ObjectiveTo assess the safety and clinical values of video-laryngoscope in spontaneous respiration tracheal intubation for emergency patients. Methods Seventy-nine patients,who needed the endotracheal intubation,were recruited in our department between January 2010 and December 2010,and were randomly ( random number) divided into two groups according to consultative sequence.Forty patients (group A ) were operated with traditional laryngoscope and thirty-nine patients (group B ) with videolaryngoscope.The operative time and success rate of tracheal intubation,Cormack-Lehane classification,as well as adverse events,were recorded.The heart rate ( HR ),mean arterial blood pressure ( MAP),respiratory rate ( RR ),and saturation of pulse oxygen ( SpO2 ) were observed pre-operation,during operation and 2 min post-operation.Results( 1 ) The Cormack-Lehane classification in group A were significantly lower than in group B. (2) The operative time of tracheal intubation in group B was significantly less than that in group A [(35.6+12.7) svs. (58.3 ± 13.5) s; P＜0.05] ; and one-time success rate of tracheal intubation in group B was higher than that in group A ( 84.6％ vs.52.5％ ; P ＜0.05).(3) Compared to group B,the HR and MAP in Group A were significantly increased at t2 and t3 ( P ＜ 0.05 ). ( 4 ) The adverse events,including restlessness,bucking and injury,were significantly decreased in group B than those in group A ( P ＜ 0.05 ).ConclusionsThe video-laryngoscope used in spontaneous respiration tracheal intubation,could improve Cormack-Lehane classification,short operative time,enhance one-time success rate and reduce adverse evevnts.

Objective To investigate the effects of constraint-induced movement therapy (CIMT) on the function of hemiplegic upper extremity in the early subacute stroke patients.Methods A prospective,single-blinded,randomized controlled study was conducted.Forty-seven stroke patients with hemiplegic upper limb dysfunction were randomly divided into 2 groups:a CIMT group and a control group.The CIMT group received constraintinduced movement therapy and the control group was treated with conventional rehabilitation therapy.Both groups were treated 3 h daily,5 d a week for 2 weeks.The Wolf motor function test (WMFT) and Fugl-Meyer Assessment (FMA) were used to evaluate upper extremity motor function,and motor activity log (MAL) was used to assess upper extremity ability in activities.WMFT,FMA and MAL were measured before and after 1 day and 6 months of treatment.Results After 1 day of treatment,FMA,WMFT,MAL-AU and MAL-HW were 53.81 ± 2.59,66.68 ± 3.54,1.89 ± 0.88 and 3.26 ± 0.65,respectively,in the CIMT group,and 48.61 ± 4.48,62.10 ± 7.97,1.25 ± 0.64 and 2.65 ± 0.93,respectively,in the control group.After 6 months of treatment,FMA,WMFT,MAL-AU and MAL-HW were 57.53 ±2.01,69.57 ± 3.00,3.00 ±0.82 and 3.84 ±0.69,respectively,in the CIMT group,and 53.30 ± 2.88,66.20 ±3.59,2.20 ± 1.06 and 3.25 ±0.64,respectively,in the control group.The scores of FMA,WMFT,MAL in the CIMT group were all higher than those in the control group at 1 day and 6 months post-treatment,and the differences were statistically significant (P ＜ 0.05).Conclusion Constraint-induced movement therapy can significantly improve the patients' hand function in the early stage of subacute stroke,which maintain up to 6 months of follow-up.

Objective To establish the method of thin layer identification of Phellodendri Chinensis Cortex and solid phase extraction - high performance liquid chromatography (SPE-HPLC) determination of the content of phellodendrine in Leshu Lotion. Methods Thin layer chromatography (TLC) was used for qualitative identification of Phellodendri Chinensis Cortex in Leshu Lotion and the Bond Elutplexa PCX strong cation-exchange solid phase was used for extraction small column purification. Platisil-NH2 column was as chromatographic column; column temperature was 30 ℃; triethylamine pH 3.82 (phosphoric acid), tetrahydrofuran, acetonitrile was 18:12:70, isocratic elution; flow rate was 1.0 mL/min; detection wavelength was 286 nm. Results TLC could obviously identify active ingredients of phellodendrine in Phellodendri Chinensis Cortex. The phellodendrine had good linear relationship between concentration and peak area within the scope of 1.22–12.2 μg (r=0.9999); the average recovery rate of phellodendrine was 101.18%, RSD was 1.71%. Conclusion This method is simple to operate, with accuracy and repeatability, and can be used for quality control of Leshu Lotion.

Objective This study was carried out in order to analyze genetic diversity in plants of Dendrobium Sw. and compare the marker index (MI) between SRAP and RAPD markers. Methods Using 40 SRAP primer combinations and 36 RAPD primers,the genetic diversity of the materials in Dendrobium Sw. was amplified. Results A total of 1 977 loci were generated by 40 SRAP primer combinations,of which 90.2% was polymorphic and the genetic similarity (GS) ranged from 0.330 2 to 0.789 2. As for RAPD,281 bands were obtained,in which 87.1% was polymorphic,and the GS was from 0.494 2 to 0.773 1. There were some differences existing in the two cluster results revealed by SRAP and RAPD markers,they were as follows:compared to that of RAPD,the result of SRAP could more comfortably reflect the kinship and geographical origin,simultaneously conform to the traditional classification. For the experiment materials,SRAP (16.46) got an MI value 4.5 folds higher than that of RAPD (3.67),which meant SRAP markers were more effective in detecting genomic polymorphisms in the plants of Dendrobium Sw. Conclusion These results show that SRAP and RAPD could be applied to detecting genetic diversity analysis of plants in Dendrobium Sw. SRAP Also shows great superiority and advantage in this study.

Objective To evaluate the relationship between smoking and p53 gene mutation in the lung cancer patients with Meta-analysis.Methods PubMed,Web of Science,ProQest and Medline were used to search all the relevant studies about the association between smoking and p53 gene mutation in the patients with lung cancer. Based on reviewing full text,the studies were selected and evaluated and the data was extracted.Statistical analysis was performed with Stata 1 2.0 software including the heterogeneity inspection, publication bias assessment, sensitivity analysis,effect consolidating and cumulative Meta-analysis.Results Totally 15 articles with 1 770 lung cancer patients were identified. 69.6% of the patients were smokers,30.4% were non-smokers.Overall,the smokers with lung cancer had a 2.70-fold higher risk for p53 gene mutation than the non-smokers with lung cancer (OR=2.70,95%CI=2.04-3.59).Conclusion p53 gene mutation is associated with smoking in the patients with lung cancer.The smokers with lung cancer have higher risk for p53 mutation than non-smokers.

AIM: To establish human umbilical vein endothelial cells (HUVECs) to express green fluorescent protein (GFP), and to study the suppression of GFP by siRNA in HUVECs. METHODS: Using lipofectamine 2000 to transform plasmid pN_3-EGFP encoding GFP into HUVECs. The HUVEC containing pN_3-EGFP, named HUVEC-GFP, was screened and selected by antibiotic G418. Using in vitro transcription T7 kit, GFPsiRNA targeting GFP mRNA and control-siRNA used as control were synthesized. The siRNAs were transfected into HUVEC-GFP with oligofectamine. 48 h later, the expression levels of GFP protein and mRNA in HUVEC-GFP were determined. RESULTS: The HUVEC-GFP was screened to express GFP in the presence of G418. The agarose gel electrophoresis analysis showed that the siRNAs prepared were integrated. 48 h after transfection with siRNAs, compared to control group, the level of GFP fluorescence was obviously decreased in the HUVEC-GFP transfected with GFPsiRNA. The results of RT-PCR detection showed that GFP mRNA expression was obviously suppressed by GFPsiRNA at the rate of 40%, and no obvious suppression of GFP mRNA expression was found in the HUVEC-GFP transfected with control siRNA. CONCLUSION: The siRNA targeting GFP mRNA, synthesized in vitro, efficiently suppresses the GFP expression in HUVECs.

AIM:To investigate the effect of caspase-8 small hairpin RNA ( shRNA) on attenuating apoptosis of human mesenchymal stem cells ( hMSCs ) .METHODS: Two recombinant plasmids for over-expression of caspase-8 shRNA, pAd-Cap8 shRNA1 and pAd-Cap8 shRNA2, were constructed.Caspase-8 mRNA was determined in pAd-Cap8 shRNA-transfected human HEK293 cells by Q-PCR.The screened pAd-Cap8 shRNA was used to construct the recombinant adenovirus plasmid , which was linearized and transfected into HEK 293 cells for packaging and amplification of the recombi-nant adenovirus rAd-Cap8 shRNA.The expression of caspase-8 at mRNA and protein levels was determined by Q-PCR and Western blotting .Annexin V/PI staining and determination of caspase-8 activity were performed to assess apoptosis of hM-SCs under the conditions of serum deprivation and hypoxia .The mRNA expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1), Bcl-2 and Bcl-xL was analyzed by Q-PCR.RESULTS:The pAd-Cap8 shRNA, which efficiently inhibited caspase-8 expression, was screened by Q-PCR.The recombinant adenovirus plasmid for caspase-8 shRNA was constructed and used to package and amplify the recombinant ad-enovirus ( rAd)-Cap8 shRNA successfully .rAd-Cap8 shRNA-mediated caspase-8 shRNA markedly inhibited caspase-8 ex-pression in hMSCs .Over-expression of caspase-8 shRNA by infection of rAd-Cap8 shRNA also efficiently decreased the ap-optotic rate and caspase-8 activity in hMSCs under the conditions of serum deprivation and hypoxia , with up-regulation of the mRNA expression of HGF, IGF-1 and Bcl-2.CONCLUSION:Caspase-8 shRNA attenuates hMSC apoptosis under the conditions of serum deprivation and hypoxia .

AIM:To construct recombinant adenovirus vector carrying human miR-133a and study its expression in human mesenchymal stem cells(hMSCs).METHODS:The PCR product containing miR-133a was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrack-CMV.Then the recombinant shuttle plasmid linearized by pmeⅠwas cotransformed into competent E.coli.BJ5183 with the adenoviral backbone plasmid pAdEasy-1 to generate the recombinant adenovirus vector rAd-mir-133a.rAd-mir-133a was then packaged and amplified in human embryonic kidney 293(HEK293) cells.The purified rAd-miR-133a was used to infect the hMSCs and the expression of miR-133a was detected by non-quantitative RT-PCR and real-time PCR.RESULTS:The recombinant adenovirus shuttle vector pAdTrack-CMV-miR-133a was constructed and verified by restriction endonuclease analysis and DNA sequence analysis.rAd-miR-133a was successfully packaged and amplified in HEK293 cells.The transcriptions of primary miR-133a and mature miR-133a were over-expressed in the hMSCs infected with rAd-miR-133a.CONCLUSION:The recombinant adenovirus vector carrying human miR-133a is successfully constructed,which lay a foundation for miR-133a function study.

AIM:To investigate the effect of microRNA-214 ( miR-214) on cardiomyocyte hypertrophy and the expression of the potential target genes .METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular cardiomyocytes ( NMVCs) .Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Western blot , respectively .RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-induced hypertrophic cardiomyocytes .Dual lu-ciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly increased in the hypertro-phic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hypertrophy-re-lated genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .

Objective To observe the effect of ultrasound-guided transversus abdominis plane block combined with diclofenac sodium suppository in anus on postoperative analgesia and early recov-ery after laparoscopic hysterectomy.Methods Sixty-six patients of laparoscopic hysterectomy,aged 40-60 years,were randomly divided into group T and group D,33 cases in each group.Group T used ultrasound-guided bilateral transversus abdominis plane (TAP)block after surgery.Group D used TAP block and diclofenac sodium suppositories in anus.All patients received PCIA.Postoperative 24 h sufentanil dosage out of analgesia pump and the effective press times were recorded.The incidence of dysphoria,nausea and vomit within postoperative 24 h,the time of first out-of-bed activity,the time of first anal exhaust and the time of removing urine tube were measured.The plasma concentra-tion of interleukin-6(IL-6),C reactive protein (CRP)and cortisol (Cor)were recorded at the follow-ing four time points:before anesthesia,just postoperative moment,8 and 12 h after surgery. Results Compared with group T,the sufentanil dosage [(18.6±3.2)μg vs (40.8±5.1)μg]and the effective press times [(9.3±2.2)times vs (20.4±3.5)times]were fewer in group D (P<0.05).The incidence of dysphoria,and nausea and vomit were less in group D compared with group T (P<0.05).The IL-6,CRP and Cor of group D at 12 h were lower than those of group T (P<0.05). But the times of first out-of-bed activity,anal exhaust and removing urine tube in two groups were not significantly different.Conclusion Transversus abdominis plane block combined with diclofenac sodium suppositories in anus can provide safe and effective postoperative analgesia for laparoscopic hysterectomy,and promote the early fast recovery.