Determination of hesperidine inErchen mixture by HPLC / 国际中医中药杂志

Objective To establish a method for the determination of hesperidine inErchen mixture. Methods HPLC method was used with Fortis C18(250 mm×4.6 mm,5μm)column and the mobile phase consisted of acetonitrile-acetic acid-water(22∶4∶74). The flow rate was 1.0 ml/min,the column temperature was 30℃, and the UV detector was set at 283 nm.Results The linear ranges of hesperidine was 2.86-57.20μg(r=0.999 8). The average recovery of Aloin was 99.4%.Conclusion The method is rapid, reliable, simple, repeatable and with high specificity. It can be an important supplement for Erchen mixture elevated standard.

Neurogranin overexpression on peripheral blood mononuclear cells from patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the mRNA expression level of neurogranin on peripheral blood mononuclear cells(PBMCs)from patients with systemic lupus erythematosus(SLE).Methods Top 20 tags of SLE PBMCs SAGE library were searched from normal lymphocytes SAGE library including navie-T,Th1,Th2, CD8+T, NK and B cells,and their abundance was compared.The mRNA expression level of neuro-granin,a differential over-expressed tag,was detected in 35 cases of SLE and 15 normal controls by reversetranscription-polymerase chain reaction (RT-PCR).Results Neurogranin tag could only be detected in SLE PBMCs SAGE library,but was hardly found in normal lymphocyte SAGE library.However,either SLE pa-tients or normal controls showed a detectable mRNA level of neurogranin on PBMCs by RT-PCR.The mRNA level of neurogranin in active SLE patients was significantly increased than those in controls(P＜0.001).but only slightly increased in inactive SLE patients (P＞0.05).Conclusion Neurogranin,as a novel proapototic factor,is overexpressed on PBMCs of SLE patients.It may be involved in the regulation of abnormal immune responses in lupus.

Expression of proliferation inducing ligand and its receptors in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the mRNA expression of a proliferation inducing ligand (APRIL) and its receptors including B cell maturation antigen (BCMA),transmembrane activator.calcium modulator and cyclophilin ligand interactor (TACI) in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SEE).Methods APRIL mRNA、BCMA mRNA and TACI mRNA in PBMCs were detected by real-time quantitative PCR in 66 SLE patients and 25 normal controls.Gene expression level was measured as 2-AACT.Results The expression levels of APRIL mRNA、BCMA mRNA and TACI-mRNA were significantly increased in both active SLE group and stable SLE group compared with those in the normal controls(P<0.01 for all).The expression levels of APRIL mRNA and TACI mRNA in active SLE group were significantly higher than those in stable SLE group(P<0.01,P<0.05,respectively).But there was no significant difierence in the expression levels of BCMA mRNA between the SLE stable and active groups-Beside,the expression levels of APRIL mRNA and TACI mRNA were significantly increased in patients with lupus nephritis (LN) compared to patients with non-LN (P<0.01 for all).Conclusion The expression levels of APRIL and its receptors are significantly elevated in SLE patients.It may suggest that APRIL and its receptors play an important role in the pathogenesis of SLE.

Study of the change of large arterial compliance and carotid artery intima-media thickness in patients with metabolic syndrome / 中国医师进修杂志

Objective To investigate the changes of compliance in large arteries and carotid artery intima-media thickness(IMT)in patients with metabolic syndrome.Methods There were 64 patients with metabolic syndrome and 56 age-matched control subjects.Their carotid-femoral pulse wave velocities(C-FPWV)were measured by the Complior SP and their carotid artery IMT were detected by B-mode ultrasound.At the same time their height,weight,waist circumstance,hip girth,blood pressure,blood glucose,blood lipid,BMI and waist to hip ratio(WHR)were measured.Results Compared with the control,the patients with metabolic syndrome had higher C-FPWV(P

Establishment of stable high expression cell line with green fluorescent protein and resistance genes / 华中科技大学学报（医学）（英德文版）

In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

Establishment of Stable High Expression Cell Line with Green Fluorescent Protein and Resistance Genes / 华中科技大学学报（医学）（英德文版）

In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

Expression of costimulatory molecules B7/CD28 in systemic lupus erythematosus / 华中科技大学学报（医学）（英德文版）

The expression of the costimulatory molecules B7/CD28 in peripheral blood mononuclear cells (PBMC) of the patients with systemic lupus erythematosus (SLE) and its relation to the pathogenesis of SLE were studied. The expression of the costimulatory molecules in PBMC in 30 patients with active SLE and 20 cases of healthy controls was detected by using the techniques of immunofluorescence and flow cytometer. The result showed that the expression percentage of CD28+, CD4+ CD28+ in T cells of PBMC from the patients with SLE decreased significantly as compared with that in healthy control group, while the expression percentage of CD80+, CD19+ CD80+ in B cells was significantly increased than that in healthy control group (P<0.01). It suggested that the abnormal expression of costimulatory molecules B7/CD28 played a role in the pathogenesis of SLE.

Leflunomide, a new disease-modifying drug for treating active rheumatoid arthritis in methotrexate-controlled phase II clinical trial / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of leflunomide in comparison with methotrexate (MTX) on patients with rheumatoid arthritis (RA) in China.</p><p><b>METHODS</b>Five hundred and sixty-six patients with active rheumatoid arthritis were randomly assigned to receive leflunomide at 20 mg once daily or MTX at 15 mg once weekly in a controlled trial. Five hundred and four patients completed the 12-week treatment and some patients continued the treatment for 24 weeks.</p><p><b>RESULTS</b>Both leflunomide and MTX could improve the symptoms, signs, and joint function, but there were no changes in X-ray observations of patients with rheumatoid arthritis. In the leflunomide group, the overall rates of effectiveness at 12 weeks and 24 weeks were 86.94% and 92.31% respectively; the rates of remarkable improvement were 64.95% and 79.81% respectively. In the MTX group, the overall rates of effectiveness at 12 weeks and 24 weeks were 84.04% and 83.15% respectively; the rates of remarkable improvement were 56.81% and 75.28% respectively. According to intent-to-treat analysis, the ACR 20% response rates at 12 weeks and 24 weeks in the leflunomide group were 62.54% and 67.18% respectively, compared with 60.08% and 61.32% respectively in MTX group. No statistical differences were shown in the efficacy between the two groups (P > 0.05). The adverse events in the leflunomide group were gastrointestinal symptoms, skin rash, alopecia, nervous system symptoms, decreased leukocyte count, and elevation of alanine aminotransferase (ALT). Most of these side effects were mild and transient. The incidence of adverse events in the leflunomide group was 16.84%, significantly lower than that in MTX group (28.17%, P = 0.002).</p><p><b>CONCLUSIONS</b>Leflunomide is effective in the treatment of RA with less adverse events than MTX. Its efficacy is similar to MTX, but the incidence of adverse events and the rate of withdrawal due to adverse events were lower in the leflunomide group than in MTX group.</p>

Autologous peripheral blood stem cell transplantation for systemic lupus erythematosns / 中华风湿病学杂志

20?10~9/L.The proteinuria decreased or disappeared.The antinuclear antibody decreased or became negative.The level of complement was increased.The following complications were ob- served:septicemia in 2 patients,cytomegalovirus infection in 2 patients,renal toxicity in 1 patient,acute left heart failure in 3 patients and cardiac arrhythmia in 3 patients.There was no transplantation related mortality. Conclusion APBSCT may improve the disease activity and the immunological markers in SLE.It is a valid therapy for refractory SLE,but the long-term effects need to be observed.

Immunophenotyping of Lymphocyte T and B in the Peripheral Blood of Systemic Lupus Erythematosus / 华中科技大学学报（医学）（英德文版）

The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus (SLE) were analyzed by using the immunofluorescence labeling-flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD+3, CD+4 and the ratio of CD+4/CD+8 in the peripheral blood of these patients were decreased (P<0.01), while the expression of CD+8, CD+20 was significantly increased (P<0.01). It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.

Immunophenotyping of Lymphocyte T and B in the Peripheral Blood of Systemic Lupus Erythematosus / 华中科技大学学报（医学）（英德文版）

The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus (SLE) were analyzed by using the immunofluorescence labeling-flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD+3, CD+4 and the ratio of CD+4/CD+8 in the peripheral blood of these patients were decreased (P<0.01), while the expression of CD+8, CD+20 was significantly increased (P<0.01). It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.

Study on the Cardiac Damage in the Patients with Polymyositis/Dermatomyositis / 华中科技大学学报(医学版)

To investigate the significance of cardiac damage in the patients with polymyositis (PM)/dermatomyositis (DM)，45 cases of PM/DM in our hospital from 1992 to 1998 were retrospectively analyzed in the aspects of the clinical data，muscle zymogram，X-ray plain films，electrocardiograph，ultrasonic cardiogram. It was found that cardiac damage (51.1 %) occurred in 23 out of 45 cases of PM/DM. Pericardium，cardiac muscle，endocardium and coronary artery were involved. Clinical manifestations were various. Among the cases of cardiac damage，3 died of heart failure. It was indicated that cardiac damage is the most common complication and also one of the common death causes of PM/DM.