Objective:To study the chemical constituents of Buddleja lindleyana.Method:Separation by chromatographic methods and identification by spectral analysis.Result:Seven compounds vanillic acid,octacosanoic acid，β-sitosterol-3-O-β-D-glucopyranoside,stigmasterol-3-O-β-D-glucopyranoside,α-spinasterol-3-O-β-D-glucopyranoside,betulin acid were isolated.Conclusion:All the compounds were obtained from this plant for the first time.

Objective To investigate expression of Human papillomavirus (HPV) 11 type E7 protein antigen in prokaryotic cells and its potential use for the serodiagnosis of condyloma acuminatum (CA).Methods The full-length gene encoding for HPV11 E7 protein was amplified by PCR,and cloned into vector pET32a(+) to form recombinant pET32a(+)/HPVll E7 plasmid.The fusion His-E7 protein was expressed and analyzed by using SDS-PAGE and Western blotting.Using ELISA assay,HPV11 E7 fusion protein were also used to screen human serum IgG antibody from 93 patients with CA,43 patients with cervix cancer and 58 healthy control subjects.Results Highly expressed fusion His-E7 protein was obtained,and purified protein served as a special diagnostic antigen to screen human serum antibody for CA serodiagnosis.It showed that CA group,cervix cancer group and healthy control human serum IgG antibody average value were 1.545?0.131,0.586?0.155 and 0.674?0.150 respectively,positive rate were 76.3% (71/93),11.6% (5/43) and 5.2% (3/58).There was significantly difference between the CA group to compare cervix cancer group and healthy control (P

OBJECTIVETo induce the differentiation of K562/MDR1 cells into dendritic cells (DC) with multidrug resistance property.

METHODSK562/MDR1 cells and K562 cells were cultured in the presence of GM-CSF and IL-4 to generate DC and matured by TNF-α. On d14 K562/MDR1-DC and K562-DC cells were harvested and the expressions of CD1a, CD83, CD80, CD86, HLA-ABC and HLA-DR were assessed by flow cytometry (FCM). The antigen presentation function of K562/MDR1-DC and K562-DC was determined by allogenic mixed lymphocyte reaction (Allo-MLR). The expression of P-glycoprotein and the intracellular accumulation of daunorubicin (DNR) were detected by FCM. The sensitivity of K562/MDR1-DC and K562-DC cell to vincristine, adriamycin was measured using MTT assay.

RESULTSBoth K562/MDR1 and K562 cells were differentiated into dendritic cells in the presence of cytokine cocktails, showing the morphologic and immunophenotypic characteristics of DC. K562/MDR1-DC more markedly enhanced proliferation of allogeneic lymphocytes in MLR than K562-DC. High level expression of P-glycoprotein and efflux of DNR were demonstrated in K562/MDR1-DC. K562/MDR1-DC showed multidrug resistance property, with higher IC(50) to VCR and ADM than that of K562-DCs.

CONCLUSIONK562/MDR1 cells can be differentiated into DC with the presence of cytokines, the induced K562/MDR1-DC cells express high level of P-glycoprotein and acquire the multidrug resistance property.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Cell Differentiation ; drug effects ; Dendritic Cells ; cytology ; Drug Resistance, Multiple ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; K562 Cells ; cytology ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo observe the functional changes of dendritic cells (DCs) after infection by recombinant retrovirus carrying human telomerase reverse transcriptase (hTERT) gene fragment.

METHODSInterleukin-12 (IL-12) levels in DC culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The abilities of DCs infected with recombinant retrovirus carrying hTERT gene (hTERT-DCs) and non-infected DCs (N-DCs) to stimulate allogeneic lymphocyte proliferation were evaluated with mixed leukocytes reaction (MLR), and the surface markers of DCs including CD80, CD83, CD86 and HLA-DR were detected by flow cytometry. Cytotoxic T lymphocyte (CTL) assay was performed with CytoTox 96 non-radioactive cytoxicity assay.

RESULTSCompared with N-DCs, hTERT-DCs showed no significant changes in IL-12 secretion and capacity to stimulate allogeneic lymphocytes reaction, but had significantly lower CD83 expression. Specific CTLs induced by hTERT-DCs resulted in higher cytotoxicity against telomerase-positive target cells than that against the negative target cells.

CONCLUSIONInfection with the recombinant retrovirus carrying hTERT fragment may jeopardize the maturation of DCs, which, however, still retain their capacity to activate and stimulate lymphocyte proliferation and to prime autologous T lymphocytes to generate specific CTL against hTERT.

Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; virology ; Genetic Vectors ; Humans ; Interleukin-12 ; biosynthesis ; Recombination, Genetic ; Retroviridae ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Telomerase ; biosynthesis ; genetics

OBJECTIVE: To investigate the effect of ziprasidone and olanzapine on glucose and lipid metabolism in first-episode schizophrenia. METHODS: A total of 260 schizophrenics were assigned randomly to receive ziprasidone or olanzapine for 6 weeks. The weight was measured at baseline, week 2, 4 and 6. Fasting blood glucose (FBS), fasting insulin, high-density lipoprotein (HDL), total-cholesterol (TC) and triglycerides (TG) were measured at baseline and the end of 6-week treatment. Low-density lipoprotein (LDL) was measured in some patients at baseline and the end of 6-week treatment. Body mass index (BMI) and insulin resistance index (IRI) were counted. RESULTS: A total of 245 patients completed the trial, including 121 ziprasidone patients and 124 olanzapine patients. The average dose was 137.5 mg/d for ziprasidone and 19.5 mg/d for olanzapine. Patients treated with olanzapine had higher weight gain than those treated with ziprasidone [(4.55±3.37) kg vs (-0.83±2.05) kg, P<0.001]. After the treatment, FBS, fasting insulin, HDL, TC, TG, LDL and IRI levels were significantly increased in the olanzapine group (all P values<0.001 ). However, in the ziprasidone group, FBS decreased significantly and HDL and TG levels increased significantly after the 6-week treatment (all P values<0.05). The mean changes of FBS, fasting insulin, TC, TG, LDL and IRI were significantly different in the two groups (all P values<0.001). CONCLUSION Ziprasidone has less glucose and lipid metabolic effect for first-episode schizophrenia patients in short-term treatment. However, olanzapine induces weight gain and dysfunction of glucose and lipid metabolism significantly, which is associated with increased risk of complications. When the doctors choose antipsychotics in the clinic, they should consider the side effects of the medication.
Adolescent ; Adult ; Benzodiazepines ; adverse effects ; therapeutic use ; Blood Glucose ; drug effects ; Female ; Humans ; Lipid Metabolism ; drug effects ; Male ; Middle Aged ; Olanzapine ; Piperazines ; adverse effects ; therapeutic use ; Schizophrenia ; drug therapy ; Thiazoles ; adverse effects ; therapeutic use ; Young Adult

OBJECTIVETo review the outcome of repeated percutaneous sperm aspiration (PESA) and testicular sperm extraction (TESE) for intracytoplasmic sperm injection (ICSI).

METHODSForty-three cycles of 31 cases of azoospermic patients which underwent at least two PESA or TESE for ICSI from January 2001 to December 2002 were collected. The sperm retrieval, fertilization, implantation and clinical pregnancy were analyzed.

RESULTSTwenty-four cases underwent PESA and 7 cases underwent TESE. There were not any complications in these patients. Compared with the first cycle of 154 cases, the fertilization rate, implantation rate and clinical pregnancy rate were 78.39% vs 73.64%, 19.68% vs 18.38% and 34.88% vs 37.91%, respectively(P > 0.05).

CONCLUSIONSRepeated PESA or TESE is safe and well tolerated in azoospermic patients. Compared with the first cycle, the differences of repeated PESA or TESE cycles in fertilization rate, implantation rate and clinical pregnancy rate were not statistically significant.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Tissue and Organ Harvesting ; methods

OBJECTIVESTo review the retrospective treatment results of the azoospermia patients during January 2001 to January 2002 in the fertility center.

METHODSOne hundred males attempted intracytoplasmic sperm injection (ICSI) cycle for treatment of azoospermia. All patients were undergone sperm retrieval by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) while their wives received conventional ovarian hyperstimulation. The hormone levels, testicular histology, the rates of sperm retrieval, fertilization, implantation and pregnancy were analysed and evaluated.

RESULTSSperm were retrieved by PESA in 76 of 100 (76%) and by TESE in 23 of 100 (23%) men of azoospermia. The fertilization rate, implantation rate and clinical pregnancy rate were 71.3%, 20.35% and 42.11% respectively in PESA group, and 75.18%, 22.05% and 41.60% respectively in TESA group. Thirty-two clinical pregnancies were achieved with 15 ongoing pregnancies and subsequent live delivery for 15 cases in PESA group, and 2 cases of miscarriage, while 10 clinical pregnancies were achieved with 6 ongoing pregnancies, 2 cases of live delivery and 2 cases of miscarriage in TESA group. One case failed to retrieve sperm by TESE and canceled.

CONCLUSIONSHormonal levels and testicular histology are unable to predict which men with azoospermia will have sperm retrieved by PESA and TESE. PESA and TESE with ICSI are effective methods to treat azoospermia. There were no significant differences in fertilization, implantation and clinical pregnancy rate between two groups.

Adult ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Oligospermia ; blood ; therapy ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods

OBJECTIVETo explore the changes of CD(14)(+) monocyte human leucocyte antigen DR (HLA-DR) and their relationship with multiple organ dysfunction syndrome (MODS) in severe sepsis.

METHODSNinety-one patients with a definite diagnosis of severe sepsis in the intensive care unit (ICU) were included. CD(14)(+) monocyte HLA-DR levels were detected by flow cytometry on the first, 4th and 7th days of the study, and Marshall scores and prognosis on day 28 were evaluated.

RESULTSThirty-four patients died within 28 days following the onset with a mortality rate of 37.4%. Persistently lowered levels of HLA-DR were detected and significantly increased Marshall scores were found in the fatal cases at all the time points (P<0.001). In the surviving patients, the levels of HLA-DR were significantly increased (P<0.01) and Marshall scores were gradually decreased (P<0.001). During the observation period, the levels of HLA-DR decreased significantly as the number of dysfunctional organs and Marshall scores increased (P<0.001). The levels of HLA-DR were significantly increased in severe sepsis patients with 2-4 dysfunctional organs and Marshall score of 5-12 (P<0.05 or P<0.001). No changes in HLA-DR levels in severe sepsis patients with 5-6 dysfunctional organs and Marshall scores of 13-22. The levels of HLA-DR showed a significant inverse correlation to Marshall scores (r=-0.368, P<0.001).

CONCLUSIONIn patients with severe sepsis, persistent low CD(14)(+) monocyte HLA-DR levels predicts high mortality. The levels of HLA-DR are significantly correlated to the severity of organ dysfunction.

Adult ; Aged ; Aged, 80 and over ; Female ; HLA-DR Antigens ; metabolism ; Humans ; Lipopolysaccharide Receptors ; Male ; Middle Aged ; Monocytes ; immunology ; metabolism ; Multiple Organ Failure ; pathology ; Sepsis ; immunology ; metabolism

OBJECTIVETo study the survival rate and secretory function of islet cells in rats under condition of three-dimensional microgravity.

METHODSIsolated islet cells were assigned to flask-cultured or bioreactor-cultured. Survival rate of islets cultured for days 3, 7, 14 in stationary flasks or microgravity bioreactors was measured by AO-PI double-staining. Cultured islets were identified by dithizone (DTZ) staining, and insulin contents of different culture liquids were measured by radioimmunoassay.

RESULTSPancreatic islets stained nacarat with DTZ were easily visualized. When islet cells were cultured for 7 days and 14 days, survival rate of bioreactor-cultured islets was (0.9000 +/- 0.0107)% and 0.8038% +/- 0.0092% and higher than flask-cultured islets (P < 0.01). Insulin level of bioreactor-cultured islets is (70.875 +/- 0.31) m micro /L on the cultured 7 days while flask-cultured islets is (41.246 +/- 0.35) m micro /L. There was statistically significant difference of insulin production between the two groups (P < 0.01). Bioreactor-cultured islet contents were higher than flask-cultured on the cultured 14, 21 and 30 days (P < 0.01).

CONCLUSIONSIslet cells survival rate and secretory function revealed that bioreactor-cultured islets functioned better compared with flask-cultured islets.

Animals ; Bioreactors ; Cell Culture Techniques ; methods ; Cell Survival ; Cells, Cultured ; Insulin ; secretion ; Islets of Langerhans ; cytology ; secretion ; Male ; Rats ; Rats, Wistar ; Weightlessness Simulation