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.

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 clinicopathological features and prognostic significance of human epidermal growth factor receptor 2 (HER2) in gastric carcinoma. Methods Pathological data of 127 patients with gastric carcinoma were retrospectively analyzed. HER2 expressions of all patients were detected by immunohistochemistry (IHC). 119 (93.7 %) patients were undergone R0 dissection and 123 (96.9 %) cases received D2 lymph nodes dissection. 51 (40.2 %) patients received adjuvant chemotherapy. The proportional differences of clinicopathological features for patients between HER2-positive and HER2-negative were compared, including the patients' survival. Results HER2 overexpression rate was 8.7 % (11/127) in gastric carcinoma patients. For the patients with HER2-positive and HER-negative, the lymph node metastasis rates were 100.0 % (11/11) and 81.9 % (95/116), respectively (P= 0.041). The 3-year overall survival (OS) rates for HER2-positive and HER2- negative patients with gastric carcinoma were 32.7 % and 42.9 % (P=0.413), and the 5-year progression free survival (PFS) rates were 27.3 % and 42.2 % (P = 0.354), respectively. Among patients with HER2-negative, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 55.3%, compared with 35.4%for patients with surgery alone (P=0.015), and the 3-year PFS rates were 53.3 % and 35.3 % (P= 0.038), respectively. Among patients with HER2-positive patients, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 0, compared with 75.0%for patients with surgery alone (P=0.002), and the 3-year PFS rates were 0 and 60.0% (P=0.004). Conclusions HER2 is expressed in gastric carcinoma tissue, related to lymph node metastasis. HER2 status are not correlated with the prognosis for gastric carcinoma patients, however, it is likely to be a predictive marker for adjuvant treatment after surgery for patients with gastric carcinoma.

OBJECTIVETo observe the effect of Ly49A transfected mouse spleen cells on graft versus host disease (GVHD) and graft versus leukemia (GVL) effect after haploidentical allogeneic bone marrow transplantation in mice.

METHODSLy49A gene was transfected into spleen cells of C57BL/6 mice by retrovirus and the expression rate of Ly49A receptor was evaluated by flow cytometry. The murine model of haploidentical allogeneic acute GVHD was established by using C57BL/6(H - 2b) mouse as donor, and (BALB/c x C57BL/6) F1(H - 2d/b) (CB(6)F(1)) mouse as the recipient which was injected EL9611 cells before transplantation. After irradiation (TBI, (60)Co 10.5 Gy), the recipient received mixed graft of spleen cells and bone marrow cells to establish a GVHD model. The effects of Ly49A transfected spleen cells on GVHD and GVL post haploidentical allogeneic bone marrow transplantation were detected with this model.

RESULTSThe expression rate of Ly49A receptor was (42.20 +/- 4.87)%, (18.67 +/- 2.48)% and (18.73 +/- 3.82)% for pLXSN-Ly49A, pLXSN transfected and untransfected spleen cells respectively. Among haploidentical allo-BMT (C57BL/6(H - 2b)-->CB6F1(H - 2d/b)) groups, the survival time was (7.80 +/- 3.36) days for irradiation group; (21.70 +/- 2.87) days for cyclophosphomide therapy group; (29.40 +/- 6.43) days for mixed bone marrow cells and spleen cells transplantation group; (29.10 +/- 7.39) days for mixed bone marrow cells and pLXSN transfected spleen cells transplantation group and (45.00 +/- 12.38) days for mixed bone marrow cells and Ly49A transfected spleen cells transplantation group, which was much longer than that of any other groups (P = 0.000).

CONCLUSIONThe Ly49A transfected spleen cell transplantation could alleviate GVHD and retain GVL effect in the acute GVHD model post haploidentical allo-BMT.

Animals ; Antigens, Ly ; genetics ; immunology ; Bone Marrow Transplantation ; Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; etiology ; immunology ; mortality ; Graft vs Leukemia Effect ; immunology ; Lectins, C-Type ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; NK Cell Lectin-Like Receptor Subfamily A ; Receptors, NK Cell Lectin-Like ; Spleen ; cytology ; metabolism ; Survival Rate ; Time Factors ; Transfection

Objective To investigate the therapeutic effects of haploidentical hematopoietic stem-cell transplantation (Haplo-PBSCT) for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy. Methods Eighty-nine cases of AML in first relapse after complete remission by standard DA/Hi-Ara-C regimens induction chemotherapy were evaluated retrospectively. Fiftythree cases were grafted by haplo-PBSCT and 26 cases were treated with iDA/Mid-Ara-C or MA/ Mid- Ara-C agents. Results The second remission rate in haplo-PBSCT group and continuous chemotherapy group was 86. 7 % (46/53 cases) and 38. 1% (9/23 cases) respectively (P＜0. 01). Survival postprogression (SPP) at 36th month was 43. 4 % (23/53 cases) in haplo-PBSCT group and 11.5 % (3/26 cases) in continuous chemotherapy group (P ＜ 0. 05). Conclusion Haplo-PBSCT could significantly increase the second remission rate and prolong the survival time of patients with acute myeloid leukernia in first relapse after complete remission by standard induction chemotherapy.

OBJECTIVETo explore the role of myeloperoxidase (MPO) and tumor necrosis factor-α (TNF-α) in myocardial injury induced by hind-limb ischemia-reperfusion (IR) in rats.

METHODSRat models of bilateral hindlimb IR established using a tourniquet were randomized into 9 groups, including a normal control group normal, 2 ischemic groups with hindlimb ischemia for 2 and 4 h, and 6 IR groups with a 4-h ischemia followed by reperfusion for 0.5, 2, 4, 6, 12, and 24 h. The plasma and myocardial levels of MPO and TNF-α in each group were measured, and the myocardial expression of TNF-α was determined with immunohistochemistry.

RESULTSCompared with the normal control group, the rats with a 2-h ischemia showed significantly increased levels of MPO and TNF-α in the plasma and myocardium. Compared with those in rats with a 4-h ischemia, the plasma and myocardial MPO levels increased significantly at 0.5 and 2 h of reperfusion, respectively; the plasma TNF-α level increased significantly at 4 h of reperfusion and myocardial TNF-α level decreased obviously at 12 h; plasma levels of MPO and TNF-α both significantly decreased at 24 h. The plasma MPO and TNF-α and myocardial TNF-α reached the peak levels at 4 h of reperfusion, and the peak myocardial MPO level occurred at 6 h. Immunohistochemistry showed that TNF-α positivity moderately increased after hindlimb ischemia, and further increased at 4 h of reperfusion but obviously reduced at 24 h.

CONCLUSIONThe activation of systemic and local neutrophils and inflammatory cytokines may play an important role in myocardial injury induced by hindlimb IR in rats.

Animals ; Disease Models, Animal ; Hindlimb ; blood supply ; Ischemia ; metabolism ; Male ; Myocardium ; metabolism ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo assess the protective effect of exogenous hydrogen sulfide (H₂S) against myocardial injury after skeletal muscle ischemia/reperfusion (IR) in rats and explore the mechanism.

METHODSThirty-one Wistar rats were randomized into normal control (n=8), IR group (n=8, with a 4-h reperfusion following a 4-h ischemia of the bilateral hindlimbs induced using a tourniquet), NaHS group (n=8, with IR and intraperitoneal injection of 14 µmol/kg NaHS), and DL-propargylglycine (PPG) group (n=7, with IR and intraperitoneal injection of 50 mg/kg PPG). The plasma levels of CK-MB and the levels of MPO, TNF-α, MDA, T-SOD, and CuZn-SOD in the plasma and myocardial tissues were measured. The expression of TNF-α in the myocardium was examined using immunohistochemistry. RESULTS Skeletal muscle IR induced significantly increased plasma CK-MB level (P<0.05) and the levels of MPO, TNF-α, and MDA in the plasma and myocardium, and significantly decreased plasma and myocardial levels of T-SOD and CuZn-SOD (P<0.05). NaHS treatment significantly decreased plasma CK-MB level (P<0.05), reduced plasma and myocardial levels of MPO, TNF-α, and MDA, and increased plasma and myocardial T-SOD and CuZn-SOD in rats with IR (P<0.05), whereas PPG treatment did not produce any obvious responses (P>0.05). Immunohistochemistry showed an obviously reduced expression of TNF-α in the myocardium in rats with NaHS treatment compared with those in IR group.

CONCLUSIONH₂S treatment can alleviate myocardial injury induced by skeletal muscle IR in rats by inhibiting the inflammatory cytokines and oxidative stress.

Animals ; Creatine Kinase, MB Form ; metabolism ; Disease Models, Animal ; Hydrogen Sulfide ; pharmacology ; Ischemia ; metabolism ; Male ; Malondialdehyde ; metabolism ; Muscle, Skeletal ; blood supply ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Oxidative Stress ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To study the characteristics of perioperative treatment of functional endoscopic nasal sinus surgery in elderly patients. METHOD: Data of 186 patients above 60 years old with chronic sinusitis were retrospectively analyzed. RESULT: After being received systematics and personal individuality treatment, the rates of curing were 72.5%, the rates of improving were 23.6% and the rates of efficiency were 96%. CONCLUSION The treatment of endoscopic sinus surgery is reliable and safe for elderly patients.
Aged ; Aged, 80 and over ; Chronic Disease ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Nasal Polyps ; surgery ; Retrospective Studies ; Sinusitis ; surgery ; Treatment Outcome

Objective Through studying the extracorporeal membrane oxygenation (ECMO)alleviating kidney damage caused by unstable circulation function following brain deaths to improve the utilization rate of donation after brain death (DBD) for renal transplantation.Methods Randomly selected 60 DBD patients with unstable circulatory function were randomly divided into 3 groups according to the time of perfusion by ECMO.The first group used ECMO perfusion for 2 h,group 24 h and group 36 h.At the same time,20 cases of stable DBD were randomly selected as the control group.The incidence of primary graft failure,delayed graft function and acute rejection,renal function recovery time,urine volume on the first day after surgery,creatinine and urea nitrogen one year after surgery were compared between two groups.Results The incidence of delayed recovery of renal function in experimental groups 1,2,3 and control group was 10％ (2/20),15％ (3/20),5％(1/20) and 10％ (2/20) respectively.The incidence of acute rejection in experimental groups 1,2,3 and control group was 30％ (6/20),25％ (5/20),35％ (7/20) and 25％ (5/20) respectively.There were no statistically significant differences between the experimental groups and the control group (P ＞0.05).The mean renal function recovery time in the experimental groups 1,2,3 and control group was (9.6 ± 2.0),(9.7 ± 2.7),(9.5 ± 1.9) and (7.3 ± 3.1) days respectively.The average urine volume in experimental groups 1,2,3 and control group was (3 405 ± 755),(3 300 ±600),(3 810±825),and (3 600 ± 80) mL respectively.In experimental groups 1,2,3 and control group,the average creatinine level was (92 ± 28),(92 ± 28),(101 ± 31),and (98 ± 30) μmol/L respectively.The average urea nitrogen in experimental groups 1,2,3 and control group was (4.2 ± 2.2),(5.6 ± 2.6),(5.2 ± 2.6) and (5.5 ± 2.5) mmol/L respectively.The recovery time of renal function in the experimental groups was longer than that in the control group (P＜0.05),but there was no significant difference among the experimental groups (P＞0.05).There was no statistically significant difference between the experimental groups and the control group in urine volume one day after surgery and kidney function one year after transplantation (P＞0.05).Only recovery time after operation of the DBD with the unstable circulation function assisted by EMCO was slightly longer than that with stable circulation function,while the rest indexes showed no statistically significant difference.Conclusion DBD with unstable circulation function with ECMO auxiliary circulatory function can protect the renal function and increase the utilization rate of donor organs.

Objective To explore the effect of depression on loneliness in medical students',and the mediating effects of introverted personality and social support.Methods EPQ,UCLA,SSRS,and POMS questionnaire were used,and surveyed data was collected from Dalian Medical University medical students.Results The scores of depressive mood,introverted personality,social support and loneliness were (17.77±4.45),(11.97±5.21),(34.52±6.15) and (38.02±6.36).Depression was positively related to loneliness and introverted personality (r=0.286,0.508,P＜0.01),and social support was negatively related to loneliness (r =-0.443,P＜0.01).Introverted personality was positively related to loneliness(r=0.401,P＜0.01).The total effect value was 0.670.The mediating effect of introversion as mediator variable was 0.221,accounting for 33％ of the total effect.Social support as intermediary variable,intermediary effect value was 0.449,accounting for 67％ of the total effect.Standard value of mediating effect between depression and introverted personality showed as 0.508 (95％CI=0.475-0.725,P＜0.0i),standard value of mediating effect between depression and social support showed as-0.150 (95％CI=0.989-1.469,P＜0.01),introverted personality was the mediating variant of depression and loneliness,standard value of mediating effect was 0.156 (95％ CI=0.080-0.232,P＜0.01),social support was the mediating variant of depression and loneliness,standard value of mediating effect was 0.317 (95％ CI=0.177-0.457,P＜0.01).Conclusions Introverted personality and social support play an intermediary role between depression and loneliness.