[Predict the effect of the number of positive preoperative serum tumor markers on the surgical method and prognosis of intrahepatic cholangiocarcinoma patients based on mediation analysis].

Objective: To investigate the effect of the number of positive preoperative serological tumor markers on the surgical approach and prognosis of patients with intrahepatic cholangiocarcinoma. Methods: This is a retrospective case-series study. Data from 548 patients with intrahepatic cholangiocarcinoma after radical resection from October 2010 to April 2019 were retrospectively collected in 10 hospitals of China. There were 277 males and 271 females with an age of (57.8±10.2)years(range:23 to 84 years). Four hundred and twenty-six patients(77.7%) had at least one positive preoperative serum tumor marker. The data collection included the results of 4 preoperative serological tumor markers,other preoperative indicators(5 prodromal symptoms, 6 medical history,8 preoperative serological indicators,5 preoperative imaging indicators,and 14 preoperative pathological examination indicators),baseline data (gender and age),surgical methods,and prognostic follow-up data. Four preoperative results of serologic tumor marker and surgical procedure were converted into categorical variables. The number of positive preoperative serum tumor markers was used as the treatment variable,the surgical method was used as the mediating variable,and the survival time was used as the outcome variable. Univariate and multivariate analysis were used to screen for other preoperative indicators which were independent factors that influenced the surgical procedure and the prognosis of patients as covariates to analyze the mediating effect. Results: Of the 548 patients included in the study, 176 patients (32.1%) underwent partial hepatectomy,151 patients(27.5%) underwent hemihepatectomy, and 221 patients(40.3%) underwent partial hepatectomy or hemihepatectomy combined with other treatments. The results of the univariate and multivariate analysis showed that the number of positive serum tumor markers,intrahepatic bile duct dilatation,portal vein invasion,pathological differentiation,pathological type,vascular invasion,T stage,N stage and maximum tumor diameter were independent factors influencing the surgical procedure(all P<0.05). Intrahepatic bile duct dilatation,pathological differentiation and T stage were independent prognostic factors for patients with intrahepatic cholangiocarcinoma(all P<0.05). Intrahepatic bile duct dilatation,differentiation and T stage were included as covariates in the mediation effect model. The results showed that the number of positive serum tumor markers before surgery had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma (ß=-0.092, P=0.039),and had a positive predictive effect on the surgical method (ß=0.244,P<0.01). The number of positive serum tumor markers had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma (ß=-0.151, P=0.002). Direct and indirect effects accounted for 71.3% and 28.7% of total effects,respectively. Conclusions: The higher the positive number of preoperative tumor markers,the worse the prognosis of patients with intrahepatic cholangiocarcinoma. The number of positive cells not only directly affects the prognosis of patients,but also indirectly affects the prognosis of patients by affecting the surgical method.

[Survival analysis of patients with intrahepatic cholangiocarcinoma treated with adjuvant chemotherapy after radical resection based on CoxPH model and deep learning algorithm].

Objective: To establish a predictive model for survival benefit of patients with intrahepatic cholangiocarcinoma (ICC) who received adjuvant chemotherapy after radical resection. Methods: The clinical and pathological data of 249 patients with ICC who underwent radical resection and adjuvant chemotherapy at 8 hospitals in China from January 2010 to December 2018 were retrospectively collected. There were 121 males and 128 females,with 88 cases>60 years old and 161 cases≤60 years old. Feature selection was performed by univariate and multivariate Cox regression analysis. Overall survival time and survival status were used as outcome indicators,then target clinical features were selected. Patients were stratified into high-risk group and low-risk group,survival differences between the two groups were analyzed. Using the selected clinical features, the traditional CoxPH model and deep learning DeepSurv survival prediction model were constructed, and the performance of the models were evaluated according to concordance index(C-index). Results: Portal vein invasion, carcinoembryonic antigen>5 µg/L,abnormal lymphocyte count, low grade tumor pathological differentiation and positive lymph nodes>0 were independent adverse prognostic factors for overall survival in 249 patients with adjuvant chemotherapy after radical resection (all P<0.05). The survival benefit of adjuvant chemotherapy in the high-risk group was significantly lower than that in the low-risk group (P<0.05). Using the above five features, the traditional CoxPH model and the deep learning DeepSurv survival prediction model were constructed. The C-index values of the training set were 0.687 and 0.770, and the C-index values of the test set were 0.606 and 0.763,respectively. Conclusion: Compared with the traditional Cox model, the DeepSurv model can more accurately predict the survival probability of patients with ICC undergoing adjuvant chemotherapy at a certain time point, and more accurately judge the survival benefit of adjuvant chemotherapy.

[A nomogram for preoperative prediction of lymph node metastasis in patients with intrahepatic cholangiocarcinoma based on inflammation-related markers].

Objectives: To construct a nomogram for prediction of intrahepatic cholangiocarcinoma (ICC) lymph node metastasis based on inflammation-related markers,and to conduct its clinical verification. Methods: Clinical and pathological data of 858 ICC patients who underwent radical resection were retrospectively collected at 10 domestic tertiary hospitals in China from January 2010 to December 2018. Among the 508 patients who underwent lymph node dissection,207 cases had complete variable clinical data for constructing the nomogram,including 84 males,123 females,109 patients≥60 years old,98 patients<60 years old and 69 patients were pathologically diagnosed with positive lymph nodes after surgery. Receiver operating characteristic curve was drawn to calculate the accuracy of preoperative imaging examinations to determine lymph node status,and the difference in overall survival time was compared by Log-rank test. Partial regression squares and statistically significant preoperative variables were screened by backward stepwise regression analysis. R software was applied to construct a nomogram,clinical decision curve and clinical influence curve,and Bootstrap method was used for internal verification. Moreover,retrospectively collecting clinical information of 107 ICC patients with intraoperative lymph node dissection admitted to 9 tertiary hospitals in China from January 2019 to June 2021 was for external verification to verify the accuracy of the nomogram. 80 patients with complete clinical data but without lymph node dissection were divided into lymph node metastasis high-risk group and low-risk group according to the score of the nomogram among the 858 patients. Log-rank test was used to compare the overall survival of patients with or without lymph node metastasis diagnosed by pathology. Results: The area under the curve of preoperative imaging examinations for lymph node status assessment of 440 patients was 0.615,with a false negative rate of 62.8% (113/180) and a false positive rate of 14.2% (37/260). The median survival time of 207 patients used to construct a nomogram with positive or negative postoperative pathological lymph node metastases was 18.5 months and 27.1 months,respectively (P<0.05). Five variables related to lymph node metastasis were screened out by backward stepwise regression analysis,which were combined calculi,neutrophil/lymphocyte ratio,albumin,liver capsule invasion and systemic immune inflammation index,according to which a nomogram was constructed with concordance index(C-index) of 0.737 (95%CI: 0.667 to 0.806). The C-index of external verification was 0.674 (95%CI:0.569 to 0.779). The calibration prediction curve was in good agreement with the reference curve. The results of the clinical decision curve showed that when the risk threshold of high lymph node metastasis in the nomogram was set to about 0.32,the maximum net benefit could be obtained by 0.11,and the cost/benefit ratio was 1∶2. The results of clinical influence curve showed that when the risk threshold of high lymph node metastasis in the nomogram was set to about 0.6,the probability of correctly predicting lymph node metastasis could reach more than 90%. There was no significant difference in overall survival time between patients with high/low risk of lymph node metastasis assessed by the nomogram and those with pathologically confirmed lymph node metastasis or without lymph node metastasis (Log-rank test:P=0.082 and 0.510,respectively). Conclusion: The prediction accuracy of preoperative nomogram for ICC lymph node metastasis based on inflammation-related markers is satisfactory,which can be used as a supplementary method for preoperative diagnosis of lymph node metastasis and is helpful for clinicians to make personalized decision of lymph node dissection for patients with ICC.

[Impact of adjuvant chemotherapy on prognosis in intrahepatic cholangiocarcinoma patients underwent radical resection].

Objectives: To investigate the clinical value of adjuvant chemotherapy(ACT) in patients with intrahepatic cholangiocarcinoma(ICC) who underwent radical resection and to explore the optimal population that can benefit from ACT. Methods: A retrospective cohort study method was adopted. The clinical and pathological data of 685 patients with ICC who underwent curative intent resection in 10 Chinese hepatobiliary surgery centers from January 2010 to December 2018 were collected;There were 355 males and 330 females. The age(M(IQR)) was 58(14) years (range: 22 to 83 years). Propensity score matching(PSM) was applied to balance the differences between the adjuvant and non-adjuvant chemotherapy groups. Log-rank test was used to compare the prognosis of the two groups of patients. A Bayesian network recurrence-free survival(RFS) prediction model was constructed using the median RFS time (14 months) as the target variable, and the importance of the relevant prognostic factors was ranked according to the multistate Birnbaum importance calculation. A survival prognostic prediction table was established to analyze the population benefiting from adjuvant chemotherapy. Results: Among 685 patients,214 received ACT and 471 did not receive ACT. A total of 124 pairs of patients were included after PSM, and patients in the ACT group had better overall survival (OS) and RFS than those in the non-ACT group(OS: 32.2 months vs. 18.0 months,P=0.003;RFS:18.0 months vs. 10.0 months,P=0.001). The area under the curve of the Bayesian network RFS prediction model was 0.7124. The results of the prognostic factors in order of importance were microvascular invasion (0.158 2),perineural invasion (0.158 2),N stage (0.155 8),T stage (0.120 9), hepatic envelope invasion (0.090 3),adjuvant chemotherapy (0.072 1), tumor location (0.057 5), age (0.042 3), pathological differentiation (0.034 0), sex (0.029 3), alpha-fetoprotein (0.028 9) and preoperative jaundice (0.008 5). A survival prediction table based on the variables with importance greater than 0.1 (microvascular invasion,perineural invasion,N stage,T staging) and ACT showed that all patients benefited from ACT (increase in the probability of RFS≥14 months from 2.21% to 7.68%), with a more significant increase in the probability of RFS≥14 months after ACT in early-stage patients. Conclusion: ACT after radical resection in patients with ICC significantly prolongs the OS and RFS of patients, and the benefit of ACT is greater in early patients.

[The analysis of long-term prognostic factors after laparoscopic liver resection for intrahepatic cholangiocarcinoma and establishment of survival Nomogram model].

Objective: To establish a survival prediction model based on the independent prognostic factors of long-term prognosis after laparoscopic liver resection(LLR) for intrahepatic cholangiocarcinoma(ICC). Methods: The clinical and pathological data of 351 consecutive patients with ICC who received radical LLR in 13 Chinese medical centers from August 2010 to May 2021 were collected retrospectively. There were 190 males and 161 females,aged(M(IQR)) 61(14)years(range:23 to 93 years). The total cohort was randomly divided into a training dataset(264 cases) and a validation dataset(87 cases). The patients were followed up by outpatient service or telephone,and the deadline for follow-up was October 2021. Based on the training dataset,the multivariate Cox proportional hazards regression model was used to screen the independent influencing factors of long-term prognosis to construct a Nomogram model. The Nomogram model's discrimination,calibration,and clinical benefit were evaluated through internal and external validation,and an assessment of the overall value of two groups was made through the use of a receiver operating characteristic(ROC) curve. Results: There was no significant difference in clinical and pathological characteristics and long-term survival results between the training and validation datasets(all P>0.05). The multivariate Cox analysis showed that CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis were independent prognostic factors for ICC patients after LLR(all P<0.05). The survival Nomogram was established based on the independent prognostic factors obtained from the above screening. The ROC curve showed that the area under the curve of 1, 3 and 5-year overall survival rates of patients in the training dataset were 0.794(95%CI:0.721 to 0.867),0.728(95%CI:0.618 to 0.839) and 0.799(95%CI:0.670 to 0.928),and those in the validation dataset were 0.787(95%CI:0.660 to 0.915),0.831(95%CI:0.678 to 0.983) and 0.810(95%CI:0.639 to 0.982). Internal and external validation proved that the model exhibited a certain discrimination,calibration,and clinical applicability. Conclusion: The survival Nomogram model based on the independent influencing factors of long-term prognosis after LLR for ICC(including CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis) exhibites a certain differentiation,calibration,and clinical practicability.

[Arthroscopic classification and management for the popliteal hiatus of the lateral meniscus tears].

OBJECTIVE: To bring forward an arthroscopic classification of the popliteal hiatus of the lateral meniscus (PHLM) tears and to assess the effects of arthroscopic all-inside repair with suture hook in management of such injuries. METHODS: This study involved 146 patients who underwent arthroscopic operation because of PHLM tears from April 2014 to October 2017, eliminating the patients who had discoid lateral meniscus. There were 81 males and 65 females, with 54 left knees and 92 right knees. The average ages were (34.7±3.7) years. Among the selected participants, there were 107 patients with anterior cruciate ligament (ACL) injuries, 39 patients with medial collateral ligament (MCL) injuries, and 48 patients with medial meniscus tears. The average preoperative Lysholm and International Knee Documentation Committee (IKDC) scores were 57.7±9.2 and 54.1±8.9, respectively. The arthroscopic classification was based on the extent and degree of PHLM tears and using the arthroscopic all-inside repair with suture hook for such injuries. For the patients associated with ACL injuries, the ipsilateral autograft hamstring tendons use as the reconstruction graft for single bundle ACL reconstructions. The suture anchors were used for treatment of MCL Ⅲ injuries, and the arthroscopic all-inside repair for medial meniscus tears. RESULTS: A total of 146 PHLM tears in 146 patients were divided into type Ⅰ (tears not involved in popliteus tendon incisura; n=86, 58.9%), type Ⅱ (tears involved in popliteomeniscal fascicles; n=36, 24.7%), and type Ⅲ (tears involved in popliteus tendon incisura; n=24, 16.4%). For type Ⅰ, there were three subtypes, including type Ⅰa: longitudinal tear (n=53, 61.6%), type Ⅰb: horizontal tear (n=27, 31.4%), and type Ⅰc: radial tear (n=6, 7.0%). For type Ⅱ, there were also three subtypes, including type Ⅱa: anterosuperior popliteomeniscal fascicle tear (n=5, 13.9%), type Ⅱb: posterosuperior popliteomeniscal fascicle tear (n=20, 55.6%), and type Ⅱc: both tears (n=11, 30.6%). For type Ⅲ, there were two subtypes, including type Ⅲa: horizontal tear (n=9, 37.5%), type Ⅲb: radial tear (n=15, 62.5%). In the follow-up for an average of 15.3±2.6 months, all the patients had done well with significantly improved Lysholm (84.6±14.3) and IKDC (83.2±12.8) scores at the end of the last follow-up relative to preoperative scores (P > 0.01). CONCLUSION: We propose that it is possible to classify lateral meniscus tears at the popliteal hiatus region for three types, which can summarize the injury characteristics of this area. The arthroscopic all-inside repair with suture hook for the PHLM tears can avoid stitching to popliteal tendon or narrowing popliteal hiatus and have satisfactory clinical results.

[A prognostic model of intrahepatic cholangiocarcinoma after curative intent resection based on Bayesian network].

Objective: To examine a survival prognostic model applicable for patients with intrahepatic cholangiocarcinoma (ICC) based on Bayesian network. Methods: The clinical and pathological data of ICC patients who underwent curative intent resection in ten Chinese hepatobiliary surgery centers from January 2010 to December 2018 were collected.A total of 516 patients were included in the study.There were 266 males and 250 females.The median age(M(QR)) was 58(14) years.One hundred and sixteen cases (22.5%) with intrahepatic bile duct stones,and 143 cases (27.7%) with chronic viral hepatitis.The Kaplan-Meier method was used for survival analysis.The univariate and multivariate analysis were implemented respectively using the Log-rank test and Cox proportional hazard model.One-year survival prediction models based on tree augmented naive Bayesian (TAN) and naïve Bayesian algorithm were established by Bayesialab software according to different variables,a nomogram model was also developed based on the independent predictors.The receiver operating characteristic curve and the area under curve (AUC) were used to evaluate the prediction effect of the models. Results: The overall median survival time was 25.0 months,and the 1-,3-and 5-year cumulative survival rates was 76.6%,37.9%,and 21.0%,respectively.Univariate analysis showed that gender,preoperative jaundice,pathological differentiation,vascular invasion,microvascular invasion,liver capsule invasion,T staging,N staging,margin,intrahepatic bile duct stones,carcinoembryonic antigen,and CA19-9 affected the prognosis(χ2=5.858-54.974, all P<0.05).The Cox multivariate model showed that gender,pathological differentiation,liver capsule invasion,T stage,N stage,intrahepatic bile duct stones,and CA19-9 were the independent predictive factors(all P<0.05). The AUC of the TAN model based on all 19 clinicopathological factors was 74.5%,and the AUC of the TAN model based on the 12 prognostic factors derived from univariate analysis was 74.0%,the AUC of the naïve Bayesian model based on 7 independent prognostic risk factors was 79.5%,the AUC and C-index of the nomogram survival prediction model based on 7 independent prognostic risk factors were 78.8% and 0.73,respectively. Conclusion: The Bayesian network model may provide a relatively accurate prognostic prediction for ICC patients after curative intent resection and performed superior to the nomogram model.

[Application value of different lymph node staging system in predicting prognosis of patients with intrahepatic cholangiocarcinoma].

Objective: To examine the value of number of metastatic lymph nodes(NMLN), lymph node ratio(LNR) and log odds of metastatic lymph nodes(LODDS) in assessing the prognosis of patients with intrahepatic cholangiocarcinoma(ICC). Methods: The clinical and pathological data of 440 ICC patients who underwent curative-intent resection in 10 of Chinese hepatobiliary surgery centers from January 2010 to December 2018 were collected, and the deadline of follow-up was April 30th, 2019. Among them, 205 were males and 235 were females, with age of (57.0±9.9) years (range:23-83 years).Eighty-five cases (19.3%) had intrahepatic bile duct stones, and 98 cases (22.3%) had chronic viral hepatitis.The Kaplan-Meier method was used for survival analysis. The univariate and multivariate analysis were implemented respectively using the Log-rank test and Cox proportional hazard model. Results: A total of 440 patients underwent curative-intent resection and lymphadenectomy.R0 resection were achieved in 424 cases (96.4%) and R1 resection were in 16 cases (3.6%). The results of postoperative pathological examination showed that high, moderate and poor differentiation was 4.2%(18/426), 60.6%(258/426) and 35.2%(150/426), respectively.Adenocarcinoma was seen in 90.2%(397/440) and non-adenocarcinoma was seen in 9.8%(43/440), respectively. T stage: 2 cases (0.5%) with Tis, 83 cases(18.9%) with T1a, 97 cases(22.0%) with T1b, 95 cases(21.6%) with T2, 122 cases (27.7%) with T3 and 41 cases(9.3%) with T4.The overall median survival time was 24.0 months, and the 1-,3-, and 5-year survival rate was 74.3%, 37.7% and 18.3%, respectively. Lymphatic metastasis occurred in 175 patients(39.8%), the median total number of TNLE(M(Q(R))) was 6(5), the median number of NMLN was 0(1), the median number of LNR was 0 (0.33) and the median number of LODDS was -0.70(-0.92). Rerults of univariate analysis showed that combined stones, pathological differentiation, vascular invasion, LODDS, margin and T staging affected the prognosis (all P<0.05). Rerults of multivariate analysis showed that pathological differentiation, LODDS, margin, and T staging were independent risk factors affecting the prognosis of ICC patients (all P<0.05). Conclusion: LODDS could be used as an optimal prognostic lymph node staging index for ICC, and it is also an independent risk factor for survival after curative intent resection.

[Clinical study of S-1 single agent adjuvant chemotherapy in the radical resection of extrahepatic biliary carcinoma].

Objective: To explore the clinical efficacy of S-1 single agent adjuvant chemotherapy for the patients undergoing radical resection of extrahepatic biliary carcinoma. Methods: The clinical data of 108 patients with extrahepatic biliary carcinoma receiving radical resection who were admitted from January 2014 to June 2017 were retrospectively analyzed. There were 62 males(57.4%)and 46 females(42.6%),with a median age of 59 years (range:26 to 79 years),10 cases(9.3%) in stage Ⅱ,85 cases(78.7%) in stage Ⅲ, and 13 cases (12.0%) in stage Ⅳ, 40 cases(37.0%) of hilar cholangiocarcinoma, 8 cases(7.4%) of middle cholangiocarcinoma, 25 cases (23.2%) of distal cholangiocarcinoma, 35 cases(32.4%) of gallbladder carcinoma.After radical resection of extrahepatic biliary carcinoma, 49 patients receiving S-1 single agent chemotherapy and 59 patients receiving non-special treatment were divided into the chemotherapy group and the operation group,respectively. All the dates of the patients were followed up and collected with the overall survival time,tumor-free survival time,1,2 and 3-year survival rate after operation,and the rate of major toxic reaction during chemotherapy of the chemotherapy group. Survival curve was drawn by the Kaplan-Meier method, and survival analysis was done using the Log-rank test. Results: There were no significant differences in the general date of two groups(sex, age, tumor size, tumor site, TNM stages, degree of differentiation). The median overall survival time and the median tumor-free survival time in the chemotherapy group were 27 months and 21 months,respectively,and in the operation group were 21 months and 17 months,respectively. There were differences between the two groups in the overall survival rates(χ(2)=3.967,P<0.05) and the 2 and 3-year survival rate(63.3%,36.6%;41.6%,20.4%;χ(2)=4.510,P<0.05;χ(2)=6.143,P<0.05),but the 1-year overall survival rate (83.4%,79.7%)was not statistically significant(χ(2)=0.286,P>0.05). There were no significant differences in the tumor-free survival time,1,2 and 3-year tumor-free survival rate(77.6%,41.4%,33.1%;62.7%,30.9%,21.2%)between the two groups(χ(2)=0.876,P>0.05;χ(2)=0.252,P>0.05;χ(2)=1.571,P>0.05;χ(2)=3.323,P>0.05,respectively). The main toxic reaction during chemotherapy were dyspepsia(28.6%, 14/49), anemia(26.5%, 13/49), and leukopenia(22.5%, 11/49), all of which were mild. Conclusion: S-1 single agent chemotherapy after radical reseetion of extrahepatic biliary carcinoma could effectly improve the survival of patients and all of the main toxic reaction during chemotherapy were mild.

[Kindlin-2 promotes gallbladder cancer metastasis and invasion by inducing epithelial-mesenchymal transition].

Objective: To investigate the effects of Kindlin-2 on malignant phenotypes of human gallbladder cancer cells and discuss the mechanisms. Methods: The expression level of Kindlin-2 in 30 cases of gallbladder cancer tissues and adjacent non-tumoral tissues collected from the First Affiliated Hospital of Zhengzhou University between September 2012 and May 2013 was assessed by real-time PCR and immunohistochemistry.Lentivirus-mediated Kindlin-2 overexpression was used in gallbladder cancer cell lines GBC-SD and SGC-996.Transwell assay and adhesion assay were investigated to explore the functional role of Kindlin-2 on gallbladder cancer cells.Western Blot was used to test the protein change of epithelial-mesenchymal transition(EMT) characteristics. The t-test was used to analyzed results. Results: The RNA and protein levels of Kindlin-2 in gallbladder cancer tissues were higher than in the non-tumoral tissues (t=4.372, P=0.001; t=7.477, P=0.000). The expression level of Kindlin-2 in gallbladder cancer tissues was correlated with Nevin stage(χ(2)=5.932, P=0.035). Compared with control groups, the cell-matrix adhesion ability of GBC-SD and SGC-996 with Kindlin-2 overexpression was obviously promoted(1.66±0.03 vs. 1.07±0.22, t=2.710, P=0.041; 2.66±0.24 vs. 1.03±0.02, t=6.610, P=0.020). The number of GBC-SD and SGC-996 cells with Kindlin-2 overexpression passing through the Transwell chamber matrix increased significantly compared with the control groups(116.1±13.9 vs. 54.7±8.4, t=3.781, P=0.019; 136.3±7.5 vs. 64.3±6.4, t=7.302, P=0.002). The wound healing rate of GBC-SD with Kindlin-2 overexpression at 12-hour and 24-hour was higher than that of the group ((42.9±2.2)% vs. (29.7±1.7)%, t=4.690, P=0.009; (65.0±2.4)% vs.(40.4±2.0)%, t=7.945, P=0.001). The wound healing rate of SGC-996 with Kindlin-2 overexpression at 12-hour and 24-hour was also higher than that of the group ((32.9±1.3)% vs. (24.1±1.5)%, t=4.518, P=0.011; (51.3±1.1)% vs. (39.2±1.1)%, t=8.001, P=0.001). The characteristics of EMT were induced in gallbladder cancer cells with Kindlin-2 overexpression, including the up-regulation of N-cadherin, Vemintin and the down-regulation of E-cadherin. Conclusion: The expression of Kindlin-2 is up-regulated in gallbladder cancer tissues and Kindlin-2 promoted the malignant phenotypes of gallbladder cancer cells partially by epithelial-mesenchymal transition.

[The survival prediction model of advanced gallbladder cancer based on Bayesian network: a multi-institutional study].

Objective: To investigate the clinical value of Bayesian network in predicting survival of patients with advanced gallbladder cancer(GBC)who underwent curative intent surgery. Methods: The clinical data of patients with advanced GBC who underwent curative intent surgery in 9 institutions from January 2010 to December 2015 were analyzed retrospectively.A median survival time model based on a tree augmented naïve Bayes algorithm was established by Bayesia Lab software.The survival time, number of metastatic lymph nodes(NMLN), T stage, pathological grade, margin, jaundice, liver invasion, age, sex and tumor morphology were included in this model.Confusion matrix, the receiver operating characteristic curve and area under the curve were used to evaluate the accuracy of the model.A priori statistical analysis of these 10 variables and a posterior analysis(survival time as the target variable, the remaining factors as the attribute variables)was performed.The importance rankings of each variable was calculated with the polymorphic Birnbaum importance calculation based on the posterior analysis results.The survival probability forecast table was constructed based on the top 4 prognosis factors. The survival curve was drawn by the Kaplan-Meier method, and differences in survival curves were compared using the Log-rank test. Results: A total of 316 patients were enrolled, including 109 males and 207 females.The ratio of male to female was 1.0∶1.9, the age was (62.0±10.8)years.There was 298 cases(94.3%) R0 resection and 18 cases(5.7%) R1 resection.T staging: 287 cases(90.8%) T3 and 29 cases(9.2%) T4.The median survival time(MST) was 23.77 months, and the 1, 3, 5-year survival rates were 67.4%, 40.8%, 32.0%, respectively.For the Bayesian model, the number of correctly predicted cases was 121(≤23.77 months) and 115(>23.77 months) respectively, leading to a 74.86% accuracy of this model.The prior probability of survival time was 0.503 2(≤23.77 months) and 0.496 8(>23.77 months), the importance ranking showed that NMLN(0.366 6), margin(0.350 1), T stage(0.319 2) and pathological grade(0.258 9) were the top 4 prognosis factors influencing the postoperative MST.These four factors were taken as observation variables to get the probability of patients in different survival periods.Basing on these results, a survival prediction score system including NMLN, margin, T stage and pathological grade was designed, the median survival time(month) of 4-9 points were 66.8, 42.4, 26.0, 9.0, 7.5 and 2.3, respectively, there was a statistically significant difference in the different points(P<0.01). Conclusions: The survival prediction model of GBC based on Bayesian network has high accuracy.NMLN, margin, T staging and pathological grade are the top 4 risk factors affecting the survival of patients with advanced GBC who underwent curative resection.The survival prediction score system based on these four factors could be used to predict the survival and to guide the decision making of patients with advanced GBC.

[A multicenter retrospective study for the prognosis of T1b stage gallbladder carcinoma underwent different surgical procedure].

Objective: To explore the prognosis of patients with T1b stage gallbladder carcinoma underwent different surgical procedure. Methods: The clinicopathological data of 97 patients with T1b stage gallbladder carcinoma came from 8 clinical centers from January 2010 to December 2016 and 794 patients who were admitted to the SEER database of USA from January 1973 to December 2014 were analyzed.There were 891 patients including 254 males and 637 females (1.0∶2.5) with age of (69.5±12.0)years. There were 380 patients who were less than 70 years old, 511 patients who were more than 70 years old. And there were 213 patients with the diameter of tumor less than 20 mm, 270 patients with the diameter of tumor more than 20 mm, 408 patients were unclear. There were 196 patients with well differentiation, 407 patients with moderately differentiation, 173 patients with poorly differentiation, 8 patients with undifferentiated, 107 patients were unclear. In the 891 patients with T1b stage gallbladder carcinoma, there were 562 cases accepted the simple cholecystectomy, 231 cases with simple cholecystectomy plus lymphadenectomy, and 98 cases with radical cholecystectomy. The time of follow-up were until June 2017. χ(2) test was used to analyze the enumeration data, rank-sum test was used to analyze the measurement data, the analyses of prognostic factors were used Cox proportional hazards model, the survival analysis was performed using Kaplan-Meier method. Results: The results of Cox proportional hazards model indicated, age, differentiation, surgical procedure were the risk factors of prognostic(1.929(1.594-2.336), P<0.01; 1.842(1.404-2.416), P<0.01; 1.216(0.962-1.538), P<0.01). The results of Kaplan Meier test indicated, the overall survival of T1b stage gallbladder carcinoma were (85.5±3.8)months, the overall survival of patients with simple cholecystectomy were (71.3±4.4)months, the overall survival of patients with cholecystectomy plus lymphadenectomy were(87.6±5.8)months, and the overall survival of patients with radical cholecystectomy were(101.7±9.3)months. The overall survival of patients with cholecystectomy plus lymphadenectomy and radical cholecystectomy were more than simple cholecystectomy(P<0.05). There were 329 patients with Lymph nodes examined in and after operations(231 patients with cholecystectomy plus lymphadenectomy, 98 patients with radical cholecystectomy). There were 265 patients with negative lymph node metastasis, the overall survival were(98.3±4.2)months. There were 64 patients with positive lymph node metastasis, the overall survival were(75.5±3.1)months. The overall survival of 38 patients with cholecystectomy plus lymphadenectomy were(62.7±2.6) months, and 26 patients with radical cholecystectomy were (82.2±3.7)months. The overall survival of patients with radical cholecystectomy were more than cholecystectomy plus lymphadenectomy(P<0.05). Conclusions: The T1b stage gallbladder carcinoma patients with cholecystectomy plus lymphadenectomy or radical cholecystectomy has improved the prognosis comparing with simple cholecystectomy, significantly. When lymph node metastasis occurs, radical cholecystectomy has improved the prognosis comparing with cholecystectomy plus lymphadenectomy.

Association between NDRG2/IL-6/STAT3 signaling pathway and diabetic retinopathy in rats.

OBJECTIVE: The aim of this study was to observe the association between N-myc downstream regulated gene 2 (NDRG2)/interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling pathway and diabetic retinopathy (DR) in rats. MATERIALS AND METHODS: The model of diabetes was successfully established in Sprague-Dawley rats. All rats were divided into diabetes model group (model group, n=10), pathway inhibitor group (CLT-005 group, n=10), and normal control group (control group, n=10). After successful modeling, blood and retinal tissues of rats were collected. The levels of blood glucose and serum IL-6 were detected. Meanwhile, oxidative and antioxidant indexes reactive oxygen species (ROS), superoxide dismutase (SOD), and malondialdehyde (MDA) were detected via enzyme-linked immunosorbent assay (ELISA). Morphological changes in retinal tissues were observed using hematoxylin-eosin (HE) staining, and the number of corneal nerve fibers was observed under a microscope. The expressions of vascular endothelial growth factor (VEGF) and NDRG2/IL-6/STAT3 pathway genes in tissues were determined via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Furthermore, the expressions of NDRG2/IL-6/STAT3 pathway proteins were determined via Western blotting. RESULTS: The level of blood glucose in model group was significantly higher than that of control group (p<0.05), suggesting successful modeling. The levels of tumor necrosis factor-α (TNF-α), IL-6, and IL-1 in model group were significantly higher than those of control group (p<0.05). The content of ROS and MDA in tissues was significantly higher in model group than the other two groups (p<0.05). However, SOD increased markedly in CLT-005 group and was close to that of control group. Besides, the number of corneal nerve fibers decreased remarkably in model group. However, it increased significantly in CLT-005 group, but was still smaller than that in control group. According to HE staining, there were significant retinal edema and telangiectasia in model group. Mild retinal edema and more ganglion cells and inner nuclear layers were observed in CLT-005 group than model group. QRT-PCR demonstrated that the mRNA expressions of VEGF, NDRG2, IL-6, and STAT3 were remarkably higher in model group than those in control group (p<0.05). However, they decreased significantly in CLT-005 group (p<0.05). Model group exhibited remarkably higher protein expressions of NDRG2, IL-6, and STAT3 than control group (p<0.05). However, CLT-005 group had decreased protein expressions of these molecules (p<0.05), which were close to those in control group. CONCLUSIONS: The activation of NDRG2/IL-6/STAT3 signaling pathway is positively correlated with the occurrence and development of DR in rats. Therefore, inhibiting the activation of NDRG2/IL-6/STAT3 signaling pathway can affect oxidation and antioxidation, thereby exerting a protective effect against retinal injury in diabetes rats.

MicroRNA-381 inhibits lung adenocarcinoma cell biological progression by directly targeting LMO3 through regulation of the PI3K/Akt signaling pathway and epithelial-to-mesenchymal transition.

OBJECTIVE: To investigate the role of miR-381 in the progression of lung adenocarcinoma (LA) and its underlying mechanism. PATIENTS AND METHODS: A total of 54 pairs of LA tissues and para-carcinoma tissues were obtained from May 2015 to April 2017 in our hospital. Four human LA cell lines (A549, SPC-A1, H1299, and PC-9) and one normal human pulmonary epithelial cell line BEAS-2B were obtained and cultured. The protein and mRNA expression levels were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Additionally, cell proliferation assays and cell migration and invasion assays were used. Furthermore, tumor xenograft model in nude mice was made in this study. RESULTS: miR-381 was notably downregulated in LA tissues. Moreover, low miR-381 expression was confirmed to be strongly correlated with poor prognosis and aggressive clinicopathological characteristics of LA patients. Exogenous miR-381 overexpression was found to notably restrict LA cell proliferation, migration, and invasion; additionally, miR-381 overexpression could significantly reduce tumor growth in vivo. Mechanistically, LMO3 was determined as a novel direct target for miR-381 in LA cells. In clinical LA tissues, the LMO3 expressions were clearly overexpressed. Furthermore, miR-381 overexpression affected the PI3K/Akt pathway and EMT in LA. CONCLUSIONS: MiR-381 played key roles in LA progression, partially via directly targeting LMO3 and regulating the PI3K/Akt signaling pathway and EMT. Thus, the miR-381/ LMO3 axis has clinical significance in the therapy of patients with LA.

Chromosomal aberrations and sister chromatid exchanges in workers exposed to lead.

The frequencies of chromosomal aberration (CA) and sister chromatid exchange (SCE) in the peripheral blood lymphocytes of 21 lead-exposed workers from a battery factory were studied, and their blood lead (Pb-B) levels were determined. The results demonstrated that when the mean Pb-B level reached 50 micrograms/dl, the CA percentage increased significantly compared with that of the nonexposed controls. The CA percentage increased as the mean Pb-B level increased, showing a dose-effect relationship. When the mean Pb-B level was as high as 80 micrograms/dl, the rate of SCE in the lymphocyte was also increased significantly, but no correlation was observed between these two parameters. On the basis of such results, lead should be considered an agent harmful to the human chromosome.

Study of development in an experimental animal model of a pediatric physeal slide-traction plate.

BACKGROUND: To develop a physeal slide-traction plate for children can provide rigid internal fixation of a bone fracture without inhibiting epiphyseal growth. METHODS: The slide-traction plates and standard plates were designed to configure to the femoral condyle. Twelve of thirty goats were used as normal controls and the rest were implanted with plates (right femurs were fixed with standard plates, and left femurs were fixed with slide-traction plates). All goats underwent X-ray examination at 1 month, 3 months and 6 months after surgery, and femoral length were measured. Histological staining and electron microscopy were performed to evaluate the development of the epiphyseal plate at 3 and 6 months after surgery. RESULTS: Compared to the standard plate, the slide-traction plate group exhibited more normal physeal growth, histologic features, safranin O staining, and electron microscopy structural features. There were significant differences in length in the femurs of goats fixed with slide-traction plates and standard plates at 1 month, 3 months, and 6 months after surgery, respectively. There was no difference between femurs of the normal control group and the femurs fixed with the slide-traction plates. Thicker epiphyseal plates were found in the left vs. the right femurs of the group fixed with plates at 3 and 6 months after surgery. In the group fixed with plates, Safranin O staining showed that the epiphyseal plates of the left femurs had more fuscous staining than the right femurs at 3 and 6 months after surgery. Electron microscopy also showed that cells in the epiphyseal plates of the left femurs were healthier in appearance than cells from the right femurs in the group fixed with plates. CONCLUSION: The physeal slide-traction plate can slide with the growth of the physis and is suitable for fixation of fractures in this region.

Effects of intratracheal instillation of fenvalerate on the ultrastructures of pulmonary alveolar macrophages in rat.

On 1 d after instillation of fenvalerate (Fen) 0.19, 0.93, 4.66, and 23.3 mg.kg-1, and on 30 min, 4 h, 1 d, 4 d, and 7 d after instillation of Fen 4.66 mg.kg-1 by a single intratracheal instillation, respectively, the ultrastructural changes in rat pulmonary alveolar macrophages (PAM) were observed, and the toxicity indices (TI) were calculated. It was found that the ineffective dose of Fen was 0.19 mg.kg-1, and the threshold dose was less than or equal to 0.93 mg.kg-1 as well as the serious intoxication dose greater than or equal to 4.66 mg.kg-1. The results suggest that Fen is quickly toxic to rat lungs and quickly biodegraded in the lungs.

[Local plantar rotatory flap for repairing of soft tissue defect of heel].

OBJECTIVE: Soft tissue defect of heel is not uncommon. Transplantation of free cutaneous flap and transfer of axial cutaneous flap have been used in treating such defect successfully, but both of them are somewhat complicated. Local plantar rotatory flap might show great importance in this field. METHODS: Since March 1993 to March 1998, 9 cases with soft tissue defect of heel were repaired by local plantar rotation flaps. The size of defect ranged from 2 cm x 4 cm to 6 cm x 8 cm, and it was designed superficial to plantar fascia. The flap was medially based, and nutrilized by proximal plantar subcutaneous plexus of blood supply as well as lateral and medial plantar nerve. RESULTS: Followed up 4 months to 2 years, all the flaps were survived. Sensation of the flap was preserved in 7 patients, who had normal sensation of the donated area preoperatively. The transferred flap was endurable to body bearing. CONCLUSIONS: The flap is easily prepared with reliable blood supply and sensation of the flap preserved. The method is worthy to be recommended for widely use because of its advantages over other methods.

Effects of drinking water from the lower reaches of the Huangpu River on the risk of male stomach and liver cancer death.

A quasi-historical cohort study method was used to collect the data of male stomach and liver cancer death and the data of exposure to relevant risk factors from 1984 to 1988 in male tap-water- and raw-water-drinking cohorts (> or = 30 years old) at both the upper and lower reaches of the Huangpu River. Total person-years observed are 172,448. The Odds Ratios of drinking water from the lower reaches for male stomach cancer and liver cancer death are 2.021 (p < 0.01) and 1.851 (p < 0.01), respectively, in unconditional logistic regression analysis after controlling possible confounding factors. The result shows that drinking water from the lower reaches of the Huangpu River is one of the important risk factors for male stomach and liver cancer death in local areas.