To study the related substances in nicergoline, electrospray positive ionization high resolution TOF/MS was used for the determination of the accurate mass and elemental composition of the related substances. Triple quadrupoles tandem MS/MS was employed for the determination of the fragmentations of the parent ions. 16 related substances were detected and identified to be eight synthetic by-products and eight degradation products, by using impurity references matching, product mass spectra fragmentations elucidation, and verified further according to synthetic processes and stress testing results. The results obtained are valuable for nicergoline manufacturing process control and quality assurance.
Chromatography, High Pressure Liquid ; Nicergoline ; chemical synthesis ; chemistry ; Quality Control ; Tandem Mass Spectrometry

Objective To investigate the value of wide-detector Revolution CTA with 70 kV tube voltage and prospective ECG-gated technique in diagnosis of congenital heart disease (CHD) in infants and children.Methods Forty-five infants and children with complicated CHD underwent echocardiography and wide-detector Revolution CTA.According to the sur gical findings,the diagnostic efficiency of Revolution CTA and echocardiography were calculated and compared.The radiation effective dose (ED) and iodine dose were calculated.The quality of CT images was also evaluated.Results There were 25 separate cardiovascular anomalies including 6 congenital cardiac structure anomalies and 19 congenital extracardiac vascular anomalies.For congenital extracardiac vascular anomalies,there was significant difference of diagnostic accuracy and the detectable rate between CTA (99.77％ [853/855],97.73％ [86/88]) and echocardiography (98.71％ [844/855],88.64％ [78/88];x2 =6.28,5.72,both P＜0.05).The average of ED was (0.20±0.05)mSv and the mean iodine dose was (2.06± 1.09)g.All CT images were qualified for diagnosis.Conclusion The wide-detector Revolution CTA,with the prospective ECG-gated technique and 70 kV tube voltage,can provide high accuracy for assessment of CHD in infants and children,which can keep good image quality,with the low radiation dose.

ObjectiveTo investigate the protective effects and mechanism of ursodeoxycholic acid (UDCA) on α-naphthylisothi (ANIT)-induced cholestatic liver injury in rats.MethodsA total of 48 Sprague-Dawley (SD) rats were selected.Fouty-two rats were gavaged with ANIT (100 mg/kg) to induce acute liver injury,six rats were sacrificed 24 hours after the liver injury and the rats left were evenly divided into control group which were gavaged with saline and UDCA group which were gavaged with UDCA (20 mg/kg).Six rats were sacrificed at 48 hours,72 hours and 96 hours after modeling.The six untreated rats were set as blank control group.Serum and liver tissues of all rats were kept after sacrificed.Serum levels of alanine transaminase (ALT),aspartate transaminase (AST),total bilirubin (TBil) and total bile acid (TBA) were tested,interleukin-10 (IL-10),interleukin-6 (IL-6),and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).The expression of multidrug resistance associated protein2 (Mrp2) at mRNA level in liver tissue was measured by quantitative real-time polymerase chain reaction (qRT-PCR) and the inflammatory reaction activity of liver tissues was inspected with Haematoidin-Eosin (HE)staining under microscope.ResultsAt 48 hours after liver injury modeling,serum TBil (143.80± 12.08) μmol/L vs.(178.50±15.19) μmol/L,TBA (13.15±3.81) μmol/L vs.(21.68±7.93)mol/L,IL-10 (44.13±3.68,37.15±6.25 ng/L),IL-6(50.80±2.09,57.32±4.63 ng/L) and TNF-α (17.53±0.84) ng/L vs,(19.10±1.64) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P ＜ 0.01 or P＜ 0.05).At 72 hours after liver injury modeling,serum ALT (721.67±97.54) U/L vs.(929.50±148.29) U/L and IL-10 (54.68±6.79)ng/L vs.(43.85±4.08) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).At 96 hours after liver injury modeling,serum ALT (156.83±14.99) U/L vs.(250.67±42.29) U/L,AST (143.67±27.45) U/L vs.(206.00±63.94) U/L and TBil (23.53±5.08) μmol/L vs.(34.02±9.98) μmol/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).The differences of Mrp2 expression at mRNA level in liver tissues between UDCA group and control group at 48 hours (0.77 ± 0.21,0.46 ± 0.25),72 hours (2.27 ±0.84,1.10 ±0.38) and 96 hours (3.64±0.54,2.75±0.69) after liver injury modeling were statistically significant (P＜0.01 or P＜0.05).ConclusionThe mechanism of the protective effects of UDCA on ANIT-induced liver injury may be related with the regulation of serum cytokines and liver Mrp2 expression.

Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.

Objective To observe adhesion and growth of Sehwann cells(SCs) on small intestinal submueosa(SIS) and study the bioeompatibility of SIS with SCs.Methods The SCs of SD neonate rat were isolated and cultured in vitro,then were seeded on prepared SIS.At different times,the adhesion,growth and proliferation of SCs on SIS were observed by phase contrast microscope,histological examination,scanning e- lectron microscope and transmission electron microscope.Results By phase contrast microscope,SCs grew well on the edge of SIS after 3 and 5 days.SCs adhered tightly on the surface of SIS after 5 days through histo- logical examination.By scanning electron microscope,on the surface of SIS,SCs grew and adhered actively, prominence of cells body were obvious.They connected end-to-end with each other or arranged in clusters. Protein granules were secreted on cells surface.By transmission electron microscope,SCs grew in good condi- tion and adhered tightly on the surface of SIS.At the interface of SCs and SIS,prominence was seen to contact with SIS in the bottom of cell body.Conclusion SCs are able to adhere and grow well on the surface of SIS.As a scaffold,SIS has good biocompatibility with SCs.

Objecttve To explore the value of electrocardiographic(ECG)Cornell criteria for detecting left ventricular hypertrophy(LVH)in elderly Chinese men.Methods Since 1990,244 autopsies were performed in our hospital in elderly men,LVH was determined in these autopsy hearts and correlated to EGG LVH signs recorded within 3 months before death according to Comer(Sv3+RIvL)and Sokolow-Lyon criteria(Svl+Rvs or RV6).The reference value of Comell criteria was obtained based on valUes from autopsied healthy hearts,the sensitivity and specificity of Cornell and Sokolow-Lyon criteria for detecting left ventricular hypertrophy in these eldedy men were talculated.Results There were significantly correlations between QRS amplitudes of Cornell and Sokolow-Lyon criteria and autopsy left ventricular wall thickness in tllese hearts.The reference value of Comer criteria(SV3+RaVL)was 2.9 mV.The sensitivity of Sokolow-Lyon and Cornell criteria for detecting LVH Wag 25.4%and 34.3%(P＜0.05 vs Sokolow-Lyon criteria),respectively.Condusion Voltage(SV3+R.RavL)≥2.9 mV might be a suitable diagnostic value for detecting left ventricular hypertrophy in Chinese elderly men.

Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

Objective To investigate the effect of microRNA-223-3p (miR-223-3p) on megakaryocytic differentiation and maturation, and explore the potential mechanism .Methods The endogenous expression of miR-223-3p during megakaryocyte ( MK) differentiation was detected by real-time PCR.Flow cytometry further indicated that alteration of miR-223-3p in human cell lines exerted effects on MK differentiation and maturation .By performing integrative bioinformatic analysis, the potential miR-223-3p target gene, MYH10,was identified.Real-time PCR, luciferase reporter assay and flow cytometry revealed that MYH10 was a direct target of miR-223-3p.Results Endogenous expression of miR-223-3p was in-creased with the differentiation and maturation of MK .The expression of megakaryocytic surface markers CD41 and CD61 and the ploidy were significantly increased in K562 and Meg-01 cells after transfection with miR-223-3p mimics.The expression of MYH10 decreased with the increase in miR-223-3p.Using a luciferase reporter assay ,we demonstrated that MYH10 was a direct target of MiR-223-3p.Furthermore, direct downregulation of MYH10 promoted MK polyploidization . Conclusion MiR-223-3p might regulate the polyploidization of MK by targeting MYH10.

OBJECTIVETo construct the recombinant adenovirus expression vector of a short hairpin RNA (shRNA) targeting phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene for gene therapy of ischemic cerebral injury.

METHODSThe U6 expression promoter and shRNA of pGenesil-1-shRNA, which was constructed and identified in our previous experiment, were subcloned to pAdTrack shuttle plasmid. The product pAdTrack-U6-shRNA was linearized by PmeI for homologous recombination with pAdEasy-1 in pAdEasy-1 competence bacteria. The positive clone was identified by enzyme digestion, PCR analysis and DNA sequence analysis. After linearization by PacI, the recombinant adenovirus DNA shuttle plasmid pAdEasy-U6-shRNA was transfected into 293 cells for packaging and amplification of Ad-U6-shRNA, which was further identified by PCR analysis and DNA sequence analysis. Western blotting was used to detect the expression of PTEN protein in the hippocampal neurons infected with the adenovirus.

RESULTSThe pAdTrack-U6-shRNA and pAd-U6-shRNA plasmids had been successfully constructed as verified by PCR analysis, enzyme digestion and DNA sequence analysis. PCR analysis and DNA sequence analysis confirmed successful packaging of the recombinant adenovirus Ad-U6-shRNA in 293 cells. PTEN protein expression decreased significantly in the hippocampal neurons after infection by the recombinant virus.

CONCLUSIONWe have successfully constructed the recombinant adenovirus Ad-U6-shRNA targeting PTEN gene, which provides a basis for investigating the role of PTEN in neuroprotection after cerebral ischemic injury using RNA interference.

Adenoviridae ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA