Objective To study the clinical pathological features of Wegener's granulomatosis (WG) in middle-aged and elderly patients,and enhance understanding of this disease.Methods Totally 21 patients with WG (11 males,10 females,aged 45 to 76 years,mean age 58.1 years) in our hospial from February 1999 to July 2012 were selected.The clinical and pathological data of WG patients were retrospectively analyzed.34 biopsies including 2 autopsies from different organs were paraffin embedded and stained by hematoxylin and eosin and histochemistry.13 renal biopsies were all examined by immunofluorescence and electron microscope.Results The average time from the onset of clinical symptoms to the diagnosis was 5.3 months (from 24 days to 11.0 months).Eyes,nose and salivary glands were the most commonly involved parts at the beginning of Wegener's granulomatosis (52.4％,11 cases).The percentages of the skin,lung and renal involvement were 14.3％ (3 cases),81.0％ (17 cases) and 71.4％ (15 cases),respectively.Among 21 patients,18 patients were examined with anti-neutrophil cytoplasmic antibody (ANCA).c-ANCA was positive in 72.2 ％ patients (13 cases,13/18),p-ANCA was positive in 16.7％ patients (3 cases,3/18),and ANCA was negative in 11.1％ patients (2 cases,2/18).3 major pathological manifestations were observed:7 kinds of vasculitis including capillaritis,acute vasculitis,chronic vasculitis,fibrinoid necrosis in vasculitis,necrotizing granulomatous vasculitis,non-necrotizing granulomatous vasculitis and cicatricial vascular changes; 4 kinds of granulomatous inflammation including scattered giant cells,palisading histiocytes,poorly formed granulomas and microabscess surrounded by granulomatousinflammation;2 kinds of parenchymal necrosis including geographic necrosis and microabscess.13 kinds of histopathologic features in 3 major manifestations were found from 2 autopsies,but various kinds histopathologic features presented in small biopsy samples.Minor manifestations such as diffuse pulmonary hemorrhage were found at the periphery of WG.Conclusions The wide variation and broad spectrum of pathologic features can occur in WG.Vasculitis,granulomatous inflammation and parenchymal necrosis are the most important histopathological features.The correct diagnosis of WG requires careful correlation of pathology with complicated clinical features.

Objective To investigate the therapeutic effect of tumor necrosis factor (TNF)-αsiRNA on typeⅡcolla-gen induced arthritis (CIA) in rats. Methods The expression vectors of siRNA against TNF-αgene were constructed suc-cessfully and were injected by tail veil into CIA rats. Twenty-four CIA rats were randomly divided into 4 groups including model group, empty vector group, TNF-α-siRNA1 group and TNF-α-siRNA2 group. CIA rats were injected with the same dose of phosphate buffered sodium (PBS) and pGFP-V-RS vector respectively in model group and empty vector group, while TNF-α-siRNA1 group and TNF-α-siRNA2 group were injected with TNF-α-siRNA1 eukaryotic expression vector and TNF-α-siRNA2 eukaryotic expression vector respectively. Another 6 rats, which were not established CIA model, were in-jected with PBS (blank control group). The serum expression levels of IL-1 were detected by ELISA on day 1, 5, 9 and 13 af-ter injection. The expression level of TNF-αmRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) on day 13. Results The expression level of IL-1 was significantly higher on day 1, 5, 9 and 13 in model group than that of blank group (P＜0.05). The expression levels of IL-1 were significantly lower on day 1, 5 and 9 in TNF-α-siRNA1 group and TNF-α-siRNA2 group than that of model group and blank group (P ＜ 0.05). The expression level of TNF-αmRNA was significantly higher on day 13 in model group than that of blank group (P＜0.05). The expression levels of TNF-αmRNA were significantly lower in TNF-α-siRNA1 group and TNF-α-siRNA2 group than those of model group and emp-ty vector group (P＜0.05). Conclusion TNF-αspecific siRNA can suppress the levels of TNF-αmRNA and IL-1, which provides experimental basis for gene therapy of rheumatoid arthritis.

Objective To investigate the expression of tumor necrosis factor alpha(TNF-α)and its relationship with infiltrating lymphocytes in lichen planus(LP).Methods Tissue specimens were obtained from the lesions of 60 patients with LP and normal skin of 20 human controls.Immunohistochemical SP method was used to detect the expression of TNF-α,and infiltrating lymphocytes were counted in TNF-α-positive tissue sections.Results TNF-α was expressed in 72％ of the LP specimens but in none of the control specimens(P ＜ 0.01).Positive staining for TNF-α was mainly located in the membrane of prickle cells,cytoplasm or membrane of dermal infiltrating lymphocytes.The expression of TNF-α in LP was uncorrelated with age,sex or disease course(all P ＞ 0.05),but was positively correlated with infiltrating lymphocyte number (rs =0.47,P ＜ 0.01).Conclusion TNF-α seems to play a certain role in the pathogenesis of LP.

Aim To discover specific telomerase inhibitors by high-throughput screening in the more than 2 000 strains of actinomycetes fermentation broth.Methods After verifying the three kinds of screening models for telomerase inhibitor by yeast PCR,the three yeast strains were inoculated overnight into yeast liquid culture which was lacking in histidine and contained 3-AT.Then concentration of yeast strains in culture liquid and dosage of screening sample were regulated for better adjustment to the high-throughput screening.At last the adjusted liquid culture containing yeast strain piped to 96-well plates,drug samples and control article were also added,then the yeast was incubated in the thermostat oscillator.At the same time OD_(595nm) of the yeast liquid was measured every 12 h by using Multifunctional Microplate Analyzer,and the samples whose inhibitory efficiency was over 0.45 were selected for secondary screening.Results The high-throughput screening method related to yeast three-hybrid system was effective in screening samples that had potential telomerase inhibitory ability.The initial concentration of yeast liquid OD_(595nm) was 0.04 and the best dosage of samples was 5 ml.14 active samples,whose inhibitory efficiency was over 0.45,were founded after primary and secondary screening.Conclusions High-throughput screening method related to yeast three-hybrid system is simple and stable in discovering telomerase inhibitors,and 14 new antitumor compounds are identified,which lays the foundation in the development of new anti-tumor agents.

Objective To study the histopathological features of hepatic portal area in autopsy specimens of elderly men with type 2 diabetes mellitus (T2DM).Methods One hundred and three autopsy cases with T2DM (diabetic group) and contemporary 48 non-diabetic cases matched by gender and age (control group) were selected in the study.The histopathological changes of hepatic portal area were observed by histochemical and immunohistochemical methods.Results (1) In diabetic group, endothelial denudation, eosinophilie deposits, eccentric intimal thickening and luminal stenosis were found in hepatic interlobular artery (arteriole, the lumen diameter < 100 μm).The medial smooth muscle cells proliferated and migrated into intima, and fibroplasia occurred in adventitia.62.1% (64/103) of the patients with T2DM had microangiopathy and there was statistically significant difference between the two groups (χ2= 27.14, P< 0.01 ).(2) Hyaline collagen fiber tissues deposited around interlobular artery, interlobular vein and interlobular bile ducts, resulting in enlargement of the portal area and the secondary atrophy and disappearance of portal triad.65.0% (67/103) of the patients with T2DM had this pathological changes in the portal area and there was statistically significant difference between the two groups(χ2= 23.27,P<0.01).(3) Terminal portal area was notable.Conclusions Mieroangiopathy can be observed in the hepatic portal areas of elderly men with T2DM, which may induce the fibrotic tissue hyperplasia, the atrophy of the portal triad and the sclerosing portal area.

Objective To observe the incidence rate of non-alcoholic fatty liver disease (NAFLD) in elderly male patients with type 2 diabetes mellitus (T2DM) and the features of pathological changes of liver. Methods The 89 cases with T2DM (T2DM group) and contemporary non-diabetic cases (control group, n=48) from 858 autopsy samples matched for gender and age were selected in the study. The histopathological changes of liver were observed by microscopy,histochemistry and immunohistochemistry. Results The incidence rates of NAFLD and nonalcoholic steatohepatitis(NASH) were 49.4% (44/89) and 14.6% (13/89) in T2DM group, 22.9 %(11/48) and 4.2% (2/48) in control group, respectively. The incidence rates of NAFLD and NASH were significantly higher in T2DM group than in control group(P＜0.01, P＜0.05). All of the NASH patients from T2DM group and one patient from control group were complicated with different stages of liver fibrosis. One patient from T2DM group was diagnosed as cirrhosis, no cirrhosis patient was found in control group. Conclusions Compared with control group, the incidence rate of NAFLD is higher and the damage of liver is more severe in T2DM group. The ratio of the progress from NAFLD to liver cirrhosis is low.

[ ABSTRACT] AIM:To observe the changes of perilipin and adipose differentiation-related protein ( ADRP) du-ring the development of diabetes mellitus and to explore the effect of perilipin and ADRP on abnormal glucose metabolism with non-alcoholic fatty liver disease ( NAFLD) .METHODS:The rat model of impaired glucose tolerance ( IGT) was in-duced by feeding high-fat diet, and the type 2 diabetes mellitus (T2DM) model was induced by feeding high-fat diet for 4 weeks and intraperitoneally injecting streptozotocin.The morphological change of the liver tissue was observed under optical microscope.The serum contents of perilipin and ADRP were measured by ELISA.The mRNA expression of perilipin and ADRP in the liver tissues was detected by real-time PCR.The protein expression of ADRP in the liver tissues was deter-mined by Western blotting.RESULTS:HE staining showed steatosis in the liver of the rats in IGT group was more serious than that in T2DM group.The biochemical and the pathological processes of rat models were consistent with the clinical feature of related diseases.The serum content of perilipin had no difference among various groups.The mRNA expression of perilipin in IGT group and T2DM group was significantly higher than that in control group.Compared with IGT group, the mRNA expression of perilipin in T2DM group was significantly increased.The serum content of ADRP in T2DM group was significantly lower than that in control group.The mRNA and protein expression of ADRP in model groups was signifi-cantly lower than that in control group.Compared with IGT group, the mRNA expression of ADRP in T2DM group was sig-nificantly reduced.CONCLUSION: The serum content of ADRP plays a role in the development and progression of T2DM.It is negatively correlated with HOMA-IR.NAFLD occurs during progression of abnormal glucose metabolism in-duced by feeding high-fat diet.The development of abnormal glucose metabolism with NAFLD is probably related to the in-creased expression of perilipin and the reduced expression of ADRP.

ObjectiveTo explore the short-term curative effect and safety of autologous bone marrow mesenchymal stem cells transplantation in patients with primary brain stem injury.MethodsFifty-four cases with primary brain stem injury were hospitalized during Jul.2007 to Jul.2010 at Liaocheng Brain Hospital,Shandong Province.All cases were randomized into transplantation group( n =30)or control group( n =24 ).The transplantation group was treated with autologous bone marrow mesenchymal stem cell transplantation by subarachnoid space injection (n =30).The control group were selected from primary brain stem injury patients without stem cell transplantation who were hospitalized at the same period with patients from the transplantation group.Respectively,National Institutes of Health Stroke Scale (NIHSS) score was employed to evaluate the condition of patients in the two groups one month after treatment,and Glasgow Outcome Scale (GOS) score was used to evaluate curative effects of the two groups at sixth months after treatment.Meanwhile,some other parameters were observed,including blood routine,clotting mechanisms,biochemicalitemsand tumor markers.ResultsThere was significant difference between the transplantation group and the control group in N IHSS score at one month after treatment [ ( 10.86 ± 7.48) vs.( 18.26 ± 8.74),t =2.681,P ＜ 0.05 ].GOS score was significantly different( Z =2.306,P ＜ 0.05 ) between the transplantation group and the control group at sixth month after transplantation.There was no significant difference between the two groups in the blood examination results during the followed-up period.Conclusion Autologous bone marrow mesenchymal stem cells transplantation is confirmed to be an effective and safe therapy in patients with primary brain stem injury in the short-term.Further evaluation still needs for its long-term efficacy on primary brain stem injury

ObjectiveTo compare the short-term efficacy and safety of autologous bone marrow mesenchymal stem cells transplantation on patients with early spinal cord injury by subarachnoid injection and by intravenous injection.MethodsNinety-six patients with early spinal cord injury were hospitalized and treated from November 2006 to March 2010.Thirty-eight cases (subarachnoid transplantation group) got transplantation by subarachnoid injection,32 cases (intravenous transplantation group) got transplantation by intravenous injection,26 cases (control group) were hospitalized in the same period but not transplanted.The motor and sensory functions of all three groups were evaluated according to the score standard developed by American Spinal Injury Association(ASIA) before treatment and at the first,the third,the sixth month after treatment.Meanwhile,routine blood test,coagulation,biochemical items and tumor markers were also examined in follow-up.ResultsThe motor and sensory function of three groups had different degree of recovery at the first month after treatment,and sensory function recovered muchsignificantly,but the comparison among three groups had no statistical significance.The scores of motor function increased in three groups at the third month after treatment,but still had no statistical significance (P＞ 0.05).The scores of sensory function of subarachnoid transplantation group[(130.9 ±41.6) scores] and intravenous transplantation group [ (131.2±22.7 ) scores ] increased obviously,and had significant difference compared with control group [ (109.3±36.4) scores] (P ＜ 0.05),but there were nosignificant difference between subarachnoid transplantation group and intravenous transplantation group (P ＞ 0.05).The scores of sensory and motor function of control group didn't increase obviously at the sixth month after treatment,while the scores of subarachnoid transplantation group and intravenous transplantation group gradually improved and had statistical significance compared with control group(P ＜ 0.05).The scores of sensory function was higher in subarachnoid transplantation group[ (151.6±46.9) scores ] than that in intravenous ransplantation group [(134.6 ±40.7) scores] (P ＜0.05).There were no obvious abnormality in the results of followed-up examination.Conclusions The safety and short-term efficacy of autologous bone marrow mesenchymal stem cells transplantation in treating early spinal cord injury by subarachnoid injection and intravenous injection is certified.The subarachnoid injection is better than intravenous injection,but the long-term efficacy need furter study.

AIM: To reproduce hypercapnic models and approach some pathophysiological changes in rats. METHODS: The mixed gases of high concentrated carbon dioxide (8% CO 2, 21% O 2, 71% N 2) were given to wistar rats 7 hours a day for 28 days. The various indexes were compared between control group (group A) and hypercapnic group (group B). RESULTS: The PaCO 2 [(55.90?4.34) mmHg] and the lipid peroxides (LPO) contents in plasma, lung tissue and right ventricle were significantly higher in group B than those in group A ( P