BACKGROUND:Diabetic acromelic gangrene(diabetic foot)has become one of the important causes for the disability and death in diabetes mellitus.OBJECTIVE:To establish model of diabetic foot in rat,and observe the interventional effect of Chinese compound on diabetic foot.DESIGN:A comparative observational experiment.SETTING:The First Affiliated Hospital of Clinical Medical College,Henan University of Science and Technology.MATERIALS:Fifty male Wistar rats of 8 weeks old,(200.0±16.3)g,were raised with common feed in separate cage at the room temperature of 18-22℃.and they were free to access of feed and water.Ten rats were randomly selected as the blank control group,and the other 40 were used for model establishment.METHODS:The experiments were carried out in the Animal Room of Henan University of Science and Technology from October 2001 to April 2002.①Grouping:The 40 rats were fasted for 6 hours,and then treated with intraperitoneal injection of streptozotocin(55.0 mg/kg),while the 10 rats in the blank control group were injected with isovolume sodium citrate buffer solution.20 models were successfully established,and they were randomly divided into model group(n=10)and treatment group(n=10).Rats in the treatment group were treated for 3 weeks with intragastric perfusion of Chinese compound(60 g/kg)at 9:00 every day after model establishment,and those in the model group were given intragastric perfusion of isovolume saline.At the end of the third week,the rats were all killed under anesthesia after fasted for 12 hours,blood samples were collected to determine the levels of fasting blood glucose,blood lipids and insulin.The daily amount of drinking water was recorded in each group during the experiment.②Scoring standards for acromelic gangrenes:The limbs rats with diabetic foot were scored by three grades,including dark skin,mild open focus on skin,and focus had invaded deep muscular tissue.The total score of each rat was calculated.The beta-cell function index (HBCI)was also evaluated.MAIN OUTCOME MEASURES:The changes of the amount of drinking water,body mass and levels of triglyceride,cholesterol,fasting blood glucose,blood lipids and insulin were observed before and after treatment.RESULTS:Totally 50 Wistar rats were used.20 of them were excluded due to unsuccessful model establishment,and the other 30 rats were involved in the final analysis of results.①The amount of drinking water was obviously higher in the model group and treatment group than in the blank control group during the experiment(P＜0.01).As the treatmentlasted,the amount of drinking water was obviously decreased in the treatment group,but gradually increased in the model group.②After treatment,the body mass was obviously lower than that before treatment in the model group(P＜0.01).but had a descending trend without obvious difference as compared with that before treatment in the treatment group(P＞0.05).③Obvious acromelic gangrenes were obvious in both groups(P＜0.01).The body mass in the treatment group was obviously higher than that in the model group(P＜0.01),and the conditions of acromelic gangrene were obviously better than those in the model group(P＜0.01).④Before treatment,the levels of fasting blood glucose in the treatment group and model group were obviously higher than that in the blank control group(P＜0.01),while the levels of insulin and HBCl were obviously lower than those in the blank control group(P＜0.01).After treatment,the levels of fasting blood glucose in the treatment group and model group were obviously higher than that in the blank control group(P＜0.01),and it was obviously lower in the treatment group than in the model group(P＜0.01),it was close to the normal value in the treatment group.⑤The levels of triglyceride and cholesterol before treatment were obviously higher in the treatment group and model group than in the blank control group(P＜0.01).After treatment,the levels of triglyceride and cholesterol in the treatment group were obviously decreased(P＜0.01), which were not obviously different from those in the blank control group (P＞0.05), while those in the model group were increased continuously,and obviously higher than those in the blank control group(P＜0.01).CONCLUSION:Increasing the serum level of insulin and decreasing the levels of blood glucose and blood lipids can prevent and treat the occurrence and development of diabetic foot to some extent.This model of diabetic foot is sensitive to drug,and can be used to investigate the pathogenesis of diabetic foot and evaluate the effect of drug therapy.

Objective To explore the distribution characteristics of monosomal karyotype (MK) in elderly patients with acute myeloid leukemia (AML).Methods The karyotype analysis was performed in 123 elderly patients with newly diagnosed AML in our center from Nov 2000 to Feb 2015.We retrospectively studied the distribution characteristics of monosomal karyotype in these patients.Results Among 123 elderly patients with AML,117 patients had enough metaphases chromosomes for analysis.Among the 117 patients,there were 16 cases with good-risk karyotype,54 cases with intermediate-risk karyotype,and 47 cases with adverse-risk karyotype.In the 47 patients with adverse-risk karyotype,43 cases had complex karyotypes (CK).In the 117 elderly AML patients,37 cases (31.6％) had monosomal karyotype (MK),22 AML cases were secondary to myelodysplastic syndrome (MDS-AML),among them 13 cases (55.0％) had MK.In the 95 cases with primary AML,the detection rate of MK was 25.3％ (24 cases).The detection rate of MK+ AML was higher in MDS-AML patients than in de novo AML patients (P=0.000).Among the 37 patients with MK+AML,35 cases had complex karyotypes.30 (81.1％) MK+AML patients had two or more distinct autosomal monosomies and 7 (18.9％) MK+ AML patients had one single autosomal monosomy in the presence of structural abnormalities,and the incidence of autosomal monosomies was higher than that of single autosomal monosomy.The presence of--5 (27.0％),-4 (18.9％),-7 (16.3％) and-6 (13.5％) chromosomes was the most common autosomal monosomy among MK+ AML patients.Conclusions The detection rate of MK is relatively high in elderly AML patients.Two or more distinct autosomal monosomies are more common.The detection rate of MK+AML is higher in patients with MDS-AML than in patients with de novo AML.

Objective:To study the effects of recombinant human platelet-derived growth factor-BB(rhPDGF-BB)combined with recombinant human transforming growth factor-β1 (rhTGF-β1 )on the expression of FAK mRNA of osteoclasts in the alveolar bone on the pressure side of orthodontic teeth in rats.Methods:Orthodontic tooth movement model was established in 160 male SD rats.The rats in experimental group were treated by injection of 10 ng rhPDGF-BB and 5 ng rhTGF-β1 in the buccal submucosal area of the mo-lar every other day from day 1 afterburner,while those in control group received equivalent volumes of PBS.The rats were sacrificed at 1,4,7,10 and 14 days(n=16)after appliance placement.Specimens were collected.Osteoclasts in the alveolar bone on the pres-sure side of the orthodontic teeth were observed by TRAP staining,the FAK mRNA expression was quantified by quantitative RT-PCR.Results:rhPDGF-BB combined with rhTGF-β1 significantly promoted an increasing number of osteoclasts on the compressing side(P<0.05),increased the expression of FAK mRNA at day 4 and 7(P<0.05),then decresed it to the control level(P>0.05) at day 14.Conclusion:Combination of rhPDGF-BB and rhTGF-β1 can increase the number of osteoclasts in the alveolar bone on compressing side,and promote FAK mRNA expression in osteoclasts.

Objective To observe the protective effects of autologous bone marrow mesenchymal stem cells transplantation alone with bone marrow stem cells mobilization on liver cells in rats with severe acute paucreatitis in order to explore their mechanism. Method After the establishment of severe acute pancreatitis in rats made by intraperitoneal injection of L-arginine, 240 SD rats were randomly divided into sham-operated group (n = 48), model control group (n = 48), bone marrow mesenchymal stem cell transplanted (MSC) group (n = 48), granu-lacyte-colony stimulating factor treated (G-CSF) group (n = 48) and MSC + G-CSF (n = 48). The rats of MSC group were prepared by injection of 1.2 mL MSC into femoral vein 6 hours after SAP. The rats of G-CSF group were prepared by subcutaneous injection of G-CSF 40 μg/kg for 3 days before SAP. The rats of MSC + G-CSF group received MSC and G-CSF together. The rats of sham-operated group were injected with equal volume of nor-real saline. The rats in each group were sabdivided into 12 h, 24 h, 48 h and 72 h sub-groups (n = 12) according to the examinations in different intervals after operation. Of different subgroups, the morality rate, pathological changes, expression levels of Bax, Bcl-2 proteins and apoptosis indexes of livers were observed respectively. The contents of serum TNF-α,IL-6,ALT,AST,LDH and CRP were simultaneously determined to compare the difference among subgroups by variance analysis. Results Compared to the respective model group, the mortality rates of all treated 72 h subgroups showed no difference (P > 0.05), and no rats died before 48 h. The pathological injuries of liver cells were rather attenuated in rats of treated group than in rats of control group. The liver cell apoptosis in-dexes of 48 h and 72 h MSC + G-CSF subgroups were 107.1 ± 7.0, 110.3 ± 8.6, respectively; the expression of Bax in livers of 24 h,48 h and 72 h subgroups was 5.60±0.Z5, 5.69±0.22, 5.73±0.27, respectively;Bcl-2 protein of 48 h,72 h subgroups was 4.61±0.28, 4.43±0.28, respectively; compared with MSC and G-CSF subgroups the differences were significant (P < 0.05). The serum TNF-α, IL-6, ALT, AST, LDH and CRP de-creased obviously in 24 h/48 h treated subgroups in comparison with control group (P < 0.05). The MSC + G-CSF group showed more significant effects on those biomarkers than MSC or G-CSF alone after 48 hours (P < 0.05). Conclusions Autologous bone marrow mesenchymal stem cells transplantation alonewith bone marrow stem cells mobilization can significantly protect livers from severe damage during the course of severe acute panere-atitis, and the probable mechanisms are likely associated with the pathological regeneration, anti-inflammatory ef-fect and apoptosis inhibition of MSC.

OBJECTIVE To investigate the incidence and pathogens of infection in 1659 consecutive cases in single center hematological unit.METHODS The incidence,pathogen,and outcome of infection in 1659 hospitalized patients with hematological disorders from 1999 to 2006 were retrospectively analyzed.RESULTS The overall incidence of infection was 24.4% according to the person-times of hospitalization,which included 22.1% of nosocomial infection and 2.3% of community acquired infection.Most of the pathogenic bacteria of the nosocomial infection were Gram-negative.The most common bacteria in the sputum samples included Enterobacter cloacae(19.3%) and Pseudomonas aeruginosa(14.8%).The most common bacteria in the blood samples were coagulase negative Staphylococcus(CNS,39.3%),the next was Pseudomonas aeruginosa and Escherichia coli.There were 4.21% bacteria resistant to most of antibiotics in nosocomial infection.There were 114 fungi isolated.Candida albicans was accounted for 35.1%.The mortality due to nosocomial infection was 7.4%.CONCLUSIONS The patients in hematology ward are susceptible to infection.The pathogens of nosocomial infection are most likely G-bacteria.Some bacteria are resistant to almost all antibiotics.The incidence of fungal infection is increasing in the near 8 years.

Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases (IRAK-4) in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope (LSCM), and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours (P<0.05), kept at a high level at 12 hours (P<0.05) , but decreased gradually to the normal oxygen level at 24 hours (P < 0.05) and to below the normal oxygen level at 48 hours (P < 0.05). Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 (r =0.84, P < 0.05 ). Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.

Objective To evaluate the feasibility of using comprehensive geriatric assessment (CGA) in estimating if standard dose treatment is fit for the elderly patients with diffuse large B cell lymphoma.Methods.Comprehensive geriatric assessments including three assessments of activity of daily living,instrumental activity of daily living and comorbidity scoring according to Cumulative Illness Rating Score for Geriatrics were adopted to assess if standard dose treatment is fit for the elderly patients in our prospective study.Thirty seven patients with diffuse large B cell lymphoma,aged ＞70 years were enrolled in the study,and grouped into fit,unfit and frail groups according to comprehensive geriatric assessment scoring and their age.The treatment protocolswere not determined by comprehensive geriatric assessment scores,but by clinical judgments made by clinicians based on their clinical experience and disease features.The clinically effective response and overall survival (OS) were analyzed in the three groups.Results According to CGA scores,patients were grouped into fit [21 cases (56.8％)],unfit [7 (18.9％)] and frail [9 (24.3％)].37 cases received 213 courses of treatment at average 5.76 courses per case.The overall response (complete / partial remission) rates were [85.7％(18/21) vs.28.6％ (2/7) vs.44.4％ (4/9),x2=9.69,P=0.008] and median survival times were (44 months vs.10 months vs.9 months;x2 =7.03,P=0.03) among fit,unfit and frail groups with statistically significant differences.Total effective rate (achieving all clinical targets) in fit group of 21 cases were 100 ％ (12/12)with receiving standard dose therapy,and 66.7％ of(6/9)with low dose therapy(P=0.06).Overall response rate(total/partial remission) [85.7％(18/21) vs.28.6％(2/7) vs.44.4％(4/9),x2=9.69,P=0.008] and median survival (44 months vs.10 months vs.9 months;x2 =7.03,P=0.03) amongfit,unfit and frail groups.In fit group,the two-year overall survival was higher in patients receiving standard dose treatment than receivingpalliativetreatment,with statistical significance [83.3 ％ (10/12) vs.33.3 ％ (3/9),P =0.032],without significant hematologic toxicity observed between the subgroups.Conclusions Comprehensive geriatric assessment can identify if elderly patients diffuse large B cell lymphoma can acquire a satisfactory curative effect from a standard dose treatment ofimmunochemotherapy.

Objective:To investigate the effect of different plasticizers on the compressive strength of injectable calcium sulfate cement (ICSC).Methods:Hydroxymethylcellulose (CMC), methylcellulose (MC), hyaluronic acid (HA), stearic acid (SA) and self-made hemihydrate calcium sulfate were used to mix them, and the compressive strength of the complex was observed after reaction with normal saline; the changes of ICSC's own properties, such as setting time and injectability, were recorded.Results:The compressive strength of calcium sulfate was 21±4.58 mPa, and the final setting time was 3.86 ± 0.09 min. Different concentrations of SA had no significant effect on the compressive strength of calcium sulfate ( F=1.593, P=0.266), but prolonged the setting time ( F=29.868, P=0.000). CMC with different concentrations significantly reduced the compressive strength of calcium sulfate ( F=23.943, P=0.000), and the setting time was prolonged to more than 120 min. Different concentrations of MC can improve the compressive strength of calcium sulfate ( F=4.808, P=0.034), and prolong the setting time ( F=191.192, P=0.000); among them, 1% and 3% MC can significantly improve the compressive strength ( P=0.007, 0.027). Different concentrations of HA can improve the compressive strength ( F=3.818, P=0.058), and prolong the setting time ( F=262.515, P=0.000), of which 3% and 5% were significantly improved ( P=0.026, 0.015), while 1% group was not significantly improved ( P=0.062). In addition, the injectable properties of HA, MC, stearic acid and CMC are better, respectively. Conclusion:SA and CMC can not be used to improve the compressive strength of calcium sulfate, while HA and MC of appropriate concentration can improve the compressive strength of ICSC, and improve the injectable performance, but MC can make the coagulation time more in line with clinical needs.

Objective:To evaluate the clinical efficiency and prognostic factors of autologous peripheral blood stem cell trans-plantation (APBSCT) in 30 cases of multiple myeloma (MM). Methods:Two of the 30 patients received the second treatment of APB-SCT because of relapse after the first treatment. Thus, a total of 32 case-times of APBSCT were studied. Combination chemotherapy was inducted regularly before APBSCT (11 patients used bortezomib as an induction drug), and chemotherapy combined with the G-CSF regimen was used to mobilize peripheral blood stem cells. Preconditioning was based on melphalan. Results:Mononuclear cells in harvest were 6.41 × 108/kg, and CD34+cells in harvest were 4.75 × 106/kg. The median times of neutrophil and platelet engraftment were 9.5 and 11 d, respectively. The complete remission (CR) and very good partial remission (VGPR) rates were 37.5%and 34.4%af-ter APBSCT, respectively. The median overall survival (OS) was 67.27 months in all patients, and the median progression-free survival (PFS) was 29.77 months. The median PFS rates were 29 and 20 months in the patients who achieved CR and PR, respectively, and the median PFS was not observed in the patients who achieved VGPR. Statistical differences in PFS were detected between the CR+VGPR and PR groups (P=0.025). The CR rates were 63.6%and 23.8%in the bortezomib (bortezomib-based chemotherapy) and non-bortezo-mib groups (P=0.034), respectively. The median OS and PFS were not obtained in the bortezomib group, whereas the median PFS was 22 months in the non-bortezomib group (P=0.045). Conclusion: MM patients treated with bortezomib-based chemotherapy followed by APBSCT had prolonged PFS. APBSCT can improve the response and survival of MM patients.

Objectives To evaluate the efficacy of rituximab combined with chemotherapy in the treatment of diffuse large B-cell lymphoma (DLBCL) and the relationship of clinical prognosis with the International Prognostic Index (IPI) by the using rituximab in autologous peripheral stem cell transplantation (APBSCT) for the patients of DLBCL. Methods 21 patients with DLBCL, 11 patients of them were at IPI low risk, and 3 patients were IPI at low intermediate risk, 3 patients were at IPI high intermediate risk, 4patients IPI high risk. Rituximab combined with CHOP regimen (cyclophosphamide, adriamycin, vincfistine and prednisone) was given for 4～8 courses. 5 patients received APBSCT. The mobilizing regimen was rituximab combined with cyclophosphamide(CTX) and etoposide(VP16). The conditioning regimen were CBV(CTX combined with VP16 and carmustine). Results In 21 patients, the complete response rate was 61.9 %,with overall response rate 90.5 %. 2-year progression free survival was (69.74±10.43)%. 2-year overall survival was (84.44:1:8.35) %. The complete response rate was 92.9 % and overall response rate was 100 % in the patients IPI≤2. The overall response rate was 71.4 % in the patients with IPI≥3. The complete response rate was higher in the patients with IPI≤ 2 (P＜0.01). The amount of mononuclear cells (M NC) in harvest were 7.34 (4.6～8.53)×108/kg. The CD+34 cells in harvest were 8.82 (2.1～10.34)×1O6/kg. The mean time of neutrephil recovering to 0.5×109/L after APBSCT was +9 day. The mean time of platelet recovering to 20×109/L after APBSCT was +12 day. The major adverse reaction were infusion related response (14.3 %) and hematological toxieities. Conclusion The efficacy of rituximab combined with chemotherapy in the treatment of DLBCL is effective, The complete response rate was higher in the patients with IPI≤2 than in the patients with IPI≥3.Using rituximab in mobilizing regimen, all patients had harvested enough CD+34 cells. Rituximab given at +1day did not affect the hematopoiesis reconstruction.