AIM:Toinvestigatetheeffectsofcalcitoningene-relatedpeptide(CGRP)onmyocardinexpres-sion and phenotypic switch in vascular smooth muscle cells ( VSMCs) .METHODS:VSMCs were obtained by aortic tissue adherent culture and treated with angiotensin Ⅱ( AngⅡ) , AngⅡ+CGRP or AngⅡ+CGRP +CGRP8-37 .The protein expression of myocardin and the phenotypic proteins of the VSMCs was detected by Western blot .RESULTS:The expres-sion of myocardin in cultured VSMCs showed downregulation along with time expansion .The protein level of myocardin was higher at 48 h and 72 h than that at baseline in the cultured VSMCs (P<0.05).However, the myocardin was lower at 48 h and 72 h than that at baseline after treatment with CGRP in cultured VSMCs (P<0.05).Furthermore, at 48 h in cul-tured VSMCs, the myocardin decreased along with α-smooth muscle actin (α-SMA) (P<0.05), and osteopontin (OPN) increased (P<0.05) in AngⅡ group compared with control group .After treatment with CGRP, the levels of myocardin andα-SMA become higher ( P <0.05 ) but OPN was lower ( P <0.05 ) in CGRP group than those in AngⅡ group. CGRP8-37 abrogated CGRP-induced increase in myocardin and α-SMA and decrease in OPN in CGRP 8-37 group compared with CGRP group .CONCLUSION: CGRP may regulate the phenotypic switch of the VSMCs and maintain the cells in contractile phenotype through the upregulation of myocardin protein , which may be accomplished by the combination of CGRP and its receptor .

OBJECTIVETo observe dynamically the development of fetal long bone and detect the expression and distribution of HIF-1alpha,to investigate the expression pattern and possible effects of hypoxia inducible factor-1alpha (HIF-1alpha) in fetal long bone development of mouse.

METHODSE12.5, E13.5, E14.5, E15.5, E16.5 and E17.5 pregnant C57BL6 mice were sacrificed. After sacrifice, the embryos were delivered by caesarean section. The development of fetal long bone was dynamically observed by stereoscopic microscope, and the distributional expression of HIF-1alpha protein was detected by using method of immunohistochemistry. The expression of HIF-1alpha mRNA and osteoblast marker gene at various stage were also detected by using methods of reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cartilaginous long bone began to form and joints outline arised at E13.5, then the primary ossification center was observed at E14.5, showing opaque ossification under stereoscopic microscope,and then the osteogenesis expanded and extended to both sides. Immunohistochemistry demonstrated lots of HIF-1alpha protein positive chondrcytes in the center of primary ossification at E14.5, then they decreased dramatically. HIF-1alpha mRNA expressed at high level from E13.5 to E15.5, and then decreased to low level.

CONCLUSIONFetal long bone development pattern appeared to be endochondral osteogenisis process, existing hypoxia microenviroment may increase HIF-1alpha mRNA expression and thus initiate the cascade of endochondral osteogenisis.

Animals ; Bone Development ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; genetics ; physiology ; Immunohistochemistry ; Male ; Mice ; RNA, Messenger ; analysis

OBJECTIVETo evaluate the effect of meatoplasty with the pedicle flap in the treatment of meatal stenosis secondary to chronic balanitis.

METHODSWe retrospectively analyzed 32 cases of meatal stenosis secondary to chronic balanitis treated by meato- plasty with the pedicle flap. All the patients had a history of chronic balanitis and had received meatal dilatation or simple ventral mea- totomy without significant effect. Their mean maximum urinary flow rate (Qmax) was (4.3 ± 2.4) ml/s. During the operation, A "/\"-shaped incision was made in the healthy epidermis and a flap was harvested from the frenulum. After complete removal of the scar, the flap was placed into the urethral wall, followed by reconstruction of the external urethral orifice.

RESULTSThe patients were fol- lowed up for 6 to 30 months, which revealed smooth urination in all the patients with Qmax of (26.7 ± 4.5) ml/s and normal erectile function and uresiesthesis.

CONCLUSIONWith little invasiveness and few complications, meatoplasty with the pedicle flap is an ideal surgical method for the treatment of meatal stenosis secondary to chronic balanitis. However, there might be some change in the normal appearance of the balanus postoperatively, and its long-term effect needs further observation.

Balanitis ; complications ; Constriction, Pathologic ; etiology ; surgery ; Dilatation ; Humans ; Male ; Postoperative Period ; Reconstructive Surgical Procedures ; Retrospective Studies ; Surgical Flaps ; Urethra ; surgery ; Urethral Stricture ; etiology ; surgery ; Urination

AIMTo determine the molecular weight and first-order structure of somatostatin.

METHODSThe molecular weight of somatostatin was determined by electrospray ionization mass spectrometry. Somatostatin was deoxidized by 2-mercaptoethanol. A series of typical fragment ions of deoxidized product were obtained by insource collision-induced dissociation (CID).

RESULTSThe m/z of quasi-molecular ion [M + H]+ of somatostatin was 1,637.8 and [M + Na]+ was 1,659.5. The m/z of double-charge ion [M + 2H]2+ was 819.5 and [M + H + Na]2+ was 830.3. It showed that the molecular weight of somatostatin was 1,636.7. The y and b series of fragment ions of deoxidized product were obtained by adjusting the fragmentor voltage. It was determined that the first-order structure of deoxidized product of somatostatin was A-G-C-K-N-F-F-W-K-T-F-T-S-C.

CONCLUSIONThe molecular weight and first-order structure of somatostatin were confirmed.

Amino Acid Sequence ; Molecular Structure ; Molecular Weight ; Somatostatin ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

AIMTo establish a RP-HPLC method for determination of cyclovirobuxine D.

METHODSCyclovirobuxine D reacted with a derivative reagent 1-naphthyl isocyanate in chloroform to form fluorescence derivatives, stopped the reaction by adding the mobile phase and then directly injected the solution into the chromatograph to seperate it by RP-HPLC. The analysis was carried out on C18 column, the mobile phase is methanol-water (85:15), the excitation wavelength was set at 305 nm, emission at wavelength 385 nm, and the flow rate was 1 mL.min-1. The effect of several factors including the reaction medium, temperature, time and amount of 1-naphthyl isocyanate on the yield of the derivatization was also investigated systematically.

RESULTSA simple and rapid RP-HPLC method for the simultaneous isolation and analysis of cyclovirobuxine D and its related substances was developed, and the absence of interference between the derivative peak responses of cyclovirobuxine D and its related substances were verified by UV diode array detecter and MS. The linearity was obtained from 0.75 microgram.mL-1 to 2.5 micrograms.mL-1 of cyclovirobuxine D derivatives with a correlation coefficient of 0.9991. The detection limit of cyclovirobuxine D derivative was 1 ng.mL-1, the repeatability of derivatization was good with relative standard derivation no more than 1.2% and derivative was stable within 48 h. The method described conforms to the validation of China Pharmacopiea compendial methods used for pharmaceutical products in general.

CONCLUSIONThe established method is proved to be reliable quantitative method for the quality control of cyclovirobuxine D.

Buxus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; analysis ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Powders ; Quality Control

OBJECTIVETo investigate the therapeutic effect of penoplasty with scrotal skin flap for the treatment of buried penis in children.

METHODSThe narrow ring was cut vertically at the ventral side of penis and the prepuce inner plate was circularly cut 0.5 cm from the coronary sulcus. The prepuce was degloved to the base of penis. The abnormal aponeurosis was removed completely. The prepuce was designed to cover the coronary sulcus. The scrotal flaps at both sides were formed and advanced to cover the penile base. Then the prepuce was sutured to reconstruct penile-scrotal angle and scrotum plasty was completed.

RESULTSFrom March 2009 to July 2011, 24 children with buried penis were treated with scrotal flaps. Adhesion at external orifice of urethra was happened in two cases which recovered after urethra expansion. There was one case of necrosis at the distal end of prepuce. All the cases were followed up for 6 months to 2 years with no penile shrinkage. The penile appearance was good without rotation or lateral curvature during erection.

CONCLUSIONSThe buried penis can be best corrected with scrotal flap. It is an ideal method with less complication.

Child ; Child, Preschool ; Humans ; Infant ; Male ; Penis ; abnormalities ; surgery ; Scrotum ; surgery ; Surgical Flaps ; Treatment Outcome

OBJECTIVETo decrease lung and esophageal radiation injuries by reducing irradiation target volume of mediastinal lymph mode drainage in conformal radiotherapy (CRT) for patients with non-small cell lung cancer (NSCLC) after thoracic surgery.

METHODSFifty-three patients with NSCLC were randomized into groups A and B to receive 3D-CRT after thoracic surgery. Patients in group A, according to conventional therapy, received preventive nodal irradiation (PNI) of the mediastinal lymph node drainage, and those in group B, according to pathological nodal staging after operation, did not have PNI of the metastasis-free area to reduce the clinical target volume (CTV). Patients in both groups were treated with conventional fractionated radiotherapy (CFRT) at 2 Gy in each fraction, and 5 fractions each week. All patients were followed up for two years to record their 2-year survival rate, local relapse of lymph node drainage and lung and esophageal radiation injuries.

RESULTSThe total 2-year survival rate was 58.5%in these patients and comparable between the two groups. The rates of local regional relapse and recurrence out of the CTV were 13.8% and 3.4% in group A and 16.7% and 8.3% in group B, respectively (P=1 and P=0.571). The incidence of radiation pneumonia and lung fibrosis were 6.9% and 62.1% in group A and 0% and 58.3% in group B (P=0.459 and P=0.782), and that of radiation esogphagitis and esophagus stricture rates were 27.6% and 6.9% in group A and 12.5% and 4.2% in group B, respectively (P=0.039 and P=1).

CONCLUSIONReduced CTV does not warrant decrease in the local control but may lower the incidence of acute esophageal radiation injury in postoperative patients with NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Drainage ; Esophagus ; pathology ; radiation effects ; Female ; Humans ; Lung ; pathology ; radiation effects ; Lung Neoplasms ; diagnostic imaging ; pathology ; radiotherapy ; surgery ; Lymph Nodes ; surgery ; Male ; Mediastinum ; Middle Aged ; Organ Size ; Radiation Injuries ; prevention & control ; Radiotherapy, Conformal ; adverse effects ; Recurrence ; Survival Rate ; Thoracic Surgery ; Tomography, X-Ray Computed

OBJECTIVE: To summarize the occurrences of adverse drug reactions (ADR) of gatifloxacin, and to guide the rational usage of antibacterial agents in clinical practice in the future. METHODS: A total of 1 077 ADR patient who received gatifloxacin were retrospectively studied in Hunan province from August 2003 to July 2007. RESULTS: Gatifloxacin could cause multi-system and multi-organ ADRs with wide variety of clinical manifestations. Of the 1 077 ADR patients, ADR incidence rate was slightly lower in males than that in females, the age of 821 (76.23%) ADR patients were 20 approximately 59 years; 905 (84.03%) were administered intravenously; and 682 (33.07%) had severe lesions of the digestive system, followed by lesions of the skin and the appendants (490 cases, 23.76%) and the nervous system (298 cases, 14.45%). CONCLUSION ADR caused by gatifloxacin should be monitored and reported so as to reduce or avoid ADR.
Adolescent ; Adult ; Adverse Drug Reaction Reporting Systems ; Aged ; Aged, 80 and over ; Anti-Infective Agents ; adverse effects ; Child ; Child, Preschool ; China ; Drug-Related Side Effects and Adverse Reactions ; Female ; Fluoroquinolones ; adverse effects ; Gatifloxacin ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo analyze the promoter methylation levels of p15, CDH1, DAPK and HICI genes of patients with myelodysplastic syndrome (MDS) and explore the relationship between the level of methylation and clinical features.

METHODSDNA methylation levels of p15, CDH1, DAPK and HICI in peripheral blood (PB) or bone marrow (BM) samples from 52 MDS patients were detected by real-time quantitative PCR. The correlation of the methylation level with clinical features and hematological findings was analyzed. 38 de novo AML patients and 46 normal individuals served as controls.

RESULTSThe methylation levels of p15, CDH1, DAPK and HICI were 16.23 ± 21.69, 6.59 ± 9.39, 0.14 ± 0.11 and 7.81 ± 9.70 in BM, and 14.96 ± 20.16, 6.00 ± 9.26, 0.12 ± 0.14 and 6.74 ± 9.72 in PB, respectively from 18 MDS patients, and the difference between BM and PB was not statistically significant (P > 0.05). The methylation levels of p15 (14.70 ± 18.17) and CDH1 (6.61 ± 8.79) genes in high risk (RAEBI/II) MDS were significantly higher than in low risk (RCMD/RARS/5q-, p15: 1.99 ± 1.59, CDH1: 1.23 ± 1.14 and RCMD, p15: 3.02 ± 3.42, CDH1:1.53 ± 2.06) MDS or control (p15: 1.69 ± 1.82, CDH1: 1.01 ± 1.12) (P < 0.05). The methylation levels of DAPK gene had no difference among subtypes of MDS, and that of HIC1 gene only differed between RAEB I/II (9.16 ± 11.95) and control (2.49 ± 2.26) (P = 0.042). The difference of methylation levels of p15, CDH1, DAPK and HICI in BM was statistically significant among subtypes of MDS (P = 0.001, 0.003, 0.039, 0.023, respectively). And so did of p15 and DAPK in PB (P = 0.013, 0.006, respectively). The methylation level of p15 and CDH1 was significantly correlated with IPSS classification and blasts percentage in BM.

CONCLUSIONSp15 and CDH1 genes are special hypermethylation genes in MDS. Methylation level of HIC1 gene showed an upward tendency from low risk to high risk MDS.

Adult ; Aged ; Aged, 80 and over ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinases ; genetics ; metabolism ; Case-Control Studies ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; metabolism ; DNA Methylation ; Death-Associated Protein Kinases ; Female ; Humans ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Young Adult