Objective To explore the role of expression of membrane Fas(mFas)antigen and serum soluble Fas(sFas)in acute leukemia.Methods Strept avidin-biotin complex method of immunohistochemical staining assay and sandwich enzyme-linked immunosorbent assay(ELISA)were used to detect the expression of mFas antigen in bone marrow cells and serum sFas in 30 patients with acute leukemia in Binzhou Medical College Affiliatd Hospital from November 2001 to September 2002,which was compared with normal control group.The relation between expression of serum sFas and chemotherapy outcome were evaluated too.Results The expression of mFas antigen in ALL(5.62?2.27)% and ANLL(8.80?4.15)% were respectively and significantly lower than that in normal control group(28.75?11.20)%(P

AIM: To explore the expressions and significance of angiopoietin-2 (Ang-2) and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains-2 (Tie-2) receptors in a rat model of acute lung injury (ALI). METHODS: Wistar rats (n=42) were divided into control group (n=12) and cecal ligation and puncture (CLP) group (n=30). Control group underwent sham operation, and CLP group underwent cecal ligation and puncture to make the model of ALI. 12 h after sham operation or CLP, 6 rats in each group were killed, and arterial blood gas analysis and lung coefficient were tested. The expressions of Ang-2 and Tie-2 receptors in lung tissue were observed by immunohistochemical method. Blood samples of the rest rats were collected from vena caudalis, and Ang-2 levels were measured by enzyme linked immunosorbent assay (ELISA). The mortality rate in each group within 36 h was compared. The lung architecture was observed under microscope. RESULTS: The lung architecture in control group was clear and intact. Alveolar septum was thicker, blood capillary was congested, and neutrophils and macrophages were infiltrated in the lung tissue in CLP group. Tie-2 receptors were expressed in bronchial epithelial cells, smooth muscle cells and endothelial cells in control group. Besides the similar expression as control group, high expression of Tie-2 on neutrophils and macrophages in CLP group was observed. In the adhesion location of Tie-2 receptors positive inflammatory cells, there was stronger staining in endothelial cells. Ang-2 was expressed in smooth muscle cells, bronchial epithelial cells and endothelial cells in control group. The Ang-2 level in CLP group were higher than that in control group [(8.14±1.74) μg/L vs (4.63±0.49) μg/L, P<0.01], and the Ang-2 level of dead rats was higher than that of survival rats within 36 h in CLP group [(8.95±1.61)μg/L vs (6.80±0.96)μg/L, P<0.01]. Oxygen partial pressure in control group was lower (P<0.01) and lung coefficient was higher (P<0.01) than that in CLP group. CONCLUSION: Ang-2 and Tie-2 receptors may participate in the pathophysiology of ALI, and Ang-2 level is correlated with mortality.

BACKGROUND: It is of great importance in improving the clinical effect of human islet allograft to study and design models of such large animals as pigs or primates preclinically.OBJECTIVE: To evaluate the effect of different doses of streptozotocin (STZ) on inducing diabetes type Ⅰ models of nonhuman primates.DESIGN, TIME AND SETTING: A contrast observational animal experiment was performed in the Cell Transplantation and Gene Therapy Center, the Third Xiangya Hospital of Central South University from October 2007 to December 2008. MATERIALS: A total of 21 adult male rhesus monkeys were divided into a 125 mg/kg STZ group (n =5), a 75 mg/kg STZ group (n=5) and a 50 mg/kg STZ group (n=11).METHODS: STZ weighed with regard to body mass of animals was prepared into 25 g/L STZ solution with buffer that was prepared in advance. After being filtered and degermed, the new-prepared STZ of 125 mg/kg, 75 mg/kg and 50 mg/kg were administered by intravenous injection into the experimental monkeys respectively, which took 1-5 minutes.MAIN OUTCOME MEASURES: Liver and renal function, glucose metabolism and histomorphological changes of animals during 1-16 weeks following administration.RESULTS: In 125 mg/kg STZ group, two rhesus monkeys died, in 8 hours following STZ administration, of serious hypoglycemia caused by severely damaged pancreas β cells; All rhesus monkeys in this group had got significantly increased liver transaminase, serum creatinine and urea nitrogen at week 1 following STZ administration, which reached a peak during 2-4 weeks; One rhesus monkey in this group showed severe shortage of endogenous trypsin and hyperglycemia irreversible by exogenous insulin following STZ administration, and finally died at day 13 following STZ administration due to the glucose metabolic disorder, ketoacidosis, liver and renal failure; The other two survivors in this group kept high level of liver transaminase,urea nitrogen and serum creatinine throughout the observation period. In 75 mg/kg STZ group, rhesus monkeys presented significantly increased liver transaminase, serum creatinine and urea nitrogen at week 1-2 following STZ administration; After 4 weeks following administration, their liver and renal function presented with abnormality of different degrees; One rhesus monkey in this group had got injured renal function, decreased power of resistance, eyelid edema, general dropsy and irreversible infected rump after injection of STZ, and finally died at the end of week 5 following administration; Another rhesus in this group presented with irreversible continuous hyperglycemia, inappetence and significantly decreased weight, and finally died ofsystemic failure at week 9 following administration. In the 50 mg/kg STZ group, renal function of monkeys were slightly affected, with a transient mild rise which return to the normal level by the end of week 4 following administration; Only 3 animals in this group appeared eyelid edema during 1-4 weeks following administration which disappeared afterwards.CONCLUSION: STZ of 50 mg/kg is possibly the optimal dose for inducing diabetes models in most rhesus monkeys.

Objective To observe clinical efficacy of NB-UVB in treating psoriasis vulgaris and its effects on expression of serum interleukin-17 (IL-17),interleukin-23 (IL-23) and interleukin-22 (IL-22) in patients with psoriasis vulgaris,and to study the underlying mechanisms of NB-UVB.Methods Ninety patients were recruited and treated with NB-UVB therapy for 8 weeks.Before and after treatment,the serum level of IL-17,IL-23,IL-22,interleukin-l0(IL-10) and transforming growth factor(TGF-β) were tested by use of ELISA method,meanwhile Psoriasis Area and Severity Index (PASI) were used to evaluate clinical efficacy.Fifty healthy volunteers were selected as control group.Results Compared to healthy controls,the level of serum IL-17,IL-23 and IL-22 was significantly higher in patients with psoriasis vulgris (P ＜ 0.01),and IL-10,TGF-β shown lower expression in psoriasis patients (P ＜ 0.01).After 8 weeks of treatment with NB-UVB,serum levels of IL-17,IL-23 and IL-22 in psoriasis patients decreased significantly (P ＜ 0.01),while IL-10,TGF-βelevated significantly (P ＜ 0.01) in contrast.The Psoriasis Area and Severity Index (PASI) results indicated significantly clinical improvement after therapy,and the total effective rate was 87.78％.Conclusion NB-UVB could down-regulate serum IL-17,IL-23,IL-22 and up-regulate IL-10,TGF-β,which may help regulate imbalance of T lymphocytes cells of psoriasis patients.The clinical data demonstrate that the treatment of NB-UVB is a safe,effective method for psoriasis vulgaris.

The animal experiment studies have demonstrated that hyperthermia is a strong teratogen to many kinds of animal with high incidence of neural tube defects(NTD).Epidermiological investigation showed that hyperthermia was closely related to the acencephaly and excenphaly in human being,but little was known about the distinct mechanism of NTD induced by hyperthermia.In order to study the developmental mechanism of NTD induced by hyperthermia,we observed the effects of hyperthermia on the neuroepithelial cells in primary culture.The neural tubes of the hamster embryos on 10 d after fertilization were obtained.the neuroepithelia were dissociated and then seeded at the density of 1×106 cells per well.The cells were divided into experimental and control groups randomly.The experimental groups were exposed to 42 C for 20 min,whereas groups exposed to 37 Cserved as control.After treatment of hyperthermia,the cells were incubated continuously at 37 C in a 95%air/5%CO2 humidified incubator,the culture was terminated after different intervals.Phase-contrast microscopy.scanning electron microscopy.transmission electron microscopy,MTT assay,agarose gel electrophoresis analysis,TUNEL detection,immunocytochemistryand image analysis were made.The results of the experimental groups indicated that the floating cells and apoptotic cells increased in number,the neurites of the cells were shortened even disappeared,the survival cells decreased in number,the ultrastructure of the cells demonstrated distinct abnormal changes,the function of mitochondria was impaired,the expressions of both bcl-2 and bax were abnormal.The above results suggest that hyperthermia may induce apoptosis of neuroepithelial cells,duringwhich bcl-2 and bax genes may play important regulatory roles.

Objective To explore and solve the problem of insufficiency and malformation due to cripple hand and defect of digital bone resulted of innate and injured. Evaluation to the methods of bone autografting combinated with flaps to reconstruct the deficient hand and digits. Methods Vascularized iliac bone or metatarsophalangeal joints (MPJ) or toes transplantation incorporated with flaps were used to reconstruct the defected bone in palm or fingers. Combined with tendon absence in such cases should be repaired or restituted by tendon autografting in one stage or by stages. Results Total 16 cases were treated by the methods metioned above. Except 1 of 16 was failure resulted in severe infection, the others were reconstructed successfully. 13 of 16 were followed up 8 months to 2 years, the evalution of the group from motion,sensation and appearance shew that the fine rate of motion was 53.8 percent, the rate of appearance and sensation were both 69 percent. Conclusion Vascularized iliac bone or metatarsophalangeal joints (MPJ) or toes transplantation incorporated with flaps are available to reconstruct the defected bone in palm or fingers, and can reduce disability effectively while obtain a satisfactory outlook and proper functions by those methods.

Purpose To quantitatively evaluate the severity of brain damage in the acute phase of CO poisoning using intravoxel incoherent motion imaging.Materials and Methods MRI scans were performed in 26 patients with CO intoxication of acute stage and 18 healthy controls with matched age and gender.The difference of D value,D* value and f value in different ROIs between the poisoning and the control groups were compared,and the best cut-off values were determined.The correlation between the consciousness state and the D value as well as f value in the vulnerable area of the brain (globus pallidus) was analyzed.Results Compared with control group,the D and F value of the poisoning group were decreased in multiple ROIs,of which statistical significance was observed in the f values in the centrum semiovale,globus pallidus and thalamus,and the D values in the centrum semiovale,lateral ventricle,globus pallidus and corpus callosum splenium (P＜0.05).The best cut-off value,sensitivity and specificity were as follows:fcs=51％,77％,73％;fga=61％,77％,83％;ft=80％,85％,64％;D,=0.69×10-3 mm2/s,85％,83％;Dlv=0.65× 10-3 mm2/s,73％,89％;Dga=0.68× 10-3 mm2/s,62％,83％;Dccs=0.70× 10-3mm2/s,81％,89％.The disturbance of consciousness was positively correlated with decrease of D value and f value (rD=0.828,rf=0.745,P＜0.05).Conclusion The D and f values of intravoxel incoherent motion imaging can quantitatively evaluate the brain damage severity in acute CO poisoning.

Objective To assess the effectiveness of pedicled iliac periosteal flap graft for treatment of old femoral neck fracture in adolescents.Methods Between June,2005 and December,2013,15 patients (15 hips) of old femoral neck fracture in adolescents treated with vascular pedicled iliac periosteal.There were 11 males and 4 females with an average age of 15.8(range 12 to 18) years.Based on the location of fracture,there was 5 cases of subcapital,8 cases of transcervical and 2 cases of basal.6 cases were treated with lower limb traction,3 cases with internal fixation and 6 just without any treatment.The average duration from injury to the second operation was 4.8 (range 1 to 19) months.There were 2 cases of femoral head necrosis after femoral neck fracture.Open reduction,pedicled iliac periosteal flap grafting and cannulate screw fixation were performed.Results All incisions healed by first intention.All patients were followed up 12 to 90 months (mean,46 months).The HHS was increased from (48.7 ± 8.3) pre-operation to (91.3 ± 6.1) at last followed-up,indicating a significant difference between before and after operation (P ＜ 0.01).One patient underwent total hip arthroplasty at 10 months after operation because of fracture nonunion and femoral head necrosis.Fracture healed successfully in 14 cases and the average time of fracture healing was 4 (range 3 to 6) months.Certain extent of remodelling and bulge of the head were observed in 2 cases of collapsed heads.Conclusion Pedicled iliac periosteal flap graft can provide good osteogenesis and vascular reconstruction for femoral head and promote fracture healing in treatment for old femoral neck fracture in adolescents.

Objective To observe the clinical efficacy of narrow-band ultraviolet B (NB-UVB) in the treatment of the patients with atopic dermatitis (AD) and the effects of NB-UVB on the expression of the CC subfamily of chemokines in AD patients. Methods Fifty-five AD patients were treated with NB-UVB with a starting dose of 50％ of the minimal erythma dose.ELISA was used to measure the serum levels of thymus and activation-regulated chemokine ( TARC),cutaneous T cell-attracting chemokine ( CTACK),macrophage-derived chemokine (MDC) and eotaxin in all of the patients before and after treatment,and the clinical efficacy was evaluated.The scores on an atopic dermatitis index (SCORAD) and a visual analogue scale were used for the clinical evaluation.Thirty healthy persons were recruited and served as normal controls. Results The serum levels of TARC,CTACK and MDC were significantly higher in patients with AD than in the normal controls.There was no significant difference between the patients and controls with regard to the average serum level of eotaxin.After treatment with NB-UVB,the serum levels of TARC,CTACK and MDC,but not eotaxin,significantly decreased in the patients.The total clinical effectiveness rate was 76.36％,and the accumulated SCORAD points and VAS scores decreased significantly. Conclusions NB-UVB is able to down-regulate significantly the serum levels of TARC,CTACK and MDC.It can affect immune function and regulate any imbalance of Th1/Th2 cells.This might be one of the mechanisms of NB-UVB treatment for AD.The clinical data demonstrate that NB-UVB is a safe and effective treatment for AD.

ObjectiveTo evaluate the primary effect of granulocyte-monocyte colony stimulating factor (GM-CSF) as an immunotherapy option for treatment of residual disease after alloHSCT.Methods Immunotherapy was performed on two patients with blood malignancy to treat residual disease after allo-HSCT. The patient one,who was diagnosed as having MDS-RAEB Ⅱ,showed bone marrow displasis and incomplete chimerism 6 months after unrelated donor HSCT.Immunosuppressive drug was withdrawn without induction of graft-versus-host disease (GVHD).The patient two B-ALL demonstrated a residual disease at molecular level 30 days post-transplantation.Both of them were given GMCSF (300 μg) subcutaneously once every two days for totally three weeks.During the whole period,skin itch and rash,liver function,subgroups of lymphocytes,and MDSCs and DCs in peripheral blood were investigated.Results In case one,grade Ⅰskin acute GVHD (aGVHD) appeared as early as one week after GM-CSF administration,as well as grade Ⅱ (skin and liver) by the end of the third weeks,and GM-CSF injection was withdrawn.One month later since the start of GM-CSF,the patient showed normal bone marrow morphology and full donor type chimerism. Cyclosporine A (CsA), mycophenolate mofetil and methylprednisolone were administered for two weeks to control GVHD.In the other case,grade Ⅰ aGVHD occurred 9 days after GMCSF administration,and whole blood CsA maintained at 0.134-0.472 μmol/L.Prednisone (30mg per day for 5 days) was used to control grade Ⅱ GVHD from the 11th day after GM-CSF,and grade Ⅰ GVHD continued without any intervention.On the 30th day after GM-CSF treatment,bone marrow aspiration showed complete molecular remission.In both of the two cases,no differences in lymphocytic subtypes were revealed before and after GM-CSF administration,while there were trends of increased DC number and decreased MDSCs in peripheral blood.ConclusionThe administration of GM-CSF as an immunotherapy option for blood malignancy may contribute to the clearance of residual disease after Allo-HSCT.