Objective To determine the effects of urapidil on L-type calcium current(I_(Ca-L))in rat cardiomyocytes.Methods Ventricular myocytes were isolated from SD rats of either sex(250-280g)by retrograde perfusion of the hearts via aorta with calcium-free Tyrode solution containing enzyme as described elsewhere.Rod shaped cells with clear borders and striations were selected.Eighteen cells were randomly divided into 3 groups(n =6 each):A urapidil group;B urapidil+methysergide group and C methysergide group.All the cells in the three groups were peffused first with Tyrode solution for 1 min(T_1).In group A and C cells were then peffused with Tyrode solution containing 0.4 ?mol?L~(-1) urapidil(A)or 40 nmol?L~(-1) methysergide(C)for 1 min(T_2) while in group B cells were perfused fwst with Tyrode solution containing 0.4 ?mol?L~(-1) urapidil for 1 min (T_2) then with Tyrode solution containing methysergide 40 nmol?L~(-1) for 1 min (T_3).Finally the cells were again perfused with regular Tyrode solution for 1 min(T_4)to wash out the drugs.The peak of I_(Ca-L) was recorded at T_(1-4) by means of the whole cell patch clamp technique with use of Axo patch 200B.Results In group A,B and C the peak of I_(ca-L) at T_2 was significantly lower than that at T_1 but there was no significant difference between the peak of I_(ca-L) at T_1 and T_4.In group B the peak of I_(Ca-L) at T_3 was significantly lower than that at T_2.Conclusion Urapidil inhibits L-type calcium current in rat isolated cardiomyoeytes.It's inhibitory effect may not be mediated by 5-H_(1A) receptor.

OBJECTIVETo analyze the clinical characteristics of elderly patients with spinal tuberculosis and explore its clinical effects with anti-TB drugs alone.

METHODSFrom January 2008 to July 2010, the data of 36 patients with spinal tuberculosis underwent conservative treatment of anti-TB drugs alone were analyzed. There were 19 males and 17 females with an average age of 73.5 years (ranged, 60 to 85). All patients were in the active phase with high ESR and CRP levels and were treated with 3HRZE/6-9HRE (course from 9 to 12 months). According to clinical symptoms, chemical examination, radiological image to adjust drug and depending on VAS score to evaluate pain.

RESULTSAll the patients were followed up from 8 to 24 months with an average of 15 months. Tuberculose of 31 patients healed after chemotherapy from 9 to 12 months and ESR and CRP recovered normally. Levofloxacin and para-amino salicylic acid were used in 4 cases because of 4 cases occurred drug fast for RFP or INH, after 15 months, their obtained healing. Symptom of 1 case got worse during chemotherapy, and surgical treatment were performed, after 3 months, ESR and CRP recovered normally, X-ray and CT showed spinal osteosclerosis and fusion without significant kyphosis and internal fixation loosening. Cobb angle was respectively(17.6+/-2.3) degrees, (18.1+/-2.7) degrees before treatment and last follow-up (P>0.05). MRI showed abscess was absorbed and spinal inflammation subsidised. VAS score was respectively 6.5+/-1.7, 1.4+/-0.5 before treatment and last follow-up (P<0.05). Seven patients had complications relating with drug adverse reaction,after discontinuation and treated with clinical symptom,the patients recovered normally.

CONCLUSIONAnti-TB drugs alone can obtain satisfactory effects in treating early senile spinal tuberculosis, but strict supervision and individual administration should not be disregardful.

Aged ; Aged, 80 and over ; Antitubercular Agents ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Tuberculosis, Spinal ; diagnosis ; drug therapy

Decompression, Surgical ; Humans ; Lumbar Vertebrae ; surgery ; Spinal Fusion ; Spondylolisthesis ; diagnosis ; physiopathology ; surgery

OBJECTIVETo evaluate the effect and safety of L-carnitine in the treatment of idiopathic oligoasthenozoospermia based on current clinical evidence.

METHODSWe searched the Cochrane Library, PubMed, MEDLINE, EMBASE, CNKI, VIP, CBM and Wanfang Database from the establishment to April 2014 for the published literature on the treatment of idiopathic oligoasthenozoospermia with L-carnitine. We conducted literature screening, data extraction, and assessment of the methodological quality of the included trials according to the inclusion and exclusion criteria, followed by statistical analysis with the RevMan 5. 2 software.

RESULTSSeven randomized controlled trials involving 751 patients with idiopathic oligoasthenozoospermia met the inclusion criteria, and 678 of them were included in the meta-analysis. L-carnitine treatment achieved a significantly increased rate of spontaneous pregnancy as compared with the control group (RR = 3.2, 95% CI 1.74 to 5.87, P = 0.0002). After 12-16 and 24-26 weeks of medication, total sperm motility (WMD = 5.21, 95% CI 2.78 to 7.64, P < 0.0001 and WMD = 9.29, 95% CI 1.28 to 17.29, P = 0.02) and the percentage of progressively motile sperm (WMD = 12.44, 95% CI 4.58 to 20.31, P = 0.002 and WMD = 9.76, 95% CI 3.56 to 15.97, P = 0.002) were remarkably higher than those in the control group, but no statistically significant differences were observed in sperm concentration between the two groups (WMD = 4.91, 95% CI -2.63 to 12.45, P = 0.2 and WMD = 0.93, 95% CI -3.48 to 5.34, P = 0.68). After 12-16 weeks of treatment, the percentage of morphologically abnormal sperm was markedly decreased in the L-carnitine group as compared with the control (WMD = -2.48, 95% CI -4.35 to -0.61, P = 0.009), but showed no significant difference from the latter group after 24-26 weeks (WMD = -4.38, 95% CI -9.66 to 0.89, P = 0.1). No statistically significant difference was found in the semen volume between the two groups after 12-16 or 24-26 weeks of medication (WMD = -0.13, 95% CI -0.43 to 0.18, P = 0.42 and WMD = 0.28, 95% CI -0.02 to 0.58, P = 0.07). No serious L-carnitine-related adverse events were reported in 4 of the randomniized controlled trials.

CONCLUSIONThe current evidence indicates that L-carnitine can improve spontaneous pregnancy and semen parameters in the treatment of idiopathic oligoasthenozoospermia, with no serious adverse reactions.

Asthenozoospermia ; drug therapy ; Carnitine ; adverse effects ; pharmacology ; Female ; Humans ; Male ; Pregnancy ; Pregnancy Rate ; Randomized Controlled Trials as Topic ; Semen Analysis ; Sperm Count ; Sperm Motility

The stem cell leukemia (SCL) gene is a new oncogene related with leukemogenesis. To explore the effects of antisense oligonucleotides of SCL on leukemic cells, SCL antisense phosphorothioate oligodeoxynucleotides (AS-PS-ODN) were used to treat K562 and CEM leukemic cell lines to observe the effects on proliferation, differentiation, apoptosis and SCL mRNA expression in the cells. The results showed that incubation of K562 or CEM cells with AS-PS-ODN at different concentrations led to inhibition of cell proliferation, and the inhibitory effects varied with the incubation time. The positive rate of benzidine staining in K562 cells increased significantly after the inhibition with AS-PS-ODN, compared with S-PS-ODN treatment. The characteristics of apoptosis were observed in K562 cells treated with AS-PS-ODN, but not in CEM cells. Expression of SCL mRNA in K562 and CEM cells and SIL-SCL mRNA in CEM cells decreased after incubation of AS-PS-ODN. It is concluded that SCL AS-PS-ODN inhibits specifically the proliferation of K562 and CEM cells, also decreases the level of SCL and SIL-SCL mRNA expression. AS-PS-ODN enhances erythroid differentiation and induces premature apoptosis in K562 cells.
Apoptosis ; drug effects ; Basic Helix-Loop-Helix Transcription Factors ; Cell Division ; drug effects ; DNA-Binding Proteins ; antagonists & inhibitors ; physiology ; Humans ; K562 Cells ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins ; antagonists & inhibitors ; physiology ; RNA, Messenger ; analysis ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Transcription Factors ; antagonists & inhibitors ; physiology

OBJECTIVETo construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration.

METHODSThe lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods.

RESULTSThe lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01).

CONCLUSIONLentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.

Fetus ; HEK293 Cells ; Humans ; Intervertebral Disc ; metabolism ; Lentivirus ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; Receptors, Cell Surface ; genetics ; Transfection

This study was aimed to explore the effects of expressing eukaryotic elongation factor 1A1 (eEF1A1) on proliferation and apoptosis in human acute T lymphocytic leukemia (T-ALL) cell line Jurkat with knocked down eEF1A1 gene and its mechanisms. eEF1A1-expressing lentivirus (LV) was constructed and used to transfect the Jurkat cells with knocked down eEF1A1 gene. Then, the expressions of eEF1A1 mRNA and protein were detected by real time PCR(RT-PCR) and Western blot respectively.Cell proliferation, apoptosis and cycle were detected by MTT method, Annexin V-APC labeling and DNA ploidy analysis respectively. The related protein expressions of phosphatidylinositol-3-kinase (PI3K)/serine/threonine kinase (Akt) signaling pathway were detected by Western blot. The results indicated that eEF1A1 mRNA and protein expressions of Jurkat cells with knocked down eEF1A1 gene were re-established by constructing eEF1A1-expression LV. Compared with negative control group (transfected with negative control LV and eEF1A1-shRNA LV), cell proliferation in eEF1A1 expression group was significantly enhanced, cell apoptosis was remarkably inhibited, percentage of cells in G0/G1 phase was significantly reduced alone with increased percentage of cells in S and G2/M phase, and the expression levels of p-Akt (Ser 473), nuclear factor kappa B (NF-κB), p-NF-κB (Ser 468), mammalian target of rapamycin (mTOR) and p-mTOR (Ser 2448) protein significantly increased. It is concluded that eEF1A1 may have a carcinogenic effect in T-ALL cells. eEF1A1 expression has noticeable effects on the proliferation enhancement and apoptosis inhibition of Jurkat cells, which may be mediated by the up-regulation of PI3K/Akt/NF-κB and PI3K/Akt/ mTOR signaling pathway.
Apoptosis ; Cell Proliferation ; Gene Expression ; Humans ; Jurkat Cells ; Peptide Elongation Factor 1 ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering ; genetics ; Signal Transduction

OBJECTIVETo investigate the preventive effect of interleukin-18 (IL-18) gene modified mature dendritic cells (mDC) vaccine on airway inflammation in mouse asthma model.

METHODSThe asthma model was induced by injection of ovalbumin (OVA) in BALB/c mice. IL-18 gene modified mouse mature dendritic cells (mDC) were detected by flow cytometry and its capacity of inducing allogeneic T cell responses was examined by mixed lymphocyte reaction (MLR). The OVA-induced asthmatic mice were randomly divided into 6 groups: PBS group, DXM group, mDC group, Ad-LacZ-mDC group, Ad-IL-18-mDC group and control group. The pathological changes in lung tissues were assayed by HE and AB-PAS staining. The numbers of inflammatory cells and percentage of eosinophils (EOS) in bronchoalveolar lavage fluid (BALF) were counted. The levels of IFN-γ IL-4 and IL-13 in culture supernatant of splenocytes were measured by ELISA method. The percentage of CD4(+)CD25(+)Foxp3(+) Treg was assessed by flow cytometry analysis.

RESULTThe vaccine was effective in decreasing the infiltration of EOS and accumulation of airway goblet cells in lung tissues, the numbers of inflammatory cells and percentage of EOS in BALF, and the levels of IL-4 and IL-13 in culture supernatant of splenocytes, and in increasing the levels of IFN-γ in culture supernatant of splenocytes and the percentage of CD4(+)CD25(+)foxP3(+) reg.

CONCLUSIONIL-18 gene modified mDC vaccine has a preventive effect on airway inflammation in OVA-induced asthmatic mice.

Animals ; Asthma ; immunology ; pathology ; prevention & control ; Dendritic Cells ; immunology ; Disease Models, Animal ; Genetic Therapy ; Interleukin-18 ; genetics ; Lung ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology

OBJECTIVETo investigate clinical outcome of short-course chemotherapy in retreating spinal tuberculosis after radical operation.

METHODSForty-six retreating patients with spinal tuberculosis were included in this series, 29 males, 17 females with the age from 27 to 61 years (average of 43.7 years). All patients were treated with radical operation and short-course anti-tuberculous chemotherapy from March 2005 to March 2008. The tuberculous focus located thoracic spine in 17 cases, thoracic-lumbar in 13 and lumbosacral vertebrae in 16 cases. Of them, 5 cases had sinuses of tuberculosis and 7 cases had incomplete palsy in lower limbs (Frankel C-D). CT or MRI showed obvious sequestra, cold abscess within spinal focus. Surgical procedures including debridement, auto-bone grafting, and one-stage internal fixation, was performed at the 4 to 6 weeks after chemotherapy. Chemotherapy regimes were 3HRZ/6-9HRE in majority of patients. Clinical effect and focus healing were evaluated at follow-up period.

RESULTSTuberculous symptoms and local pain of vertebral volume were obvious in all patients before chemotherapy,with average ESR 65.3 mm/h and average CRP 37.4 mg/L. After 4-6 weeks chemotherapy, tuberculosis symptoms and vertebral pain improved in all patients, and the average ESR decreased to 38.3 mm/1h, the average CRP decreased to 17.2 mg/L. Two to three months after operation, tuberculous symptoms and local pain relived in all patients,ESR and CRP became normal in 37 cases. Six to twelve months after operation, bonegraft complex in each patient became stable and there were no instrument loosening or deformity correction loss. Six patients with incomplete palsy recovered and 1 case improved from Frankel C to D grade. Focus healing was achieved in 44 cases (95.7%) after short-course chemotherapy (3HRZ/6-9HRE), and there were no resurgence in 2 to 4 years follow-up period. Drug fast 2 cases for RFP+INH cured at the 15 months after chemotherapy.

CONCLUSIONSRemoved tubercular focus for the treatment of retreating spinal tuberculosis can improve clinical effect and shorten chemotherapy course.

Adult ; Antitubercular Agents ; therapeutic use ; Combined Modality Therapy ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Thoracic Vertebrae ; surgery ; Treatment Outcome ; Tuberculosis, Spinal ; drug therapy ; surgery

OBJECTIVETo study the curative effect of surgical treatment of drug-resistant spinal tuberculosis.

METHODSFrom March 2005 and April 2009, the clinical data of 60 patients with drug-resistant spinal tuberculosis were retrospectively analyzed. Including 36 males and 24 females; aged from 5 to 79 years with an average of 47.3 years. Thirty-four patients had neurological deficits, among them, 2 cases were grade A, 5 cases were grade B, 13 cases were grade C, 14 cases were grade D according to ASIA standard. According to the severity and location of the infection, the patients underwent anterior, posterolateral costotransversectomy or posterior debridement and bone grafting and internal fixation. The antituberculous chemotherapy for a total of 12 to 18 months was guided by conventional and genotypic drug susceptibility testing. Tubercular relapse, neurological function, spinal fusion were observed by ASIA grade, X-ray and CT scan.

RESULTSAll cases were followed up from 1 to 5 years with an average of 3.1 years. Recurrence was found in 2 cases who were cured after second operation. 34 cases with neurological deficits recovered totally or partially. X-ray or CT films showed spinal fusion in 57 patients.

CONCLUSIONThe therapeutic effect of individuall operative options is good in treating drug-resistant spinal tuberculosis after antituberculous chemotherapy based on conventional and genotypic drug susceptibility testing.

Adolescent ; Adult ; Aged ; Antitubercular Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Middle Aged ; Mycobacterium ; drug effects ; genetics ; Radiography ; Retrospective Studies ; Spine ; Tuberculosis, Multidrug-Resistant ; diagnostic imaging ; drug therapy ; microbiology ; surgery ; Tuberculosis, Spinal ; diagnostic imaging ; drug therapy ; microbiology ; surgery ; Young Adult