Objective To investigate the alteration of the cytoskeleton of airway smooth muscle cells in young asthmatic rats with airway remodeling and the effect of RhoA/ROCK signal pathway.Methods Airway smooth muscle cells (ASMCs) were primary cultured and purified from Sprague-Dawley(SD) rats that were induced by ovalbumin (OVA) inhalation for 8w,then incubated by Pho kinase inhibitor Y27632.Real time quantitative polymerase chain reaction (RT-PCR),Western blot,and immunohistochemistry were used to measure the alteration of F-actin,and α-tubulin in the cytoskeleton of airway smooth muscle.Results (1) The asthma group showed a high average gray value of F-actin in ASMC than control groups,especially 8 weeks;and were significantly down in the group after adding Y27632(P ＜0.01).(2) The intension and intensity of fluorescence signal of α-tubulin in asthma groups in 8 weeks were higher than control greup(P ＜0.01),and were significantly decreased in Y27632 group.(3) A higher expression of α-tubulin protein was shown in the asthma group in 8 weeks relative to control group(P ＜0.01),and was significantly down-regulated in Y27632 group(P ＜0.05).Conclusions Alteration of the cytoskeleton of airway smooth muscle exists in young asthmatic rats and the RhoA/ROCK signal pathway possibly plays a significant role.

Cholesteatoma ; diagnosis ; Ear, Middle ; Humans ; Otitis Media ; diagnosis

Objective:Facial nerves can be dissected using anterograde and retrograde approaches. The optimal technique for the facial nerve dissection of a patient with benign parotid tumor has not yet been determined. This study focused on facial nerve dysfunc-tion and recovery rate after anterograde and retrograde facial-nerve dissections. Methods:The data of 110 patients with benign carotid adenoma from the Head and Neck Department of this hospital who were hospitalized between January 2011 and January 2013 were col-lected. These patients were divided into groups A (n=52) and B (n=58). Anterograde and retrograde dissections of the facial nerve were performed on group A and group B patients, respectively. Based on the preferential order of dissection, group B was divided into groups B1, B2, and B3 representing the zygomatic, buccal, and marginal mandibular branches, respectively. The patients were postoperatively observed to check for potential symptoms, such as facial paralysis along with its severity and recovery. The House-Brackmann grading system was used to assess all patients. Results:The operation could be successful, with better nerve exposure, using these approaches. Statistical differences were observed in the nerve injury and recovery rates between the groups, with group A better than group B, and group B2 better than the other two groups (P<0.05). Conclusion:Anterograde facial nerve dissection should be routinely used in be-nign parotid tumor, and the buccal branch of facial nerve dissection should be preferentially considered when no other option apart from retrograde dissection is available.

AIMTo determine Danshensu in urine and study its pharmacokinetics in human.

METHODSA solid phase extraction-HPLC method was used for determination of Danshensu in urine of human. HPLC separation is performed on a Shim-pack CLC-ODS column (150 mm x 6.0 mm ID, 5 microns) with a mobile phase composed of acetonitrile -0.01 mol.L-1 KH2PO4 (adjusted to pH 2.8 with phosphoric acid). The flow rate was 1.0 mL.min-1 and the UV detector was set at 280 nm. The linear range of Danshensu was 0.2-50 mg.L-1 (r = 0.9999), and its limit of detection was 1.5 ng. The mean recovery was 99.4% (RSD = 2.9%).

RESULTSThe pharmacokinetics of Danshensu after p.o. administration of two kinds of pharmaceutical preparations containing Danshen (with 20 mg of Danshensu) were investigated in 6 healthy human volunteers by determining the Danshensu in urine samples. The elimination half lives (T1/2) of Danshensu after p.o. administration of compound granule preparation A and decoction of Danshen were (0.92 +/- 0.16) h and (0.94 +/- 0.21) h, respectively. Their excretions of Danshensu in urine were (6.2 +/- 2.8)% and (14 +/- 4)% of the dose in 8 hours, respectively.

CONCLUSIONUnder normal doses, Danshensu can be eliminated from kidney. There is no evident difference on elimination half lives of Danshensu after p.o. administration of the two doses, but the excretions of Danshensu by urine after p.o. administration of compound granule preparation A were lower than that of decoction of Danshen.

Adult ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Humans ; Lactates ; pharmacokinetics ; urine ; Male ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry

AIMTo separate and quantitatively determine six alkaloids: aconitine, mesaconitine, hypaconitine, beiwutine, benzoylaconine and benzoylmesaconine in the Chinese traditional medicine Radix Aconiti Lateralis Preparata (Fuzi).

METHODSA RP-ion-pair HPLC method was established. An AichromBond-1 C18 column was used at a column-temperature of 35 degrees C. The mobile phase was CH3CN5 mmol x L(-1) NaH2PO4(50:50) containing 7 mmol x L(-1) SDS at a flow-rate of 1.0 mL x min(-1). The detector was set at UV 235 nm.

RESULTSThese six alkaloids can be completely separated and determined quantitatively.

CONCLUSIONThis method is accurate and suitable for the determination of six alkaloids in Fuzi.

Aconitine ; analogs & derivatives ; analysis ; isolation & purification ; Aconitum ; chemistry ; Alkaloids ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Objective To investigate the relationship between concentration change of serum 5‐hydroxy tryptamine and clinical symptoms improvement in primary premature ejaculation with the treatment of paroxitine .Methods 81 cases of lifelong PE and an intra‐vaginal ejaculation latency time (IELT ) ≤60 s were included in this study .Subjects were divided into 2 groups according to the IELT ,group A (IELT≤30 s) and group B (30 s

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

AIM: To study and evaluate the changes of two main kinds of voltage-gated K+ currents in human atrial fibrillation (AF) and to discuss the role of these changes in the atrial electrical remodeling (AER) caused by AF. METHODS: Specimens of human atrial appendage were obtained from 36 RHD patients (18 with chronic AF and 18 without AF). Single atrial myocytes were acutely dissociated by tissue chunk enzymatic digestion. I_~to1 and I_~Kur in the two groups were measured respectively with the patch-clamp technique in a whole-cell configuration and the I-V curves were compared. RESULTS: I_~to1 and I_~Kur amplitudes in AF groups were significantly reduced and the current densities of both I_~to1 and I_~Kur in AF patients were lower than those in NAF patients. CONCLUSION: The reduction of I_~to1 and I_~Kur may be related to changes in atrial conduction, refractory period and may constitute two main parts of the major mechanisms in the AER of chronic AF. Whether exists a relation between changes of the above K+ currents and that of other ionic currents and the AF initiation and perpetuation deserves further investigation. [

BACKGROUND: Growth differentiation factor 5 (GDF-5) is an important factor to regulate the formation and development of the cartilage and bone, it plays a crucial role on the promotion of repairing bone, cartilage and tendon ligament injury. OBJECTIVE: To transfect eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 to bone marrow stroma stem cells of mouse and to check the expression of extracellular matrix and proteoglycan which relates with the cartilage formation and differentiation. DESIGN, TIME AND SETTING: An in vitro observation regarding cells was performed in the central laboratory of Wuhan Union Hospital between March and December in 2008.MATERIALS: Twenty Kunming specimen male mice were offered by Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology. The eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 was preserved at the laboratory.METHODS: The man-ow stroma stem cells were isolated from mouse bone marrow and cultured in vitro with whole bone marrow adherence method. Passage 3 cells were incubated on 6-well plate and began to transfect when they were 90% confluent. Experiment was assigned into three groups: trensfection group underwent transient transfection of liposome-mediated pcDNA 3.1(+)/GDF-5 using LipofectamineTM2000; blank plasmid group was transfected with blank plasmid pcDNA 3.1(+); control group was added with equal volume of liposome and other protocols were the same as above. MAIN OUTCOME MEASURES: The transfection efficacy was identified success by the expression of GDF-5 gene and protein using RT-PCR and immunocytochemistry at 72 heurs following transfection, cartilage matrix Ⅱ collagen expression was determined as above methods. Then marrow stroma stem cells were cultured for additional two weeks to check expression of proteoglycan with alcian blue staining.RESULTS: In the transfection group, a 219-bp specific amplification band was visible, there were brown positive stain in the cytoplasm of marrow stroma stem calls; In blank plasmid group and control group, no GDF-5 trensfection, specific amplification band or obvious stain of cytoplasm was observed. In the transfection group, the collagen Ⅱ gene was detected to express at 225 bp, with brown yellow stain in cytoplasm; in the blank plasmid group and control group, no collagen Ⅱ gene expression or SP ～ stain was observed. Alcian blue staining results showed the transfected cells were stained blue while those in the blank plasmid group and control group were not metachromasia stained.CONCLUSION: Gene trensfection of pcDNA 3.1 (+)/GDF-5 to marrow stroma stem cells can significantly raise expression of collagen II and proteoglycan, and promote the chondrogenic differentiation of marrow stroma stem cells.

OBJECTIVETo analyse a special kind of Schisandra chinensis with the white fruit using ITS2 barcode at molecular levels.

METHODITS2 regions were sequenced bidirectionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner, MEGA 5.0 software was used to align the sequences. The ITS2 secondary structure was predicted using ITS2 web server, BLAST 1 method was used to identify the S. chinensis with the white fruit.

RESULTThe length of the ITS2 sequence was 231 bp. And the sample was identified as S. chinensis using the method of BLAST 1. Their mean interspecific genetic distance (K2P distance) among the populations of the S. chinensis with the white fruit and S. chinensis was far lower than the mean interspecific genetic distance between the S. chinensis and S. sphenanthera.

CONCLUSIONBy using ITS2 the S. chinensis with the white fruit was identified as S. chinensis, and the ITS2 barcode could be used to identify S. chinensis and S. sphenanthera.

DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Fruit ; chemistry ; classification ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Schisandra ; chemistry ; classification ; genetics ; Sequence Analysis, DNA ; Software