Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P＜0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.

OBJECTIVETo explore the characteristics of cellular immunity in drug abusers with pulmonary tuberculosis.

METHODSSixty drug abusers with pulmonary tuberculosis and 60 non-drug abusers with pulmonary tuberculosis (control) were enrolled in this study. Three days after establishment of a definite diagnosis, peripheral blood was taken from the patients for lymphocyte subgroup (CD3(+), CD3(+)/CD4(+), CD3(+)/CD8(+) T lymphocyte subgroups and NK cells) examination by flow cytometry, and the CD4(+)/CD8+(+) ratio was calculated. The difference of cellular immunity between the drug abusers and control group was analyzed statistically.

RESULTSCD3(+) and CD3(+)/CD4(+) T lymphocytes subgroups and NK cells of the drug abusers were significantly lower than those of the control patients (P=0.037, 0.028 and 0.015), and the former patients had also significantly lower CD4(+)/CD8(+) ratio (P=0.021). The pulmonary tuberculosis types and CD3(+)/CD8(+) T lymphocyte subgroup were not significant different between the two groups (P=0.053 and 0.85).

CONCLUSIONDrug abuse might depress cellular immunity in patients with pulmonary tuberculosis, which further complicate the treatment of this disease.

Adolescent ; Adult ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Substance-Related Disorders ; complications ; immunology ; T-Lymphocytes ; immunology ; metabolism ; Tuberculosis, Pulmonary ; complications ; immunology ; prevention & control ; therapy ; Young Adult

Objective To investigate the effect of electroacupuncture on hypothalamic insulin receptor substrate 1 (IRS-1) in a rat model of type 2 diabetes mellitus (T2DM). Methods Sixty Wistar rats were randomized to a normal group (15 rats) and an observation group (45 rats). In the observation group, a rat model of T2DM was made by high-energy diet induction. After the model was successfully made 8 weeks later, the observation group was randomized to model making, treatment and blocker groups, 15 rats each. The treatment group received electroacupuncture and the blocker group, electroacupuncture plus intraventricular perfusion of phosphatidylinositol 3-hydroxyl kinase (PI3K) blocker. After 8 weeks of treatment, fasting plasma glucose (FPG) was measured using a glucometer, fasting insulin (Fins) was determined by ELISA, insulin resistance index (IRI) was calculated and IRS-1 expression was examined by SABC immunohistochemistry assay in every group of rats. Results FPG and Fins increased significantly (both P<0.01) and IRI and IRS-1 expression decreased significantly (P<0.01) in the model making group compared with the normal group. FPG and Fins decreased significantly (both P<0.01) and IRI and IRS-1 expression increased significantly (P<0.01) in the treatment group compared with the model making group. FPG and Fins decreased significantly (P<0.05, P<0.01) and IRI and IRS-1 expression increased significantly (P<0.01) in the treatment group compared with the blocker group. Conclusion Electroacupuncture can improve FPG, Fins and insulin sensitivity by regulating hypothalamic IRS-1 expression in T2DM rats.

OBJECTIVETo develop a colon-specific prodrug of Indomethacin microbially triggered, carry out in vitro/in vivo evaluation of drug release, and appraise its inhibitory effect on liver metastasis from colon cancer.

METHODSIndomethacin prodrugs were synthesized and characterized by FTIR and NMR, and dissolution test simulating gastrointestinal tract was employed to screen the colon-specific prodrug. Then, the pharmacokinetic profile of portal vein and peripheral blood in Sprague-Dawley rats was studied. Lastly, the inhibitory effect on liver metastasis from colon cancer in nude mice was observed.

RESULTSThe chemical structure characterized by FTIR and NMR demonstrated that six kinds of indomethacin-block-amylose with different drug loading (IDM-AM-1-6) were synthesized, among which IDM-AM-3 was degraded 1.3%, 9.3% and 95.3%, respectively, in simulated gastric fluid for 4 h, small intestine for 6 h, and colon for 36 h. The pharmacokinetic test of IDM-AM-3 showed that absorption was delayed significantly (P < 0.01), peak time [(11.35 + or - 2.45) h], elimination half-life [(16.74 + or - 4.04) h] and mean residence time [(22.27 + or - 0.52) h] were significantly prolonged (P < 0.01), as well as peak serum concentrations [(9.69 + or - 2.40) mg/L] and AUC(0-t) [(236.7 + or - 13.1) mg x L(-1) x h] were decreased markedly (P < 0.01) as compared with those of IDM regarding to portal vein. Additionally, its AUC(0-t) in peripheral blood was remarkably lower than that in Portal vein (P < 0.01). The tumor suppression observation showed that it could remarkably reduce the number of liver metastases in contrast to IDM (P < 0.05).

CONCLUSIONColon-specific IDM-AM-3 possesses advantage of sustained release in portal vein providing some experimental basis for colon-specific delivery system applied to sustained release in the portal vein.

Amylose ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Animals ; Colon ; metabolism ; Colonic Neoplasms ; pathology ; Delayed-Action Preparations ; Drug Delivery Systems ; HT29 Cells ; Humans ; Indomethacin ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Prodrugs ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the regulatory effect of IFN-gamma on recognition of target cells by human natural killer (NK) cells.

METHODSThe cytotoxic activity of human NK cell lines (NK92, NKL) was detected by MTT method. Expression of NK cell receptors (NKG2D, NKG2A/B, KIR2DL1 and KIR2DS1) and MICA on target cells (the ligand of NKG2D) was measured by RT-PCR.

RESULTSBoth NK92 and NKL cells exerted higher cytotoxicity to tumor cells with MICA expression, while tumors without MICA expression could resist NK cell lysis. IFN-gamma (> 1000 U/ml) inhibited NK lysis of tumor cells with MICA expression through down-regulating the expression of NKG2D, but up-regulating the expression of NKG2A/B and KIR2DL1.

CONCLUSIONIFN-gamma has a negative effect on activation and cytotoxicity of human NK cells by altering the balance between the expression of activating and inhibitory receptors on NK cells in favor of inhibition. This may serve to limit NK cell over-activation in vivo.

Cell Division ; drug effects ; Cytotoxicity, Immunologic ; drug effects ; Histocompatibility Antigens Class I ; analysis ; physiology ; Humans ; Interferon-gamma ; pharmacology ; Killer Cells, Natural ; immunology ; NK Cell Lectin-Like Receptor Subfamily C ; NK Cell Lectin-Like Receptor Subfamily K ; Receptors, Immunologic ; metabolism ; Receptors, KIR2DL1 ; Receptors, Natural Killer Cell ; Recombinant Proteins ; Tumor Cells, Cultured

OBJECTIVETo study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.

METHODSImmunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.

RESULTSThe expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).

CONCLUSIONSThe decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.

Cicatrix ; metabolism ; pathology ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Serpins ; metabolism

OBJECTIVETo evaluate and improve the diagnosis and surgical treatment of the retroperitoneal fibrosis (RPF).

METHODSThe medical records of 26 patients with the RPF (21 men and 5 women with mean age 54 years) were analyzed retrospectively. They were been treated from January 1996 to May 2007. Fourteen cases received double-J inter-ureter drainage or pricking pyelostomy and 9 of 15 cases who received open surgery were performed bilateral ureterolysis with their ureters translocated intraperitoneally.

RESULTSFor masses in retroperitoneal space, the diagnostic rate of B mode ultrasonography, CT and MRI was 12% (3/26), 86% (18/21) and 57% (8/14) respectively. The patients were followed up from 1 to 106 months. After drained by double-J inter-ureter stent or pricking pyelostomy, the mean serum creatinine level decreased from 373.9 micromol/L to 157.1 micromol/L of 14 patients. Those patients who underwent ureterolysis with ureteral intraperitoneal translocation had good results and their mean serum creatinine level decreased from 171.0 micromol/L before operation to 139.6 micromol/L after operation. Four patients had normal B-ultrasound and intravenous urogram findings with at least 24 months of follow-up.

CONCLUSIONSCT scan has better accuracy for diagnosis of the RPF than B mode ultrasonography and MRI. Prompt and appropriate relief of urinary obstruction with surgical intervention can effectively protect the renal function in patients with the RPF, and the ureterolysis with ureteral intraperitoneal translocation is an effective surgical procedure to treat this disease.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retroperitoneal Fibrosis ; diagnosis ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the ultrastructural variation of the facial nerve of rabbit with different dosage of (125)I seed brachytherapy.

METHODSFifty-four big ear rabbits were divided into 3 groups randomly and given 40 Gy, 80 Gy, 120 Gy respectively. Radioactive seeds were implanted in one side of parotid gland, the other side was implanted with vacant shell as a control group. The facial nerves were obtained 2, 4, 6 months respectively after operation and the histological ultrastructural changes observed by electromicroscope.

RESULTSIn the control group, epineurium was continuous, there was slight pitting edema under the epineurium, and axonal myelin was loose. In the test groups, there was slight pitting edema under the epineurium, and axonal myelin sheath was loose at 4th month. Macrophage and regenerated fibers were found in the 80 Gy group and myelin sheath lamellar separation, regeneration of nerve in the 120 Gy dosage. The myelin sheath lamellar was separated and axonal myelin loose in the test group at 6th month. Myelin sheath amellar separation and edema under the epineurium were found in the group of 80 Gy and 120 Gy.

CONCLUSIONSThe ultrastructure of the facial nerve is damaged by the dosage of 40 Gy, 80 Gy brachytherapy with (125)I seeds. The higher dosage the nerve receives, the more serious the damage will be. Both of the epineurium and axonal myelin sheath are integral and continuous 6 months after operation with dosage of 120 Gy.

Animals ; Brachytherapy ; Dose-Response Relationship, Radiation ; Facial Nerve ; radiation effects ; ultrastructure ; Female ; Iodine Radioisotopes ; administration & dosage ; radiation effects ; Male ; Rabbits ; Radiation Injuries, Experimental ; pathology ; Random Allocation

Objective To investigate the chemotherapeutic sensitivity of glioma cells influenced by RNA interference of pituitary tumor transforming gene (PTTG). Methods The glioma U251 cell line was divided into normal control group, TMZ treatment group, PTTG shRNA infection group and PTTG shRNA combined with TMZ treatment group. Cells in the later 2 groups were treated with PTTG shRNA and PTTG shRNA combined with TMZ. MTT assay and flow cytometry were employed to detect the cell proliferation and apoptosis of U251 cell line, respectively. Results The outcome of MTT assay showed that the growth and survive abilities were influenced after treating with PTTG shRNA and PTTG shRNA combined with TMZ; the optical density (OD) value of the control group, TMZ group,PTTG shRNA treatment group and PTTG shRNA combined with TMZ treatment group were (0.85±0.07),(0.58±0.06), (0.55±0.07) and (0.41 ±0.05), respectively. The inhibitory rate of cell proliferation in the TMZ treatment group, PTTG shRNA treatment group and PTTG shRNA combined with TMZ treatment group were (31.56±5.51 )%, (35.53±4.60)%, (51.49±6.74)%, respectively; statistical significance between control group and both PTTG shRNA group and TMZ group was noted (P＜0.05); and statistical significance between group PTTG shRNA combined with TMZ and both PTTG shRNA group and TMZ group was noted too. The apoptosis rate in the control group, TMZ group, PTTG shRNA group and PTTG shRNA combined with TMZ group were (6.29±0.78)%, (33.63±4.88)%, (39.61 ±4.95)% and (66.23±7.60)%, respectively, 48 h after the treatment; significant differences were found between the control group and both PTTG shRNA group and TMZ group (P＜0.05); and statistical increased apoptosis rate in the PTTG shRNA combined with TMZ group was noted as compared that in the PTTG shRNA group and TMZ group (P＜0.05). Conclusion PTTG RNA interference, by inhibiting the cell proliferation and inducing the apoptosis of glioma cells, up-regulates the chemotherapeutic sensitivity and improve the effectiveness of chemotherapy.

OBJECTIVETo conduct molecular and prenatal diagnosis for a couple with β thalassemia.

METHODSBlood routine examination and hemoglobin analysis were used for screening of thalassemia. Seventeen common Chinese mutations of β thalassemia were detected for the carriers with β thalassemia using PCR/RDB. The unknown mutation of β thalassemia was identified by DNA sequencing and DHPLC analysis.

RESULTSThe husband was heterozygote of CD41/42 (-TCTT). The wife carried a mutation IVS-I-110 (G→A) of β thalassemia having not been reported in Chinese so far. The fetus was a double mutated heterozygote of IVS-I-110 (G→A) and CD41/42 (-TCTT). The pregnancy was terminated.

CONCLUSIONMutation IVS-I-110 (G→A) of β thalassemia in Chinese is of importance to the genetic counseling and prenatal diagnosis of thalassemia.

Base Sequence ; DNA Mutational Analysis ; Female ; Humans ; Male ; Mutation ; Pregnancy ; Pregnancy Complications, Hematologic ; genetics ; Prenatal Diagnosis ; beta-Thalassemia ; diagnosis ; genetics