgeon was (99±20.4) min, and the average blood loss was (11.6±6.2) ml. The overall average operation time of the three surgical methods was (206.9±50.1) min, the overall average operating time of the chief surgeon was (136.5±51.1) min, the overall average blood loss was (21.0±9.2) ml, and the overall average incision size was (3.5±0.5) cm, all added a 12 mm auxiliary channel, and the overall average hospital stay was (8.1±2.7) days. All operations were successfully completed, and there was no conversion to open surgery during the operation, and no operation holes were added. There was no Clavien-Dindo≥grade 3 complication after operation. Conclusions:The novel single-port robot could safely and effectively perform radical prostatectomy, partial nephrectomy and adrenalectomy which are common in urology through extraperitoneal approach.

Objective:To investigate the preoperative risk factors affecting early extremity blood supply after repair of major arterial injury so as to provide clues for prevention of limb ischemia.Methods:The clinical data were retrospectively analyzed of the 139 patients (140 extremities) with major extremity arterial injury who had been admitted to Department of Microsurgery, Orthopaedic Trauma and Hand Surgery, The First Hospital Affiliated to Sun Yat-sen University from January 2003 to December 2019. There were 112 males and 27 females, with a mean age of 30 (20, 44) years. The primary outcome was the early status of blood supply to the injured extremity (48 hours after surgery). Univariate analysis was conducted of such factors as gender, age, ischemia time, injury mechanism, injury site, fracture, soft tissue lesion, and duration of surgery. The significant factors ( P<0.1) were then analyzed by logistic regression, and P<0.05 was considered statistically significant. Results:Ischemia happened in 44 (31.4%, 44/140) extremities within 48 hours after surgery. There were significant differences in injury mechanism, ischemia time, fracture, and soft tissue lesion between patients with and without postoperative extremity ischemia ( P<0.05). Logistic regression analysis indicated that blunt injury ( OR=5.639, 95% CI: 1.068 to 29.761, P=0.042) and soft tissue lesion ( OR=12.568, 95% CI: 3.402 to 46.431, P<0.001) were significant preoperative risk factors affecting the early blood supply after repair of major extremity arterial injury. Conclusion:As blunt injury and soft tissue defect are preoperative risk factors for early extremity ischemia after repair of major extremity arterial injury, surgeons should pay more attention to them when assessing patients and making repair protocols.

Objective:To assess the influence of an artificial intelligence (AI) -assisted diagnosis system on the performance of endoscopists in diagnosing gastric cancer by magnifying narrow banding imaging (M-NBI).Methods:M-NBI images of early gastric cancer (EGC) and non-gastric cancer from Renmin Hospital of Wuhan University from March 2017 to January 2020 and public datasets were collected, among which 4 667 images (1 950 images of EGC and 2 717 of non-gastric cancer)were included in the training set and 1 539 images (483 images of EGC and 1 056 of non-gastric cancer) composed a test set. The model was trained using deep learning technique. One hundred M-NBI videos from Beijing Cancer Hospital and Renmin Hospital of Wuhan University between 9 June 2020 and 17 November 2020 were prospectively collected as a video test set, 38 of gastric cancer and 62 of non-gastric cancer. Four endoscopists from four other hospitals participated in the study, diagnosing the video test twice, with and without AI. The influence of the system on endoscopists′ performance was assessed.Results:Without AI assistance, accuracy, sensitivity, and specificity of endoscopists′ diagnosis of gastric cancer were 81.00%±4.30%, 71.05%±9.67%, and 87.10%±10.88%, respectively. With AI assistance, accuracy, sensitivity and specificity of diagnosis were 86.50%±2.06%, 84.87%±11.07%, and 87.50%±4.47%, respectively. Diagnostic accuracy ( P=0.302) and sensitivity ( P=0.180) of endoscopists with AI assistance were improved compared with those without. Accuracy, sensitivity and specificity of AI in identifying gastric cancer in the video test set were 88.00% (88/100), 97.37% (37/38), and 82.26% (51/62), respectively. Sensitivity of AI was higher than that of the average of endoscopists ( P=0.002). Conclusion:AI-assisted diagnosis system is an effective tool to assist diagnosis of gastric cancer in M-NBI, which can improve the diagnostic ability of endoscopists. It can also remind endoscopists of high-risk areas in real time to reduce the probability of missed diagnosis.

Report a case sustained Gustilo type III C open fracture of the left humerus with brachial artery injury who has limb ischemia and wound infection after operation in June, 2014. To salvage the limb, performed cross limb vessel transfer to restore blood supply at one-stage. After multiple debridement, Flow-through flap transfer was performed for definitive reconstruction of the arterial injury and repair the wound in secondary stage. In the 3rd stage, cutting the pedicle of transposition vessels. Follow-up at 1 year after surgery, the patient's left upper limb had survived with limited movement and confirmed Flow-through the vessel reconstruction using CTA.

Objective: To investigate the effects of metformin on mitochondrial pathway of apoptosis and oxidative stress in cell model of nonalcoholic fatty liver disease. Methods: An in vitro cell model of nonalcoholic fatty liver disease was established using 0.6 mmol/L oleic acid to induce lipid accumulation in HepG2 cells. HepG2 cells were divided into control (Con) group, oleic acid (OA) group, and metformin-low (1mmol/L) and high (10mmol/L) dose group. Oil Red O stain was used to detect intracellular lipid droplet distribution. The levels of alanine aminotransferase and aspartate aminotransferase in the culture supernatant were detected by assay kits. DCFH-DA method was used to detect the reactive oxygen species of HepG2 cells. Double staining flow cytometry was used to detect the apoptosis rate of HepG2 cells. Western blot was used to detect caspase-3, B-lymphocyte lymphoma-related protein, B-cell lymphoma 2, and cytochrome c protein. One-way analysis of variance was used to compare the data between groups. Results: Oleic acid-induced HepG2 cells were significantly increased with lipid droplets. Low and high-dose metformin had reduced intracellular lipid droplets accumulation. The effect of metformin in the high-dose group was more significant than that in the low-dose group. Aspartate aminotransferase and alanine aminotransferase in HepG2 cells of OA group were significantly increased, which were (43.41 ± 7.11) U/L and (29.56 ± 4.11) U/L, respectively. The intracellular aspartate aminotransferase and alanine aminotransferase were decreased significantly after the treatment with low and high-dose metformin, which were (32.44 ± 4.08)U/L, (19.31 ± 3.03) U/L, (26.00 ± 3.11) U/L and (15.11 ± 4.11) U/L, respectively and the differences were statistically significant (P < 0.05). DCFH-DA test results showed that the fluorescence intensity of reactive oxygen species in the oleic acid group was 41.21% ± 4.23%, while the fluorescence intensity of reactive oxygen species in the low and high-dose metformin groups were reduced to 27.44% ± 3.91%, and 17.55% ± 5.11%, respectively and the differences between the groups were statistically significant (P < 0.05). The results of flow cytometry analysis showed that the cell apoptosis rate of the OA group was significantly higher than that of the Con group (12.12% ± 0.72% vs. 3.04% ± 0.57%, P < 0.05).The apoptosis rate of HepG2 cells was significantly reduced after metformin treatment at low and high doses (8.71% ± 0.71%, 5.71% ± 0.61%, P < 0.05). Western blot results showed that compared with the Con group, the expressions of B-lymphocyte lymphoma-related protein, cytochrome c, and caspase-3 were increased in the OA group, while the B-cell lymphoma 2 were decreased (P < 0.05). The expression of B-lymphocyte lymphoma-related protein, cytochrome c, and caspase-3 protein in HepG2 cells was decreased after treatment with low and high-dose metformin, while B-cell lymphoma 2 was increased (P < 0.05). Conclusion Metformin can effectively alleviate steatosis and improve the HepG2 function in cell model of nonalcoholic fatty liver disease. The mechanism of metformin may be related to the reduction of oxidative stress injury, the regulation of protein expression related to mitochondrial apoptosis pathway and the inhibition of cell apoptosis.

Objective:For Bayne and Klug type Ⅲ radial longitudinal deficiency, the aim of the surgical methods is to create a centralized stable wrist and restore the length of the radius. The purpose of the current study was to validate the use of the staged procedures with vascularized proximal fibular epiphyseal transfer, based on a single vascular pedicle of the inferior lateral genicular artery to treat radius defects.Methods:Between June 2007 and June 2012, five children with an averaged age of 4.3 years (range, 3.3 to 5.8 years) who had a type Ⅲ radial longitudinal deficiency were identified. The staged procedures we performed for this patient consisted of: (1) soft tissue distraction of the wrist with an external distraction device to centralize the wrist and create a space for placement of the proximal fibular graft, ulnar osteotomy, and internal fixation to correct the bowing deformity of the ulna if necessary; (2) vascularized proximal fibular epiphysis transferred to repair the radius defect and to reconstruct the wrist; and (3) pollicization or thumb reconstruction to treat thumb deficiency. In all patients, the range of motion of the digits, wrist, forearm, and elbow; the length of the forearm; and the deviation of the wrist were evaluated.Results:The averaged duration of follow-up was 52.6 months (range, 42 to 72 months). All transplants survived and united with the host bone within 3 months postoperatively. An averaged correction of 28 degrees in the hand-forearm angle was obtained. At the final follow-up, the average ulna and radius lengths were 14.4 cm and 12.1 cm, respectively; the percentage length compared with the normal side was 64.0 and 56.3 percent, respectively. The overall range of wrist motion was approximately 56 percent of that of the contralateral extremity. The averaged pronation and supination were 75 and 45 percent of the normal side, respectively. The overall range of elbow motion was similar to the normal side.Conclusions:The results of the present study indicated that vascularized proximal fibular epiphyseal transfer, based on the inferior lateral genicular artery, is a technically feasible method for treatment of Bayne and Klug type Ⅲ deficiency, which restores the length of the radius and stabilized the wrist.

Objective:For Bayne and Klug type Ⅲ radial longitudinal deficiency, the aim of the surgical methods is to create a centralized stable wrist and restore the length of the radius. The purpose of the current study was to validate the use of the staged procedures with vascularized proximal fibular epiphyseal transfer, based on a single vascular pedicle of the inferior lateral genicular artery to treat radius defects.Methods:Between June 2007 and June 2012, five children with an averaged age of 4.3 years (range, 3.3 to 5.8 years) who had a type Ⅲ radial longitudinal deficiency were identified. The staged procedures we performed for this patient consisted of: (1) soft tissue distraction of the wrist with an external distraction device to centralize the wrist and create a space for placement of the proximal fibular graft, ulnar osteotomy, and internal fixation to correct the bowing deformity of the ulna if necessary; (2) vascularized proximal fibular epiphysis transferred to repair the radius defect and to reconstruct the wrist; and (3) pollicization or thumb reconstruction to treat thumb deficiency. In all patients, the range of motion of the digits, wrist, forearm, and elbow; the length of the forearm; and the deviation of the wrist were evaluated.Results:The averaged duration of follow-up was 52.6 months (range, 42 to 72 months). All transplants survived and united with the host bone within 3 months postoperatively. An averaged correction of 28 degrees in the hand-forearm angle was obtained. At the final follow-up, the average ulna and radius lengths were 14.4 cm and 12.1 cm, respectively; the percentage length compared with the normal side was 64.0 and 56.3 percent, respectively. The overall range of wrist motion was approximately 56 percent of that of the contralateral extremity. The averaged pronation and supination were 75 and 45 percent of the normal side, respectively. The overall range of elbow motion was similar to the normal side.Conclusions:The results of the present study indicated that vascularized proximal fibular epiphyseal transfer, based on the inferior lateral genicular artery, is a technically feasible method for treatment of Bayne and Klug type Ⅲ deficiency, which restores the length of the radius and stabilized the wrist.