Pentamer guided HLA-restricted epitope identification for mucoprotein 4 antigen of pancreatic ductal adenocarcinoma / 中华外科杂志

<p><b>OBJECTIVES</b>To identify HLA-restricted epitope of mucoprotein 4 (MUC4) antigen as a tumor associated antigen of pancreatic ductal adenocarcinoma (PDAC), and to validate its natural presentation in PDAC patient peripheral blood.</p><p><b>METHODS</b>Two epitope prediction databases (SYFPEITHI and ProPred-I) were used to predict HLA-A*0201 restricted MUC4 epitope, T2 cell assay was used to determine the peptide binding affinity with HLA-A*0201 molecule. Dendritic cells (DCs) were induced from the HLA-A* 0201-positive healthy individuals' peripheral blood mononuclear cells (PBMC). Mature DCs were pulsed with synthesized peptides. Autologous CD8(+) T cells from the HLA-A* 0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase release assay and IFN-γ released by enzyme-linked immunospot assay. Pentamer was synthesized for HLA-A* 0201 restricted epitope P1126, then was used to detect specific CTL in PBMC of PDAC patients.</p><p><b>RESULTS</b>Five candidate HLA-A*0201 epitopes were predicted, LLLGVGTFV (P1125) and LLGVGTFVV (P1126) were determined as the two with more HLA-A*0201 affinity. Mature DCs could be induced from PBMCs. CTL induced by peptide P1126 could lyses T2 cells pulsed with peptide P1126 and HCT-116 cells [MUC4(+), HLA-A2(+)]. The number of CTL induced by peptide P1126 which could secret IFN-γ (130.3 ± 6.6) was obviously higher than that in the negative group. By Pentamer assay, P1126-pentamer and CD8 double positive CTL could be detected in PBMC of PDAC patients with MUC4(+) than patients with MUC4(-), but no significant difference of CTL frequency between patients with HLA-A2(+) and with HLA-A2(-) in MUC4(+) PDAC patients.</p><p><b>CONCLUSIONS</b>Tumor associated antigen MUC4-derived HLA-A* 0201-restrictive cytotoxic T lymphocyte (CTL) epitope P1126 can induce CTL reaction. The CTL can secret immunologic active material to induce the specific target cells lysis. P1126 epitope can be naturally presented in PBMC of PDAC patients, but its HLA-restriction may not be perfect.</p>

Diagnosis and surgical treatment for non-functional islet cell tumor: a retrospective analysis of 44 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the methods of diagnosis and surgical treatment for nonfunctional islet cell tumor (NICT).</p><p><b>METHODS</b>Forty-four patients with non-functional islet cell tumor treated at the First Affiliated Hospital of Nanjing Medical University during January 1968 to June 2008 were analyzed retrospectively. There were 9 males and 35 females, aged from 7- to 70-years-old. Clinical manifestation: 15 cases (34.1%) of abdominal masses, 17 patients (38.6%) with epigastric or back pain, 5 cases of jaundice, 5 cases (11.4%) for upper abdominal fullness or vomiting, 10 cases (22.7%) of pancreatic tumor noticed by routine health checkups or imaging examinations. Imaging examination: CT scan, sonography, ERCP, MRI, upper GI series were performed in 33 (75.0%), 16 (36.4%), 6 (13.6%), 2 (4.5%), and 10 cases (22.7%) respectively. Operation methods: 39 patients (88.6%) underwent surgical resection and the other 5 patients did not.</p><p><b></b>RESULTS</p><p><b>COMPLICATIONS</b>pancreatic fistula in 7 patients (15.9%), intra-abdominal bleeding in 4 (9.1%), gastrojejunal anastomosis outlet obstruction in 1 (2.3%), biliary fistula in 2 (4.5%) and incisional infection in 3 (6.8%). Surgery related mortality happened in 2 patients (4.5%), both treated before 1999. Twenty-five patients underwent operation between January 1999 and June 2008 were followed up for 6 to 108 months. All survive except one died 75 months after the surgery for unknown reason.</p><p><b>CONCLUSIONS</b>No specific clinical manifestation is recognized for non-functional islet cell tumor. Spiral CT is an optimal diagnostic method, while surgery is the first choice for treatment. Middle segmental pancreatectomy has become an alternative surgical protocol for NICT.</p>

Evaluating liver functional reserve by combining D-sorbitol with indocyanine green measurement / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To noninvasively evaluate hepatic functional blood flow, intrahepatic shunt rate and hepatic functional reserve in both normal and cirrhotic liver using D-sorbitol and indocyanine green measured by high performance liquid chromatography (HPLC).</p><p><b>METHODS</b>Male Sprague-Dawley (SD) rats were divided into normal control and cirrhotic group in which the rats were administrated with tetrachloride. Then the isolated perfused liver models were established. The pharmacokinetic indexes of D-sorbitol and indocyanine green (ICG) were measured by the traditional spectrophotometry (SPEC) and HPLC respectively.</p><p><b>RESULTS</b>(1) HPLC showed that ICG contained genuine ICG (ICGg) and ICG degraded products (ICGdp), which had similar spectrum but metabolic kinetics different with the retention time of 8.9 minutes and 24.2 minutes respectively. (2) Hepatic intrinsic metabolic capacity (QINT, I) was (36.57+/-13.03) ml/min in control group and (14.39+/-5.13) ml/min in cirrhotic group (t=7.08, P<0.01). (3) Hepatic functional blood flow (QFUNC) in cirrhotic group declined, compared with that in control group (34.06 ml/min+/-5.12 ml/min vs. 17.54 ml/min+/-7.02 ml/min, t=8.41, P<0.01), while intrahepatic shunt rate (QIHS) increased markedly (9.9%+/-1.4% vs. 47.5%+/-20.9%, t=8.35, P<0.01).</p><p><b>CONCLUSION</b>(1) HPLC method is superior to SPEC in measuring ICG, because it can avoid the disturbance from ICGdp, so that ICG measured by HPLC is valid for QINT, I evaluation. (2) The hepatic clearance of D-sorbitol measurement is a noninvasive and reliable method for evaluating the total blood flow in normal liver, and hepatic functional blood flow and intrahepatic shunt rate in cirrhotic liver. (3) Combining D-sorbitol with indocyanine green measurement is helpful for assessment of liver functional reserve.</p>