Objective To study the effect of curcumin on rat model of N-methylnitrosourea ( MNU) -induced bladder cancer and its mechanism. Methods One hundred SD rats were randomly divided into four groups:control group (n=10), model group (n=10), intervention group (n=40) and treatment group (n=40). Rats in the control group re-ceived intravesical infusion of distilled water. Rats in the other three groups were given MNU (1 mg/mL) in 2 mL saline at 2nd, 4th, 6th and 8th weeks to induce bladder cancer. In the model group, the rats were injected with distilled water in the bladder. The rats in the intervention group received 2 mL curcumin solution (400 μmol/L) at the 1st, 3rd, 5th, 7th and 9th weeks, and were sacrificed at the 11th week. In the model group, the rats were injected with distilled water in the bladder. In the treatment group, the rats had intravesical instillation of curcumin in the bladder (400 μmol/L, 2 mL) at 10, 12, 14, 16, and 18 weeks, and sacrificed at the 19th week. Bladder tissue samples were taken for pathological exami-nation using hematoxylin and eosin ( HE) staining. TUNEL staining assay was used to detect the apoptosis in tumor tissue. The expression of apoptosis-related proteins was detected by Western blot. Results The incidence of bladder cancer was 90% (9/10) in the model group, 12. 5% (5/40) in the intervention group and 92. 5% (37/40) in the treatment group at the 10th week, showing a significant difference between the intervention group and model group (P<0. 05), indicating an obvious interventional effect of curcumin on the bladder cancer. The incidence rate of bladder cancer in the treatment group was 78. 4% (30/37) at the 19th week, and compared with the 10th week before treatment, showing that curcumin can de-lay the recurrence of bladder cancer. TUNEL staining assay confirmed that curcumin significantly promoted the apoptosis in bladder cancer cells and inhibited their proliferation. The Western blot analysis showed that curcumin inhibited the activa-tion of NF-κB and effectively down-regulated the expression of NF-κB-regulated gene product. Conclusions Curcumin has a significant interventional effect on MNU-induced bladder cancer in the rat models. The mechanism may be through inhibi-tion of NF-κB activation and effective down-regulated NF-κB regulation of the gene products, and to regulate the expression of related proteins in bladder cancer, i. e. , inhibition of proliferation, induction of apoptosis, and further play a role of an-ti-cancer intervention and prevention of bladder cancer recurrence.

Objective To investigate the effect of bacillus Calmette-Guérin(BCG)on bladder cancer cells and their metabolites, and to preliminarily explore the possible mechanisms of BCG in the treatment of bladder cancer. Methods The rat model of bladder cancer was induced by intravesical instillation with N-methylnitrosourea(MNU). Bladder cancer cells and normal transitional epithelial cells were isolated and primarily cultured, and were divided into 5 groups according to the different components of the culture medium. The concentration of tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10)in the supernatant of each group was detected by enzyme linked immunosorbent assay (ELISA). The concentration of BCG to inhibit the cancer cell growth was determined by MTT assay. Apoptosis of bladder cancer cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL). Results Among the 15 rats,2 rats died after 2 times of instillation, and 3 rats died after 3 times of instillation, without obvious tumors found at autopsy. The other 10 rats were killed after completion of the intravesically instillation of MNU, and obvious tumors were found in 8 of them after dissection. The results of MTT assay showed that BCG had an inhibitory effect on the growth of bladder cancer cells,and the inhibitory rate was positively correlated with the concentration of BCG. The results of ELISA showed that the concentrations of TNF-α in the supernatant of groups B and D were(160.654 ± 5.775) ng/L and(124.443 ± 4.972)ng/L, respectively, with significant differences from those of the other three groups. The concentrations of IL-10 in the groups B and E were(16.973 ± 3.428)ng/L and(20.327 ± 2.721)ng/L, significantly higher than those of the other three groups. Apoptosis of cancer cells was not found in all groups. HE staining of the primary bladder cancer cells showed that the volume of cell nucleus was increased, and the nucleo-cytoplasmic ratio was increased. The number of nucleoli in some cells was increased and some nuclei appeared like ink drops with prominent nucleoli. Conclusions BCG has an inhibitory effect on the growth of rat bladder cancer cells. IL-10 and TNF-α secreted by the tumor cells might be involved in this regulatory process. However,apoptosis does not show an obvious effect on this inhibitory process.

OBJECTIVETo investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer.

METHODSRecombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined.

RESULTSThe viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group.

CONCLUSIONThe combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; blood supply ; pathology ; Transfection

OBJECTIVESTo investigate whether the human PC-3 cell infected with recombinant Ad-PTEN and Ad-p27Kip1 can steadily produce PTEN and p27Kip1 protein and change the biologic behaviors such as cell proliferation, cell cycle and apoptosis. The synergistic effect of PTEN and p27Kip1 on the therapy for prostate cancer has also been investigated.

METHODSWe constructed recombinant adenovirus vector of human tumor suppressor gene PTEN and p27Kip1. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and p27Kip1 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-p27Kip1 were determined by RT-PCR and Western blot respectively. MTT assay was used to determine the effect of PTEN and p27Kip1 on growth and proliferation of PC-3 cell; the change of cell cycle and apoptosis was examined by flow cytometry, and to compare between the combined therapy group and single gene therapy group.

RESULTSThe viral titers of Ad-PTEN and Ad-p27Kip1 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. After infected by adenovirus, it had been verified that the mRNA and protein expression of PTEN and p27Kip1 were steady in human PC-3 cell. Ad-PTEN and Ad-p27 Kip1 inhibited the growth and proliferation of PC-3 cells. The progression of cell cycle of PC-3 cell was arrested in G(0)-G(1) phase, meanwhile the apoptosis rate of PC-3 was also affected after Ad-PTEN or/and Ad-p27 Kip1 infected. There was significant difference between combined therapy group and single gene therapy group.

CONCLUSIONThe recombinant Ad-PTEN and Ad-p27Kip1 vector were constructed successfully and the expression of specific PTEN and p27Kip1 was high, steadily in PC-3 cell line. These results suggested that combination of PTEN with p27Kip1 has an application value in treatment of prostate cancer in future.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Chromosomes, Human, Pair 10 ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; Gene Deletion ; Genetic Therapy ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; pharmacology ; Male ; PTEN Phosphohydrolase ; genetics ; pharmacology ; Prostatic Neoplasms ; genetics ; physiopathology ; therapy ; Transfection

OBJECTIVEAllogeneic marrow transplantation is a curative therapy for thalassemia, but no more than 30% of patients have HLA-indentical sibling marrow donor. The selection of alternative donors of unrelative marrow and the study on the probability of treating thalassemia major with unrelated donor bone marrow transplantation are of importance.

METHODSNine children with thalassemia were included in the study, and their gene mutational type were homozygote of thalassemia and double heterozygote, respectively. All of them were finally diagnosed of thalassemia major, and treated with unrelated donor bone marrow transplantation. To high-resolution HLA typing, two patients were matched, five had one unmatched isoform and two had two unmatched isoforms. The erythrocyte blood type was not matched in six patients. The preparative regimen included busulfan (oral use, 16 mg/kg, divided for 4 days), cyclophosphamide (intravenous use, 200 mg/kg, divided for 4 days), antithymocyte immunoglobulin (intravenous use, 30 mg/kg, divided for 3 days), and fludarabine (intravenous use, 125 mg/m(2), divided for 3 days). Ciclosporin A and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis.

RESULTSAll patients had allergen reactions. One had hypotension. Five patients experienced I degrees approximately III degrees acute GVHD in the skin, while one had II degrees acute GVHD in liver. One patient had III degrees GVHD of intestines and gradually developed chronic GVHD in the skin, lungs and brain. One patient died of pulmonary hemorrhage. The duration when peripheral blood neutrophil count exceeded 0.5 x 10(9)/L was 12 - 26 days. The recovery time of WBC was as long as 23 - 110 days. Thrombocytes exceeded 50 x 10(9) within 61 approximately 142 days. The time when hemoglobin reached 100 g/L varied from 23 to 116 days. The last blood transfusion was on 13 - 62 days. Eight patients were fully grafted, while one was not grafted. During the 6 - 24 months of follow-up, seven patients' genotype of thalassemia major became normal. The erythrocyte blood type of five patients also changed into the same as that of donor. The hemoglobin was kept over 110 g/L without blood transfusion.

CONCLUSIONThe transplantation of unrelated donor bone marrow for thalassemia major was successful. Unrelated donor bone marrow transplantation could cure thalassemia major, which expanded the marrow donor source for the transplantation of thalassemia major.

ABO Blood-Group System ; Bone Marrow Transplantation ; adverse effects ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Follow-Up Studies ; Graft Rejection ; Graft Survival ; Histocompatibility Testing ; Humans ; Infant ; Male ; Transplantation Tolerance ; Transplantation, Homologous ; adverse effects ; Treatment Outcome ; beta-Thalassemia ; diagnosis ; therapy

To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

OBJECTIVETo investigate the clinical therapeutic results of allograft tendon for anatomical reconstruction of medial patellofemoral ligament (MPFL) for the treatment of patellar dislocations.

METHODSFrom September 2008 to June 2013, 16 patients with patellar dislocation underwent MPFL reconstructions. There were 2 males and 14 females, aged 11 to 27 years old (16 years old on average). Patellar dislocations occurred in 11 left and 5 right knees. The disease course ranged from 3 to 10 years. The frequency of dislocation ranged from 9 to 33 times (19 times on average). Affected knee joints showed patellar instability; the range of action for patella obviously increased. The X-ray films showed patellar dislocation. The preoperative Q angle was (36 ± 9)°, and the congruence angle was (63 ± 18)°. Reconstruction was performed via allograft tendon. Allograft tendon was fixed through the superomedial pole of the patella, and the other end was fixed at the natural MPFL insertion site near the medial femoral condyle with an interference screw in a bone tunnel. All the patients were evaluated postoperatively; Kujala patellofemoral scores, objective knee function, complications, and reoperations were assessed.

RESULTSPrimary healing was achieved in all cases. No infection or necrosis and absorption of grafts was observed. All the patients were followed up for an average of 16.4 months (ranged, 10 to 24 months) postoperatively. At the latest follow-up, all the patients had no pain, swelling and patellar instability; neither patella redislocation nor fracture occurred. The X-ray films showed good position of tunnel 6 months after operation, and the congruence angle was (5 ± 9)°, showing statistically significant difference when compared with preoperation (P < 0.05). The postoperative Q angle was (17 ± 8)°, the Kujala knee function score improved significantly from 45.20 ± 9.20 to 89.30 ± 6.40 at the latest follow-up, showing statistically significant difference (P < 0.05).

CONCLUSIONMPFL reconstruction improves clinical symptoms. Anatomical MPFL reconstruction is effective for patellar dislocation, and it offers good recovery of the premorbid patella mechanics. The interference screw provides firm fixation. Allograft can avoid the graft harvest site morbidity, but it increases the cost of the surgery.

Adolescent ; Adult ; Allografts ; Child ; Female ; Humans ; Ligaments, Articular ; surgery ; Male ; Patellar Dislocation ; surgery ; Patellofemoral Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Tendons ; transplantation

OBJECTIVETo evaluate the integrity of the resected mesentery specimen after total mesorectal excision (TME) for low rectal cancer using methylene blue perfusion via the superior rectal artery.

METHODSTwenty patients with low rectal cancer were randomly divided into the methylene blue group (n=10) and the control group (n=10). All the patients received TME and macroscopic examination of the mesorectal surface was performed to evaluate the quality of the surgical specimen. The methylene blue was injected into the specimen postoperatively via superior rectal artery.

RESULTSThe mesorectal surface of all the specimens was intact on macroscopic examination. However, after methylene blue perfusion, 2 specimens were found to be incomplete. The number of lymph nodes in the methylene blue group were significantly larger (17.3+/-2.4 vs 12.4+/-5.4, P=0.016).

CONCLUSIONSIntegrity evaluation of TME specimen is necessary. Methylene blue perfusion is a convenient and effective method to identify subtle incompleteness of specimen and can improve the detection of lymph node.

Adult ; Aged ; Digestive System Surgical Procedures ; Female ; Humans ; Infusions, Intra-Arterial ; Male ; Mesenteric Artery, Inferior ; Mesentery ; pathology ; surgery ; Methylene Blue ; Middle Aged ; Postoperative Period ; Prognosis ; Rectal Neoplasms ; surgery ; Rectum ; blood supply

OBJECTIVETo investigate the clinical characteristics and prognosis of second primary tumor of tongue (SPTr) after nasopharyngeal carcinoma (NPCR) treated with radiotherapy.

METHODSClinical data of 53 patients with SP7T after NPCR (group A) and 252 patients with primary tongue carcinoma (group B) were analyzed retrospectively with regard to clinical characteristics and survival rate (Kaplan-Meier); and multivariate analysis was performed using Cox proportional hazards model.

RESULTSThere was no significant difference between group A and group B ( P > 0. 05) in the presenting age, sex, tumor size, cTNM stage, tumor differentiation and the rate of distant metastasis. The overall 5-year survival rates were 41.6% in group A and 56.3% in group B (chi2 = 4.40, P = 0.0359) with a statistically significant difference between two groups. The differences of tumor location (chi2 = 61.18, P = 0.000) and rate of clinical (cN+, chi2 = 6.846, P = 0.009) or pathological lymph node metastasis (pN+, X2 = 3.993, P = 0.046) were also statistically significant between group A and group B, respectively. Multivariate analysis showed that age at presence, cTNM stage and with or without neck lymph node dissection were independent risk factors affecting survival.

CONCLUSIONSecond primary tongue carcinoma after radiotherapy for nasopharyngeal carcinoma is likely to occur on the dorsal aspect of the tongue with worse prognosis but with a lower rate of lymph node metastasis than that of primary tongue carcinoma. However, radiotherapy history is not an independent influencing factor on prognosis. Surgical resection or combined modality therapy may give a better prognosis.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; statistics & numerical data ; Female ; Follow-Up Studies ; Glossectomy ; methods ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Nasopharyngeal Neoplasms ; radiotherapy ; Neck Dissection ; Neoplasms, Radiation-Induced ; etiology ; pathology ; therapy ; Neoplasms, Second Primary ; etiology ; pathology ; therapy ; Prognosis ; Proportional Hazards Models ; Radiotherapy ; adverse effects ; Retrospective Studies ; Tongue Neoplasms ; etiology ; pathology ; therapy

OBJECTIVETo establish a mouse model of humoral immune response by immunization with rabbit red blood cells (RRBCs).

METHODSThe mice were immunized with RRBCs and the serum hemolysin level was measured by micro-hemolysis spectrophotometry.

RESULTSThe peak time needed for hemolysin production against RRBCs was 6 days after the immunization, and 20% RRBCs in a total volume of 0.2 ml was optimal for intraperitoneal injection. Hydrocortisone (25 mg/kg) and cyclophosphamide (20 mg/kg) inhibited hemolysin production. Mannatide (4 mg/kg) produced no significant effect on serum hemolysin level in normal mice, but significantly potentiated hemolysin production in immunosuppressed mice induced by cyclophosphamide (20 mg/kg).

CONCLUSIONIntraperitoneal RRBC injection is feasible for establishing mouse models of humoral immune response.

Animals ; Erythrocytes ; immunology ; Female ; Guinea Pigs ; Hemolysin Proteins ; blood ; Immunity, Humoral ; Immunization ; Male ; Mice ; immunology ; Models, Animal ; Rabbits