Objective:To investigate the preoperative administration time and dose of indo-cyanine green (ICG) for extrahepatic biliary tract imaging in the laparoscopic cholecystectomy (LC).Methods:The retrospective study was conducted. The clinical data of 252 patients with gallbladder diseases who were admitted to The Affiliated Hospital of Anhui Medical University from December 2022 to December 2024 were collected. There were 137 males and 115 females, aged (45±4)years. All patients underwent LC after injection of 1.25 mg or 2.50 mg ICG, with ICG fluore-scence navigation during the operation. Observation indicators: (1) effective fluorescence imaging during surgery; (2) the ratio of fluorescence intensity between gallbladder duct and liver, and the ratio of fluorescence intensity between common bile duct and liver; (3) the imaging effect of extra-hepatic biliary tract. Comparison of measurement data with skewed distribution among groups was conducted using the Kruskal-Wallis H test, and the Bonferroni method was used for pairwise com-parison. The consistency evaluation was conducted using the Kendall test. Results:(1) Effective fluorescence imaging during surgery. The Kendall coefficient index was 0.83, indicating high consis-tency in evaluation of fluorescence imaging of extrahepatic biliary tract between doctors. The effective fluorescent imaging sites during surgery were located in the liver, cystic duct, common bile duct, cystic duct-common bile duct junction, hepatic duct, and gallbladder. The intraoperative effective fluorescence imaging of patients who received intravenous injection of 1.25 mg and 2.50 mg ICG before surgery showed that as the interval between ICG injection and surgery increased, the proportion of fluorescence imaging in the liver and gallbladder gradually decreased. The proportion of fluorescence imaging in the gallbladder duct, common bile duct, cystic duct-common bile duct junction, common hepatic duct showed a trend of first increasing and then decreasing. (2) The ratio of fluorescence intensity between gallbladder duct and liver, and the ratio of fluorescence intensity between common bile duct and liver. Results of Kruskal Wallis H test showed that there were significant differences in the fluorescence intensity ratios of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver among patients who received intravenous injection of 1.25 mg ICG at different time intervals to surgery ( H=73.22, 77.17, P<0.05). Results of pairwise comparison showed that there were significant differences in the fluorescence intensity ratio of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver between patients who received intravenous ICG injection 4.0-<6.0 hours before surgery and those who received ICG injection <0.5 hours, 0.5-<2.0 hours, 2.0-<4.0 hours, 6.0-<8.0 hours, 8.0-<10.0 hours, and 10.0-<12.0 hours before surgery, respectively ( P<0.002). There were significant differences in the fluorescence intensity ratio of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver among patients who received intravenous injection of 2.50 mg ICG at different time intervals to surgery ( H=127.06, 126.39, P<0.05). Results of pairwise comparison showed there were significant differences in the fluorescence intensity ratio of gall-bladder duct to liver and the fluorescence intensity ratio of common bile duct to liver between patients who received ICG injection 8.0-<10.0 hours before surgery and those who received ICG injection <0.5 hours, 0.5-<2.0 hours, 2.0-<4.0 hours, 4.0-<6.0 hours, 6.0-<8.0 hours, 12.0-<14.0 hours, and 14.0-<16.0 hours before surgery ( P<0.001). (3) The imaging effect of extrahepatic biliary tract. Among 102 patients who received preoperative intravenous injection of 1.25 mg ICG, the number of patients with grade A extrahepatic biliary system imaging increased and then decreased as the interval time extending, reaching a peak at 4.0-<6.0 hours. Among 150 patients who received preoperative intravenous injection of 2.50 mg ICG, the number of patients with grade A extrahepatic biliary tract imaging increased and then decreased as the interval time extending, reaching a peak at 8.0-<10.0 hours. Conclusion:Prolonging the time interval between ICG administration and surgery can effectively reduce the fluorescence intensity of the liver background, thereby increasing the fluorescence intensity ratio of gallbladder duct to liver and common bile duct to liver to obtain the best development effect. Intravenous injection of 1.25 mg ICG 4.0-<6.0 hours before surgery or 2.50 mg ICG 8.0-<10.0 hours before surgery provide better results for intraoperative extrahepatic biliary tract imaging.

Objective:To investigate the preoperative administration time and dose of indo-cyanine green (ICG) for extrahepatic biliary tract imaging in the laparoscopic cholecystectomy (LC).Methods:The retrospective study was conducted. The clinical data of 252 patients with gallbladder diseases who were admitted to The Affiliated Hospital of Anhui Medical University from December 2022 to December 2024 were collected. There were 137 males and 115 females, aged (45±4)years. All patients underwent LC after injection of 1.25 mg or 2.50 mg ICG, with ICG fluore-scence navigation during the operation. Observation indicators: (1) effective fluorescence imaging during surgery; (2) the ratio of fluorescence intensity between gallbladder duct and liver, and the ratio of fluorescence intensity between common bile duct and liver; (3) the imaging effect of extra-hepatic biliary tract. Comparison of measurement data with skewed distribution among groups was conducted using the Kruskal-Wallis H test, and the Bonferroni method was used for pairwise com-parison. The consistency evaluation was conducted using the Kendall test. Results:(1) Effective fluorescence imaging during surgery. The Kendall coefficient index was 0.83, indicating high consis-tency in evaluation of fluorescence imaging of extrahepatic biliary tract between doctors. The effective fluorescent imaging sites during surgery were located in the liver, cystic duct, common bile duct, cystic duct-common bile duct junction, hepatic duct, and gallbladder. The intraoperative effective fluorescence imaging of patients who received intravenous injection of 1.25 mg and 2.50 mg ICG before surgery showed that as the interval between ICG injection and surgery increased, the proportion of fluorescence imaging in the liver and gallbladder gradually decreased. The proportion of fluorescence imaging in the gallbladder duct, common bile duct, cystic duct-common bile duct junction, common hepatic duct showed a trend of first increasing and then decreasing. (2) The ratio of fluorescence intensity between gallbladder duct and liver, and the ratio of fluorescence intensity between common bile duct and liver. Results of Kruskal Wallis H test showed that there were significant differences in the fluorescence intensity ratios of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver among patients who received intravenous injection of 1.25 mg ICG at different time intervals to surgery ( H=73.22, 77.17, P<0.05). Results of pairwise comparison showed that there were significant differences in the fluorescence intensity ratio of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver between patients who received intravenous ICG injection 4.0-<6.0 hours before surgery and those who received ICG injection <0.5 hours, 0.5-<2.0 hours, 2.0-<4.0 hours, 6.0-<8.0 hours, 8.0-<10.0 hours, and 10.0-<12.0 hours before surgery, respectively ( P<0.002). There were significant differences in the fluorescence intensity ratio of gallbladder duct to liver and the fluorescence intensity ratio of common bile duct to liver among patients who received intravenous injection of 2.50 mg ICG at different time intervals to surgery ( H=127.06, 126.39, P<0.05). Results of pairwise comparison showed there were significant differences in the fluorescence intensity ratio of gall-bladder duct to liver and the fluorescence intensity ratio of common bile duct to liver between patients who received ICG injection 8.0-<10.0 hours before surgery and those who received ICG injection <0.5 hours, 0.5-<2.0 hours, 2.0-<4.0 hours, 4.0-<6.0 hours, 6.0-<8.0 hours, 12.0-<14.0 hours, and 14.0-<16.0 hours before surgery ( P<0.001). (3) The imaging effect of extrahepatic biliary tract. Among 102 patients who received preoperative intravenous injection of 1.25 mg ICG, the number of patients with grade A extrahepatic biliary system imaging increased and then decreased as the interval time extending, reaching a peak at 4.0-<6.0 hours. Among 150 patients who received preoperative intravenous injection of 2.50 mg ICG, the number of patients with grade A extrahepatic biliary tract imaging increased and then decreased as the interval time extending, reaching a peak at 8.0-<10.0 hours. Conclusion:Prolonging the time interval between ICG administration and surgery can effectively reduce the fluorescence intensity of the liver background, thereby increasing the fluorescence intensity ratio of gallbladder duct to liver and common bile duct to liver to obtain the best development effect. Intravenous injection of 1.25 mg ICG 4.0-<6.0 hours before surgery or 2.50 mg ICG 8.0-<10.0 hours before surgery provide better results for intraoperative extrahepatic biliary tract imaging.

Objective:To investigate the application value of augmented reality navigation combined with indocyanine green(ICG) fluorescence imaging technology in laparoscopic anatomical segment 8 liver resection.Methods:Clinical and pathological data from 8 patients with hepatocellular carcinoma located in segment 8 of the liver admitted to the First Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University from October 2021 to October 2022 were collected restrospectively. Among them,there were 5 males and 3 females,aged between 40 and 72 years. During the operation,the self-developed laparoscopic augmented reality surgical navigation system was used to integrate the three-dimensional liver model with the laparoscopic scene,and ICG fluorescence imaging technology was used to guide the anatomical liver resection of segment 8. The predicted liver resection volume and actual liver resection volume,related surgical indicators and postoperative complications were analyzed.Results:Among the 8 patients, 4 underwent laparoscopic anatomical segment 8 liver resection,1 underwent laparoscopic anatomical ventral subsegment of segment 8 liver resection,2 underwent laparoscopic anatomical ventral subsegment combined with medial subsegment of segment 8 liver resection, and 1 underwent laparoscopic anatomical dorsal subsegment of segment 8 liver resection. All operations were completed under the guidance of augmented reality navigation combined with ICG fluorescence imaging,without conversion to open surgery. The operation time was (276.3±54.8)minutes(range:200 to 360 minutes). Intraoperative blood loss was (75.0±35.4)ml(range:50 to 150 ml). No blood transfusion was performed during the operation. The length of postoperative hospital stay was (7.6±0.8)days(range:7 to 9 days). There were no deaths or postoperative complications such as bleeding or biliary fistula during the perioperative period.Conclusion:Augmented reality navigation combined with ICG fluorescence imaging technology can guide the implementation of laparoscopic anatomical segment 8 liver resection.

Objective:To investigate the application value of augmented reality navigation combined with indocyanine green(ICG) fluorescence imaging technology in laparoscopic anatomical segment 8 liver resection.Methods:Clinical and pathological data from 8 patients with hepatocellular carcinoma located in segment 8 of the liver admitted to the First Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University from October 2021 to October 2022 were collected restrospectively. Among them,there were 5 males and 3 females,aged between 40 and 72 years. During the operation,the self-developed laparoscopic augmented reality surgical navigation system was used to integrate the three-dimensional liver model with the laparoscopic scene,and ICG fluorescence imaging technology was used to guide the anatomical liver resection of segment 8. The predicted liver resection volume and actual liver resection volume,related surgical indicators and postoperative complications were analyzed.Results:Among the 8 patients, 4 underwent laparoscopic anatomical segment 8 liver resection,1 underwent laparoscopic anatomical ventral subsegment of segment 8 liver resection,2 underwent laparoscopic anatomical ventral subsegment combined with medial subsegment of segment 8 liver resection, and 1 underwent laparoscopic anatomical dorsal subsegment of segment 8 liver resection. All operations were completed under the guidance of augmented reality navigation combined with ICG fluorescence imaging,without conversion to open surgery. The operation time was (276.3±54.8)minutes(range:200 to 360 minutes). Intraoperative blood loss was (75.0±35.4)ml(range:50 to 150 ml). No blood transfusion was performed during the operation. The length of postoperative hospital stay was (7.6±0.8)days(range:7 to 9 days). There were no deaths or postoperative complications such as bleeding or biliary fistula during the perioperative period.Conclusion:Augmented reality navigation combined with ICG fluorescence imaging technology can guide the implementation of laparoscopic anatomical segment 8 liver resection.

Objective To evaluate the effect of nucleolar protein 14 (NOP14) on angiogenesis in melanoma.Methods Melanoma tissues were collected from 40 patients with pathologically diagnosed melanoma in Guangzhou First People's Hospital from January 2016 to December 2018,and immunohistochemical study was conducted to determine the expression of NOP14 and CD31 (expressed as microvessel density [MVD]).Melanoma cell lines A375 and SK-MEL-1 were both divided into 4 groups:empty vector group transfected with the empty vector,NOPI4 group transfected with a NOP14-overexpressing vector,siNOP14 group transfected with the siRNA targeting NOP14,and siNC group transfected with a negative control siRNA.Fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of NOP14 respectively,and Western blot analysis and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in cells and their culture media.Coculture models of human umbilical vein endothelial cells (HUVECs) and A375/SK-MEL-1 cells in the above groups were established in Transwell chambers,and cell counting kit-8 (CCK8) assay,Transwell migration and invasion assays and Matrigel-based vasculogenic mimicry assay were performed to evaluate the cellular proliferative,migratory,invasive activity and tube formation capacity respectively.A linear regression model was used to analyze the relationship between NOP14 expression and MVD in melanoma tissues,multi-way analysis of variance to analyze the difference in cellular proliferative activity,and independent-sample t test to compare other experimental indices between 2 groups.Results The expression of CD31 (MVD) was 44 ± 13 in the group with high NOP14 expression (n =20),58 ± 16 in that with moderate NOP14 expression (n =17),and 62 ± 11 in that with low NOP14 expression (n =3).The NOP14 expression was negatively correlated with MVD (r =-0.525,P =0.017).Compared with the empty vector group,the expression of VEGF and VEGFR in A375 and SK-MEL-1 cells and their culture media significantly decreased in the NOP14 group (all P ＜ 0.05).Compared with the siNC group,the expression of VEGF and VEGFR in the A375 and SK-MEL-1 cells and their culture media significantly increased in the siNOP14 group(all P ＜ 0.05).In the co-culture models of A375 cells and HUVECs,the NOP14 group showed significantly decreased proliferative activity of HUVECs (F =131.85,P ＜ 0.05),and numbers of migratory cells (22 ± 5 vs.63 ± 8,t =7.07,P =0.002),invasive cells (14 ± 5 vs.45 ± 10,t =4.94,P =0.008) and branch points (8 ± 2 vs.14 ± 3,t =5.06,P ＜ 0.001) compared with the empty vector group;compared with the siNC group,the siNOP14 group showed significantly increased proliferative activity of HUVECs (F =79.92,P ＜ 0.01),and numbers of migratory cells (152 ± 30 vs.59 ± 4,t =5.36,P =0.006),invasive cells (134 ± 21 vs.50 ± 8,t =6.40,P ＜ 0.001) and branch points (27 ± 3 vs.15 ± 4,t =6.10,P ＜ 0.001).In the co-culture models of SK-MEL-1 cells and HUVECs,the 4 groups showed the same trend of changes in the cellular proliferative,migratory,invasive activity and tube formation capacity of HUVECs as the above groups in the co-culture models of A375 cells and HUVECs.Conclusion The NOP14 expression is negatively correlated with MVD in melanoma tissues,and NOP14 can inhibit angiogenesis in melanoma.

Objective To establish a method for rapid preparation of antiserum against influenza virus (H7N9) hemagglutinin,and to study the possibility of using it in single radial immunodiffusion (SRID) assay for quantitative detection of antigen in H7N9 influenza vaccine.Methods Hemagglutinin proteins expressed in eukaryotic cells were used to immunize sheep.Serum samples were collected to detect antibody titers by ELISA and double immunodiffusion assay.Different concentrations of antiserum were used in SRID assay to get the optimized concentration.Results After 4 times of immunization,the antiserum titers achieved 1 ∶ 1 000 000 and 1 ∶ 32 as indicated by ELISA and double immunodiffusion assay,respectively.The antiserum could form a clear precipitation line in SRID assay.The detection of antigen in the range of 10 to 40 μg/ml showed good linearity in the standard curve.The antigen titers in six batches of H7N9 vaccine detected by this SRID assay were identical with those by SDS-PAGE assay.Conclusion The antiserum against H7N9 hemagglutinin for SRID assay was developed successfully,and could be used as a reagent for the quantitative detection of antigen in H7N9 influenza vaccine.