China ; Congresses as Topic ; Drug Therapy, Combination ; Humans ; Medicine, Chinese Traditional ; Physicians ; Societies, Medical

China ; History, 20th Century ; Humans ; Integrative Medicine ; history ; trends ; Medicine, Chinese Traditional ; history ; trends

In the heart, stimulation of beta-adrenergic receptors (betaAR) serves as the most powerful means to increase cardiac contractility and relaxation in response to stress or a "fight-or-flight" situation. However, sustained beta-adrenergic stimulation promotes pathological cardiac remodeling such as myocyte hypertrophy, apoptosis and necrosis, thus contributing to the pathogenesis of chronic heart failure. Over the past decade, compelling evidence has demonstrated that coexisting cardiac betaAR subtypes, mainly beta(1)AR and beta (2)AR, activate markedly different signaling cascades. As a result, acute beta(1)AR stimulation activates the G(s) -adenylyl cyclase-cAMP-PKA signaling that can broadcast throughout the cell, whereas beta(2)AR-evoked cAMP signaling is spatially and functionally compartmentalized, due to concurrent G(i) activation. Chronic stimulation of beta(1)AR and beta(2)AR elicits opposing effects on the fate of cardiomyocytes: beta(1)AR induces hypertrophy and apoptosis; but beta(2)AR promotes cell survival. The cardiac protective effect of beta(2)AR is mediated by a signaling pathway sequentially involving G(i), G(betagamma), PI3K and Akt. Unexpectedly, beta(1)AR-induced myocyte hypertrophy and apoptosis are independent of the classic cAMP/PKA pathway, but require activation of Ca(2+)/calmodulin-dependent kinase II (CaMK II). The outcomes of cardiac-specific transgenic overexpression of either beta AR subtype in mice have reinforced the fundamentally different functional roles of these betaAR subtypes in governing cardiac remodeling and performance. These new insights regarding betaAR subtype stimulation not only provide clues as to cellular and molecular mechanisms underlying the beneficial effects of beta AR blockers in patients with chronic heart failure, but also delineate rationale for combining selective beta(1)AR blockade with moderate beta(2)AR activation as a potential novel therapy for the treatment of chronic heart failure.
Adenylyl Cyclases ; metabolism ; Animals ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; GTP-Binding Proteins ; metabolism ; Heart ; physiology ; Heart Failure ; physiopathology ; Humans ; Myocardium ; metabolism ; Receptors, Adrenergic, beta ; classification ; physiology ; Receptors, Adrenergic, beta-1 ; physiology ; Receptors, Adrenergic, beta-2 ; physiology ; Signal Transduction

OBJECTIVETo explore clinical outcomes and advantages of anterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal tuberculosis.

METHODSFrom February 2010 to February 2014, totally 82 patients with lumbar spinal tuberculosis were treated by posterior individual fixation with small-incision focus debridement,including 50 males and 32 females with an average of 50.5 years old. All patients were divided into two groups according to different procedures. Forty-nine patients in group A were treated with anterior small-incision focus debridement with posterior internal fixation through muscle spa ring at stage I ; and 33 patients in group B were treated with focus debridement with posterior internal fixation by extraperitoneal approach at stage I . Postoperative mechanical ventilation time, preoperative and postoperative Cobb angle, visual analogue scale (VAS), erythrocyte sedimentation rate (ESR) and Frankel grading were observed and compared. Postoperative complications, stability of internal fixation and bone union were compared.

RESULTSAll patients were followed-up from 15 to 36 months with an average of 23.7 months. Psoas abscess of three patients in group A and 1 patient in group B on the opposite side increased and were healed by the secondary apocenosis. The other 78 cases were healed at stage I, and no sinus tract formation, incisional hernia, leakage of cerebrospinal and occurrence of spinal tuberculosis were occurred. Fracture healing time ranged from 3 to 7 months with an average of 4.6 months. Postoperative mechanical ventilation time and VAS score in group A was better than group B. There were no statistical differences in Cobb angle, ESR and Frankel grading at the final following-up between two groups.

CONCLUSIONAnterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal according to degree of damage is a safe and effective method.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Debridement ; methods ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Treatment Outcome ; Tuberculosis, Spinal ; surgery ; Young Adult

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

Using matchmaker yeast two-hybrid system, it has been demonstrated that there exists an interaction between the cellular C terminal of alpha(1A)-adrenergic receptor (alpha(1A)-AR) and a segment of bone morphogenetic protein-1 (BMP-1). In the present study binding between the two proteins was further determined in human embryonic cell 293 (HEK293), a mammalian expression system. Mammalian expression vector PCP3HA was constructed by PCR and consisted of segments of BMP-1 cDNA, and vector PDT-alpha(1A) consisted of the full-length cDNA of human alpha(1A)-AR. They were transfected to HEK293 cells and examined by Western blot. alpha(1A)-AR and the segment of BMP-1 could be detected in the lysis of transfected cells. Then binding between alpha(1A)-AR and the segment of BMP-1 in HEK293 cell was determined by enzyme-linked immunosorbent assays (ELISA) and co-immunoprecipitation. In ELISA experiment, the ELISA microwell plate was first coated with anti-FLAG M2 antibody, which recognizes the FLAG-tagged alpha(1A)-AR, then the cell lysis, anti-HA rabbit polyclonal antibody and HRP conjugated anti-rabbit antibody were added in turn. The OD(490) values among the control group, PDT-alpha(1A) transfection group and PCP3HA transfection group, exhibited no significant difference (0.034+/-0.027, 0.042+/-0.019, 0.030+/-0.0096), but the OD(490) values of PDT-alpha(1A) and PCP3HA co-transfection group (0.57+/-0.12) were significantly higher than those of the other groups (P<0.001, respectively). In co-immunoprecipitation experiments, HEK293 cells expressing alpha(1A)-AR or/and segment of BMP-1 were lysed and incubated with anti-FLAG M2 antibody, then the immunoprecipitation pellet was immunoblotted with either the HRP conjugated anti-FLAG antibody or the anti-HA antibody, which recognizes the HA-tagged segment of BMP-1. Segment of BMP-1 was present in the pellet immunoprecipitation of PDT-alpha(1A) and PCP3HA co-transfected group. In conclusion, the results indicate that alpha(1A)-AR and the segment of BMP-1 are present in the same complex in HEK293 cells.
Adrenergic alpha-Agonists ; pharmacology ; Bone Morphogenetic Protein 1 ; Bone Morphogenetic Proteins ; genetics ; metabolism ; Cells, Cultured ; Embryo, Mammalian ; Enzyme-Linked Immunosorbent Assay ; Humans ; Kidney ; cytology ; metabolism ; Metalloendopeptidases ; genetics ; metabolism ; Protein Binding ; Receptors, Adrenergic, alpha-1 ; genetics ; metabolism ; Transfection ; Two-Hybrid System Techniques

OBJECTIVETo examine whether Chinese medical herb Pueraria crude extract (CP) and standard of pure puerarin (SP) possess the same neuroprotective effects during concomitant ethanol (EtOH) treatment.

METHODSHippocampus cultures were prepared from mice at gestational age of 18 day. Cell viability was measured by MTT assay. RT-PCR was employed to determine mRNA expression of superoxide dismutase (SOD).

RESULTSAs measured by MTT assay, supplementation with 15 mg/L CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50, 200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at mRNA level.

CONCLUSIONThis study suggests that CP and SP could decrease oxidative stress induced by ethanol treatment by the decreased expression of SOD at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and SP against developmental ethanol exposure in vitro.

Animals ; Antiviral Agents ; pharmacology ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Ethanol ; toxicity ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Hippocampus ; cytology ; embryology ; metabolism ; Isoflavones ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Plant Extracts ; pharmacology ; Pregnancy ; Pueraria ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Superoxide Dismutase ; genetics

A simple and sensitive Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) method was established to provide a new alternative for clinical diagnosis of Avian influenza A H5N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the target for amplification of nucleic acid under isothermal conditions. In current study, fifty-one experimentally infected animal specimens and viral cultures that had been tested were analyzed by RT-LAMP for NA gene and HA gene, respectively. The amplification process of LAMP was monitored in real-time by the addition of SYBR Green dye. Meanwhile, the result showed high correlation between nested PCR and RT-LAMP. The specificity of the RT-LAMP assay was confirmed by restriction enzyme digestion analysis and sequencing of the amplified product. When the sensitivity of this assay was tested by serial 10-fold dilutions of RNA molecules from specimens, it was found that the RT-LAMP method achieved theoretically a sensitivity of 10 RNA molecules. Thus, we concluded that the RT-LAMP assay has potential usefulness for rapid detection of the Avian influenza A H5N1 virus.
Animals ; Birds ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; diagnosis ; virology ; Nucleic Acid Amplification Techniques ; methods ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

This study was aimed to determine the in vivo signal transduction pathway responsible for isoproterenol (ISO)-induced cardiac hypertrophy or remodeling. Mice were treated with ISO (15 mg/kg body weight) or vehicle by intraperitoneal injection (i.p.). Activation of mitogen-activted protein kinase (MAPK), NF-kappaB and JAK/STAT pathway in the left ventricular myocardium was measured by Western blot analysis. ISO significantly activated MAPK (ERK1/2 and p38) at early phase (5 min); biphasic activation of NF-kappaB was observed in our in vivo study; and ISO caused a delayed STAT3 activation (at 60 to 240 min) in mouse myocardium. Taken together, these results indicate that ISO activates these signal transduction pathways in different time course.
Animals ; Heart ; drug effects ; Isoproterenol ; pharmacology ; Janus Kinase 1 ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mitogen-Activated Protein Kinases ; metabolism ; Myocardium ; metabolism ; NF-kappa B ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Time Factors

The expression of beta-adrenergic receptor subtypes and its effect on neonatal rat cardiac fibroblast proliferation were investigated by radioligand binding assay and [(3)H]-thymidine incorporation analysis, respectively. The results indicated that there was no significant difference in the beta-adrenergic receptor density (B(max)) and affinity (K(D)) between cardiomyocytes and cardiac fibroblasts. The [(125)I]-pindolol competitive inhibition curves (ICI 118551 and CGP 20712A) were significantly better fit in a one-site model in membrane preparation of cardiac fibroblasts. In cultured cardiac fibroblasts, 0.1 micromol/L isoproterenol-induced [(3)H]-thymidine incorporation was completely inhibited by a selective beta (2)-AR antagonist ICI 118551, or a non-selective beta-AR antagonist propranolol, but not by CGP 20712A, a selective beta(1)-AR antagonist. These results suggest that isoproterenol-induced cardiac fibroblast proliferation is mediated by beta(2)-AR, the preponderant beta-AR subtype in cardiac fibroblasts.
Animals ; Animals, Newborn ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Rats ; Rats, Wistar ; Receptors, Adrenergic, beta-2 ; genetics ; metabolism ; physiology