BACKGROUND: The effect of Chinese traditional medicine 3'-meisoindigo as well as indimbin derivatives on normal immunocytes is less reported while it is used for antitumor.OBJECTIVE: To investigate the effect of 3'-meisoindigo on the proliferation of the splenocytes and thymocytes of C56BL/6 mouse.DESIGN, TIME AND SETTING: The randomized cytopathology observation was performed between August 2007 and January 2008 at Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong Province, China.MATERIALS: C57BL/6 mice, clean, male, 6-8 weeks old, weighing (20±2) g.METHODS: The thymus gland and the spleen of C57BL/6 mice were ground to get the single-cell suspension and cells were treated by 5, 10, 15, 20 and 25 μ mol/L 3'-meisoindigo,respectively. Cells without any drug treatment were used as blank control and cells treated by concanavalin A were used as positive control.MAIN OUTCOME MEASURES: Proliferation of splenocytes and thymocytes detected using MTT method; IL-12 activity in culture supematant detected using ELISA method; the cell cycle, apoptosis rate, cell death rate and intracellular reactive oxygen species level detected using flow cytometry; the mRNA level of bcl-2 and cdk2 detected using reverse transcription-polymerase chain reaction; the expression rate of Bcl-2, CDK2 and Bax detected using fluorescence microscope.RESULTS: After treating for 24 hours, 15, 20 and 25 μ mol/L 3'-meisoindigo can significantly inhibit the proliferation of thymocytes and splenocytes (P < 0.05) and the inhibition was dose-dependent and time-dependent. Cells resumed proliferation after removing the 3'-meisoindigo, although they had been treated by 25 μ mol/L 3'-meisoindigo for 72 hours. The secretion of IL-12 was markedly reduced in all 3'-meisoindigo groups versus control groups at each time point (P < 0.05). 15 μ mol/L 3'-meisoindigo could decrease the mRNA expression level of apoptosis-related protein bcl-2 and cyclin cdk2 gene, decrease the expression level of BCL-2 protein and CDK protein, increase Bax expression level, decrease Bcl-2/Bax ratio markedly and started apoptosis.15 μ mol/L 3'-meisoindigo arrested cells in the G2/M stage of the thymocytes and splenocytes, and intracellular reactive oxygen species level elevated dose-dependently and time-dependently.CONCLUSION: In certain concentration range, 3'-meisoindigo can reversibly inhibit the proliferation of thymocytes and splenocytes of C57BL/6 mouse and can inhibit IL-12 secretion in parallel, and start apoptosis.

ObjectiveTo study the cardioprotective effects of ischemic post-conditioning on elderly patients with ST-elevation acute myocardial infarction (STEM1).MethodsConsecutive 215 patients with STEMI undergoing emergency percutaneous coronary intervention(PCI) were randomly assignedto receive ischemicpost-conditioningorconventional PCItreatment.The ischemic postconditioning (n=38) were conducted by 3 episodes of 30-second occlusion followed by 30-second reperfusion, while the control group (n= 46) was without any intervention after PCI.Reperfusion arrhythmias, corrected TIMI frame count (cTFC) and TIMI myocardial perfusion grade (TMPG)were compared between the two groups, respectively.Results The incidence of reperfusion arrythmias was less frequent in ischemic postconditioning group (21.1％ ,8/38) than in control group (45.7％ ,21/46) after PCI (x2 = 5.571, P＜0.05). The TIMI grade 3 flow was similar between two groups [(94.7％(36/38) vs. 82.6％( 38/46), x2= 2.919, P＞0.05], the cTFC levels (23.6±3.7vs. 26.1 ±5.9) and TMPG 3 perfusion [ 89.5％ (34/38) vs.69.6％ (32/46)] were significantly different (t= 5.434, P＜0.05; x2 = 4.899, P＜0.05, respectively) between two groups.ConclusionsIschemic postconditioning may reduce myocardial reperfusion injury in elderly patients with STEMI undergoing emergent PCI.

Objective To investigate the value of modified pancreatoduodenectomy in the treatment of ampullary carcinoma by comparing the efficacy of pancreatoduodenectomy with preservation of ascending portion of duodenum and that of Whipple procedure. Methods The clinical data of 133 patients with ampullary carcinoma who had been admitted to the Qilu Hospital from 1990 to 2006 were retrospectively analyzed. Of the 133 patients, 88 received Whipple procedure (group A) and 45 received pancreatoduedenectomy with preservation of ascending portion of duodenum (group B). The preoperative condition of the patients and short- and long-term efficacy of the 2 procedures were compared by Fisher exact probability, t test, chi-square test, Kaplan-Meier survival curve and Log-rank test, and the differences between the efficacy of the 2 procedures were compared. Results The operation time, time of gastrointestinal function recovery, hospital stay and ratio of patients with blood transfusion were (366±111) minutes, (7.4±1.4) days, (24±9) days and 76% (67/88) in group A, and (325±32)minutes, (4.3±1.4)days, (31±14)days, 42% (19/45) in group B, with significant difference between the 2 groups (t = 2.34, 2.08, 1.98 ; χ2 = 14.99, P < 0.05). The medium survival time of patients in groups A and B were (19.0±1.5)months and (16.9±1.7) months, with no significant difference between the 2 groups (χ2 = 0.46, P > 0.05). There was no significant difference in the occurrence of postoperative complications between the 2 groups (P > 0.05). Conclusions Pancreateduodenectomy with preservation of ascending portion of duodenum can be applied for patients with ampullary carcinoma.

Objective To explore the role of integrin αvβ6 in mediating the tolerance of gastric cancer AGS cells to 5-fluorouracil, and to determine whether direct β6-extracellular signal-regulated protein kinase(ERK) binding is involved. Methods Gastric cancer AGS cells in the logarithmic phase were incubated with either 5-fluorouracil for 24 hours ( control group), with 0.1 g/L of mouse anti-αvβ6 monoclonal antibody 10D5 for 6 hours and then with 5-fluorouracil for24 hours ( 10D5 group), with IgG2a and 5-fluorouracil ( IgG2a group), or with 5-fluorouracil and 20 μnol/L of ERK inhibitor PD98059 for 24 hours ( PD98059 group). Cell proliferation,apoptosis and the expression of Bcl-2 and caspase-3 protein were detected by MTT assay, flow cytometry and western blotting, respectively. All data were analysed by one-way analysis of variance and LSD test. Results The cell inhibition rates of the control group, 10D5 group, IgG2a group and PD98059 group were 28.1% ±2.7%,84.5% ± 1.6%, 31.4% ±5.2%, 86.7% ±5.2%, respectively, with a significant difference ( F = 342. 5, P ＜0.05). The apoptosis rates of the control group, 10D5 group, IgG2a group, and PD98059 group were 6.6% ±1.4%, 30.6% ± 2.4%, 8.1% ± 1.3%, 36.0% ±4.0%, respectively, with a significant difference among the four groups (F = 105.4, P ＜0.05 ). There was a significant difference in the expression of caspase-3 and Bcl-2 among the four groups (F=292.1, 181.6, P＜0.05). Conclusion Integrin αvβ6 can mediate the tolerance of gastric cancer AGS cells to 5-fluorouracil through direct β6-ERK binding.

Objective To study the effect of pelvic autonomic nerve preservation(PANP)on urination and sexual function in total mesorectal excision(TME). Methods Two hundred and forty cases of male rectal cancer patients,divided into the PANP who accept the pelvic autonomic nerve preservation in TME,and the control group of 120 patients who do not.The urination and sexual function were observed and compared.3-year-survival rate,local recurrence rates of the two groups were recorded. Results The urinary disorder rates,erective disorder rates and ejaculation disorder rates of PANP group were 30.8%,28.3%and 34.2%,while values of control group were 55.0%、60.0%and 62.5%.The difference between them had statistical significance(P<0.05).The 3-year-survival rate and local recurrence rate of PANP group were 9.4%and 75.0%.The 3-year-survival rate and local recurrence rate of control group were 9.0%and 65.0%.There was no significant difference between them(P>0.05). Conclusion The PANP technique in TME could improve the urinary and sexual function of male patients without affect the prognosis.

Objective To study the expression and significance of EGFR, PDCD4, TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and oral squamous cell carcinomas(OSCC).Methods Serum levels of EGFR, PDCD4 were measeured with ELISA in Chinese hamster during the oral mucosa carcinogenesis;The expressions of TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and OSCC tissues were examined by immunohistochemistry.Results In the process of oral carcinogenesis,the expression level of EGFR increased significantly,while the expression of PDCD4 was decreased, the negatively correlation was evident between expression of these two proteins in ED and SCC.The expression of TGF-β1, Smad7 was higher in OSCC than in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia(P < 0.05), while the expression of Smad3 was decreased.Further analysis showed the expression of TGF-β1 was correlated with the expression of Smad7.Conclusions The expressions of EGFR, PDCD4, TGF-β1, Smad3, Smad7 are closely related with the biological behaviors of oral squamous cell cancinoma.This experiment provides a theoretical basis for the study of oral squamous cell carcinomas mechanism research.So it has important significance for the development and prognosis of oral squamous carcinomaare.

Chinese hamster is an important laboratory animal in medical and biological researches,but the molecular genetic markers research was rarely reported.In our study the base composition,gene structure,genetic evolution and other characteristics of mitochondrial genome of Chinese hamster were analyzed using the methods of bioinformatics and comparative genomics,genetic quality detection system of Chinese hamster were also established.These results would supply genome data for animal models of human diseases,and lay the foundation for scientific evaluation and reasonable utilization.

OBJECTIVETo observe the influence of decreasing conditioning regimen intensity on the engraftment of HLA haplotype peripheral blood stem cell transplantation.

METHODTwelve patients with leukemia, including 4 in complete remission, whose HLAs were full matched with donors, and 8 with refractory leukemia, whose HLAs were mismatched, were transplanted with G-CSF mobilized allogeneic peripheral blood stem cells after conditioned with a regimen consisting of fludarabine (30 mg/m(2) x 6 days), busulfan (4 mg/kg x 2 days) and cyclophosphamide (30 approximately 60 mg/kg x 2 days) (FBC). Donor lymphocytes were infused at day + 30, + 60 and + 90 after transplantation, respectively. Hematopoietic reconstitution was observed. Engraftment was documented by the analysis of short tandem repeats with polymerase chain reaction (STR-PCR).

RESULTPatients in HLA haplotype group received a mean number of 4.87 x 10(8)/kg donor mononuclear cells (MNC), with CD(34)(+) cells of 4.58 x 10(6)/kg and patients in HLA identical group a mean number of 4.85 x 10(8)/kg MNC with CD(34)(+) cells of 4.47 x 10(6)/kg. The mean time of white blood cell count more than 1.0 x 10(9)/L was 14 (10 approximately 18) days in HLA matched patients and 29 (11 approximately 90) days in HLA haplotype group. One three locus mismatched patient failed to engraft, but auto-hematopoiesis was recovered on day + 50. Full donor chimerism was observed in all patients except one with mixed chimera. The mixed chimera was converted into full donor chimera after three times donor lymphocyte infusion. One each died from severe acute GVHD, severe VOD and severe chronic GVHD in HLA haplotype group, and one from chronic GVHD in HLA identical group.

CONCLUSIONPatients survived engraftment was not influenced by decreasing conditioning intensity as in this regimen. Haplotype stem cells could be engrafted durable in recipients by this regimen combined with donor lymphocyte infusion.

Adolescent ; Adult ; Busulfan ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Female ; Graft Survival ; drug effects ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Humans ; Immunosuppressive Agents ; therapeutic use ; Leukemia ; therapy ; Male ; Middle Aged ; Transplantation Conditioning ; Transplantation Tolerance ; drug effects ; Vidarabine ; analogs & derivatives ; therapeutic use

BACKGROUND: It is easy to established human solid tumor nude mouse model, but for leukemia which is difficult. We inhibited immune system further by radioactive ray or CTX, to decrease cost and increase the stability.OBJECTIVE: To establish a human acute myeloblastic leukemia M2 Kasumi-1 models containing AML/ETO positive genes in BALB/c nude mouse. METHODS: Nude mice were randomly divided into three groups: CTX group was injected CTX 2 mg/day in abdominal cavity for two days, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein next day; irradiation group was exposed to total body irradiation, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein that day; untreated group was inoculated with 8×10~5/mouse Kasumi-1 cells by caudal vein injection. Three additional mice were considered as the normal control group. The blood smearing and bone morrow slides were detected, immunity type of BMC was detected using flow cytometry, loading of leukemic cellular tumor was detected using RT-PCR, and positive ratio of AML/ETO fusion gene was detected using FISH method. RESULTS AND CONCLUSION: After inoculated into untreated nude mice by caudal vein injection for 14 days, the ratio of leukemia cell in blood smearing was 3.5%, and over 40% in bone marrow slides, which was equal to the results of FISH and FCM. The increasing of tumor loading was time-dependent. For irradiation group and CTX treated group, the tumor loading was higher that untreated group, and the cells also survived more than 60 days. AML/ETO band was observed by RT-PCR in all experimental groups, for normal mice it was negative. The results indicated that the systemic disseminated leukemia model was established successfully by caudal vein injection 8×10~5/mouse Kasumi-1 cells in the three experimental groups.

Objective To explore the curative effect of tirofiban treatment on high-risk acute coronary syndromes (ACS) in elderly patients receiving an early percutaneous coronary intervention (PCI) treatment. Methods The 162 elderly cases including unstable angina pectoris and non-ST -segment elevation myocardial infarction (NSTEMI) undergoing early PCI were enrolled in this study.And they were assigned to early treatment group (n=82) and deferred selective group (n=80)according to the time of using tirofiban (Gp Ⅱ b/Ⅲ a inhibitor) treatment. The effectiveness of either strategic option on tissue-level perfusion was evaluated using the TIMI myocardial perfusion grade (TMPG) before and immediately after PCI. The corrected TIMI frame count (cTFC) was also used to assess coronary artery flow and myocardial perfusion. Bleeding complications and the composite end point events at 30 days were also evaluated. Results Of all the 162 patients, the TMPG 0-1 perfusion was observed in 65 patients (40.1%). The TMPG 0-1 perfusion was significantly less frequent in early treatment group (32.9%) than in deferred selective group (47.5%) before PCI (x2=3.58, P＜0.05); while the results of TIMI grade 0-1 flow (26.8% vs. 25.0%) and cTFC levels (34.2±11.8 vs. 34. 9±12. 7) before PCI were similar between the two groups (x2 =0. 07, P=0.47; t= 0.13, P=0.71, respectively). No differences were seen both in composite end point events at 30 days and bleeding complications (x2 = 0.31, P＞0.05; x2=0.004, P＞0. 05). Conclusions High -risk ACS patients treated with an early invasive strategy, routine upstream use of tirofiban are associated with improved tissue-level perfusion before PCI and does not increase bleeding complications when bleeding risks are carefully evaluated before enrollment.