Objective To investigate the epidemiological characteristics,clinical features,diagnosis,treatment and prognosis of benign tumor of the bile ducts.Methods The retrospective cross-sectional multicenter study was conducted.The clinical data of 136 patients with benign tumor of the bile ducts who were admitted to the eight hospitals between January 2007 and December 2016 were collected,including 70 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,19 in the First Affiliated Hospital of Zhengzhou University,15 in the First Affiliated Hospital of Xi'an Jiaotong University,11 in the First Affiliated Hospital of Dalian Medical University,7 in the Affiliated Hospital of North Sichuan Medical College,6 in the Southwest Hospital of the Third Military Medical University,4 in the Cancer Center of Sun Yat-Sen University and 4 in the Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine.All the patients received laboratory and imaging examinations,and then underwent corresponding surgery when treatment planning was respectively determined by comprehensive hospitals according to clinical features and results of examinations.Surgical procedures were performed based on the results of intraoperative frozen section in rapid pathological diagnosis.Observation indicators:(1) epidemiological characteristics;(2) clinical features;(3) results of laboratory and imaging examinations;(4) treatment situations;(5) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the survival of patients up to March 2017.Measurement data with normal distribution were represented as-x±s.Results (1) Epidemiological characteristics:Of 136 patients,the male to female ratio was 1.78 ∶ 1.The incidence of whole bile duct tumors was from high to low,including 52 patients with duodenal papilla adenoma,32 with extrahepatic bile duct adenoma,24 with intrahepatic biliary cystadenoma,11 with intrahepatic papillary adenoma,9 with intrahepatic bile duct epithelial tumor,7 with epithelial tumor of duodenal papilla and 1 with neuroendocrine tumor of duodenal papilla.Among 136 patients,adenomas (including cystadenoma) was detected in 108 patients,papillomas in 11 patients,intraepithelial neoplasias in 16 patients,neuroendocrine tumor in 1 patient;intrahepatic bile duct benign tumors in 44 patients and extrahepatic bile duct (including duodenal papilla) benign tumors in 92 patients.(2) Clinical features:of 44 patients with intrahepatic bile duct benign tumors,29 had abdominal pain,fever and abdominal masses,4 had jaundice,11 had no obvious clinical symptoms and were diagnosed by physical examination.Among 92 patients with extrahepatic bile duct benign tumors,76 developed obstructive jaundice,68 were accompanied by abdominal pain or colicky pain,8 were combined with pancreatitis and 2 deveIoped hemobilia,some patients were combined with multiple clinical symptoms.(3) Results of laboratory and imaging examinations:82 patients received CA19-9 test,results of 22 patients were abnormal,with a level of (148-± 126)U/mL.Ninety-seven patients received carcinoembryonic antigen (CEA) test,with a level of test of (2.7±2.0) μg/L,and a level of CEA in 1 patient was slight abnormal,with a level of 11.2 pμg/L.One hundred and thirty-six patients underwent preoperative ultrasound examinations,showing unqualified hepatic and bile duct space occupying lesions and bile duct dilatation.Ninetyfive patients underwent preoperative computed tomography (CT),80 underwent preoperative magnetic resonance imaging (MRI) or magnetic resonanced cholangio-pancreatography (MRCP),and 13 underwent preoperative endoscopic ultrasonography (EUS).Twenty-nine patients were considered for intrahepatic bile duct benign tumors;76 were diagnosed with obstructive jaundice,with uncertain benign or malignant tumors;other patients had bile duct space occupying,considering bile duct tumor (including cancer).Twelve patients with bile duct obstruction underwent ERCP,showing obstruction site and morphology.(4) Treatment situations:among 136 patients,65 underwent pancreaticoduodenectomy,17 underwent right hemihepatectomy + cholecystectomy,16 underwent cholecystectomy + hepatic left lateral lobectomy,11 underwent left hemihepatectomy + cholecystectomy,11 underwent duodenal papillary local excision+papilla reconstruction,11 underwent Roux-en-Y choledochojejunostomy anastomosis,4 underwent cholecystectomy + extrahepatic bile duct local excision + end-to-end bile duct anastomosis and 1 underwent endoscopic mucosal resection of duodenal papillary adenomna.Of 136 patients with postoperative complications,25 were complicated with pancreatic leakage,11 with bile leakage,2 with postoperative hemorrhage and 1 with hepatic failure.Two patients with pancreatic leakage died of massive hemorrhage caused by abdominal infection,1 died of hepatic failure and other patients were discharged from hospital after symptomatic treatment.(5) Follow-up situations:47 of 136 patients were followed up for 3-123 months,with a follow-up rate of 34.6％.During follow-up,2 patients undergoing duodenal papillary local excision + papilla reconstruction had canceration,and other patients had good survival.Conclusions There is a low clinical incidence of benign tumor of the bile ducts,which is more common in male than in female,and in adenomas (including cystadenoma) and papillomas.The preoperative imaging examinations or ERCP biopsy pathological examination can increase an accuracy of preoperative diagnosis.Benign tumors with high canceration rates need positive surgical treatment.

The 53rd annual meeting of the American Society of Clinical Oncology (ASCO) was held in Chicago,United States between June 2 and 6,2017.The latest advances in biliary tract cancer research from this meeting were summarized and analyzed in this paper.The adjuvant therapy in biliary tract cancer made a breakthrough in this meeting,the findings could provide the basis for a new standard of changing the current management model in the disease.The precision medicine and targeted therapy will be the development direction in the future.Doctors should attach great importance to the adjuvant and comprehensive therapy in biliary tract cancer and initiate high level multi-center clinical trials to improve the overall the diagnostic and treatment levels of biliary tract cancer.

Objective To investigate the effects of the extent of regional lymph node dissection on the prognosis of patients with T4 gallbladder carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 64 patients with T4 gallbladder carcinoma who underwent radical cholecystectomy in the 4 medical centers between January 2013 and December 2016 were collected,including 31 in the Eastern Hepatobiliary Surgery Hospital of Naval Medical University,16 in the First Affiliated Hospital of Xi'an Jiaotong University,11 in the Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine and 6 in the Affiliated Hospital of North Sichuan Medical College.There were 27 males and 37 females,aged from 35 to 77 years,with a median age of 59 years.Sixty-four patients underwent radical cholecystectomy and regional lymph node dissection.According to the extent of intraoperative lymph node dissection,25 patients (13 in the Eastern Hepatobiliary Surgery Hospital of Naval Medical University,6 in the First Affiliated Hospital of Xi'an Jiaotong University,4 in the Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine and 2 in the Affiliated Hospital of North Sichuan Medical College) whose extent of lymph node dissection involved lymph nodes next to cystic duct,hepatoduodenal ligament,back of head of pancreas,next to common hepatic artery and celiac trunk were allocated into the extended dissection group,39 patients (18 in the Eastern Hepatobiliary Surgery Hospital of Naval Medical University,10 in the First Affiliated Hospital of Xi'an Jiaotong University,7 in the Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine and 4 in the Affiliated Hospital of North Sichuan Medical College) whose extent of lymph node dissection involved lymph nodes next to cystic duct and hepatoduodenal ligament were allocated into the control group.Observation indicators:(1) postoperative complications;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative overall survival up to January 2018.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was evaluated with the independentsample t test.Count data were represented as absolute number or percentage,and comparison between groups was analyzed using the chi-square test and Fisher exact probability.The survival curve was drawn using the KaplanMeier method,and the comparison of survival rates was done by the Log-rank test.Results (1) Postoperative complications:64 patients with T4 gallbladder carcinoma underwent successful radical cholecystectomy and regional lymph node dissection,without intraoperative death.Twelve patients had different degrees of postoperative complications.Four of 7 patients undergoing extended radical cholecystectomy had postoperative complications.Twenty-five patients in the extended dissection group were cured by conservative treatment,including 4 with intraperitoneal infection and 2 with pancreatic leakage,with a complication incidence of 24.0％ (6/25).Thirtynine patients in the control group were cured by conservative treatment,including 5 with intraperitoneal infection and 1 with gastric retention,with a complication incidence of 15.4％ (6/39).There was no statistically significant difference in the complication incidence between the two groups (x2=0.284,P＞0.05).(2) Follow-up and survival situations:64 patients were followed up for 1-60 months.The postoperative overall median survival time was l l months.The postoperative median survival time,1-,3-and 5-year cumulative survival rates were respectively 18 months,80％,16％,9％ in the extended dissection group and 8 months,21％,4％,0 in the control group,with a statistically significant difference in the prognosis between the two groups (x2=14.744,P＜ 0.05).Conclusions On the premise of practiced surgical skill,extended regional lymph node dissection cannot increase incidence of surgical complication in patients with T4 gallbladder carcinoma after radical resection.Actively extending lymph node dissection up to near common hepatic artery,peri-celiac trunk and back of head of pancreas can improve long-term survival and prognosis.

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. χ² 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.

Objective To analyze the pathological results and current treatment situation of patients with unexpected gallbladder carcinoma from multi-centers in China,and explore the diagnosis and treatment of unexpected gallbladder carcinoma.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 223 patients with unexpected gallbladder carcinoma who were admitted to the 8 clinical centers from January 2010 to December 2016 were collected,including 86 in the First Affiliated Hospital of Xi'an Jiaotong University,41 in the First Affiliated Hospital of Zhengzhou University,30 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,27 in the Xinhua Hospital of Shanghai Jiaotong University,13 in the First Affiliated Hospital of Dalian Medical University,11 in the Tianjin Medical University Cancer Institute & Hospital,9 in the First Affiliated Hospital of Army Medical University (Third Military Medical University) and 6 in the Affiliated Hospital of North Sichuan Medical College.Treatment of patients with unexpected gallbladder carcinoma who were diagnosed by intraoperative frozen section biopsy and postoperative pathological examination followed guideline for the diagnosis and treatment of gallbladder carcinoma (2015 edition).According to tumor staging and patients' decision,postoperative adjuvant treatment was selectively performed.Observation indicators:(1) diagnosis and treatment of unexpected gallbladder carcinoma;(2) followup and survival.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to June 2017.Measurement data with normal distribution were represented as (x)±s.Measurement data with skewed distribution were described as M (range).The survival time was calculated using the Kaplan-Meier method.Results (1) Diagnosis and treatment of unexpected gallbladder carcinoma:of 223 patients with unexpected gallbladder carcinoma,80 were initially diagnosed using intraoperative frozen section biopsy [20 received T stage results (intraoperative T stage of 14 patients had not matched postoperative results),and 60 didn't receive T stage results],and 143 were initially diagnosed using postoperative pathological examination (13 were initially diagnosed with gallbladder benign disease by intraoperative frozen section biopsy and 130 didn't intraoperatively receive frozen section biopsy).Of 223 patients,209,10,3 and 1 were respectively confirmed as adenocarcinoma,adenoma canceration,neuroendocrine tumor and squamous cell carcinoma;6,16,32,73,75,12 and 9 were respectively detected in Tis,T1a,T1b,T2,T3 and T4 stages and undefined stage;140 underwent reoperations,including 106 with radical resection of gallbladder carcinoma and 34 with extended radical resection of gallbladder carcinoma;operation of 126 patients reached the standard and operation of 97 patients didn't reach the standard.Of 27 patients with postoperative complications,12 with postoperative hemorrhage received successful hemostasis by reoperations (7 with cystic artery hemorrhage and 5 with blood oozing from gallbladder bed);8 with suppurative cholangitis received endoscopic retrograde cholangiopancreatography and choledochotomy with drainage,including 2 deaths and 6 with improvement;2 with common bile duct injury were improved by reoperation of choledochojejunostomy + T tube drainage;2 were complicated with bile leakage induced to peritonitis and underwent bile duct repair with drainage,including 1 death and 1 with improvement;2 with hepatic failure died of treatment failure;1 with colonic injury was improved by reoperation of anastomosis.Of 223 patients,207 didn't receive postoperative adjuvant treatment and 16 received postoperative adjuvant treatment,including 8 with chemotherapy,4 with radiotherapy,2 with immunologic therapy and 2 with Chinese medicine treatment.(2) Follow-up and survival:of 223 patients,193 were followed up for 6-90 months,with a median time of 33 months.Of 193 patients with follow-up:① The operation of 2 patients in stage Tis reached the standard,including 1 with cholecystectomy and 1 with radical resection of gallbladder carcinoma,and the postoperative survival time of them were respectively 28 months and 52 months.② The operation of 14 patients in stage T1a reached the standard,including 8 with cholecystectomy and 6 with radical resection of gallbladder carcinoma,and the postoperative survival time of them were respectively (74±5)months and (79±6)months.③ Of 26 patients in stage T1b,13 and 13 received respectively cholecystectomy and radical resection of gallbladder carcinoma (reaching the standard),and postoperative survival time of them were respectively (66±4)months and (76±8)months.④ Of 68 patients in stage T2,25,37,4 and 2 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard),extended radical resection of gallbladder carcinoma (reaching the standard) and palliative resection,and postoperative survival time of them were respectively (42±7) months,(66±6) months,(42±3) months and (26±3) months.⑤ Of 71 patients in stage T3,20,48 and 3 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard) and extended radical resection of gallbladder carcinoma (reaching the standard),and postoperative survival time of them were respectively (39±8) months,(48± 11) months and (10±6) months.⑥ Of 12 patients in stage T4,3,1,5 and 3 patients received respectively cholecystectomy,radical resection of gallbladder carcinoma (reaching the standard),extended radical resection of gallbladder carcinoma (reaching the standard) and palliative resection,and postoperative survival time of them were respectively (10±4) months,12 months,(9± 5) months and (11±3) months.Conclusions The intraoperative frozen section biopsy and pathological results are the key points for diagnosis and treatment of unexpected gallbladder carcinoma.Patients in stage Tis and T1a should undergo cholecystectomy,while patients in stage T1b and above should undergo radical resection of gallbladder carcinoma or extended radical resection of gallbladder carcinoma.

Objective To analyze the clinicopathological features and lymph node metastases characteristics of intrahepatic cholangiocarcinoma (ICC).Methods The retrospective case-control study was conducted.The clinicopathological data of 1 321 ICC patients who were admitted to 12 hospitals from April 2007 to November 2017 were collected,including 182 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,173 in the First Affiliated Hospital of Xi'an Jiaotong University,156 in the First Hospital Affiliated to Army Medical University (Third Military Medical University),139 in the Cancer Center of Sun Yatsen University,128 in the Zhongshan Hospital of Fudan University,113 in the Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine,109 in the First Affiliated Hospital of Dalian Medical University,84 in the Tianjin Medical University Cancer Institute and Hospital,65 in the First Affiliated Hospital of Zhengzhou University,62 in the Affiliated Hospital of North Sichuan Medical College,58 in the Affiliated Drum Tower Hospital of Nanjing University Medical School and 52 in the Second Affiliated Hospital of Zhejiang University School of Medicine.Treatment planning was respectively determined by comprehensive hospitals according to clinical features and results of related examinations.Surgical procedures were performed based on the results of intraoperative examinations and patients' conditions.Observation indicators:(1) laboratory examination and treatment situations;(2) relationship between clinicopathological features and primary tumor location and diameter;(3) diagnosis of lymph node dissection and metastases;(4) relationship between clinicopathological features and lymph node metastases.Measurement data with normal distribution were represented as x±s,and comparison between groups was done by the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the chi-square test.Results (1) Laboratory examination and treatment situations:of 1 321 patients,cases and percentages of positive hepatitis B virus (HBV),positive serum alpha-fetoprotein (AFP) (＞ 20 μg/L),positive serum carcinoembryonic antigen (CEA) (＞ 5 μg/L),positive serum alanine transaminase (ALT) (＞75 U/L),positive serum CA19-9 (＞37 U/mL) and positive serum total bilirubin (TBil) (＞20 μmol/L) were respectively 202,80,329,207,590,300 and 15.586％ (202/1 296),7.339％(80/1 090),26.299％ (329/1 251),16.587％ (207/1 248),49.789％(590/1 185),24.000％ (300/1 250).Of 1 321 patients,1 206 underwent surgery,including 904 with hepatectomy,193 with transcatheter arterial chemoembolization (TACE),72 with percutaneous transhepatic biliary drainage (PTBD),3 with endoscopic retrograde cholangiopancreatography (ERCP),3 with PTBD + ERCP and 31 with other treatments;115 had missing registration of surgical procedures.(2) Relationship between clinicopathological features and primary tumor location and diameter:① Relationship between clinicopathological features and primary tumor location:cases with CA19-9 level ≤ 37 U/mL,38-200 U/mL and ＞ 200 U/mL were respectively 227,91,146 with primary tumor located in left liver and 282,134,137 with primary tumor located in right liver.Cases combined with hepatitis and intrahepatic bile duct stone were respectively 67,73 with primary tumor in left liver and 111,47 with primary tumor in right liver,with statistically significant differences in above indictors (x2 =6.710,5.656,12.534,P＜0.05).The results of further analysis showed that incidence age of ICC was (63± 10)years old in patients with hepatitis and (59± 10) years old in patients without hepatitis,with statistically significant differences (t =4.840,P＜0.05).② Relationship between clinicopathological features and primary tumor diameter:cases with primary tumor diameter ≤ 3 cm,with 3 cm ＜ primary tumor diameter ≤ 5 cm and with primary tumor diameter ＞ 5 cm were respectively 159,250,229 with CEA level ≤ 5 pg/L and 40,65,95 with CEA level ＞ 5 μg/L and 165,258,286 with ALT level ≤ 75 U/L and 34,57,36 with ALT level ＞ 75 U/L and 148,242,281 with TBil level ≤ 20 μmol/L and 51,73,43 with TBil level ＞ 20 μmol/L,and 37,70 and 131 patients had satellite loci of tumor,with statistically significant differences in above indictors (x2=8.669,6.637,15.129,34.746,P＜0.05).(3) Diagnosis of lymph node dissection and metastases:of 904 patients with hepatectomy,346 received lymph node dissection,total number,number in each patient and median number of lymph node dissected were respectively 1 894.0,5.5 and 4.0 (range,1.0-26.0);157 had lymph node metastases,with a rate of lymph node metastasis of 45.376％ (157/346),number and number in each patient of positive lymph node were respectively 393.0 and 2.5.Of 346 patients with lymph node dissection,114 had lymph node metastasis by preoperative imaging examination,postoperative pathological examinations confirmed 64 with lymph node metastasis and 50 without lymph node metastasis;232 didn't have lymph node metastasis by preoperative imaging examination,postoperative pathological examinations confirmed 93 with lymph node metastasis and 139 without lymph node metastasis.The sensitivity,specificity and diagnostic accuracy by imaging examination were respectively 40.8％,73.5％ and 58.7％.(4) Relationship between clinicopathological characteristics and lymph node metastases:lymph node metastasis rate was respectively 51.397％ (92/179) in patients with primary tumor located in left liver,39.103％ (61/156) in patients with primary tumor located in right liver,34.615％(18/52) in patients with primary tumor diameter ≤ 3 cm,42.500％(51/120) in patients with 3 cm ＜ primary tumor diameter ≤ 5 cm,52.980％(80/151) in patients with primary tumor diameter ＞ 5 cm,48.790％(121/248) in patients with CEA ≤ 5 μg/L,59.302％ (51/86) in patients with CEA ＞ 5 μg/L,40.234％ (103/256) in patients with TBil ≤ 20 μmol/L,35.106％ (33/94) in patients with TBil ＞ 20 μmol/L,with statistically significant differences in above indictors (x2 =5.078,6.262,9.456,5.156,P ＜ 0.05).Conclusions The related etiological factors may be different due to different primary location of ICC.Primary tumor in left liver is significantly associated with intrahepatic bile duct stone of left liver,and primary tumor in right liver is associated with combined hepatitis,while combined hepatitis could lead early occurrence of ICC.At present,there is a low rate of lymph node dissection in ICC patients and a high lymph node metastasis rate in patients receiving lymph node dissection.Lymph node metastasis is associated with primary tumor location and diameter,levels of CEA and TBil.Lymph node metastasis rate in left liver is higher than that in right liver,and is getting higher with increased primary tumor diameter.

Objective:To establish and optimize the in vitro enrichment method for human coronavirus NL63 reference strain (HCoV-NL63-NC_005831).Methods:HCoV-NL63 was cultured in rhesus monkey kidney cell line (LLC-MK2). Then at 135 000× g, the virus culture supernatant was centrifuged for 4 h, 6 h, 8 h, 10 h and 12 h to enrich the virus, and compared with the enrichment method of commercial PEG Precipitation Kit. Real time reverse transcription polymerase chain reaction (qRT-PCR), TCID 50 assay and plaque assay were used to quantitatively detect the viral nucleic acid and viral biological activity of the original virus solution and the enriched virus solution respectively. The virus morphology before and after enrichment was observed after negative staining under a transmission electron microscope. Results:After 100∶1 volume enrichment by ultracentrifugation and PEG precipitation, both of the concentration of virus nucleic acid and live virus were higher than that of the original virus solution ( P < 0.001), and before and after the enrichment of the two method, the normal negative staining morphology of virus particles can be observed under the electron microscope. After 4 h, 6 h, 8 h, 10 h and 12 h ultracentrifugation, the concentration of live virus was (7.35±1.62) times, (13.98±1.71) times, (36.36±10.41) times, (48.16±9.38) times, (48.16±9.38) times and (54.26±7.02) times that of the original virus solution, respectively; the concentration of live virus after PEG precipitation was (3.39±0.16) times that of the original virus solution, and the nucleic acid concentration and live virus concentration of concentrated virus solution obtained by ultracentrifugation were significantly higher than those obtained by PEG precipitation ( P < 0.05). After 8 h, 10 h or 12 h ultracentrifugation, the virus concentration was significantly higher than that of 4 h or 6 h ( P < 0.05). However, there was no significant difference in virus content between 8 h, 10 h and 12 h ( P >0.05). Conclusions:Compared with commercial PEG precipitation method, ultracentrifugation method can enrich HCoV-NL63 virus more effectively; when the relative centrifugal force is 135 000× g, the better enrichment effect in vitro can be obtained by selecting the super separation time of 8 h.

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.

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( Q R)) 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 na?ve 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(χ 2=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 na?ve 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.

Objective:To explore the clinical value of adjuvant therapy in patients with T3 gallbladder cancer (GBC) who have undergone R0 resection.Methods:Clinical and pathological data from 415 patients with T3 GBC who underwent surgical treatment in 7 tertiary centers in China from January 2013 to December 2018 were collected,including 251 males and 164 females,aged (61±11)years (range: 26 to 88 years). Depending on whether to receive adjuvant therapy after radical resection,the patients were divided into the radical resection group alone (group A, n=358) and the radical resection combined with the postoperative adjuvant therapy group (group B, n=57). The general data of the two groups were matched 1∶1 by propensity score matching method,and the caliper value was 0.02.Clinicopathological characteristics,overall survival and disease-free survival of the two groups were compared.The Cox regression model was used for multivariate analysis,and patients with at least one or more independent risk factors were classified as high-risk clinicopathological subtypes. Subgroup analysis was performed to assess the clinical value of adjuvant therapy after radical resection in patients with high-risk clinicopathological subtypes. Results:After the matching,there were 42 patients in each of the two groups. The incidence of gallbladder cancer and the number of dissected lymph nodes in group B after cholecystectomy were higher than those in group A ( χ 2=9.224,2.570,both P<0.05). There were no significant differences in overall survival rate and disease-free survival rate between the two groups before and after matching (all P>0.05). The results of the univariate and multivariate analysis showed that CA19-9>39 U/ml,nerve invasion,tumor location (liver side or bilateral),TNM stage ⅢB to ⅣB ,poorly differentiated tumor were independent prognostic factors of overall survival and disease-free survival of patients with T3 stage gallbladder cancer (all P<0.05).Three hundred and twenty-nine patients(79.3%) had high-risk clinicopathological subtypes,and the median survival time after curative resection with and without adjuvant therapy was 17 months and 34 months respectively,and the 3-year and 5-year overall survival rates were respectively 40.0%,21.3% and 46.0%,46.0% ( χ 2=4.042, P=0.044);the median disease-free survival time was 9 months and 13 months,and the 3-year and 5-year disease-free survival rates were 23.4%,13.6% and 30.2%,18.2% ( χ 2=0.992, P=0.319). Conclusions:Postoperative adjuvant therapy following radical surgery did not yield significant improvements in the overall survival and disease-free survival rates of patients diagnosed with T3 gallbladder cancer. However, it demonstrated a significant extension in the overall survival rate for patients presenting high-risk clinicopathological subtypes.