Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

Objective:To observe the effects of electroacupuncture (EA) of three different frequencies (2 Hz,80 Hz and 2 Hz/80 Hz) on the free radicals in hippocampus of vascular dementia (VD) model mice.Methods:A total of 100 Kunming mice were randomly divided into a sham operation group,a model group,a 2 Hz EA group,an 80 Hz EA group and a 2 Hz/80 Hz EA group,with 20 mice in each group.The ischemia-reperfusion VD model was established by repeated blockade of bilateral common carotid arteries.Mice in EA groups began EA treatment on the 4th day after the operation.Baihui (GV 20),Dazhui (GV 14),Geshu (BL 17) and Zusanli (ST 36) were punctured and then connected to EA instrument,with different waves of 2 Hz,80 Hz or 2 Hz/80 Hz (10 min/time) applied accordingly,once a day.During the jumping stand experiment,the learning performance,memory performance and hippocampal calcitonin gene-related peptide (CGRP),nitric oxide synthase (NOS),malondialdehyde (MDA),changes in superoxide dismutase (SOD) and true choline esterase (TChE) were observed.In hippocampus,the CGRP level was determined by radioimmunoassay;the MDA level was determined by thiobarbituric acid colorimetric method;the activities of NOS and TChE were determined by spectrophotometry;the activity of SOD was determined by xanthine oxidase method.Results:Compared with the sham operation group,the performances of learning and memory decreased significantly in the model group (P＜0.01);in hippocampus,the CGRP level decreased,the MDA level increased,the activities of NOS and TChE increased,and the activity of SOD decreased in the model group.Compared with the model group,the learning and memory performances of the EA groups were significantly improved (P＜0.05 or P＜0.01);in hippocampus,the CGRP level increased,the MDA level decreased,the NOS and TChE activities decreased,and the SOD activity increased (P＜0.05 or P＜0.01).Among EA groups,the 2 Hz/80 Hz EA group was superior to the 2 Hz EA group and the 80 Hz EA group (P＜0.05 or P＜0.01).Conclusion:EA can improve the cognitive impairment of mice with ischemia-reperfusion VD.The mechanism may be related to the improvement of cerebral blood circulation,regulation of the central neurotransmitters,fighting lipid peroxidation and promoting nerve cell repair.The therapeutic effects of EA with different frequencies were different,and the intervention effect by EA at 2 Hz/80Hz is the most significant.

Clinical transplantation has indicated that cord blood (CB) can be used in the hematopoietic reconstitution in the children, but not well used in the adult patients because of the low cell amount. The present study aimed to explore the capability of proliferation and differentiation of the hematopoietic stem/progenitor cells derived from human mature placenta tissue (PT) in vitro, and to find a new source of hematopoietic/progenitor cells for clinical transplantation. CD34(+) cells in human mature placenta tissue were isolated and characterized by using enzyme-digestion method and flow cytometry. A long culture system without cytokines was established with human mature placenta tissue-derived mononucleated cells and cord blood mononuclear cells. The number of nucleated cells was weekly counted in culture for 14 weeks. The number of CFC was counted in culture for 2 weeks. The results showed that the CFC yields (CFU-GM, 186.90 +/- 24.52; BFU-E, 101.40 +/- 13.35) and the percentage of CD34(+) cells (2.74 +/- 0.61%) and CD34(+)/CD38(-) cells (2.46 +/- 0.42%) in placenta tissue (PT) were higher than CFC (CFU-GM, 136.90 +/- 25.15; BFU-E, 49.20 +/- 8.13), CD34(+) cells (1.73 +/- 0.32%) and CD34(+)/CD38(-) cells (0.80 +/- 0.25%) in cord blood (CB). The MNCs from PT have shown more survival ability than the cells from CB in the long-term cell culture condition; and the cells from PT increased by 2 times. It is concluded that the placenta may be another hematopoietic organ in ontogeny. The cells from placenta were more juvenile, and may be favorable source for clinical stem cell transplantation.
Antigens, CD34 ; analysis ; CD36 Antigens ; analysis ; Cell Differentiation ; physiology ; Cells, Cultured ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Placenta ; cytology

The present study was to investigate the effects of diltiazem, a ghrelin receptor agonist, on food intake and gastrointestinal functions in rats. Rats were intragastrically administered with diltiazem solution (daily 16 mg/kg, 30 mg/kg or 80 mg/kg, 30 d), and the rats with saline as control. To detect the effects of diltiazem on food intake and body weight, the average daily food intake and body weight were recorded, and the serum metabolic hormones of plasma growth hormone (GH) and neuropeptide Y (NPY) were tested by radioimmunoassay. By means of the spectrophotometer and the modified Mett's method, the effects of diltiazem on rat's gastrointestinal function and pepsin activity were tested, respectively. In addition, the gastric juice's acidity of rats was detected by titration and the secretion amount was calculated. The results showed that the food intake and body weight were maximally promoted by diltiazem at the dose of 30 mg/kg daily (30 d). The average daily food intake and body weight were significantly increased, and the serum concentrations of GH and NPY were also remarkably increased in diltiazem-treated groups compared with those in control group. The results also showed that the gastric emptying rate, gastric acid secretion and the activity of pepsin were significantly increased in diltiazem-treated group compared with those in control group. These results suggest that diltiazem induces enhancement of eating, in the same time, it can also stimulate the gastrointestinal function and regulate growth of rat.
Animals ; Body Weight ; drug effects ; Diltiazem ; pharmacology ; Eating ; drug effects ; Female ; Gastric Emptying ; drug effects ; Gastrointestinal Motility ; drug effects ; Gastrointestinal Tract ; physiology ; Growth Hormone ; blood ; Neuropeptide Y ; blood ; Rats ; Rats, Sprague-Dawley ; Receptors, Ghrelin ; agonists

OBJECTIVETo investigate the correlation between clinicopathologic factors and peritoneal dissemination in colorectal cancer, and the impact of surgery on the prognosis of patients with peritoneal dissemination.

METHODSBased on the clinical database built in 1994, the clinicopathologic data and the result of follow-up of all colorectal cancer patients were analyzed retrospectively.

RESULTOne hundred and fifty cases (7.40%) in all 2019 patients with primary colorectal cancer were found complicated with peritoneal dissemination. The clinicopathologic factors in patients with peritoneal dissemination were significantly correlated with tumor penetrating through serosa, lymph node metastasis, undifferentiated carcinoma, ascites, different pathological type, circumference of tumor, neoplastic intestinal obstruction, and Dukes staging. Peritoneal dissemination was associated with tumor penetrating through serosa, different pathological type on multivariate analysis. The 1-, 3-, 5-year survival rate of the patients with peritoneal dissemination were 70.4%, 38.1%, 30.2%; The 1-, 3-, 5-year survival rate of the patients undergoing radical resection were significant better than those in the cases undergoing palliative operation or palliative resection (P < 0.05).

CONCLUSIONSColorectal cancer complicated with peritoneal dissemination has poorer clinicopathologic characteristics. Those with local peritoneal dissemination has rather better prognosis than those with wide peritoneal dissemination. Radical resection of the disseminated tumor can improve the prognosis of the patients.

Colorectal Neoplasms ; pathology ; surgery ; Follow-Up Studies ; Humans ; Neoplasm Invasiveness ; Neoplasm Seeding ; Peritoneal Neoplasms ; secondary ; Prognosis ; Retrospective Studies ; Survival Analysis

OBJECTIVETo investigate the clinical effect of surgery following systemic targeted therapy of tyrosine kinase inhibitors (TKIs) in patients with metastatic gastrointestinal stromal tumors (GIST).

METHODSFrom June 2007 to December 2009, data of 15 consecutive patients with metastatic GIST treated with imatinib/sunitinib followed by surgery were retrospectively analyzed.

RESULTSDisease responses to TKI treatment was categorized into controlled disease (including partial response and stable disease) (6, 40.0%), limited progression (4, 26.7%), and generalized progression (5, 33.3%), respectively. Surgeries were performed after mean 12 months following TKI therapies. Gross complete resection or optimal debulking with minimal residual disease were managed to performed in 8/10 patients with disease controlled and limited progression, while optimal debulking only achieved in 2/5 patients with generalized progression. Surgical morbidity was 20.0% (3/15). After operation, patients with disease controlled and limited progression had a median progression-free survival of 25.0 months and 2-year overall survival rate of 100%. In contrast, for patients with generalized progression, the median progression- free survival was 3 months (P<0.01), and median overall survival 10.5 months.

CONCLUSIONSPatients with metastatic GIST who have controlled disease or limited progression to TKI therapy can benefit from surgical resection. Surgery should be selective in patients with generalized progression since surgery hardly improves survival in these patients.

Adult ; Aged ; Benzamides ; Disease-Free Survival ; Female ; Gastrointestinal Stromal Tumors ; pathology ; therapy ; Humans ; Imatinib Mesylate ; Indoles ; therapeutic use ; Intraoperative Period ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Protein Kinase Inhibitors ; therapeutic use ; Pyrimidines ; therapeutic use ; Pyrroles ; therapeutic use ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo explore the impact of clinicopathological features and extent of lymph node dissection on the prognosis in early gastric cancer (EGC) patients.

METHODSA total of 142 EGC cases screened from database of gastric cancer of Sun Yat-sen University, from Aug. 1994 to Jan. 2010, were included in this study. According to the lymph node metastasis status, they were divided into lymph node negative (n = 116) and lymph node positive (n = 26) groups. The clinicopathological features of the two groups and the impact of extent of lymph node dissection on the prognosis were analyzed.

RESULTSThere were no significant differences in age, gender, tumor size and location, Borrmann typing, WHO TNM staging, histological typing, and CEA value between the two groups (P > 0.05). The TNM stages in the lymph node positive group were higher than that in the lymph node negative group (P < 0.001). Between the cases who underwent D1 (n = 21) and D2 (n = 121) dissection, there were no significant differences in postoperative hospital days, blood transfusion volume, and operation time (P > 0.05). The median numbers of LN dissected in D1 and D2 cases were 4 (0 to 16) and 20 (12 to 30), with a significant difference (P = 0.000), but the number of positive LN without significant difference (P = 0.502). The postoperative complication rates were 9.5% in the D1 and 3.3% in the D2 dissection groups, without a significant difference (P = 0.128). The median survival time of the lymph node negative and positive groups was 156 vs. 96 months (P = 0.010). In cases who received D2 and D1 lymph node dissection, the median survival time (MST) was 156 vs. 96 months (P = 0.0022). In the lymph node positive group, D2 dissection prolonged survival time significantly than D1 dissection (96 vs. 27months) (P = 0.001). Cox regression analysis showed that the extent of lymph node dissection and LN metastasis were independent prognostic factors for EGC patients.

CONCLUSIONSIt is not able to accurately assess the LN metastasis status preoperatively according to the routine clinicopathological features. For the patients with unknown LN metastasis status, D2 dissection should be the first choice. Comparing with D1 dissection, the morbidity of D2 dissection are not increased, but survival time is prolonged.

Adenocarcinoma ; drug therapy ; pathology ; surgery ; Adenocarcinoma, Mucinous ; drug therapy ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Signet Ring Cell ; drug therapy ; pathology ; surgery ; Chemotherapy, Adjuvant ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Leucovorin ; administration & dosage ; Lymph Node Excision ; methods ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Retrospective Studies ; Stomach Neoplasms ; drug therapy ; pathology ; surgery ; Survival Rate

OBJECTIVETo summarize the clinicopathological characteristics and analyze the prognostic factors of young patients with gastric cancer.

METHODSA total of 99 patients with the age less than or equal to 40 were admitted in The First Affiliated Hospital of Sun Yat-sen University from August 2001 to December 2009. Their clinicopathological and follow-up data were compared with middle-aged and elderly patients with the age more than 40.

RESULTSThere were statistically significant differences in gender, tumor location, Borrmann type, histological type, differentiated histology, depth of invasion, peritoneal metastasis between young patients and elder ones. The 5-year survival rates of young and elder patients were 49.1% and 44.4% respectively, and the difference was not statistically significant (P>0.05). Univariate and multivariate analyses showed that TNM stage (P=0.014) and surgical methods (P=0.012) were independent predictive factors of survival for young patients. For the young patients, the 5-year survival rate was 56.7% after curative resection, 11.1% after palliative resection. Those who underwent palliative surgery or biopsy alone died within 1 year after surgery. The difference between difference surgical procedures in survival were statistically significant (P<0.05).

CONCLUSIONSAs compared to elder patients, young patients with gastric cancer have special clinicopathological features. However, no significant difference of survival rate is found between the young and the elder patients. TNM stage and surgical methods are independent prognostic factors of young patients with gastric cancer. Radical resection appears to confer the only chance of prolonged survival.

Adult ; Age Factors ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Male ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; epidemiology ; pathology ; surgery ; Survival Rate

OBJECTIVETo evaluate the efficacy and influencing factors of imatinib in patients with advanced gastrointestinal stromal tumor(GIST).

METHODSFrom April 2004 to January 2010, clinicopathological data of 73 adult patients with advanced GIST treated with imatinib at the First Affiliated Hospital of Sun Yat-sen University were retrospectively analyzed. The treatment outcomes and associated factors were investigated.

RESULTSTreatment outcomes included complete response in 1(1.4%) patients, partial response in 53(72.6%), stable disease in 14(19.2%), and primary resistant in 5(6.8%). All the patients had routine followed up, the length of which ranged from 12 to 76 (median 32) months. The median progression-free survival was 45.0 months(95% confidence interval, 34.2-55.8). The progression-free survival(PFS) rate was 87.7% in 1 year, 63.6% in 3 year, and 39.6% in 5 years. On multivariate analysis, both mutation status and patient performance were independent factors influencing the efficacy of imatinib treatment(both P<0.01). PFS was significantly better in patients with c-kit exon 11 mutations than those with exon 9 mutations, and better in lower ECOG scales than in higher ones.

CONCLUSIONImatinib is effective in treating patients with advanced GIST, c-kit exon 9 mutations and poor performance status predict an adverse survival benefit of imatinib therapy.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; therapeutic use ; Exons ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; drug therapy ; genetics ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Mutation ; Piperazines ; therapeutic use ; Proto-Oncogene Proteins c-kit ; genetics ; Pyrimidines ; therapeutic use ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVESTo understand the role cellular immunology plays in the pathogenesis of chronic hepatitis B (CHB) through analysis of T cell receptor (TCR) beta chain variable region gene (BV) family dominant usage and beta chain complementarity determining region3 (CDR3) sequences of peripheral blood mononuclear cells of the patients.

METHODSTCR BV families were amplified by inverse polymerase chain reaction (RT-PCR), and the dominant usage of BV families and CDR3 repertoire were analyzed by immunoscope technology for 8 CHB patients during their acute exacerbations and for 4 healthy blood donors who served as controls. The clonality of the T cells suspected by immunoscope was further confirmed by CDR3 sequencing.

RESULTSThe TCR BV CDR3 repertoire of the 4 healthy blood donors showed a Gaussian distribution. In the 8 CHB patients, however, the clonal expansion of T cells showed different TCR BV families with each patient. The T cells of the clonal expansion shared different CDR3 sequences.

CONCLUSIONThe peripheral blood T cells of CHB patients during their acute exacerbation showed significantly a clonal expansion and their T cell clonal expansion may be stimulated by several HBV epitopes. These results indicate that cellular immunology is involved in the pathogenesis of the liver inflammation process of CHB.

Adult ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; genetics ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Receptors, Antigen, T-Cell ; genetics