Objective To investigate the clinical significance of serum lactate dehydrogenase (LDH) and its isoenzymes,M or H subunits in the retreated patients with multiple myeloma(MM).Methods 141 medical records of retreated patients with multiple myeloma were collected,and the relationship of LDH and its isoenzymes,subunits with DS staging system,ISS staging system,creatinine,bone marrow plasma cell percentage,immunoglobulin and microglobin was analyzed.Results Serum total LDH activity was not significantly related to DS staging system and ISS staging system.LDH1 percentages was negatively correlated with ISS staging system (P＜0.01),and LDH4 percentages positively associated with ISS staging system (P＜0.05).LDH activity,absolute value of LDH1 ,LDH2,LDH3,LDH4,and H subunits were positively linked to serum creatinine in myeloma kidney disease patients with renal inadequacy (P＜0.05).Both percentage and absolute value of LDH5 were negatively correlated with bone marrow plasma cell percentage (P＜0.01 and P＜0.05 respectively).Conclusion The detection of serum LDH isoenzyme is more valuable than total LDH activity in retreated patients with multiple myeloma.

It is reported by current investigations,that BNP can suppress heart hypertrophy but it can also depress compensatory hypertrophic response.BNP is an useful marker in the early diagnosis,prognosis,therapy guiding of heart failure and coronary heart disease,but comorbid conditions should be considered in the diagnosis of heart failure.The polymorphism in the 5'-flanking region of the NPPB gene is found to be related with essential hypertension.On the basis of some investigation,it is supposed that the instillation of BNP at the early stage of hypertension will contribute to therapy of hypertension.

Objective To study the effect of astragaloside on TGF-β1,SMAD2/3,and α-SMA expression in the kidney tissue of diabetic KKAy mice,and evaluate its potential role in renal interstitial fibrosis.Methods 20 type 2 diabetic KKAy mice were randomly divided into model group and astragaloside group,while 10 male C57BL/6J mice were selected as the control.Astragaloside at 40 mg · kg-1 · d-1 was given when the KKAy mice fed with high-fat diet to 14 weeks old.The mice in the control and model group received normal saline at 40 mg · kg-1 · d-1.Blood glucose meter was used to detect the blood glucose value of each mice at 16th,20th and 24th week.The mice were killed at 24 weeks old and the kidney tissue samples were collected.Pathology morphological changes were observed.Results (1) blood glucose value:cmpared with the control group,the blood glucose value of KKAy mice at 14 week increased significantly,and that of model group also increased significantly at 16th,20th and 24th week (P＜0.05);the blood glucose value of astragaloside group decreased compared with control group (P＜0.05).(2) Morphology of kidney:in the control group,the glomerular and tubular had clear structure,there was no renal interstitial fibrosis;in the model group,the renal glomerular mesangial matrix had broaden,mesangial cell had increased,renal tubular epithelial cell cytoplasm showed vacuole degeneration,renal interstitial inflammatory cell had increaised.In astragaloside group,there were few renal tubular epithelial cell cytoplasm,and there was no obvious fibrosis.(3)TGF-β1,SMAD2/3,and α-SMA expression levels of the kidney issuse:compared with control group,mice in model group up-regulated TGF-β1,SMAD2/3 and α-SMA expression (P＜ 0.05).TGF-β1,SMAD2/3,and α-SMA expression levels in astragaloside group were significantly lower than those in the model group (P＜0.05).There was few phosphorylated SMAD2/3 expression in renal tubular and glomerular nuclei,while that of model group increased (P＜0.01),and compared with model group,that of the astragaloside group decreased (P＜0.05).Conclusion Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1 and α-SMA expression,thus to relieve renal fibrosis in diabetic mice.

Objective To analyze the clinical manifestation of diffusive alveolar hemorrhage in acute leukemia induction therapy.Methods Clinical data of two diagnosed cases of diffusive alveolar hemorrhage secondary to acute leukemia were collected.Clinical data of eight cases of diffusive alveolar hemorrhage secondary to acute leukemia which were published were also collected by searching in Medline database.The clinical manifestation,diagnosis,strategy of differential diagnosis and treatment of diffusive alveolar hemorrhage secondary to acute leukemia were analyzed.Results Diffusive alveolar hemorrhage was a rare but fatal complication of acute leukemia.The common clinical manifestations included hemoptysis,progressive dyspnea and progressive decrease in concentration of hemoglobin.The analysis of blood gas showed type Ⅰ respiratory failure.The manifestations of chest computed tomography included diffusive ground glass opacity and infiltration of parenchyma.The bronchoalveolar lavage fluid was bloody.And lung biopsy showed congestion of alveoli and capillaritis.The detection for pathogens,vasculitis related antibodies,brain natrium peptide were negative.The mortality of those cases was 40 ％ (4/10).Corticosteroids therapy was effective.The mortality of patients received corticosteroids therapy was 25 ％ (2/8).Conclusion Diffusive alveolar hemorrhage is a rare but fatal complication of acute leukemia.The mortality is high.The key points of therapy are early diagnosis and corticosteroids therapy.

Objective To explore the role and possible mechanism of CD28/B7 molecules in T cell ab-normal activation by establishing a mouse model of the autoimmune aplastic anemia. Methods Unmanipulated B6D2F1 or CByB6F1 hybrid mice were infused with about 40 × 106 lymph node (LN) cells from their C57BL/6 (B6) parent. Distribution of the injected T cells was assayed by CFDA-SE fluorescent staining. Anti-D80 and anti-CD86 monoclonal antibodies were used to block CD28/B7 signal transduction pathways and to test the change of peripheral blood of F1 mice at different times. Damage was assessed by histological staining. Bone marrow (BM) cells and LN iymphocytes were cultured to observe the effect of different number of lymphocytes in the LN on BM cells' hematopoiesis by the count of hematopoietic colony-forming cells, and to test the effect of cyclosporine A of different concentration on BM cells' hematopoiesis. Results Infusion of about 40 × 106/mouse B6 LN cells led to the development of BM failure in the fifth day: anemia, neutropenia and thrombocytopenia. At 21st day recipients began to appear death. Frozen section revealed the injected lymph node major in myeloid tissue. Pathological sec-tion revealed obvious immune-induced marrow destruction and tissue destruction. There was similar performance of the above in the recipients infused with anti-D80 and anti-CD86 monoclonal antibodies. B6 LN five times the num-ber of lymphocytes in the blood cells F1, CFU-E and CFU-G colony formation of a blank group difference was not significant (P >0.05); When B6 LN 10 times the number of lymphocytes in the blood cells F1, CFU-E colony forming significantly reduce the number of (P < 0.05) ; When B6 LN lymphocytes 50 times in F1 hematopoietic cells, not observed CFU-E colony formation. CFU-E and CFU-G colony formation reducing the number of lympho-cytes showed a dose-dependent. Cyclosporine A can significantly increase the CFU-E and CFU-G colony forming rate. Conclusion This mouse model indicates that activated lymphocytes play important roles in marrow destruc-tion in lymphocyte infusion-induced BM failure. Only blocking the CD28/B7 signal transduction can not block the abnormal T-cells activated.

Objective To assess the concordance of MRI diagnosis for patients suspected of lumbar disk herniation by using Kappa statistic.Methods One hundred patients(48 males and 52 females)with lumbosaeral radicular pain,aged from 17 to 86(average 61).All patients underwent fast spin-echo T1 and T2 weighted imaging on a 3.0 T MR scanner and spine surface coil.Two radiologists(doctor A and doctor B)evaluated the lumbar disks from L3-4,L4-5.and L5-S1 in 50 out of the 100 patients independently.The presence of a bulging disk or a herniation was reported.Images were interpreted twice:once before and once after disclosure of clinical information.And disks of 52 patients out of the 100 samples were interpreted by the two radiologists independently without clinical information as well.The Kappa statistics was employed to assess the concordance of each radiologist's diagnoses as well as the observer variation of the two radiologists.Results Diagnoses before and after disclosure to clinical information were concordant in 114 disks for doctor A and in 109 for doctor B.respectively.Diagnoses before and after disclosure to clinical information were not concordant in 36 disks for doctor A and in 41 disks for doctor B,respectively.The Kappa values were 0.60±0.06 and 0.57±0.06 for doctor A and doctor B,respectively.The concordance was moderate.After disclosure to elinical information.the numbers of reported bulging disks increased significantly.by 10 and 31 for doctor A and doctor B,respectively.Without clinical information,the diagnoses of the two radiologists were concordant in 77 disks,while not concordant in 79 disks.The interobserver agreement was poor(Kappa=0.24±0.06).The diffcrence on diagenoses made between with and without clinical information mainly happened on the differential diagnosis of normal disks and bulging disks.The different,diagnoses made between with and without clinical information were on 20 disks and on 30 disks for doctor A and doctor B,respectively;that accounted for 55.6%(20/36)and 73.2%(30/41)of total variation respectively.The diagnostic difference between the 2 doctors happened mainly on differentiation of bulging disks and normal disks,which happened in 56 disks,aceountiong for 70.9%(56/79)of total variation.Conclusion Variation on diagnoses of the same radiologist or between tworadiologists was mainly caused by disagreement on bulging disks.

AIM:To study the effects of transforming growth factor-β_1 (TGF-β_1) on cell apoptosis,cell cycle,production of endogenous TGF-β_1,expressions of P27~(Kip1),cyclin E and bcl-2 mRNA levels in NB4 cells. METHODS:Apoptotic morphological changes were observed by Wright-Giemsa staining. Cell cycle and apoptosis were detected with flow cytometry. Semiquantitative RT-PCR was used to examine the mRNA levels of endogenous TGF-β_1,P27~(Kip1),cyclin E and bcl-2. RESULTS:TGF-β_1 significantly restrained the growth and promoted the apoptosis of NB4 cells. The blockage of NB4 cells treated by TGF-β_1 at concentration of 5 μg/L was in G1 phase. Endogenous TGF-β_1 mRNA expression in NB4 cells was up-regulated when the concentration of exogenous TGF-β_1 was <5 μg/L. Meanwhile,the expression of endogenous TGF-β_1 mRNA was down-regulated when the concentration of exogenous TGF-β_1 was 10 μg/L. After treated with TGF-β_1 at concentration of 5 μg/L,P27~(Kip1) mRNA expression in NB4 cells was up-regulated,cyclin E and bcl-2 were reduced. CONCLUSION:TGF-β_1 is able to induce apoptosis and cell cycle distribution abnormally in NB4 cells by (1) Up-regulation of endogenous TGF-β_1,so that NB4 cells was induced into apoptosis through consequently high expression of P27~(Kip1). (2) TGF-β_1 may lead to cell cycle arrest by inhibiting the expression of cyclin E directly,or by inhibiting the activity of cyclin E through the increased expression of P27~(Kip1). (3) Down-regulation of bcl-2 induces apoptosis of NB4 cells.

Objective To optimize several key factors in preparing protein microarray,and establish microarrays to detect autoantibodies.Methods Four factors,including incubation time of the microarray after spotting,the blocking reagent,the length of blocking time,the length of time conjugating with the second-an- tibody,were selected to carry out orthogonal optimization,then we determined the optimal spotting concentra- tion of antigen and the best dilution of serum with the optimized conditions obtained from the above.On the basis of the optimized conditions,we prepared microarray to detect six autoantibodies simultaneously,which were compared with IIF and ELISA.Results The best conditions we determined from the experiment were: incubating the spot microarray for 120 min,PBST containing 1% BSA and 2.5% saccharose as the blocking reagent and blocking for 30 min,reacting with the second-antibody for 30 min,the appropriate spotting con- centrations of six antigens differed from 100?g/ml to 300?g/ml and the best serum dilution was 1:2.Com- pared with IIF for ANA detection,the sensitivity of the microarray was 88.6%,the specificity was 83.3%,and compared with ELISA in detecting the other five autoantibodies,the sensitivity of the microarray was between 80.0% and 88.2%,and the specificity was between 90.2% and 98.6%.Conclusion The optimization condi- tions we obtained are suitable for preparing protein microarray,and the detection sensitivity and specificity of autoantibodies by microarray are consistent with the conventional methods in general.The microarray for de- tecting autoantibodies is applicable in the clinical diagnosis of autoimmune disease.

The secondary structures of fusion protein pp150/MDBP, including alpha-helix, beta-sheet, turn regions, were analyzed by Garnier-Robson's and Chou-Fasman's methods; the antigenic epitopes of B cells were analysed by using hydrophilicity plot. The results showed that the fusion protein pp150/MDBP might have less alpha-helix, but be rich in beta-sheet and turn regions. The epitopes recognized by B cells may be at 7-56 amino acid residues or adjacent to 137-192 amino acid residues.
Amino Acid Sequence ; Binding Sites ; Cytomegalovirus ; chemistry ; Epitopes ; Humans ; Molecular Sequence Data ; Phosphoproteins ; chemistry ; immunology ; Protein Structure, Secondary ; Viral Fusion Proteins ; chemistry ; immunology ; Viral Matrix Proteins ; chemistry ; immunology

We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solution, spotting concentration of the 12 antigens and the dilution of serum. We prepared a protein chip that could detect 12 autoantibodies simultaneously using the optimized antigen concentration. We established a new method to determine the cutoff of each autoantibodies by evaluation of 678 positive and 120 negative serum of clinical sample. We also evaluated the sensitivity and specificity of our new detection system. The optimal spotting solution was 0.1% TBST, the dilution of serum was 1:4 and the best spotting concentration of the 12 antigens were ANA 520 microg/mL, Ro-60/SSa 465 microg/mL, La/SSb 530 microg/mL, Jo-1 530 microg/mL, Scl-70 525 microg/mL, Sm 520 microg/mL, Ro-52/SSa 615 microg/mL, RF 340 microg/mL, CCP 465 microg/mL, ulRNP 410 microg/mL, CENP-B 490 microg/mL and dsDNA 580 microg/mL respectively. It had higher coincidence rate compared to current clinical used methods. We have developed a 12 antigens protein chip for the detection of autoantibodies based on the NBT/BCIP color reaction system. This system was fast, convenient, efficient, and cost-effective.
Autoantibodies ; blood ; immunology ; Autoimmune Diseases ; blood ; immunology ; Humans ; Protein Array Analysis ; instrumentation ; methods ; Sensitivity and Specificity