Aim To compare the cytotoxicity of geni-poside (GS)and its metabolite genipin (GP)on hu-man hepatocelluar HepG2 cells and explore the sub-stance and mechanism of hepatotoxicity induced by Fructus Gardeniae.Methods The cytotoxic effect of GS and GP was analyzed by MTT method;the antioxi-dant enzyme activities of manganese superoxide dis-mutase (Mn-SOD),catalase (CAT)and levels of glu-tathione (GSH)were detected by respective kits;the change of intracellular reactive oxygen species (ROS ) was measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA)staining method;multiparameter cytotox-icity analysis (cell loss,nuclear size and morphologi-cal changes,DNA content,cell membrane permeabili-ty,mitochondrial membrane potential changes,cyto-chrome C release ) were measured simultaneously by high content screening (HCS)assays.Results No cytotoxicity was found in GS (20~1 000 μmol·L-1 ) groups (P>0.05 ),but GP was found to exert obvi-ous cytotoxic effect,and 50μmol·L-1 GP could obvi-ously inhibit HepG2 cell proliferation (P<0.05 ),and the IC50 value was (450.00 ±26.15)μmol·L-1;GS showed no obvious effects on Mn-SOD, CAT, GSH ,ROS (P >0. 0 5 ),GP could significantly decrease the activity of Mn-SOD,CAT and the level of GSH,and obviously increase the content of ROS (P<0.05 or P <0.01 );treatment with 50,500,1 000μmol·L-1 GP resulted in loss of mitochondrial mem-brane potential,cytochrome C release and increased cell permeability (P<0.05 or P<0.01),500,1 000μmol· L-1 GP could also show abvious nuclear con-densation,increased total nuclear intensity and cell loss (P<0.0 1 ).Opposed with GP,GS had no signif-icant effect on the cytotoxic parameters (P>0.05 ). Conclusion GP is the direct substance of hepatotox-icity induced by Fructus Gardeniae,and the mecha-nism might be associated with oxidative stress,mito-chondria injury and apoptosis.

OBJECTIVE:To study in vitro bacteriostatic activity of the extracts from Miao medicine Polygonum runcinatum. METHODS:Using chloramphenicol and fluconazole as positive control,the inhibitory effects of water extract,95％ ethanol extract and its ethyl acetate and n-butanol fraction on 9 kinds of strains were determined by cup plate method, such as Staphylococcus aureus,Escherichia coli,Shigella flexneri,Salmonella typhi,Bacillus subtilis,Pseudomonas aeruginosa,type B Hemolytic streptococcus,Candida albicans and Cryptococcus neoformans. The parts with bacteriostatic activity and trial strains sensitive to drug were screened. Micro-broth dilution method and agar culture medium plate method were used to determine MIC and MBC of 95％ ethanol extract and its fractions of P. runcinatum to sensitive strains. RESULTS:The water extract of P. runcinatum almost had no inhibitory effect. 95％ ethanol extract showed different degrees of bacteriostatic activity to strains;bacteriostatic effects of it to 6 kinds of bacteria as S. typhi was better than that of type B H. streptococcus and 2 kinds of fungus. The ethyl acetate and n-butanol fraction of 95％ ethanol extracts showed good bacteriostatic effect and the ethyl acetate fraction had stronger effect,while all the fractions of 95％ ethanol extracts showed no inhibitory effect on fungus. MIC and MBC of 95％ethanol extract to above 6 kinds of bacteria were 6.25-12.5 and 12.5-25 mg/mL,respectively;MIC and MBC of ethyl acetate fraction were 3.13-6.25, 6.25-12.5 mg/mL; MIC and MBC of n-butanol fraction were 6.25-12.5, 12.5-25 mg/mL. CONCLUSIONS:95％ ethanol extract,ethyl acetate and n-butanol fractions of Miao medicine P. runcinatum have obvious in vitro bacteriostatic activity to 6 common bacterias as S. typhi.

Objective:To investigate the value of robot-assisted laparoscopic indocyanine green sentinel lymph node (SLN) tracing in treating endometrial carcinoma.Methods:Thirty-two patients with early-staging endometrial carcinoma were operated with laparoscopic comprehensive staging laparotomy from January 2019 to December 2021. At the same time, the SLN detection was performed by near-infrared fluorescence imaging tracer technology, in which the tracer was indocyanine green. Sixteen cases were injected with indocyanine green before laparoscopic surgery, and 16 cases were injected with indocyanine green before robot-assisted laparoscopic surgery. The operation index, postoperative complications, prognosis, and lymph node dissection were compared between the two groups.Results:(1) The mean age of patients in the robot group was (54.7±8.1) years old, and was (54.9±8.8) years old in the laparoscopic group. There were no significant difference between the two groups ( t=0.06, P=0.951). (2) Intraoperative blood loss [(131±40) vs (169±57) ml], hemoglobin difference before and after surgery [(11.2±5.4) vs (15.5±5.7) g/L], the length of stay after operation [(6.2±1.3) vs (8.6±1.4) days] between the robot group and the laparoscopic group were compared, and the differences were statistically significant (all P<0.05). (3) SLNs were detected in all 16 patients in the robotic group, and a total of 41 SLNs were detected. SLNs were detected in 15 of the 16 patients in the laparoscopy group, and a total of 40 SLNs were detected. Compared with the laparoscopic group (15/16), the total detection rate of SLN in the robotic group (16/16), there were no statistical significance ( χ2=1.03, P=0.310). Compared with the laparoscopic group (7/15), the SLN bilateral detection rate in the robotic group (10/16), there were also no significant difference ( χ2=0.78, P=0.376). The number of lymph nodes detected in surgery group (16.6±4.1) were lower than those in the laparoscopy surgery group (21.0±7.1), while there were no statistically difference between the two groups ( χ2=2.01, P=0.054). There was no tumor metastasis in the resected lymph nodes and SLN between the two groups. The false negative rate of SLN in diagnosing endometrial cancer postoperative lymph node metastasis was 0, and the negative predictive value was 100%. (4) The pelvic and retroperitoneal lymph nodes were divided into five regions, which were the left pelvis, the right pelvis, the presacral region, the deep inguinal region, and the abdominal aorta. The numbers of SLN of unilateral detection and bilateral pelvic detection between two groups showed no significant differences (all P>0.05). The left pelvis had the most SLN imaging in both groups, followed by the right pelvis, para-aortic, and deep groin. (5) There was one patient in both robotic group and laparoscopic group with postoperative complications, which were urinary retention and pelvic lymph node cyst respectively. There were no significant differences in the incidence of complications between the two groups ( χ2=0.97, P=1.000). The median follow-up time after operation was 14 months (range 6-24 months). During the follow-up period, no local recurrence or distant metastasis was found between the two groups of endometrial cancer patients. Conclusions:Compared with the laparoscopic group, the robot group has less intraoperative blood loss and shorter postoperative hospital stay. The bilateral detection rate of SLN in the group was better than that of laparoscopy.

Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID₅₀, respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 μg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 μg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10⁴ and 1×10⁵. The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.