Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; pathology ; Biopsy ; Biopsy, Fine-Needle ; Child ; Cytodiagnosis ; methods ; Diagnosis, Differential ; Female ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; pathology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Lymphoma, T-Cell ; pathology ; Male ; Middle Aged ; Pleural Effusion ; pathology ; Young Adult

To examine the feasibility of low-charge electrotherapy (LCE) in treating geriatric major depressive disorder (MDD) patients. Bi-temporal LCEs (approximately 25 mC) were performed with an electroconvulsive therapy (ECT) instrument three times per week. We used the Hamilton Depression Scale 17 (HAMD-17) and the Hamilton Anxiety Scale (HAMA) to assess the effects of LCE and the Mini-Mental State Examination (MMSE) to evaluate the cognitive function change before and after LCE. Six visits occurred at the baseline, after LCE sessions 3, 6, and 9, after the last session, and at the end of the one-month follow-up period. Four patients were enrolled in the study. Two patients completed all LCE sessions. Two patients withdrew during the trial, one due to the adverse event of uroschesis potentially caused by atropine and the other due to her own will. All four patients completed the follow-up sessions. The HAMD-17 and HAMA scores were reduced significantly at the last LCE session and the end of the follow-up period compared with the scores at the baseline. As measured by the MMSE, cognitive impairment showed no significant changes at the last LCE session and the end of the follow-up period compared with that at the baseline. In this case series, LCE showed potential as an alternative current-based treatment for treating geriatric MDD patients. Further research is needed to assess the efficiency and safety of LCE.
Anxiety ; Atropine ; Cognition ; Cognition Disorders ; Depression ; Depressive Disorder, Major ; Electric Stimulation Therapy ; Electroconvulsive Therapy ; Follow-Up Studies ; Humans

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

AIMTo study the histologic changes of the vas deferens following Nd: YAG laser irradiation.

METHODSIntravasal laser irradiation was given to (i) 52 segments of rabbit (laser dosage: 2 seconds at 40 W approximately 50 W) and 16 segments of human (3 seconds at 45 W approximately 55 W) vas deferens in vitro, (ii) 25 rabbit vasa (2 seconds approximately 2.5 seconds at 40 W approximately 45 W) in vivo and (iii) 2 human vasa (3 seconds at 55 W) in vivo. Segments of vasa were removed from the in vivo irradiated vasa deferentia 15 days approximately 180 days (rabbit) or 15 days (man) after the exposure. All vas segments were embedded in methacrylate resin. Serial sections (thickness 25 microm approximately 30 microm) were obtained and observed under a light microscope.

RESULTS(i) Laser-induced damage reached the muscularis layer in 27% and 94% of the rabbit and human vas segments in vitro, respectively. (ii) Fourteen of the 25 in vivo rabbit vasa were completely occluded by fibrous tissue and the longer the time interval after treatment, the more likely was the vas occluded. Those unoccluded vasa had either a normal histology or a mucosal damage. (iii) One in vivo human vas was almost completely occluded by the fibrous tissue but the other had a relatively large lumen packed with sperm granulomatous tissue and partial destruction of the smooth muscle layer.

CONCLUSIONLaser irradiation can induce long-term vas occlusion; for rapid occlusion, laser doses just completely destroying the mucosal layer will be advisable.

Animals ; Humans ; Laser Coagulation ; Male ; Rabbits ; Sterilization, Reproductive ; methods ; Vas Deferens ; anatomy & histology ; Vasectomy

OBJECTIVETo investigate the factors that affect the prognosis of primary gastrointestinal non-Hodgkin's lymphoma (PGI-NHL).

METHODSThe clinical data of 116 patients with pathologically confirmed PGI-NHL we treated from January 1993 to December 2003 were analyzed retrospectively. Kaplan-Meier survival analysis was used for analyzing the survival of the patients, and Log-rank test was performed to compare the survival rates in relation to different prognostic factors.

RESULTSThe 3-year and 5-year survival rates of the patients were 63.8% (74/116) and 48.2% (40/83), respectively. Univariate analysis revealed that the factors affecting the prognosis of the patients included the presence of B symptom, tumor size, clinical stage, pathological type, depth of invasion, and treatment methods. The patients with B symptom, tumor size no less than 10 cm, advanced clinical stage (stages III(E) and IV(E)), T-cell type, and invasion beyond the serosa who received only surgical management had poorer prognosis than those free of B symptom with tumor size <10 cm, early clinical stage (stages I(E) and II(E)), B-cell type, and submucosal or serosal invasion managed with chemotherapy alone or in combination with surgery. Multivariate analysis showed that B symptom, tumor size no less than 10 cm, advanced clinical stage (stages III(E) and IV(E)), T-cell type, invasion beyond the serosa, and surgery alone were independently associated with poor prognosis.

CONCLUSIONThe tumor size, clinical stage, pathological type, treatment methods are the independent factors affecting the prognosis of patients with PGI-NHL.

Adolescent ; Adult ; Aged ; Child ; Female ; Gastrointestinal Neoplasms ; diagnosis ; mortality ; pathology ; Humans ; Kaplan-Meier Estimate ; Lymphoma, Non-Hodgkin ; diagnosis ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Young Adult

To explore the mechanisms of suppression growth and induction apoptosis of curcumin on human multiple myeloma cell line RPMI8226, the suppressive effect of curcumin on RPMI8226 was examined by MTT assay; the induction apoptosis and cell cycle arrest of curcumin on RPMI8226 were determined by flow cytometry (FCM); the changes of survivin, Bcl-2, Bax mRNA levels were detected by RT-PCR. The results showed that curcumin obviously suppressed the proliferation of RPMI8226 in both time- and dose-dependent manners, and the IC(50) were 12.15 micromol/L, 4.9 micromol/L for 24 and 48 hours respectively. FCM indicated that the apoptosis ratio rose from 10.6% of untreated cells up to 36.9% of treated cells (p < 0.05), and curcumin arrested cell cycle of RPMI8226 at G(2)/M phase. RT-PCR showed that RPMI8226 cells expressed survivin, Bcl-2 strongly and Bax slightly; while RPMI8226 cells were treated with curcumin 10 micromol/L for 24 hours, the expressions of survivin, Bcl-2 mRNA were apparently down-regulated, and the expression of Bax mRNA was markedly up-regulated. It is concluded that curcumin can suppress the proliferation of human multiple myeloma cell line RPMI8226, and induce their apoptosis. The mechanism of antitumous effect of curcumin may be related to down-regulation of survivin, Bcl-2 mRNA and up-regulation of Bax mRNA.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Curcumin ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism

OBJECTIVETo assess the therapeutic efficacy and adverse effects of endogenetic field hyperthermia (EFH) in combination with L-OHP /LV / 5-FU in the treatment of advanced gastric cancer.

METHODSThis study included 147 surgical patients with stage II-IV gastric cancer, who received postoperative chemotherapy with FOLFOX (L-OHP 85 mg /m square, 3 h intravenous infusion, followed by infusion of LV at 200 mg /m square in 2 h, intravenous injection of 5-Fu at 400 mg /m square, and intravenous infusion of 5-FU at 3000 mg /m square in 48 h). Eight treatment cycles (each lasting for 14 days) were administered. In 68 cases randomly selected from the cohort, EFH was performed on the first and third days (treatment group), but not in the other 79 cases (control group).

RESULTSThe response rate was 68.4% in the treatment group and 36.4% in the control group, showing significant difference between them (P<0.05). The 1-year survival rate was 88.2% in the treatment group, similar to the rate of 81.0% in the control group (P< 0.05), but the 3, 5-year survival rates in treatment group (67.6% and 30.9%) was significantly higher than those in the control group (47.6% and 15.4%, P<0.05). The adverse effects were similar between the two groups.

CONCLUSIONEFH combined with the chemotherapeutic regimen FOLFOX might improve the therapeutic effect of stage II-IV gastric cancer without obviously increasing the adverse effects.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Combined Modality Therapy ; Female ; Fluorouracil ; therapeutic use ; Humans ; Hyperthermia, Induced ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Organoplatinum Compounds ; therapeutic use ; Stomach Neoplasms ; drug therapy ; pathology ; surgery ; therapy ; Treatment Outcome

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Self-directed learning ( SDL) skill is one of the necessary capabilities of an excellent doctor .SDL train-ing has become the important content in the top creative talents training .SDL training has been carried out for 6 years since 2011 by physiology department of Tongji University .Feedback from medical students had been collect-ed, which was used to make modifications and supplementary including direct to learning ( D2L) platform to im-prove SDL.SDL training is proved to be a widely accepted model in medical students and receive good learning out-come.D2L platform is helpful in progressing SDL training .