Objective To investigate the relationship between the level of tumor necrosis factor (TNF)? and the clinical features of multiple sclerosis(MS). Methods The TNF? level of 58 MS patients (MS group) was determined by means of sandwich enzyme-linked immunosorbent assay (ELISA). The correlative analysis between level of TNF? and the Expanded Disability Status Scale (EDSS), the clinical data (including the duration of disease,the number of relapses and the age of onset) were take in MS patients. Results (1)TNF? level was significantly increased in MS acute stage compared with those in MS remission stage and healthy controls (P

[ ABSTRACT] AIM:To explore the effect of sterol regulatory element-binding protein 2 ( SREBP-2) on tunicamy-cin-induced endoplasmic reticulum stress ( ERS) in chondrocytes.METHODS:After isolation of human normal chondro-cytes and osteoarthritis ( OA) chondrocytes, the normal cells were cultured and treated with tunicamycin and SREBP-2 siR-NA.After 24 h treatment, fluorescent quantitative RT-PCR ( RT-qPCR) was applied to quantify microRNA-185 ( miR-185) levels.The cell apoptotic rate was determined by flow cytometry.The expression of SREBP-2 and ERS-related pro-teins, C/EBP homologous protein (CHOP), phosphorylated eukaryotic initiation factor-2α(p-eIF2α) and activating tran-scription factor 4 (ATF4), and the expression of apoptosis-related proteins, Bcl-2, Bax and caspase-3, were determined by Western blot.The caspase-3 activity kit was used to determine the caspase-3 activity.RESULTS: Compared with hu-man normal chondrocytes, both SREBP-2 up-regulation and miR-185 down-regulation were observed in OA chondrocytes (P<0.05).SREBP-2 siRNA transfection enhanced tunicamycin-inhibited miR-185 level (P<0.05).miR-185 overex-pression reduced tunicamycin-induced SREBP-2 expression ( P <0.05 ) .OA control group and tunicamycin treatment group consistently resulted in ERS and cell apoptosis with concomitant enhancement of CHOP, p-eIF2αand ATF4 proteins, increases in Bax and caspase-3 proteins, and reduction of Bcl-2 (P<0.05).However, SREBP-2 silencing significantly re-versed these effects ( P<0.05) .The apoptotic rates were consistent with the expression tendency of apoptosis-related pro-teins (P<0.05).SREBP-2 siRNA transfection markedly down-regulated tunicamycin-induced caspase-3 activity, which was notably blocked by miR-185 inhibition (P<0.05).CONCLUSION:SREBP-2 silencing may inhibit tunicamycin-in-duced ERS and cell apoptosis via up-regulating miR-185 expression.

ObjectiveTo investigate the effects of caffeine citrate (CC) on the neuronal proliferation and apoptosis and long-term learning ability in neonatal rats with hypoxia-ischemic brain damage (HIBD).MethodsForty-eight 7-day-old Sprague-Dawley neonatal rats were randomly divided into 3 groups: sham operation group (n=16), HIBD group (n=16), HIBD + caffeine citrate group (CC group,n=16). Rats in HIBD and CC groups received ligation of left common carotid artery followed by 2 hours of hypoxia to establish HIBD model. Rats in CC group were injected intraperitoneally with CC (20 mg/kg) before and at 0 min, 24 h, 48 h, and 72 h after hypoxia-ischemic (HI), and rats in the other two groups were injected intraperitoneally with an equal volume of normal saline at the corresponding time. Meanwhile, from postnatal day 10, each rat was injected intraperitoneally with 5-bromo-2’-de-oxyuridine (BrdU) (50 mg/kg) for 5 consecutive times, once every 12 h. On postnatal day 12, BrdU in the hippocampal dentate gyrus and cleaved caspase-3 in the hippocampal CA1 area were detected by immunohistochemistry, and neuronal apoptosis in hippocampal CA1 area were detected by TUNEL staining. On postnatal day 28, long-term learning and memory ability of rats was tested by Y maze.ResultsThere was signiifcant difference in the number of BrdU-positive cells in brain tissues of rats among three groups (F=101.38,P<0.01). The BrdU-positive cells in HIBD group and CC group were signiifcantly more than those in sham operation group (P<0.05). There was signiifcant difference in the number of cleaved caspase-3-positive cells in hippocampal CA1 area among three groups (F=379.77,P<0.01). The cleaved caspase-3-positive cells in CC group were signiif-cantly fewer than those in HIBD group but signiifcantly more than those in sham operation group (P<0.05). The TUNEL-pos-itive cells in hippocampal CA1 area were signiifcantly different among three groups (F=505.92,P<0.01) which was most in HIBD group and fewest in sham operation group and signiifcant difference was found through multiple comparison (P<0.05). The total learning number of avoiding electric shock tested by Y maze was signiifcantly different among three groups (F=32.05, P<0.01) which was most in HIBD group. Correct response rate was significantly different among three groups (F=24.99, P<0.01) which was lowest in HIBD group.ConclusionsCaffeine citrate can improve the ability of long-term learning and memory in neonatal rats with hypoxia-ischemic brain damage, the mechanism of which may be related to reducing the neuronal apoptosis after hypoxia ischemia.

Objective To study the changes of the T-lymphocyte subsets in patients with Guillain-Barré syndrome(GBS)before and after intravenous immunoglobulin treatment(IVIG),and to explore the possible mechanism of the IVIG curing GBS further.Methods Chose 31 cases of clinically confirmed GBS were enrolled and compared before and after the treatment.According to the effect of the therapy,31 cases of the total were sub-divided into effective and ineffective groups.Relative counting of peripheral blood T-lymphocyte subsets was preformed with flow cytometry.Results ①The percentage of CD8+ T and CD4+CD29+ T cell was significantly lower(CD8+T:28.77%±11.02% vs 31.84%±12.35%,CD4+CD29+T:56.71%±12.44% vs 62.40%±12.72%,t=2.995,3.919,P＜0.05)after therapy,while the rate of CD4+/CD8+T and the percentage of CD4+CD45RA+T cell increased notably(t=2.368,3.860,P＜0.05);but there was no notable difference in the percentage of CD3+T and CD4+T cell.②The percentage of CD8+T and CD4+CD29+ T cell was significantly lower(t=2.144,3.343,P＜0.05)after the treatment,while the rate of CD4+/CD8+T and the percentage of CD4+CD45RA+T cell increased notably(t=2.159,3.277,P＜0.05)in the good curative effect group,but there was no change in the bad curative effect group.③61.29%(19/31)of the patients significantly improved by IVIG,and there was no death case.Conclusions T-lymphocyte subsets change in a varing degree after IVIG treatment in acute GBS patients,which lays an immunological foundation for the further study of pathogenesis and mechanism of IVIG curing GBS;effective on GBS,IVIG can actively suppress pathogenetic condition and promote the recovery of nrevous function.

OBJECTIVE: To study the mechanism and clinical significance of specific immunotherapy (SIT) on the expression changes of GM-CSF and IL-5 in the tissue samples of recurrent nasal polyps. METHOD: Perennial allergic rhinitis patients with recurrent nasal polyps were randomly divided into 2 groups. The experimental group of 19 patients was treated by SIT and standardized treatment (glucocorticoid nasal spray) , and the control group of 17 patients was only treated by standardized treatment (glucocorticoid nasal spray). We measured the expression levels of GM-CSF and IL-5 in the tissue samples of the nasal polyps by ELISA, and compared the results obtained before treatment with expression levels detected at 6 months and 1 year after the treatment. RESULT: The expression of GM-CSF and IL-5 in the recurrent nasal polyps reduced significantly (P < 0.05) in both groups after 6 months and 1 year post-treatment compared with pre-treatment, and the expression of GM-CSF and IL-5 in the experimental group was much lower than that of the control group. CONCLUSION SIT decreases the expression of GM-CSF and IL-5 and reduces the inflammatory reaction in the tissue samples of recurrent nasal polyps.
Enzyme-Linked Immunosorbent Assay ; Granulocyte-Macrophage Colony-Stimulating Factor ; metabolism ; Humans ; Immunotherapy ; Inflammation ; drug therapy ; Interleukin-5 ; metabolism ; Nasal Mucosa ; pathology ; Nasal Polyps ; drug therapy ; metabolism ; Rhinitis, Allergic, Perennial ; drug therapy ; metabolism

Background and purpose: Langerhans cell histiocytosis (LCH) is a rare complex reticulocndotheliai disease that often invades the head and neck. There in no consensus of treatment. Radiation is one of the treatment options for the localized lesions. Our aim in the study was to analyze the radiotherapeutic efficacy for LCH in the head and neck region. Methods: 8 patients with eosinophilic granuloma (EG) and 1 case with Hand-Schuller-Christian disease (HSC) were treated with radiotherapy in our hospital from July 2000 to May 2007. Their clinical data were retrospectively analyzed. Results: 5 cases of EG were treated with tumor partial excision and radiation, while the other 3 cases of EG were given radiation alone after biopsy. The HSC was administered with radiation and endocrine therapy. All the patients were followed up for 1.5-8 years. The response rate was 89%. Conclusion: Radiotherapy of LCH is an effective modality, but the optimal dose needs to be further studied.

Objective To study the constituents in Melastoma dodecandrum.Methods The constituents were isolated by chromatographic methods,and their structures were elucidated by spectroscopic evidences.Results Five compounds were purified and their structures were identified as: daucosterol(Ⅰ),oleanolic acid(Ⅱ),avicularin(Ⅲ),3,7,4′-trimethoxyquercetin(Ⅳ),and atractylenolidone(Ⅴ).Conclusion Compound Ⅴ is a new chemical constituent named atractylenolidone.Compound Ⅳ is isolated from M.dodecandrum for the first time.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

In order to study the constituents and pharmacology of Tripterygium plants (Tripterygium willfordii Hook.f), a variety of chromatography methods were used. Four compounds were isolated from Tripterygium plant and their structures were elucidated by UV, IR, MS, HR-MS, 1H NMR, 13C NMR and 2D-NMR techniques. The isolated compounds were named as triptonide (1), neo-triptetraolide (2), 2alpha-hydroxytriptonide (3), and 15-hydroxytriptonide (4), separately. Compounds 3, 4 belong to new diterpenoids, which can inhibit the growth of K562 cells (leukemia cells) and HL60 cells (acute myeloid leukemia cells).

Objective To explore the relationship between immune functions of patients with acute leukemia (AL) and invasive fungat infection (IFI). Methods T lymphocyte subpopulations and natural killer (NK) cells in 61 AL patients complicated with IFI at first visit, AL remission, the time of IFI onset and 4 weeks after antifungal treatment were measured by flow cytometry. Meanwhile,levels of IgG, IgM and IgA were detected by immunoturbidimetry. The statistical analysis was done using ANOVA, t test and chi-square test. Results CD3+ , CD3+ CD4+ , CD8+ CD28+ T lymphocyte as well as CD4+/CD8+ ratio at the time of IFI onset in AL patients were all lower than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 25.6,26.6,13. 1,167.9; all P＜0.05), while CD8+ CD28- T lymphocyte were higher than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 220.2,P＜0.01). CD3+ , CD3+ CD4+ and CD4 + /CD8+ ratio of patients who responded effectively to antifungal treatment were all higher than those of non-responders (t=3.75,8. 61,3.17; all P＜0.05). The serum levels of IgG, Igm and IgA at first visit, ALremission, the time of IFI onset and 4 weeks after treatment were similar (F=0.78,0.72,0.81; all P ＞0.05). The effect rate of antifungal therapy in AL remission group was higher than that in nonremission group (87% vs 53%,x2 = 7.62, P＜0.05). Conclusions The cellular immune functions are impaired severely in AL patients complicated with IFI, while the levels of IgG, IgM and IgA are similar during IFI. Therefore, the efficacy of antifungal therapy may partly depend on the recovery of cellular immune functions and remission of AL.