OBJECTIVE To explore the effect and potential mechanisms of berberine on formation of biofilms of clinical methicillin-resistant Staphylococcus aureus(MRSA)isolates.METHODS Totally 95 clinical MRSA iso-lates were collected from Shanghai Eighth People's Hospital from Jan.2023 to Dec.2023.The 14 biofilm forma-tion-related genes in the strains were detected by polymerase chain reaction(PCR)and multiplex PCR,the mini-mum inhibitory concentration(MIC)of berberine was determined by microbroth dilution method,the effect of berberine on resistance of biofilm formation was evaluated by crustal violet staining,fluorescence microscope,Congo red agar plate and extracelluar DNA(eDNA).The transcription levels of 9 biofilm formation-related genes were detected by real-time fluorescent quantitative reverse transcription PCR.RESULTS All of the 95 strains of MRSA carried eno,clfA,clfB and icaA genes,the most widespread gene profile was bbp-eno-ebpS-fnbA-fib-clfA-clfB-icaA-sasG,and 29 strains had the phenotypes with strong capability of biofilm formation.The MIC score of berberine ranged between 64 and 1 024 μg/ml.Berberine with the concentration of 1/2 MIC could inhibit the biofilm formation of MRSA(P＜0.001),and the inhibiting rate of biofilm formation of the MIC ≥512 μg/ml group was higher than that of the MIC≤128 μg/ml group and the MIC 256 μg/ml group(all P＜0.05).The re-sult under the fluorescence microscope showed that the fluorescence intensity of biofilms decreased with the rise of berberine concentration.Berberine could reduce the formation of amyloid fibrils and the release of eDNA,down-regulating the transcription levels of ica A,sasG,ebpS,fib,eno,clfA,clfB,bbp and fnbA genes(P＜0.05).CONCLUSION Berberine may inhibit the biofilm formation of MRSA by downregulating expression levels of related genes,interfering the formation of amyloid fibrils and blocking the release of eDNA,which may provide experimental bases for development of drugs resisting to MRSA biofilms.

Objective:To explore the prognostic differences and influencing factors between pancreatic cancer originated from intraductal papillary mucinous neoplasm (IPMN)-termed IC-Ds-and pancreatic cancer concomitant with IPMN (C-PDACs).Methods:Clinical data of 382 patients with pathologically confirmed IPMN who underwent surgical resection in the Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Naval Medical University from January 2016 to January 2022 were collected. According to pathological diagnosis, patients were divided into the IC-Ds group ( n=288) and the C-PDACs group ( n=94). The IC-Ds group was further divided into the colloid carcinoma subgroup and the ductal adenocarcinoma subgroup based on pathological typing. Data including age, gender, preoperative CA19-9 level, surgical margin status, lymph node metastasis, pathological grade, T stage, postoperative adjuvant chemotherapy, and survival follow-up were recorded. The median follow-up time was 35.00 months for IC-Ds patients and 29.00 months for C-PDACs patients. Clinicopathological characteristics and prognostic factors were compared between the IC-Ds and C-PDACs groups, as well as between the colloid carcinoma and ductal adenocarcinoma subgroups. Kaplan-Meier curves for overall survival were generated. Results:There were no significant differences in age, gender, R1 resection margin between the IC-Ds group and the C-PDACs group. However, in the C-PDACs group, 70 cases (74.47%) had elevated preoperative CA19-9, 40 cases (42.55%) had lymph node metastasis, 25 cases (26.60%) were pathologically confirmed as poorly differentiated carcinoma after surgery, and 54 cases (57.45%) received postoperative adjuvant chemotherapy; the proportions of all these indicators were higher than those in the IC-Ds group (90/288, 31.25%; 72/288, 25.00%; 32/288, 11.11%; 105/288, 36.46%). In contrast, the proportion of T1 stage in the IC-Ds group was higher (40.97% vs 20.21%), and all these differences were statistically significant (all P value <0.05). Among the 288 patients in the IC-Ds group, 97 cases (33.68%) were colloid carcinoma and 191 cases (66.32%) were ductal adenocarcinoma. There were no significant differences in age, gender, R1 resection margin, proportion of poorly differentiated carcinoma between the two subgroups. However, in the ductal adenocarcinoma subgroup, 67 cases (35.08%) had elevated preoperative CA19-9, 56 cases (29.32%) had lymph node metastasis confirmed by postoperative pathology, and 80 cases (41.88%) received postoperative adjuvant chemotherapy; all these proportions were significantly higher than those in the colloid carcinoma subgroup (23/97, 23.71%; 16/97, 17.02%; 25/97, 26.60%). In addition, the ductal adenocarcinoma subgroup had higher proportions of T2 and T3/T4 stages, while the proportion of T1 stage in the colloid carcinoma subgroup (60/97, 61.86%) was significantly higher than that in the ductal adenocarcinoma subgroup (58/191, 30.37%), with all differences being statistically significant (all P value <0.05). The median survival time was 47.00 months (95% CI 42.91-51.09) in the IC-Ds group and 34.00 months (95% CI 29.67-38.33) in the C-PDACs group. For the IC-Ds subgroups, the median survival time was 59.00 months (95% CI 50.79-67.21) in the colloid carcinoma subgroup and 42.00 months (95% CI 35.15-48.85) in the ductal adenocarcinoma subgroup. Significant differences in median survival time were observed between the IC-Ds and C-PDACs groups, between the colloid carcinoma and ductal adenocarcinoma subgroups, and between the ductal adenocarcinoma subgroup and the C-PDACs group (all P value <0.01). Conclusions:IC-Ds has a better prognosis than C-PDACs, and there is significant heterogeneity within IC-Ds, indicating different biological behaviors between the two types, which requires the development of targeted diagnosis and treatment strategies.

Objective:To explore the prognostic differences and influencing factors between pancreatic cancer originated from intraductal papillary mucinous neoplasm (IPMN)-termed IC-Ds-and pancreatic cancer concomitant with IPMN (C-PDACs).Methods:Clinical data of 382 patients with pathologically confirmed IPMN who underwent surgical resection in the Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Naval Medical University from January 2016 to January 2022 were collected. According to pathological diagnosis, patients were divided into the IC-Ds group ( n=288) and the C-PDACs group ( n=94). The IC-Ds group was further divided into the colloid carcinoma subgroup and the ductal adenocarcinoma subgroup based on pathological typing. Data including age, gender, preoperative CA19-9 level, surgical margin status, lymph node metastasis, pathological grade, T stage, postoperative adjuvant chemotherapy, and survival follow-up were recorded. The median follow-up time was 35.00 months for IC-Ds patients and 29.00 months for C-PDACs patients. Clinicopathological characteristics and prognostic factors were compared between the IC-Ds and C-PDACs groups, as well as between the colloid carcinoma and ductal adenocarcinoma subgroups. Kaplan-Meier curves for overall survival were generated. Results:There were no significant differences in age, gender, R1 resection margin between the IC-Ds group and the C-PDACs group. However, in the C-PDACs group, 70 cases (74.47%) had elevated preoperative CA19-9, 40 cases (42.55%) had lymph node metastasis, 25 cases (26.60%) were pathologically confirmed as poorly differentiated carcinoma after surgery, and 54 cases (57.45%) received postoperative adjuvant chemotherapy; the proportions of all these indicators were higher than those in the IC-Ds group (90/288, 31.25%; 72/288, 25.00%; 32/288, 11.11%; 105/288, 36.46%). In contrast, the proportion of T1 stage in the IC-Ds group was higher (40.97% vs 20.21%), and all these differences were statistically significant (all P value <0.05). Among the 288 patients in the IC-Ds group, 97 cases (33.68%) were colloid carcinoma and 191 cases (66.32%) were ductal adenocarcinoma. There were no significant differences in age, gender, R1 resection margin, proportion of poorly differentiated carcinoma between the two subgroups. However, in the ductal adenocarcinoma subgroup, 67 cases (35.08%) had elevated preoperative CA19-9, 56 cases (29.32%) had lymph node metastasis confirmed by postoperative pathology, and 80 cases (41.88%) received postoperative adjuvant chemotherapy; all these proportions were significantly higher than those in the colloid carcinoma subgroup (23/97, 23.71%; 16/97, 17.02%; 25/97, 26.60%). In addition, the ductal adenocarcinoma subgroup had higher proportions of T2 and T3/T4 stages, while the proportion of T1 stage in the colloid carcinoma subgroup (60/97, 61.86%) was significantly higher than that in the ductal adenocarcinoma subgroup (58/191, 30.37%), with all differences being statistically significant (all P value <0.05). The median survival time was 47.00 months (95% CI 42.91-51.09) in the IC-Ds group and 34.00 months (95% CI 29.67-38.33) in the C-PDACs group. For the IC-Ds subgroups, the median survival time was 59.00 months (95% CI 50.79-67.21) in the colloid carcinoma subgroup and 42.00 months (95% CI 35.15-48.85) in the ductal adenocarcinoma subgroup. Significant differences in median survival time were observed between the IC-Ds and C-PDACs groups, between the colloid carcinoma and ductal adenocarcinoma subgroups, and between the ductal adenocarcinoma subgroup and the C-PDACs group (all P value <0.01). Conclusions:IC-Ds has a better prognosis than C-PDACs, and there is significant heterogeneity within IC-Ds, indicating different biological behaviors between the two types, which requires the development of targeted diagnosis and treatment strategies.

OBJECTIVE To explore the effect and potential mechanisms of berberine on formation of biofilms of clinical methicillin-resistant Staphylococcus aureus(MRSA)isolates.METHODS Totally 95 clinical MRSA iso-lates were collected from Shanghai Eighth People's Hospital from Jan.2023 to Dec.2023.The 14 biofilm forma-tion-related genes in the strains were detected by polymerase chain reaction(PCR)and multiplex PCR,the mini-mum inhibitory concentration(MIC)of berberine was determined by microbroth dilution method,the effect of berberine on resistance of biofilm formation was evaluated by crustal violet staining,fluorescence microscope,Congo red agar plate and extracelluar DNA(eDNA).The transcription levels of 9 biofilm formation-related genes were detected by real-time fluorescent quantitative reverse transcription PCR.RESULTS All of the 95 strains of MRSA carried eno,clfA,clfB and icaA genes,the most widespread gene profile was bbp-eno-ebpS-fnbA-fib-clfA-clfB-icaA-sasG,and 29 strains had the phenotypes with strong capability of biofilm formation.The MIC score of berberine ranged between 64 and 1 024 μg/ml.Berberine with the concentration of 1/2 MIC could inhibit the biofilm formation of MRSA(P＜0.001),and the inhibiting rate of biofilm formation of the MIC ≥512 μg/ml group was higher than that of the MIC≤128 μg/ml group and the MIC 256 μg/ml group(all P＜0.05).The re-sult under the fluorescence microscope showed that the fluorescence intensity of biofilms decreased with the rise of berberine concentration.Berberine could reduce the formation of amyloid fibrils and the release of eDNA,down-regulating the transcription levels of ica A,sasG,ebpS,fib,eno,clfA,clfB,bbp and fnbA genes(P＜0.05).CONCLUSION Berberine may inhibit the biofilm formation of MRSA by downregulating expression levels of related genes,interfering the formation of amyloid fibrils and blocking the release of eDNA,which may provide experimental bases for development of drugs resisting to MRSA biofilms.