Objective To evaluate the clinical efficacy and safety of tapering the dosage of recom-binant human tumor necrosis factor receptor-Fc fusion protein (rhTNFR-Fc) combined with DMARDs in the treatment of peripheral joints involvement of ankylosing spondylitis. Methods Sixty patients who met the classification criteria of ankyloding spondylitis were enrolled. Meanwhile, all patients had one or more of the following joint involvement: hip, knee, ankle, and shoulder. Their BASDAI was higher than 4, joint pain VAS≥4, ESR ≥30 mm/1 h and CRP≥8 mg/L. Tuberculosis, hepatitis B, hepatitis C infection or other microorgan-isms infections were excluded. All enrolled patients had no serious heart,liver,kidney, or other internal organ involvement. During the first stage (The first eight weeks patients were matched by age and, disease activity, then randomly divided into the rhTNFR-Fc (the control group) treatment group in which patients were treated with 25 mg rhTNFR-Fc subcutaneous injection twice per week for 4 months) and rhTNFR-Fc dosage tapering group in which 25 mg rhTNFR-Fc were subcutaneously injected once per week for 4 weeks and then followed by 12.5 mg per week for 4 weeks, then once every 10 days for 6 times. Then the dosage of rhTNFR-Fc dosage of the dosage tapering group (the experimental group) was changed to 12.5 mg subcutaneous injection once every 15 days for another 4 times combined with methotrexate 7.5 mg per week and Salfasalazine 2 g daily and thalidomide 100 mg per night. The second stage started from week 9 to 24. In addition to the 30 cases at the first stage, 42 cases were included based on the same inclusion criteria for stage one. Patients' clinical and laboratory parameters were evaluated at week 0, 4, 8, 16 and 24. Results During the first four weeks, all patients of both control group and experimental group reached ASAS20, 97% (29/30) patients reached ASAS50 in the control group, 83% (25/30) patients reached ASAS50 in the experimental group. At week 8, patients in both groups maintained at 100% ASAS20 improvement, 100% (13/13) patients in the control group reached ASAS50, and that of the experimental group was 97% (29/30), the differences between the two groups were not statistically significant (P>0.05). In the second stage, 72 cases (100%) could achieve ASAS20, 63 cases (88%) achieved ASASSO at week 16. At week 24, 72 cases (100%) remained to achieve ASAS20, 71 cases (99%) achieved ASAS50. The safety and compliance of the two groups were good. Two cases developed infection, one patient had mild elevation of serum transaminase. Conclusion Tapering the dosage combined with DMARDs is an effective and safe approach in the treatment of peripheral joints involvement of ankylosing spondylitis. The compliance of this strategy is good and only few patients have serum transaminase elevation. But attention should be paid to the increased rate of infection.

Objective To investigate the relationship between plasma homocysteine (Hey) level and ankylosing spondylitis (AS).To analyze the association between the NS,N10 methylenetetrahydrofolate reductase (MTFHR) gene polymorphism and AS.Methods One hundred patients with AS and 60 healthy controls were included in the study.The plasma Hey level was examined by enzyme-linked immunoadsorbent assay and MTHFR gene polymorphism was analyzed by the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).Results Compared with heahhy controls,the plasma Hey level in AS patients was significantly higher than that of the controls (P＜0.01).There was no significant difference in the frequen-cies of MTHFR genotype and alleles between AS and the controls (P＞0.05),But the ratio of T/T genotype mutation was different between AS and the controls (P＜0.05).The plasma Hey level of T/T genotype was significantly higher than that of C/T or C/C genotype in AS and the controls (P＜0.01).Logisticalregression analysis indicated that Hey was an independent risk factor for AS (P＜0.01,0R=4.582,95%CI=1.984～10.585).Conclusion The plasma homocysteine level is significantly increased in AS patients.Hyperhomo-cysteinemia is an independent risk factor for AS.MTHFR T/T genotype mutation is an important mechanism of hyperhomocysteinemia and may be related with AS.

OBJECTIVESThe effect of vascular active peptides on the development of pulmonary remodeling and pulmonary hypertension due to left to right shunt congenital heart diseases is the focus of today's studies. The present study was conducted to investigate the roles of adrenomedullin (ADM) and adrenotensin (ADT) in pulmonary remodeling due to left to right shunt in rat lungs.

METHODSTwenty-one male Wistar rats were divided into two groups randomly. A right common carotid artery to external jugular vein shunt operation was performed on experimental rats (n = 9) to establish a left to right shunt animal model. Meanwhile, the common carotid artery and external jugular vein of the control group rats (n = 12) were just isolated without connection. Twelve weeks later, the mean pulmonary artery pressure (mPAP), the right ventricle to left ventricle plus septum ratio [weight, RV/(LV + SP)], the percentage of media wall thickness (MT%) were calculated. The distributions and relative protein contents of ADM and ADT in lungs were measured by immunohistochemical staining and Western blotting analysis. The relative gene expression for ADM, ADT, p46-p54 stress-actived protein kinase (SAPK) and p44 extracellular signal-regulated protein kinase 1 (ERK(1)) were investigated by RT-PCR.

RESULTSThe muscular and the tunica intimae layer of pulmonary artery were thicker in experiment group rats than those of control group, and the mPAP increased significantly in shunt group [(27.10 +/- 6.67) mm Hg (1 mm Hg = 0.133 kPa)] compared with that in control group [(14.32 +/- 3.14) mm Hg] (t = 5.5507, P < 0.001). The ratios of RV/(LV + SP) and MT% increased significantly in experimental group in contrast to the control group (P < 0.001). ADM and ADT positive granules distributed mainly over vascular smooth muscle cells, and Western blotting and integrated optical density analysis showed that the content of ADM increased in shunt group rats (P < 0.001), however, ADT content decreased (P < 0.001). The mRNA expression of ADM, SAPK and ERK(1) up-regulated in experiment group compared with the control group (P < 0.01, and P < 0.001 respectively), however, the ADT mRNA expression decreased in experimental rats in contrast to the control group (P < 0.001).

CONCLUSIONSThe phenomenon of intramolecular regulation of ADM and ADT, which both derived from proadrenomedullin, existed in the development of pulmonary remodeling and pulmonary hypertension due to left to right shunt. The mitogen-activated protein kinases (MAPKs) signal transduction pathway has been activated in the formation of left to right shunt pulmonary remodeling and pulmonary hypertension, and ADM may slow down the occurrence of pulmonary hypertension through cutting off MAPKs signaling pathway.

Adrenomedullin ; biosynthesis ; Airway Remodeling ; Animals ; Hypertension, Pulmonary ; metabolism ; Lung ; metabolism ; physiopathology ; Male ; Mitogen-Activated Protein Kinases ; metabolism ; Peptide Fragments ; biosynthesis ; Pulmonary Artery ; metabolism ; Pulmonary Circulation ; Rats ; Rats, Wistar ; Signal Transduction

OBJECTIVETo study the inhibitive effects of gastric cancer cell-dendritic cell fusion vaccine on tumor cells of proliferation cycle.

METHODSPeripheral blood mononuclear cells were separated from gastric cancer patients and co-cultured with granulocyte macrophage colony stimulating factors(GM-CSF), interleukin-4(IL-4) and tumor necrosis factor-alpha(TNF-alpha) to generate mature dendritic cells. The dendritic cells and SGC7901 cells were fused by polyethylene glycol, and the pure fusion cells were screened out by selective culture systems. The inhibitive effects of gastric cancer cell-dendritic cell fusion vaccines on tumor cell proliferation cycle in vivo and in vitro were detected by flow cytometry.

RESULTSTreated with the fusion vaccine in vitro, the percentages of G(0)/G(1), S and G(2)/M cells of tumor cells were (76.77+/- 4.38)%, (16.50+/- 2.90)% and (6.73+/- 1.59)% respectively. There were significant differences in the percentages of different cell cycle tumor cells between the tumor cells treated with the fusion vaccine and those co-cultured with dendritic cell or T cells alone(P< 0.01). The proliferative index of the tumor cells treated with the fusion vaccine was 23.34+/- 3.51, significantly lower than those co-cultured with dendritic cell and controls (P< 0.05).

CONCLUSIONSFusion vaccines can affect cell cycle of the tumor cells, thus inhibit tumor cell proliferation and growth.

Cancer Vaccines ; immunology ; pharmacology ; Cell Cycle ; drug effects ; Cell Fusion ; Cell Proliferation ; drug effects ; Dendritic Cells ; immunology ; Humans ; Stomach Neoplasms ; immunology ; Tumor Cells, Cultured

AIMTo design and synthesize new phenyloxyisobutyric acid analogues as antidiabetic compounds.

METHODSEight new target compounds were synthesized by combination of lipophilic moieties and acidic moiety with nucleophilic replacement or Mitsunobu condensation. The eight compounds were confirmed by 1H NMR, 13C NMR, IR and MS.

RESULTSIn vitro insulin-sensitizing activity (3T3-L1 adipocyte) demonstrated, that the cultured glucose concentration of up-clear solution detected with GOD-POD assay were 5.942, 6.339, 6.226 and 6.512 mmol x L(-1), respectively, when rosiglitazone, pioglitazone, compounds A and B were added to the insulin-resistant system.

CONCLUSIONIn vitro insulin-sensitizing activity of target compound A is in between that of rosiglitazone and pioglitazone, and activity of target compound B is slightly less than that of pioglitazone.

3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Butyrates ; chemical synthesis ; chemistry ; pharmacology ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Insulin ; pharmacology ; Mice ; Molecular Structure ; PPAR gamma ; agonists ; pharmacology

OBJECTIVETo explore the effect of microRNA-21 (miR-21) antisense oligonucleotide on the biological characteristics of human cervical squamous carcinoma cell lines SiHa in vivo and in vitro.

METHODSSpecific phosphorothioate antisense oligodeoxynucleotides targeting miR-21 were synthesized and transfected into cervical cancer cells in vitro. Expression of miR-21 in SiHa after transfection was detected by real-time RT-PCR. The cell proliferation was evaluated by MTT assay and colony formation experiment. The cell apoptosis was analyzed by annexin V-FITC/PI analysis. The inhibitory effect of miR-21 antisense oligonucleotide on tumor growth was evaluated by tumor growth curves and immunohistochemistry (MaxVision method). H-E staining was used to document morphological changes and fluorometric TUNEL assay was to detect the apoptotic activity.

RESULTSAfter the transfection of antisense miR-21, the expression of miR-21 decreased along with an obvious growth inhibition, compared with that of the control groups (P < 0.05). Colony formation of both cell lines was markedly inhibited with antisense miR-21 (55.6% ± 1.4%), as compared with that in the negative group (98.3% ± 2.0%, P < 0.05). Flow cytometry assay showed that antisense miR-21 expression significantly enhanced the cell apoptosis (6.7% ± 1.3% and 29.4% ± 1.7%, P < 0.05). The tumor-forming rates of miR-21 transfected group, and negative control groups were 3/8 and 6/8, respectively (P < 0.05). Ki-67 proliferative marker staining decreased significantly (42% vs 90%) in the transfected group compared with negative control groups. Extensive dead tumor cells were seen in the miR-21 transfected cells along with a marked increase of apoptosis (P < 0.05).

CONCLUSIONTargeted antisense oligonucleotide miR-21 effectively suppresses the growth of cervical carcinoma SiHa cells both in vitro and in vivo through an induction of apoptosis.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Humans ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; MicroRNAs ; genetics ; metabolism ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; metabolism ; Transfection ; Uterine Cervical Neoplasms ; genetics ; metabolism ; pathology

In recent years, Bacterial resistance is more and more serious for the irrational use of antibiotics produces resistant strains and other reasons. Human are trying to solve the problem from different ways, including the study of antimicrobial peptides. Defensin is one of the most important of antimicrobial peptides. A novel antimicrobial peptide, human beta-defensin 3, was isolated and demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes. The total RNA was extracted from human tonsil and the hbetaD-3 specific cDNA sequence was amplified with RT-PCR. After sequenced, the target DNA fragment was cloned into pQE-80L vector together with the DNA fragment encoding carrier protein DHFR. The recombinant vectors were transformed into E. coli M15 and the expression was induced based on the optimal values of the IPTG concentration incubation temperature and induction time determined in the previous section. The expressed proteins were analyzed by SDS-PAGE and Western-blotting. The mass of the recombinant protein was about 40% of total bacteria protein. Isolate and purify the target protein. The recombinant hbetaD-3 fusion proteins possess the antimicrobial activity to staphylococcus aureus, multiresistant staphylococcus aureus (only vancomycin-sensitive) and Candida albicans in the assay of drug susceptibility. Advanced study can be continued based on our experiments.
Cloning, Molecular ; Escherichia coli ; genetics ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Staphylococcus aureus ; drug effects ; Tetrahydrofolate Dehydrogenase ; genetics ; beta-Defensins ; biosynthesis ; genetics ; pharmacology

OBJECTIVESTo evaluate the feasibility and efficacy of transcatheter closure of perimembranous ventricular septal defects (pmVSD) with aneurysmatic formation and muscular ventricular septal defects (mVSD) with Amplatzer duct occluder II.

METHODSThis retrospective analysis included 48 cases received transcatheter closure of pmVSD aneurysmatic formation or mVSD from February 2011 to March 2012 in our hospital (42 pmVSD with aneurysmatic formation and 6 mVSD). Median age was 5.2 years (range: 1.8 - 15 years), and median weight was 20.2 kg (range: 12 - 44 kg). Amplatzer duct occluder II was selected depending on the condition of ventricular septal defect. The device was implanted by antegrade or retrograde approach. Complications such as residual shunt, valvular regurgitation and arrhythmia were evaluated by echocardiography or angiography. Median follow-up was 9.5 months (range: 1 - 13 months).

RESULTSThe mean ratio of pulmonary (Qp) to systemic (Qs) blood flow was 1.35 ± 0.15 before transcatheter closure. The diameter of exit hole of ventricular septal defects was (2.46 ± 0.53) mm measured by transthoracic echocardiography, and (2.35 ± 0.40) mm by angiography. Successful implantation of the device was achieved in 46 patients (96%) and unsuccessful in two cases due to acute aortic insufficiency. Forty-two (92%) patients were closed successfully, and trivial residual leak was evidenced in four patients and remained unchanged during follow-up. One patient with mVSD still had trivial residual shunt at 6 months post procedure. New trivial tricuspid insufficiency was observed in 1 patient (2.1%) during follow-up. Two patients developed procedural related left anterior fascicular block and remained unchanged during follow-up.

CONCLUSIONSpmVSD with aneurysm and mVSD could be successfully treated with Amplatzer duct occluder II. However, the long waist and large disc of the device could interfere with tricuspid valve function and cause tricuspid insufficiency.

Adolescent ; Cardiac Catheterization ; instrumentation ; methods ; Child ; Child, Preschool ; Female ; Heart Septal Defects, Ventricular ; surgery ; Humans ; Infant ; Male ; Retrospective Studies ; Septal Occluder Device ; Treatment Outcome

BACKGROUNDPulmonary thromboembolism (PTE) has become a common disease that severely endangers people's health. This study analysed the changes in proportion and mortality of PTE in hospitalized patients to provide data for prevention and management of the disease.

METHODSThe data of 763 hospitalized patients with PTE from 1974 to 2005 in Fuwai Hospital were analysed.

RESULTSDuring the 1970s, 0.27% of patients in a cardiovascular hospital had PTE (< 5 cases per year); while so far this century the proportion is 0.94% (48 to 113 per year). The mortality of hospitalized PTE patients fell from 20.00% in the 1970s to 4.10% this century. Prior to 1990, the mortality of hospitalized PTE patients was 12.50%, and in the years after 1990 only 3.40%. The difference was statistically significant (P < 0.005). People with this disease were mostly between the ages of 30 and 69 years. Men were most susceptible between the ages of 30 and 69 years, while women between the ages of 40 and 69 years. Men contracted PTE 10 years earlier than women. The mortality of male PTE patients was 4.70%, not significantly different from female patients, 5.06% (0.50 < P < 0.75). There were not any significant differences between the mortality of patients in the different age groups overall (< or = 39, 40 - 49, 50 - 59, and > or = 60 years, P > 0.1). More people contracted the disease in winter than in other seasons (P < 0.05). There was no obvious difference between the mortality in different seasons overall (0.75 < P < 0.90).

CONCLUSIONPTE is an increasingly significant disease and deserves adequate attention.

Adult ; Age Factors ; Aged ; Female ; Hospitalization ; Humans ; Male ; Middle Aged ; Pulmonary Embolism ; epidemiology ; mortality ; Seasons ; Time Factors

OBJECTIVETo investigate the effects of microRNA-383 (miR-383) on PRDX3 gene expression, cell proliferation and apoptosis of human medulloblastma.

METHODSPRDX3 and miR-383 RNA expression was detected by real-time quantitative RT-PCR in human medulloblastoma tumor tissue samples, Daoy cell line and normal brain tissue samples. Western blot was used to detect protein expression of PRDX3. Synthetic miR-383 mimics were transfected into Daoy cells by lipofectamine. Using Cell Counting Kit-8 (CCK-8) method, flow cytometry was used to investigate the cell proliferation and apoptosis, cells reactive oxgen species(ROS), mitochondrial membrane potential changes in each experimental groups.

RESULTSOf 15 cases of human medulloblastoma tumor, 13 cases had miR-383 expression levels significantly lower than that of normal brain tissue, and 14 had PRDX3 mRNA expression levels significantly higher than that of normal brain tissue. The expression levels of miR-383 and PRDX3 in Daoy cells were 0.353 and 1.315 times than those of normal brain tissue, respectively. The protein expression levels of PRDX3 were higher in human medulloblatoma tumors and Daoy cells than that of normal brain tissue. Transfected miR-383 mimics increased the expression level of miR-383 after 24 h and 48 h was significantly higher than that of the control. In contrast, PRDX3 gene mRNA and protein expression levels were significantly decreased at 48 h compared with the control group. Using CCK-8 assay, the cell proliferation rate in the experimental group was significantly lower than that of the control group (P < 0.05). Annexin V-FITC assay demonstrated that early apoptosis rate of the experimental group (11.60 ± 0.30)% was significantly higher than those of the control group (2.3 ± 0.20)% and negative control group (10.37 ± 0.25)% (P = 0.000) after 48 h of transfection. The intracellular ROS levels after transfection at 24 and 48 h significantly increased than those of the control group. Mitochondrial membrane potential level at 24 h after transfection significantly decreased, comparing with the blank control group and the negative control group.

CONCLUSIONSCompared with normal brain tissue, decreased expression of miR-383 but elevated expression of PRDX3 are medulloblastoma tumour and Daoy cell lines. Up-regulation of miR-383 knockdowns the expression of PRDX3, inhibits proliferation and promotes apoptosis of Daoy cells, leading to increased intracellular ROS and decreased levels of mitochondrial membrane potential.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; genetics ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Medulloblastoma ; genetics ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; MicroRNAs ; genetics ; metabolism ; Peroxiredoxin III ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Reactive Oxygen Species ; metabolism ; Transfection