Insufficient alveolar bone thickness increases the risk of periodontal dehiscence and fenestration, especially in orthodontic tooth movement. Abaloparatide (ABL), a synthetic analog of human PTHrP (1-34) and a clinical medication for treating osteoporosis, has recently demonstrated its potential in enhancing craniofacial bone formation. Herein, we show that intraoral submucosal injection of ABL, when combined with mechanical force, promotes in situ alveolar bone thickening. The newly formed bone is primarily located outside the original compact bone, implying its origin from the periosteum. RNA sequencing of the alveolar bone tissue revealed that the focal adhesion (FA) pathway potentially mediates this bioprocess. Local injection of ABL alone enhances cell proliferation, collagen synthesis, and phosphorylation of focal adhesion kinase (FAK) in the alveolar periosteum; when ABL is combined with mechanical force, the FAK expression is upregulated, in line with the accomplishment of the ossification. In vitro, ABL enhances proliferation, migration, and FAK phosphorylation in periosteal stem cells. Furthermore, the pro-osteogenic effects of ABL on alveolar bone are entirely blocked when FAK activity is inhibited by a specific inhibitor. In summary, abaloparatide combined with mechanical force promotes alveolar bone formation via FAK-mediated periosteal osteogenesis. Thus, we have introduced a promising therapeutic approach for drug-induced in situ alveolar bone augmentation, which may prevent or repair the detrimental periodontal dehiscence, holding significant potential in dentistry.
Osteogenesis/drug effects* ; Periosteum/cytology* ; Parathyroid Hormone-Related Protein/administration & dosage* ; Animals ; Focal Adhesion Protein-Tyrosine Kinases/metabolism* ; Alveolar Process/drug effects* ; Cell Proliferation/drug effects* ; Phosphorylation ; Rats ; Male ; Humans ; Focal Adhesion Kinase 1/metabolism* ; Cell Movement/drug effects*

Objective:To explore genomic features associated with gemcitabine sensitivity, patient-derived organoid models of biliary tract cancer (BTC) were established and characterized.Methods:This is an experimental study. The tissue specimens of BTC were collected from patients who underwent surgical resection at the Department of Hepatobiliary and Pancreatic Surgery,the Second Affiliated Hospital of Zhejiang University School of Medicine between January 2020 and December 2023. The tumor organoids were cultured in vitro and histologically characterized. Drug sensitivity testing was performed using gemcitabine,cisplatin,paclitaxel,fluorouracil,and lenvatinib etc. to evaluate cell viability. The correlation between the drug sensitivity of organoids and clinical therapeutic response was analyzed.Results:Thirty-eight patient-derived organoids (PDO) models were successfully established from 43 biliary tract malignancy patients with complete follow-up data,including gallbladder cancer PDO 14 cases,distal bile duct cancer PDO 16 cases,intrahepatic cholangiocarcinoma PDO 8 cases,achieving an overall success rate of 88.4%. Drug sensitivity testing (DST) was performed on the successfully generated PDO,with 35 models successfully completing DST experiments. The overall consistency rate between drug responses in PDOs and clinical survival outcomes in corresponding patients was 8/14. Transcriptomic analysis of gemcitabine-sensitive vs. gemcitabine-resistant PDO identified 71 differentially expressed genes in the resistant group,the significantly up-regulated genes including GLDC, LINC01595, IL-27, ANGPTL3, CYP7A1,and AKR1C1;the significantly down-regulated genes including P2RY2,LIPC,and ECHDC3. Conclusion:A biobank of patient-derived organoids of BTC has been established,which demonstrates its potential as preclinical models and tools for predicting chemotherapy responses for BTC patients.

Objective:To explore genomic features associated with gemcitabine sensitivity, patient-derived organoid models of biliary tract cancer (BTC) were established and characterized.Methods:This is an experimental study. The tissue specimens of BTC were collected from patients who underwent surgical resection at the Department of Hepatobiliary and Pancreatic Surgery,the Second Affiliated Hospital of Zhejiang University School of Medicine between January 2020 and December 2023. The tumor organoids were cultured in vitro and histologically characterized. Drug sensitivity testing was performed using gemcitabine,cisplatin,paclitaxel,fluorouracil,and lenvatinib etc. to evaluate cell viability. The correlation between the drug sensitivity of organoids and clinical therapeutic response was analyzed.Results:Thirty-eight patient-derived organoids (PDO) models were successfully established from 43 biliary tract malignancy patients with complete follow-up data,including gallbladder cancer PDO 14 cases,distal bile duct cancer PDO 16 cases,intrahepatic cholangiocarcinoma PDO 8 cases,achieving an overall success rate of 88.4%. Drug sensitivity testing (DST) was performed on the successfully generated PDO,with 35 models successfully completing DST experiments. The overall consistency rate between drug responses in PDOs and clinical survival outcomes in corresponding patients was 8/14. Transcriptomic analysis of gemcitabine-sensitive vs. gemcitabine-resistant PDO identified 71 differentially expressed genes in the resistant group,the significantly up-regulated genes including GLDC, LINC01595, IL-27, ANGPTL3, CYP7A1,and AKR1C1;the significantly down-regulated genes including P2RY2,LIPC,and ECHDC3. Conclusion:A biobank of patient-derived organoids of BTC has been established,which demonstrates its potential as preclinical models and tools for predicting chemotherapy responses for BTC patients.

Objective:To investigate the clinical value of the novel approach,radical resection of the retroperitoneal lipo-lymphatic layer (RRRLLL),in the surgical treatment of resectable pancreatic head cancer.Methods:Between June 2020 and June 2022,a total of 221 patients with pancreatic head cancer underwent surgical treatment using the RRRLLL approach(RRRLLL group),while 107 patients received traditional surgical treatment(traditional group) in five high-volume pancreatic centers in China. Data from surgical technique and clinical perioperative outcomes,including lymph node harvested,surgical time,and complications,were analyzed. The RRRLLL group consisted of 144 males and 77 females with an age of (67.5±9.0) years(range:41.3 to 81.1 years). The traditional group included 71 males and 36 females,with an age of (66.3±8.1) years(range:45.1 to 79.2 years). Statistical analysis was performed using the K-S test, Z test,or χ 2 test, respectively. Results:Pancreaticoduodenectomy was performed successfully in all patients,achieving R0 resection. RRRLLL group surgery required mobilization of retroperitoneal adipose and lymphatic tissues starting from the right edge of the inferior vena cava and extending to the left side,up to the superior mesenteric artery,down to the inferior mesenteric artery,and left to the left side of the aorta,including the perineural and lymphatic tissues around the superior mesenteric artery and the sheath of the mesenteric artery. However,the traditional group did not include the areas mentioned above in the scope of clearance. There were no statistically significant differences between the RRRLLL group and the traditional group in terms of age,sex,tumor size,T stage,and vascular invasion (all P>0.05). However,the number of lymph nodes harvested in the RRRLLL group was significantly higher at 28.7±9.0 (range: 18 to 39) compared to 18.2±8.0 (range: 12 to 21) in the traditional group ( Z=-10.691, P<0.05). There were no statistically significant differences in the number of positive lymph nodes,N staging,and postoperative complications between the two groups. Conclusion:The RRRLLL approach improved lymph node dissection compared to the traditional approach,potentially leading to reduced recurrence rates.

Objective:To investigate the clinical value of the novel approach,radical resection of the retroperitoneal lipo-lymphatic layer (RRRLLL),in the surgical treatment of resectable pancreatic head cancer.Methods:Between June 2020 and June 2022,a total of 221 patients with pancreatic head cancer underwent surgical treatment using the RRRLLL approach(RRRLLL group),while 107 patients received traditional surgical treatment(traditional group) in five high-volume pancreatic centers in China. Data from surgical technique and clinical perioperative outcomes,including lymph node harvested,surgical time,and complications,were analyzed. The RRRLLL group consisted of 144 males and 77 females with an age of (67.5±9.0) years(range:41.3 to 81.1 years). The traditional group included 71 males and 36 females,with an age of (66.3±8.1) years(range:45.1 to 79.2 years). Statistical analysis was performed using the K-S test, Z test,or χ 2 test, respectively. Results:Pancreaticoduodenectomy was performed successfully in all patients,achieving R0 resection. RRRLLL group surgery required mobilization of retroperitoneal adipose and lymphatic tissues starting from the right edge of the inferior vena cava and extending to the left side,up to the superior mesenteric artery,down to the inferior mesenteric artery,and left to the left side of the aorta,including the perineural and lymphatic tissues around the superior mesenteric artery and the sheath of the mesenteric artery. However,the traditional group did not include the areas mentioned above in the scope of clearance. There were no statistically significant differences between the RRRLLL group and the traditional group in terms of age,sex,tumor size,T stage,and vascular invasion (all P>0.05). However,the number of lymph nodes harvested in the RRRLLL group was significantly higher at 28.7±9.0 (range: 18 to 39) compared to 18.2±8.0 (range: 12 to 21) in the traditional group ( Z=-10.691, P<0.05). There were no statistically significant differences in the number of positive lymph nodes,N staging,and postoperative complications between the two groups. Conclusion:The RRRLLL approach improved lymph node dissection compared to the traditional approach,potentially leading to reduced recurrence rates.

Objective:To explore the feasibility and clinical value of a new classification for resectable intrahepatic cholangiocarcinoma (IHCC) according to the actual anatomy.Methods:The data of 135 patients with IHCC who were admitted to the Department of Hepatopancreatobiliary Surgery ,Second Affiliated Hospital of Zhejiang University School of Medicine from November 2011 to November 2020 after discussion by a multidisciplinary team and planned to undergo radical resection were analyzed retrospectively. There were 77 males and 58 females,with a median age of 61 years (range:26 to 86 years),of which 38 cases had vascular invasion. This new classification was carried out independently by two hepatobiliary surgeons. First,a preliminary classification was made based on the location of the tumor,and then the final classification was based on vascular invasion. All patients were followed up by telephone,and the follow-up was as of November 2020. Survival time is defined as the time after surgery to follow-up or death. Log-rank test was used to compare patients′ median recurrence-free survival and overall survival time. The Cox proportional hazard model was used to analyze the prognosis factors of the overall survival time of patients with IHCC. Results:Among the 135 patients,129 underwent R0 resection and 6 underwent R1 resection. According to the actual anatomy,28 cases (20.7%) belonged to segmental type, 43 cases (31.9%) belonged to branch type, 64 cases (47.4%). The median survival time of all patients was 35.2 months(95% CI:21.3 to 70.5 months),the 1-year cumulative survival rate was 75.1%,the 3-year cumulative survival rate was 45.8%,and the 5-year cumulative survival rate was 39.0%. After grouping according to the classification,the median survival time of segmental patients was 36.9 months (more than 50% of patients reached the median survival time),and the median survival time of branched patients was 33.8 months (95% CI:16.8 to 38.5);The median survival time of lobe patients was 25.0 months (95% CI:13.6 to 58.7). The result of Log-rank test between groups indicated that the median survival time of patients with segmental type was better than that of patients with branch and lobe type( HR=2.03,95% CI:1.24 to 3.64, P=0.006);There was no significant difference in survival time between patients with branch type and lobe type ( P=0.685). The results of the multivariate analysis of the Cox risk ratio model suggested that the actual anatomical location classification ( HR=2.32,95% CI:1.10 to 4.92, P=0.028) and the postoperative lymph node metastasis rate ( HR=2.06,95% CI:1.24 to 3.45, P=0.005) were independent factors related to survival after radical resection of IHCC patients. Conclusion:It is simple and convenient to classify resectable IHCC by actual anatomy,which can be used to preliminarily judge the prognosis of patients and provide a feasible classification scheme for the clinic.

Objective:To explore the feasibility and clinical value of a new classification for resectable intrahepatic cholangiocarcinoma (IHCC) according to the actual anatomy.Methods:The data of 135 patients with IHCC who were admitted to the Department of Hepatopancreatobiliary Surgery ,Second Affiliated Hospital of Zhejiang University School of Medicine from November 2011 to November 2020 after discussion by a multidisciplinary team and planned to undergo radical resection were analyzed retrospectively. There were 77 males and 58 females,with a median age of 61 years (range:26 to 86 years),of which 38 cases had vascular invasion. This new classification was carried out independently by two hepatobiliary surgeons. First,a preliminary classification was made based on the location of the tumor,and then the final classification was based on vascular invasion. All patients were followed up by telephone,and the follow-up was as of November 2020. Survival time is defined as the time after surgery to follow-up or death. Log-rank test was used to compare patients′ median recurrence-free survival and overall survival time. The Cox proportional hazard model was used to analyze the prognosis factors of the overall survival time of patients with IHCC. Results:Among the 135 patients,129 underwent R0 resection and 6 underwent R1 resection. According to the actual anatomy,28 cases (20.7%) belonged to segmental type, 43 cases (31.9%) belonged to branch type, 64 cases (47.4%). The median survival time of all patients was 35.2 months(95% CI:21.3 to 70.5 months),the 1-year cumulative survival rate was 75.1%,the 3-year cumulative survival rate was 45.8%,and the 5-year cumulative survival rate was 39.0%. After grouping according to the classification,the median survival time of segmental patients was 36.9 months (more than 50% of patients reached the median survival time),and the median survival time of branched patients was 33.8 months (95% CI:16.8 to 38.5);The median survival time of lobe patients was 25.0 months (95% CI:13.6 to 58.7). The result of Log-rank test between groups indicated that the median survival time of patients with segmental type was better than that of patients with branch and lobe type( HR=2.03,95% CI:1.24 to 3.64, P=0.006);There was no significant difference in survival time between patients with branch type and lobe type ( P=0.685). The results of the multivariate analysis of the Cox risk ratio model suggested that the actual anatomical location classification ( HR=2.32,95% CI:1.10 to 4.92, P=0.028) and the postoperative lymph node metastasis rate ( HR=2.06,95% CI:1.24 to 3.45, P=0.005) were independent factors related to survival after radical resection of IHCC patients. Conclusion:It is simple and convenient to classify resectable IHCC by actual anatomy,which can be used to preliminarily judge the prognosis of patients and provide a feasible classification scheme for the clinic.

In this study, a new combined enzyme immunoassay(NRAg ELISA) for detection of HBV PreS1 and core antigens which was highly consistent with serum HBV DNA test was established. The serial serum dilution test indicated that the average sensitivity of the assay was 10(3.2) genome copies/mL (95% CI: 10(2.2-4.2) genome copies/mL), which was notably higher than the test performed on Pre S1 or core antigen alone. The test with sera from 994 blood donors whose HBsAg were negative demonstrated that the specificity of this assay was 99.7% (95% CI: 99.1%-99.9%). 271 serum samples from chronic hepatitis patients were also examined and the result showed that the total consistent rate between NRAg ELISA and HBV DNA was 96.3% (95% CI: 93.3%-98.2%). The NRAg ELISA S/CO(signal/cutoff) was closely correlated with HBV genome copies (R = 0.9158, n=231). Furthermore,by using this assay,we found a patient whose HBsAg was negative but HBV DNA was positive. Sequencing result showed that HBV genome from this patient had a point mutation in the "a"epitope of S gene. Our results indicate that HBV NRAg ELISA has a high relativity with HBV DNA test, and can effectively detect the mutation of HBsAg,it is expected to be a potent tool for screening HBsAg mutant and is a convenient method for substituting HBV DNA test.
DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Hepatitis B Antigens ; blood ; Hepatitis B Core Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results