Objective To study the effect of 99Tc-MDP on receptor activator of nuclear factor-κB ligand/osteoprotegerin (RANKL/OPG) system of collagen-induced arthritis (CIA) rat. Methods The CIA was established by subcutaneous injection with type Ⅱ collagen and complete Freud's adjuvant to rats, except the group (n=8, treated with 99Tc-MDP). The histopathological changes of joints were scored based on the extent of infiltration of inflammatory cells, cartilage destruction and bone erosion. The serum levels of RANKL/OPG were tested with ELISA, and the synovial membrane levels of RANKL/OPG were tested with Western blot.Repeated ANOVA and one way ANOVA were used for statistical analysis. Results The MTX group relative value of RANKL/OPG in serum [35 d (1.076±0.016), 42 d (1.140±0.005), 49 d (1.155±0.023)], and in synovial membrane (1.156±0.014), and the histopathological scores [synovial membrane (1.8±0.5), cartilage (1.9±0.6), bone (1.8±0.5) ] were lower than model group ( 1.269±0.025, 1.296±0.015, 1.340±0.011 ), (2.9±0.4, 2.6±0.5, 2.6±0.5), ( 1.340±0.013 )(P＜0.01 ). And all above index of 99Tc-MDP group ( 1.035±0.034,0.986±0.019, 0.991±0.020), (1.5±0.5, 1.4±0.5, 1.2±0.5), (0.098±0.026) were notably lower than the MTX treat ment group (P＜0.01). Conclusion 99Tc-MDP may retard the progression of the disease by decreasing the relative RANKL/OPG expression and it can initiate its effect earlier than MTX.

This study was aimed to detect the expression of c-Jun N-terminal kinase (JNK) in renal tissues of lupus nephritis (LN) mice with chronic graft-versus-host disease (cGVHD) and to investigate that LN could be intervened by fluvastatin at different doses through the inhibition of JNK expression.LN models with cGVHD in mice were established first,and then diseased mice were randomly divided into four groups:normal control,fluvastatin intervention at high-dose group (10 mg/kg),fluvastatin intervention at low-dose group (5 mg/kg) and models without treatment.After killing the mice sixteen weeks later,total urine protein of every mouse was determined by a Biuret colorimetric assay.The protein and mRNA levels of JNK and p-JNK in kidneys were measured by immunohistochemistry,Western blot and RT-PCR,respectively.Compared with normal control,otal urine protein,JNK and p-JNK expressions at both mRNA and protein levels were significantly increased in cGVHD group (P ＜0.01),but their expressions were suppressed by fluvastatin treatment (P ＜0.01).JNK may play an important role in the pathogenetic progress of LN in mice,and fluvastatin is able to prevent LN via inhibition of JNK expression in renal tissues in cGVHD mice.

Objective:To establish a bacterial endotoxins testing method for L-oxiracetam injection. Methods: According to the gel-clot technique for bacterial endotoxin inspection and the related regulations in Chinese Pharmacopoeia (2015 edition, volume Ⅳ, general rule 1143), tachypleus amebocyte lysate (TAL) from two different manufacturers were used for bacterial endotoxins test on 3 batches of L-oxiracetam injection with methodology studies. Results:Diluted to 8. 3 mg·ml-1 , the samples showed no interference a-gainst the bacterial endotoxins test. Conclusion:The bacterial endotoxins test method for L-oxiracetam injection is applicable with the endotoxin limit of 3 EU·ml-1 .

This study is to prepare the pantoprazole sodium enteric-coated tablet which is compacted by pellets. The enteric-coated pantoprazole sodium pellets were prepared by fluid bed coating technology. The pantoprazole sodium enteric-coated tablets were prepared by direct compression of the enteric-coated pellets and suitable excipients. In vitro dissolution method and scanning electron microscope method were used for the observation of the drug release behavior before and after compression of the pellets. The optimized formulation is: the coating level is 55%, the plasticizer content is 20%, the ratio of Eudragit L30D-55/NE30D is 8 : 2, enteric-coated pellets/excipients (MCC/PPVP/PEG 6000 = 2 : 1 : 1) is 5 : 5, the enteric-coated tablets release in artificial gastric fluid in 2 h is less than 10%, while in artificial intestinal fluid in 1 h is more than 85%. The release behavior of pantoprazole sodium enteric-coated pellets-type tablet is quite well. And it may be used in industrial production.

The present study was designed to investigate the ultrastructural changes of Sertoli cells of rat testis following gossypol administration at a dosage of 30 mg/kg/day for 4～6 weeks. Results indicate that 4 weeks after gossypol treatment, a series of ultrastructural changes concerning with phagocytic activity such as increased in amount of lysosomes, lipid droplets, ring or cup shaped mitochondria as well as the multiform changes of mitochondria and lysosomes were evident in the Sertoli cell. However, by 6 weeks after treatment, the Sertoli cells became "inactive" and began to show degenerative changes: distension and vesiculization of endoplasmic reticulum, accumulation of lipid droplets, cellular debris and lysosomes in varying sizes and phases, and the occurrence of atrophic changes of mitochondria. The nature of these changes in Sertoli cells following gossypol treatment and its significance was discussed.

Objective To investigate interleukin 10 (IL 10,Th2 type cytokines) and interferon gamma (IFN ?,Th1 type cytokines) secreting capacity by peripheral blood mononuclear cells (PBMCs) in 30 patients with systemic lupus erythematousus (SLE) and 10 healthy individuals,and try to understand the role of Th1/Th2 balance in the pathogenesis of SLE.Method Sandwich ELISA was used to measure IL 10 and IFN ? level in the supernatant of culture from PBMCs.Results The level of IL 10 produced spontaneously by PBMCs in active stage was significantly higher than in static phase and healthy person ( P 0 05).Conclusion These data suggest that the Th2 type cytokine (IL 10) was increased in patients with SLE,and an imbalance towards Th2 predominance may play a significant role in the pathogenesis of SLE.Regulating IL 10 level may provide a new approach for the treatment of SLE.

Objective To investigate whether monocyte chemoattractant protein-1 mediates lupus nephritis (LN) in BXSB mice and whether methylprednisolone ameliorates LN by inhibiting MCP-1 expression. Methods 18-week-old male BXSB mice (n=6) displaying clinical symptoms of glomerulonephritis were treated (i.p.) for 3 weeks with MPS (25 mg?kg-1?d-1) dissolved in N.S. BXSB controls were age-and sex-matched BXSB mice (n=6) that received N.S. alone. Age- and sex-matched BALB/C mice (n=6) were used as normal controls that were treated in the same way as the BXSB controls. MCP-1 expression was investigated by RT-PCR and immunohistochemical examination. The heaviness of proteinuria was also evaluated. Medical imaging analysis was performed to detect the relationship between MCP-1 expression and proteinuria. Results MCP-1 was strongly expressed in kidneys of all BXSB mice, stronger staining was found in cytoplasm of tubular epithelial cells than glomerular, while no expression was found in kidneys of BALB/C mice. MCP-1 expression in tubular cells in BXSB control group was closely correlated with proteinuria. MPS significantly reduced proteinuria and MCP-1 expression and down-regulation of MCP-1 expression in tubular cells was also closely correlated with that of proteinuria. Conclusion MCP-1 over-expression may mediate LN in BXSB. MPS ameliorates LN partially by inhibiting MCP-1 expression.

Objective:To explore whether different phospholipid peptides could induce different pathological features of EAE in Lewis rats.Methods:Lewis rats were immunized with myelin basic protein 82-99 (MBP82-99),MBP68-86,and myelin oligodendroglia glycoprotein 35-55 (MOG35-55),respectively,and evaluated everyday for neurological scores.The cerebrum,brain stem,cerebellum and spinal cord of every rat were removed and pathological features observed.Results: Rats immunized with the two MBP peptides exhibited neurological signs of EAE wherein the central nervous system had extensive inflammation infiltration.The number of infiltrates in spinal cords in the two MBP peptides groups was higher than that in the cerebrums,brain stems and cerebellums.In spinal cords, there was no statistical difference of infiltrates between MBP82-99 and MBP68-86 groups;however,the both groups had more infiltrates than MOG35-55 group.Inflammatory cells were observed only in spinal cords of animals immunized with MOG35-55.Conclusion:This study provides certain evidence for understanding the diversity of pathological manifestations of EAE in Lewis rats.

Objective To explore the optimal time and sequence for getting the best magnetic resonance (MR) imaging when MR image of synovium macrophages was used for the diagnosis of collageninduced arthritis (CIA) in a rat model,and whether it can be used to monitor the efficacy of drug treatment.Methods CIA was induced by subcutaneous injection of chicken type Ⅱ collagen and complete Freund's adjuvant (CFA).Arthritis rats were randomly divided into the model group,the leflunomide group and the control group.Knees of the model group rats were imaged before and 24 h,48 h,72 h after USPIO intravenous administration (300 μmol Fe/kg) on day 28,29,30,31,respectively.From day 28,the leflunomide group was given a gauge of drug at a dose of 8 mg/kg.Then they were imaged before and 24 hours after USPIO administration on day 42,43 respectively.MR sequences included SE T1WI,SE T2WI,GRE T2 * WI.After the completion of MR imaging,rats were sacrificed to obtain histopathologic samples of synovial membrane.LSD-t test and one-way analysis of variance (ANOVA) were used for statistical analysis.Results No distinct signal enhancements were observed on the 24 h,48 h,72 h post-contrast enhancement on T1WI.On T2WI,signal intensity ratio of synovium (SNR) pre-contrast and 24 h,48 h post-contrast were 24.13±1.96,17.09± 1.23,19.14±0.91,respectively.On T2 * WI,SNR pre-contrast and 24 h,48 h post-contrast were 22.28±0.92,11.40±0.53,17.18±0.63,respectively.Distinct signal changes were observed on 24 h,48 h post-contrast on T2WI and T2 * WI (P＜0.05).The changes between SNR at 24 h,48 h,72 h post-contrast and pre-contrast were-29.09±2.42,-20.83±2.90,-6.2±2.9 respectively on T2WI,which were-48.4±1.3,-22.9±0.8,-8.2±1.6 respectively on T2 * WI.Changes were more obvious at 24 h post-contrast than 48 h post-contrast on both T2WI and T2 * WI (P＜0.05).The quantitative analyses were coinci-dent with the visual differences in signal changes between pre-contrast and post-contrast images.Difference between △SNR of leflunomide group and the control group on T1WI was not significant,while that on T2WI and T2 * WI were significantly different (P＜ 0.01).Histological examination confirmed the uptake of iron in the macrophages of arthritic knees.Signal intensity changed more on GRE T2 * WI than SE T2WI in all arthritis rats.Conclusion GRE T2 * WI is more sensitive for the diagnose of rat CIA,and 24 h post-contrast is better than 48 h and 72h post-contrast to get better images.We successfully observed the effects of leflunomide through signal changes of synovium,and the technique maybe contribute to diagnosis and therapeutic monito-ring of rheumatoid arthritis.

Objective To evaluate whether the timing of maternal hypoxia during pregnancy could myocardial remodeling in adult offspring.Methods Twenty-four pregnant rats were assigned to the maternal hypoxia group starting from the early period of pregnancy (G1,day 3 to 21 of pregnancy),the group starting from the middle period of pregnancy (G2,day 9 to 21 of pregnancy),the group starting from the late period of pregnancy (G3,day 15 to 21 of pregnancy),or the control group (G0).Six rats were included in each group.Rats in the maternal hypoxia groups (G1,G2 and G3) were subjected to hypoxia for 3 hours in a low pressure cabin with an oxygen concentration of (10± 1) ％.Systolic blood pressure,wet weight of the left ventricle normalized for body weight (left ventricular weight/body weight,LVW/BW),diameter of cardiomyocytes,and the expression levels of collagen Ⅰ and collagen Ⅲ were measured in adult male offspring at the age of 3 and 5 months.One-way ANOVA and SNK test were used for statistical analysis.Results Maternal hypoxia from day 3 to 21 of pregnancy induced higher systolic blood pressure in male offspring at the age of 3 months and 5 months [3 months:G1 (122.0± 11.7) mmHg,G0 (108.3±9.5) mmHg,F=2.97; 5 months:G1 (128.5±7.9) mmHg,G0 (114.6±-10.0) mmHg,F=3.17,both P＜0.05].Maternal hypoxia also led to a higher LVW/BW ratio (G1 1.99±0.03,G0 1.80±0.02,P＜0.05),and increased collagen Ⅰ and collagen Ⅲ expression (collagen Ⅰ /β-actin:G1 1.76±0.07,G0 0.48±0.04,P＜0.01; collagen Ⅲ /β-actin:G1 0.68±0.05,G0 0.34±0.03,P＜0.01) in adult male offspring at the age of 5 months.Maternal hypoxia from day 9 to 21 of pregnancy induced a higher LVW/BW ratio (3 months:G2 2.15 ± 0.05,G0 1.98 ± 0.02,P＜0.05; 5 months:G2 1.96±±0.05,G0 1.80±0.02,P＜0.05),and increased collagen Ⅰ and collagen Ⅲ expression (3 months:collagen Ⅰ /β-actin G2 0.98±±0.02,G0 0.87±0.02,P＜0.05; collagen Ⅲ /β-actin:G2 0.87±±0.01,G0 0.82±0.01,P＜0.05; 5 months; collagen Ⅰ /β-actin G2 1.61±0.05,G0 0.48±0.04,P＜0.01; collagen Ⅲ / β-actin:G2 0.61 ±0.04,G0 0.34±0.03,P＜0.01) in male offspring at the age of 3 months and 5 months,but did not affect systolic blood pressure.Maternal hypoxia from day 15 to 2l did not have any effect.The diameter of cardiomyocytes in male offspring was not significantly different between the groups.Conclusions Maternal hypoxia during different periods of pregnancy has different effects on cardiac structure in adult rat offspring.Maternal hypoxia started in early or mid-pregnancy leads to cardiac collagen deposition without an increase in myocyte size in adult offspring,which may be independent of the change in blood pressure.