Objective To investigate the possibility that the construction and expression of a eukaryotic expression vector system of rat NN-NAT gene. Methods The full-length cDNA fragment of rat AA-NAT gene was amplified by RT-PCR method.After retrieving the PCR products,ligating it with pTARGET~(TM) vector,transformating ligation reaction to JM109 huge efficiency competent cells and identifying the recombinant plasmid,the recombinant eukaryotic expression vector pTARGET~(TM)-AANAT was transfected into rat L6 myoblasts with lipofectamine.Accordingly,engineered cells selected by antibiotic G418 were detected by the methods of RT-PCR and Westem blotting. Results It was revealed that,amplified AA-NAT cDNA confirmed by agarose gel electrophoresis could ligate with pTARGET~(TM) vector and subcloned into JM109 cells.L6 cells transfected with pTARGET~(TM)-AA-NAT survived well after G418 selection and expressed AA-NAT protein. Conclusion Our results suggest that we have prepared rat AA-NAT stable eukaryotic expression system successfully although it was just a primary result.This system can be used for the transfection of L6 myoblast.

Cardiomyopathies ; diagnosis ; etiology ; therapy ; Humans

OBJECTIVE:To systemically review the efficacy and safety of Ambroxol hydrochloride injection by intravenous drip combined with aerosol inhalation in the treatment of neonatal pneumonia,and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from PubMed,Medline,CBM,CJFD,VIP and Wanfang Database,randomized controlled trials (RCT)about Ambroxol hydrochloride injection by intravenous drip combined with aerosol inhalation(test group)versus Ambroxol hydrochloride injection alone (control group) in the treatment of neonatal pneumonia. Meta-analysis was performed by using Rev Man 5.2 software after data extracting and quality evaluating by Cochrane 5.1.0. RESULTS:Totally 14 RCTs were enrolled,involv-ing 1 407 patients. Results of Meta-analysis showed the clinical efficacy[RR=1.19,95% CI(1.13,1.24),P<0.001] in test group was significantly higher than control group,the time of cough disappearance[MD=-1.75,95%CI(-1.91,-1.58),P<0.001], asthma disappearance[MD=-1.38,95%CI(-1.67,-1.08),P<0.001] and pulmonary rales disappearance [MD=-1.44,95%CI (-1.77,-1.11),P<0.001] and hospitalization days[MD=-2.04,95%CI(-2.25,-1.82),P<0.001] were significantly shorter than control group,the differences were statistically significant;and there was no significant difference in the incidence of adverse reactions between 2 groups[RR=1.00,95%CI(0.49,2.05),P=1.00]. CONCLUSIONS:Both the efficacy and safety of Ambroxol hydrochloride injection by intravenous drip combined with aerosol inhalation are good in the treatment of neonatal pneumonia.

Batch fermentation of lycopene by recombinant E. c ol i ZYL-2 at various temperatures ranging between 23℃～37℃ were studied in 7L fermentor. At 33℃, cell specific growth rate in earlier stage of culture was higher, and the time obtained maximum cell dry weight was shorter at other tempe rature. while after 9 h, lycopene specific production rate was higher at 28℃. Based on these results, a two-stage temperature control strategy was developed in which 33℃ was used for fermentation for the first 9 h and the temperaturew as switched to 28℃ after 9h. Using this temperature-shift strategy, the maxim al lycopene content and productivity reached 605.25?g L-1 and 28.82?g L-1h-1. The lycopene fermentation level obtained by the strategy was higher than those in single temperature-control experiments.

OBJECTIVE To d e f i n e t h e microsurgical anatomy and exposure obtained by the maxillary swing approach. METHODS Ten adult cadaver specimens fixed by formalin solution were dissected under 5 to 20 times magnification. The microsurgical anatomy and measurement were performed by mimicking the maxillary swing approach. The anatomic relationship of the muscles, arteries,veins,nerves were carefully recorded. RESULTS The maxillary swing approach provides wide exposure extending from the central base to the lateral cranial base,including turkish saddle,the ipsilateral cavernous sinus,the clivus and C1,the nasopharynx,the pterygopalatine and infratemporal fossa,and the parapharyngeal space,etc. CONCLUSION The maxillary swing approach provides a suitable route for accessing lesions involving both the central and lateral cranial base. The sphenoid pterygoid process,the Eustachian tube,and the lateral pharyngeal wall separates the central cranial base from the lateral cranial base and limits anterolateral access to the central cranial base. Removing the pterygoid process provides exposure extending from the central to the lateral cranial base and allows for wide anterolateral access to the clivus and upper cervical spine. The maxillary swing approach is difficult and limited to reach the poststyloid compartment of the parapharyngeal space,the lateral region of styloid process,the horizontal segment of the petrous carotid.

Antihypertensive Agents ; administration & dosage ; therapeutic use ; Humans ; Hypertension ; drug therapy ; genetics ; Molecular Biology

Objective To study the anticoagulation effects of Supplemented Salvia Beverage (SSB) on myocardial ischemia in rats. Methods Rat myocardial ischemia model was induced by occluding the left anterior descending coronary artery,the effects of SSB (8.24,4.12,2.06 g/kg) on the activated partial thromboplastin time (APTT),prothrombin time (PT),thrombin Time (TT),platelet adherence rate,platelet aggregation,length of thrombus and the dry and wet weight of thrombus,as well as amount of Fibrinogen (Fg) were observed. Results Compared with model group,the shortening of PT and TT,and the decrease of Fg were obviously suppressed in high dosage SSB group (P

Transient receptor potential vanilloid 1 (TRPV1) is a non-selective cation channel,which can be activated by multiple pathways during the course of the diseases.Recent studies indicate that primary sensory neurons of the pancreas express TRPV1 receptor and the activation of TRPV1 receptor promotes pancreatic inflammation.Moreover,blockade of these transient receptor potential channels can greatly ameliorate the pain response in experimental pancreatitis.

Objective:To explore potential differentiation from antigen-1 positive stem cells (Sca-1 +cells) in murine fetal liver tissue into cardiomyocytes in vivo. Methods: Sca-1 + cells from the murine fetal liver of 14.5 d were separated by Magnetic cell sorting (MACS).The sequence of the sex determination region of the Y chromosome was determined by PCR.Sca-1 + cells (2?10 3) of male mice were transfused to female mice receiving 8-12 weeks irradiation at a lethal dose of ?-ray(10 Gy) from a 60Co source.Two months later,the mice were killed and the hearts were removed to make slices. Fluorescence in situ hybridisation (FISH) tests with a Y chromosome probe were used to detect sex of the cells.Immunohistochemistry with antibodies of desmin,Flt-1,white blood cell common antigen CD45 and macrophage F 4/80 were used to detect tissue characteristics of cardiomyocytes. Results: Cells with Y chromosome were found in the cardiomyocytes of female mice transplanted with Sca-1 + cells,and showed phenotypic characteristics of Desmin +/Flt-1-/CD45-/F- 4/80 at the same time.Conclusion: Sca-1 + cells from the murine fetal liver are mostly hematopoietic stem cells,having the potential of differentiating into cardiomyocytes.

AIM: To study the effect of acute renal failure (ARF) on the differentiated frequency of Sca-1+ cells from murine fetal liver in irradiated mice. METHODS: The Sca-1+ cells from murine fetal liver were isolated with magnetic cell sorting (MACS) technique, the sex of which was identified by PCR. The 2?104 Sca-1+ cells were transplanted into a lethally irradiated ([ 60Co], 8 Gy) inbred female mouse. After 8 weeks, these recipient mice were divided to A, B, and C groups at random (A group: irradiated; B group: ARF; C group: ARF and Sca-1+). The mice in B and C groups were induced to ARF with 50% (V/V) glycerin (11.6 mL/kg). 72 hours later, the mice in C group were injected with the fresh prepared Sca-1+ cells again. 8 weeks later, mice were sacrificed, and their kidneys were taken out, fixed and slices were prepared. Fluorescence in situ hybridization (FISH) of renal slices was performed and the pictures of them were taken and analyzed. RESULTS: The cells containing Y chromosome were found in renal slices from the mice in A, B and C groups, which located in epithelial cells of renal tubules, interstitium, glomeruli, and glomerular margin and increased gradually. The double and encircle zone of Y chromosome cells were found in the slices from the mice in B and C groups separately, which was consist of new renal tubules. The differentiation frequency of Sca-1+ cells in kidney in A, B and C groups were (1.65?0.18)%, (8.58?1.34)% and (18.13?1.91)%, respectively, which showed significant difference between former group and later group (P