Basic fibroblast growth factor (bFGF) is a multi-potential growth factor whose biological activities depend on its receptor's intrinsic tyrosine kinase activity and second messengers such as the mitogen activated protein kinases. Heparin sulfate proteoglycans (HSPGs) have been demonstrated to enhance or inhibit bFGF activity. The response elicited by HSPG is related to the relative concentrations and binding kinetics for bFGF of the various pools of HSPG. The type of cellular response might depend on the specific HSPG and FGF receptor expressed on the cell surface. The specific core protein of HSPG, and tissue specific differences in heparin sulfate modification result in altered bFGF regulation.

AIM: To screen the proteins interacting with human augmenter of liver regeneration (hALR) by yeast two-hybrid system and to study the mechanism of hALR action. METHODS: hALR bait plasmid was constructed by ligating the gene of hALR into pGBKT7, then transformed into yeast AH109. The yeast strain AH109 containing pGBKT7-hALR was mated with yeast Y187 containing human liver cDNA library plasmid. Diploid yeast was plated on SD/-trp-leu-his-ade (QDO) for screening and on QDO containing X-?-gal for further selection.The AD/library inserts were amplified by PCR and the PCR products were characterized by digesting with Sau3AⅠ and HaeⅢ restriction enzyme to eliminate the duplicates. After sequencing, the positive clones were analysed by bioinformatics. RESULTS: Several positive clones were obtaind. The sequencing and analysis shown that one of them is 669 bp DNA fragment encoding ? subunit of Na +,K +-ATPase. The 224 bp 3′terminal DNA fragment is non-encoder region, and the 445 bp 5′terminal DNA encodes C-terminal 147 amino acid residues of Na +, K +-ATPase ? subunit. CONCLUSION: The results of screening proteins using yeast two-hybrid system showed that hALR could interact directly with Na +, K +-ATPase in the yeast cell.

Objective To develop a better probe preparation method of gene chips for the further gene chips fabrication and its application in the gene expression. Method Using human leukemia K562 cells as material,applying a new kind of technology for gene differential display restriction display(RD PCR),we isolated a lot of cDNA fragments as probes of gene chips. Results This method has overcome the shortcoming of more pseudopositives in DD PCR,and was easy to be manipulated.Compared with the chip fabricated with full cDNA probes,the probe length is comparatively homogeneous,and the hybridization dynamics is apt to be controlled.Conclusion\ RD PCR technology could provide a new strategy for the probe preparation of gene chips.\;[

Objective:To evaluate the application value of small IT knife pre-cut in assistance to duodenal papillary cannulation compared with conventional guidewire cannulation.Methods:A retrospective analysis was performed on the data of 90 patients with choledocholithiasis including 52 patients with small IT knife pre-cut assisted intubation (small IT knife group) and 38 patients with conventional guidewire intubation (conventional group) in endoscopic retrograde cholangiopancreatography (ERCP) from May 2016 to July 2019 in the digestive endoscopy center of Nanjing Drum Tower Hospital. The basic data, curative effect and complications of the two groups were collected and compared.Results:There was statistically significant difference in gender composition between the small IT knife group and the conventional group ( χ2=5.679, P=0.017), but no significant difference in other baseline data between the two groups (all P>0.05). The median intubation time of the small IT knife group was significantly shorter than that of the conventional group (141.5 s VS 270.0 s, Z=1 268.0, P=0.022). There were no significant differences in the success rate of intubation [98.1% (51/52) VS 94.7% (36/38), χ2=0.760, P=0.571], the incidence of intraoperative bleeding [15.4% (8/52) VS 7.9% (3/38), χ2=1.148, P=0.345], postoperative pancreatitis [5.8% (3/52) VS 7.9% (3/38), χ2=0.159, P=0.694], and postoperative cholangitis [1.9% (1/52) VS 5.3% (2/38), χ2=0.760, P=0.571] between the two groups. No perforation occurred in the two groups. After stratifying according to the operator′s proficiency, the median intubation time was significantly different between the small IT knife expert group and the conventional expert group (116.0 s VS 258.0 s, Z=276.0, P=0.038), while there was no significant difference in the intubation time among other groups (all P>0.05). Conclusion:The small IT knife is safe and effective to pre-cut and assist intubation in ERCP, and it may shorten the intubation time.

Objective To investigate the efficacy and safety of endoscopic ultrasound-guided biliary drainage ( EUS-BD ) on obstructive jaundice patients who had failed in the endoscopic retrograde cholangiography ( ERC ) . Methods Seventeen obstructive jaundice patients who underwent EUS-BD at Nanjing Drum Tower Hospital from October 2015 to July 2017 were enrolled in the study. The types of biliary drainage, technical success rate, clinical efficacy, post-procedure adverse events, and follow-up were analyzed. Results EUS-BD was successfully performed in 16 out of 17 patients, with technical success rate of 94. 1％. Operation pathway included 12 EUS-guided hepaticogastrostomy, 2 EUS-guided antegrade stenting, 1 EUS-guided rendezvous, and 1 EUS-guided choledochoduodenostomy. All the 16 patients′bilirubin decreased at different levels after operation, and the clinical success rate was 94. 1％ (16/17). Post-operation complications included 5 cases of cholangitis, 2 of bile leakage, and 1 of pneumoperitoneum and duodenal perforation. The patients were followed up for 14-390 days after procedures. Two patients died of underlying disease one month after operation. Jaundice in another 2 patients worsened during follow-up and were improved after replacement of previous stent. Conclusion EUS-BD has a relatively high technical success rate, and can be considered as an alternative choice for patients with obstructive jaundice after failure of ERC. Operation pathway should be decided according to patient′s condition.

Objective To investigate the diagnostic value of intraductal ultrasonography ( IDUS ) with or without brush cytology on distinguishing malignant from benign biliary stricture. Methods The data of patients with biliary stricture, who underwent endoscopic retrograde cholangiography( ERC) and IDUS in Drum Tower Hospital from September 2007 to October 2015 were retrospectively analyzed. Features of IDUS images and results of brush cytology were compared with postoperative histological results to evaluate the diagnostic yield of IDUS and brush cytology on differential diagnosis of malignant biliary stricture. Results A total of 230 patients with biliary stricture underwent ERC and IDUS, and 206 patients meanwhile received brush cytology, including 84 cases of malignant biliary stricture. Of the 230 patients, 87 cases underwent surgery, and 75 revealed malignant results, including 65 cases of cholangiocarcinoma, 8 cases of pancreatic adenocarcinoma, and 2 cases of periampullary adenocarcinoma. Using postoperative pathologic results as the gold standard, 72 malignant and 9 benign biliary strictures were correctly diagnosed by IDUS. The sensitivity, specificity, positive predictive value, and negative predictive value of IDUS for diagnosis of malignant biliary stricture was 96. 0％, 75. 0％, 96. 0％, and 75. 0％, respectively. Sixty-five patients underwent IDUS with brush cytology during ERC before surgery, and 56 out of 65 showed malignant results. Thirty-one malignant and 9 benign biliary stricture were correctly diagnosed by brush cytology. The sensitivity, specificity, positive predict value, and negative predict value of brush cytology for diagnosis of malignant biliary stricture was 55. 4％, 100％, 100％, and 26. 5％, respectively. If brush cytology combined with IDUS to diagnose malignant biliary stricture, the sensitivity rose up to 94. 6％, while the specificity went down to 77. 8％, and the positive and negative predictive value was 96. 4％ and 70. 0％, respectively. Conclusion IDUS exhibited excellent sensitivity and specificity, while brush cytology revealed perfect specificity and positive predictive value on diagnosis of malignant biliary stricture. If IDUS combined with brush cytology, the sensitivity and negative predictive value could rise significantly, while the specificity and positive predictive value mildly went down, which suggested that IDUS remained a certain degree of over judgment and false positive rate.

Objective:To investigate the efficacy and safety of a new China-made direct visualization system of peroral cholangiopancreatography in the diagnosis and treatment of biliopancreatic diseases.Methods:Clinical data of 37 patients who underwent endoscopic examination through the direct visualization system of peroral cholangiopancreatography at Digestive Endoscopy Center of Nanjing Drum Tower Hospital from April 2020 to June 2021 were retrospectively analyzed. Technical success rate and complications were analyzed.Results:The examination was completed in 37 patients through the system. The technical success rate was 100.0%. The nature of biliary stricture was confirmed in 24 cases, presenece or absence of bleeding or residual stones in the bile duct was confirmed in 6 cases, neoplasm or residual stones in the pancreatic duct was determined in 2 cases, biliary stricture was passed assisted with visualized guidewire in 2 cases, and lithotripsy was performed assisted with biliary laser in 3 cases. Nine patients were pathologically diagnosed as having malignant biliary stricture, and 8 of them were confirmed malignant by the system. Drainage was performed in 34 cases after the examination. There were 3 cases of cholangitis, 4 cases of bacteremia and 2 cases of postoperative pancreatitis after the operation, which were relieved in a short time after conservative treatment. Bleeding occurred in 1 case which was improved after two times of endoscopic hemostasis.Conclusion:The new direct visualization system of peroral cholangiopancreatography is safe and effective in the diagnosis and treatment of biliopancreatic diseases.

[摘 要] 目的：探究小核核糖核蛋白多肽A（SNRPA）在肝细胞癌（HCC）组织和细胞中的表达及其调控HCC细胞HepG2和Hep3B恶性生物学行为的作用及其机制。方法: 数据库分析SNRPA在泛癌组织中的表达及其与病理分期、HCC患者预后的相关性。常规培养HepG2和Hep3B细胞，将si-NC，si-SNRPA#1、si-SNRPA#2转染HepG2和Hep3B细胞，实验分为si-NC组、si-SNRPA#1组和si-SNRPA#2组；将SNRPA-vector和SNRPA-oe载体转染LO2细胞，分为SNRPA-vector组和SNRPA-oe组。qPCR法检测正常肝细胞和肝癌细胞以及转染各组HepG2和Hep3B细胞中SNRPA mRNA的表达，MTT法、Transwell法和WB法分别检测转染后各组HepG2和Hep3B细胞的增殖、迁移和侵袭能力以及EMT相关蛋白表达的变化。结果: 数据库分析显示，SNRPA mRNA在多数肿瘤组织中均呈高表达（均P<0.001）且与病理分期有关联（P<0.05或P<0.01）。SNRPA在HCC组织和细胞中均呈高表达（P<0.05或P<0.01），且与HCC患者的预后有关联（P<0.01）。敲减SNRPA表达明显抑制HepG2和Hep3B细胞增殖（P<0.05或P<0.01）而过表达SNRPA则能促进LO2细胞增殖（P<0.01），敲减SNRPA表达明显抑制HepG2和Hep3B细胞的迁移和侵袭能力（均P<0.01），明显促进E-cadherin的表达上调（P<0.01），而抑制N-cadherin、vimentin的表达（P<0.01）。结论: SNRPA在HCC组织及细胞中呈明显高表达，其可能通过调控上皮间质转化（EMT）进程进而促进HepG2和Hep3B细胞的增殖、迁移和侵袭。