This article introduces the proteomic study of the cell lines,tissue,pancreatic juice and serum from well-diagnosed patients of pancreatic cancer,benign pancreatic disease,normal control,etc.The highly specific and sensitive biomarkers and their significant roles in the mechanism of development and progress of pancreatic cancer,early diagnosis,therapy monitoring,prognosis,follow-up,and so on,are presented and discussed.

Pancreatic cancer is a commonly malignant gastrointestinal tumor with an significantly increasing incidence.Those patients without nonspecific symptoms at early stage had mostly lost the opportunity of surgical therapy when pancreatic cancer was detected at advanced stage,and its prognosis is poor.Therefore,it is rather important to improve the early diagnosis of pancreatic cancer.In recent years,proteomics is developing rapidly.Proteomics technologies have been widely used in clinical research.Using proteomics technology screening pancreatic cancer tumor markers becomes the research focus,thus we try to find a kind of or a group of pancreatic tumor markers,so as to improve the diagnosis of pancreatic cancer.

Objective A comparative proteomic method was utilized to analyze serum proteins among pancreatic cancer patients,pancreatic benign tumor group,chronic pancreatitis group and normal control group to discover a new potential specific early diagnostic marker.Methods Comparative analysis on the pancreatic peripheral blood protein profiling from 40 pancreatic cancer patients,10 benign tumor patients,10 chronic pancreatitis patients and 40 cancer-free controls from May 2009 to April 2011 was carried out by 2D differential gel electrophoresis (2D-DIGE) and differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).Results Three differentially expressed proteins,Hemopexin (Hpx),Ficolin 3 (FCN3) and Serum amyloid P-component (SAP) was identified.Those proteins were higher expression in pancreatic cancer group compared with benign tumor group,chronic pancreatitis group and normal control group.Each point in pancreatic cancer expression were 1.57,1.99,1.63 times than normal control expression,respectively (P ＜0.05).Conclusions In this study,the identified proteins,Hpx,FCN3 and SAP may be as potential specific early diagnostic markers of pancreatic carcinoma.2D-DIGE and MALDI-TOF-MS technology in screening specific serum tumor markers of pancreatic cancer has a well repeatability and stability.

Objective To determine the expression of brain-derived neurotrophic factor (BDNF) in pancreatic ductal adenocarcinoma and its clinical significance.Methods SP immunohistochemical staining method was used to detect the expression of BDNF in 46 cases of pancreatic ductal adenocarcinoma,38 cases of benign pancreatic diseases and 20 cases of normal pancreatic tissue.Real time PCR and Western blot was used to detect the protein and mRNA expression levels.The relationship between BDNF expression and clinicopathological parameters of pancreatic cancer was determined.Results The positive expression rate of BDNF was 52.2％ (24/46) in pancreatic ductal adenocarcinoma pancreatic ductal adenocarcinoma,7.8％ (3/38) in benign pancreatic diseases,and none of the normal pancreatic tissue was BDNF positive.The BDNF protein expression levels in pancreatic ductal adenocarcinoma,benign pancreatic diseases and normal pancreatic tissue were 0.38± 0.01,0.56± 0.01,0.97± 0.01,respectively,and the BDNF mRNA expression levels were 0.85 ± 0.14,1.67 ± 0.21,3.45 ± 0.67,respectively,and the expression levels in pancreatic ductal adenocarcinoma and benign pancreatic diseases were significantly higher than that in normal pancreatic tissue,while the expression level in pancreatic ductal adenocarcinoma was significantly higher than that in benign pancreatic diseases (P ＜0.05).Positive BDNF expression was correlated with nerve infiltration and lymph node metastasis of pancreatic ductal adenocarcinoma,but it was not related to age,sex,tumor size,location and differentiated degree (P ＞ 0.05).Conclusions BDNF is involved in the development and growth of pancreatic cancer,and it may be related with patient's prognosis.

Objective To investigate the clinicopathological features of patients with serous cystadenomas of the pancreas (SCAP).Methods The clinical and pathological features of 21 cases of SCAP were retrospectively analyzed.Results The mean age of the 21 cases was 61 years old,male:female ratio was 1∶ 1.33,18 (85.7％) patients presented with abdominal pain,bloating,abdominal mass,weight loss,and 3 (14.3％) patients were found during check-up.The tumors were located in pancreatic head in 9 patients,in pancreatic body and tail in 12 patients.The clinical manifestations were pancreatic cystic lesions.All patients underwent surgery.Histologically,the cyst wall was complete and lined with flat or cuboidal epithelium,cytoplasm was translucent,nucleus were round or oval with similar size,no significant nuclear atypia and mitotic activity was found.The pathologic diagnosis was micro-cyst type in 15 cases,single-cyst type in 6 cases.Immunohistochemistry method showed EMA,CK7,CK19 positive and PAS staining positive.The positive expression rate of Ki 67 was between 1％ and 3％.After follow-up of 19 cases ranging from 3 months to 7 years,no recurrence and metastasis was detected.Conclusions SCAP is seen predominantly in elderly female patients with significant symptoms.A majority of tumors are located in the pancreatic body and tail.SCAP presents with characteristics of pancreatic ductal epithelial,and the prognosis is excellent.

ObjectiveA comparative proteomic method was used to analyse serum proteins between pancreatic cancer patients and control group,and to find a new protential specific marker.MethodsComparative analysis on the pancreatic peripheral blood protein profiling from 40 pancreatic cancer patients,10 chronic pancreatitis patients,10 benign tumor patients and 40 cancer-free controls was carried out by 2D differential gel electrophoresis,and differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry.ResultsTwo differentially expressed proteins:transthyretin and apolipoprotein E were identified.Those proteins were highly expressed in pancreatic carcinoma group compared with normal control group,chronic pancreatitis group and benign tumor group.Conclusion2D differential gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry technology in screening specific serum biomarkers of pancreatic cancer has a well repeatability and stability.The identified protein transthyretin in this study may be as specific serum biomarkers of pancreatic carcinoma.

OBJECTIVEThe purposes of this study were to clone and sequence the major histocompatibility complex type I (MHC I) molecular antigen recognizing gene (H-2Kk) of 615 mice, and to provide the functional gene for transgenic therapy.

METHODSThe 1.4 kb full-length fragment of H-2Kk gene complementary DNA (cDNA) was amplified from the total RNA of 615 mouse liver by using reverse transcription polymerase chain reaction (RT-PCR). The cDNA was inserted into PGEM3Zf(+) vector directionally, and the competent E. coli JM109 was transformed with the ligated product. The recombinant PGEM3Zf(+)-H-2Kk cDNA plasmid was obtained using restricted enzyme analysis of the transfectants. The complete sequence of 615 mouse H-2Kk cDNA was determined by using Sanger's method.

RESULTSThe sequences of 615 mouse H-2Kk cDNA were 99% similar with those of H-2Kk cDNA which were reported by other researchers, and the sequences encoding antigen recognizing regions (ARS) were identical with each other.

CONCLUSIONThe authors cloned the MHC I molecular antigen recognizing gene (H-2Kk) of 615 mice successfully and got the functional gene of MHC I.

Animals ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Genes ; genetics ; Genes, MHC Class I ; genetics ; Genetic Therapy ; H-2 Antigens ; genetics ; Mice ; Mice, Inbred C57BL ; Point Mutation ; Sequence Analysis, DNA ; Transgenes

clues for interpretation of myocardial ischemia.

Background It is thought in recently that photodynamic therapy (PDT) is an effective treatment method for chronic central serous chorioretinopathy (CSC), but the dosage of verteporfin and its long-term efficacy and complications is rarely elucidated ever before.Objective This study was to observe the long-term efficacy and safety of 60％ dose verteporfin PDT for chronic CSC.Methods This is a retrospective study and a self-controlled design was used.The clinical data of 25 eyes of 21 chronic CSC patients who received 60％-dose verteporfin PDT in Henan Eye Institute from January 2009 to May 2010 were reviewed, with the male 18 (85.71％) and female 3 (14.29％) as well as monocular CSC 17 patients and binocular CSC 4 patients.The average ages of the patients were (43±5) years.Fundus fluorescein angiography (FFA) , indocyanine green angiography (ICGA), optical coherence tomography(OCT) and best corrected visual acuity (BCVA) were examined in all the patients before and after treatment.PDT with the 60％-dose verteporfin (3.6 mg/m2) was carried out on the CSC eyes.The treated eyes were examined 2 weeks, 1 month and 3 months after PDT.The BCVA,subfoveal choroid thickness,FFA and ICGA findings before and after PDT were compared.The following-up duration was 5 years or more.Results The BCVA before and 3 months after PDT were 0.5 ±0.1 and 0.9±0.2, respectively, with a statistically significant difference between them (t =19.17,P =0.00).The subfoveal choroidal thickness value 3 months after PDT was (326.56±39.47) μm,which was significantly reduced in comparison with (486.24 ±47.53) μm before PDT (t =25.17, P =0.00).FFA and ICGA showed that the leakage of fluorescein (hyperfluorescence) was disappeared in all the treated eyes.No systemic or local adverse effects and recurrence were observed during the follow-up period.Conclusions On the basis of the results of this study and available information,60％-dose verteporfin PDT seems to have a better long-term efficacy and safety than full-dose verteporfin in treating chronic CSC.

Objective To observe the cortical functioning of healthy volunteers during acupuncture as a way of exploring acupuncture's neural mechanisms.Methods Twenty healthy volunteers received acupuncture applied to the left hegu and waiguan acupoints while their cortical activity was examined using blood oxygenation level-dependent functional magnetic resonance imaging (BOLD-fMRI).Brain activation,especially of the regions related to motor function,were observed and analyzed.Results Acupuncture applied to the left hegu and waiguan acupoints was observed to significantly activate the left middle frontal gyrus and the inferior frontal gyrus,with many activated regions in the left insula and a few in the left cerebellum,the left precentral gyrus,the left postcentral gyrus,the left inferior parietal lobule,the left medial frontal gyrus,the left precuneus,the left anterior cingulate gyrus and the left claustrum.The right side of the brain was excited mainly in the right middle frontal gyrus and the right medial frontal gyrus.The right inferior parietal lobule and the right precentral gyrus were also activated to some extent.There was slight activation of the right middle temporal gyrus,the right superior temporal gyrus,the right insula,the right inferior frontal gyrus and the right postcentral gyrus.The negatively activated regions were mainly located on both sides of the limbic lobe,including the hippocampus,the parahippocampal gyrus and the cingulate gyrus.The left superior temporal gyrus,the left middle temporal gyrus and the right middle frontal gyrus also had small negative activation zones.Conclusions In brain regions associated with motor function,in addition to partial activation of the contralateral primary sensorimotor area,acupuncture at these two points clearly generates bilateral activation of secondary motor areas with some activation in the ipsilateral cerebellum.This may serve as a neuropathological basis for acupuncture treatment of motor dysfunction.BOLD-fMRI imaging displays the neural effects of acupuncture in an intuitive way.It can be a useful technique for further study of the neural effects of acupuncture on pathological conditions.