A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

Objective To study the effects of duration of migration from sea-level to high-altitude on cerebral hemodynamic parameters and their ethnic differences.Methods This randomized comparative study recruited 5 groups of healthy male subjects which were native Hans at sea-level (NHS),Han migrants from sea-level to high-altitude (HMSH,where HMSH-a for residence duration of 4 days,HMSH-b for 1 year,and HMSH-c for 5 years),and native Tibetans at high-altitude (NTH).Color duplex Doppler sonography (CDDS) was used to measure bilateral vertebral and internal carotid arterial diameters,mean flow velocities.The heart rate,arterial blood pressure,and arterial oxygen saturation were also recorded simultaneously,and in combination,hemoglobin was measured to assess total cerebral blood flow (TCBF),total cerebrovascular resistance (TCVR),and total cerebral oxygen supply (TCOS).Results After migration to high-altitude,Hans' TCVR and TCBF maintained the level of NHS after transient changes during early stage (P<0.05),and the TCBF was markedly higher than that of NTH (P<0.05);while the blood pressure and TCOS increased abruptly (P<0.05) at the beginning and kept for a relatively long time at a constant level which were higher than those of NTH all the time (P<0.05).Conclusions Han migrants can acclimate to high-altitude by adaptive change of cerebral blood flow over a short time and maintain a constant adaptability which is always weaker than that of native Tibetans.CDDS can be used for non-invasive measurement of actual cerebral blood flow,and is of good value in the study on high-altitude-related cerebral hemodynamics.

Objective To investigate the expression of ki-67 and the development of the intimal hyperplasia(IH) of the irradiated human umbilical artery incorperated with nofloxacin and silver(IHUAINS) grafts into the carotid arteries of the rabbit. Methods Twenty IHUAINSs were sterilely produced. Thirty rabbit were performed bilateral carotid bypass grafting. The IHUAINS(experimental group)and the left carotid arteries (control group) were implanted in the left and right carotid arteries respectively. Graft patency was checked at the 2nd and 6th week after implantation, and the grafts were studied with standard histological techniques and immunohistochemieal method for meas-urement of intimal thickness and the expression of ki-67. Results The total patency rate of the grafts was 89.6%. Light microscopic exami-nation of the grafts revealed intimal and media proliferation, cellular in-filtration. The endothelial cells covered the vascular lumen. There was no significant difference of the intimal thickness between two groups at the 2nd week after grafting (P>0.05). The intimal thickness of the experimental group was larger than that in control group at the 6th week after implantation without statistical significance (P>0.05). At the same time, immunocytochemical analysis showed that the expression of ki-67 in the experimental group was stronger than that in control group without statistical significance(P>0.05). Conclusion The IH of the IHUAINS was larger than that of the autologous artery, but there is no difference between these two groups. Thus, IHUAINS may be an ideal graft in the field of coronary surgery.

ObjectiveTo improve the long term outcomes of the surgery for Stafford type A aorticdissection, we performed ascending aorta and total aortic arch replacement combined with transaorticstented graft implantation into the descending aorta for acute type A aortic dissection.MethodsFrom May 2005 to February 2011,36 consecutive patients with acute Stanford type A aorticdissection underwent this procedure.Right axillary artery cannulation was routinely used forcardiopulmonary bypass and selected cerebral perfusion.The stented elephant trunk was implanted through the aortic arch under hypothermic circulatory arrest.The stented elephant trunk was a 10 cm long selfexpandable graft.34 patients were followed up for 2 ～36 months.ResultsCardiopulmonary bypass time was (160 ± 31)min, average cross clamp time was (101 ±26)min, and average selective cerebral perfusion and lower body arrest time was (31 ± 16)min.The in-hospital mortality was 5.5％ (2/36).One patient died of multi-organ failure postoperatively and another died of cerebral infarction 9 day after surgery.No one suffered from spinal cord injury perioperatively.There was no late death during follow up.ConclusionsAscendingaorta and total aortic arch replacement combined with transaortic stented graft implantation into the descending aorta is an effective way in closing the residual false lumen of the descending aorta and might contribute to better long term outcomes of type A aortic dissection.

OBJECTIVETo identify the human hematopoietic stem cells from the human/goat xenogeneic model with molecular techniques.

METHODSDNA and total RNA were extracted from 11 transplanted goat peripheral blood cells. Human CD(34), GPA and SRY genes were amplified with PCR in these samples, and CD(34), GPA mRNA transcripts were detected using RT-PCR in 5 and 6 goat peripheral blood cells, respectively. Southern blot analysis was performed in 8 goat DNAs to detect the human specific alpha-satellite sequence. Meanwhile FISH was also performed to detect the human cells in goat blood with a probe of human Y chromosome.

RESULTSHuman CD(34) and GPA genes could be detected with PCR in all the 11 goats, and SRY gene did in 5 goats transplanted with hematopoietic stem cells derived from male human babies. Southern blot showed that human specific alpha-satellite sequence was present in 8 goats. By RT-PCR, human CD(34) mRNA was detected in 5 experimental goats, GPA mRNA was found in the other 6 experimental goats and FISH assay showed that some peripheral blood cells of the human/goat xenogeneic model were positive.

CONCLUSIONExistence of human cells in the recipient goats was identified by molecular detection, which was feasible for the examination of human/goat xenogeneic models.

Animals ; Antigens, CD34 ; genetics ; Blotting, Southern ; Female ; Genes, sry ; genetics ; Glycophorin ; genetics ; Goats ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymerase Chain Reaction ; Transplantation Chimera ; genetics ; Transplantation, Heterologous

Objective To evaluate the efficacy of Disposcope endoscope(DE)-guided nasotrache-al intubation in patients with difficult airway by comparing with fiberoptic bronchoscope(FOB). Methods One hundred and twenty American Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both se-xes, with body mass index<25 kg∕m2, aged 18-64 yr, with mouth opening<3 cm, of Mallampati classifi-cation Ⅲ or Ⅳ, undergoing maxillofacial surgery requiring nasotracheal intubation were divided into DE group(n=60)and FOB group(n=60)using a random number table.Nasotracheal intubation was per-formed under the guidance of DE or FOB after induction of anesthesia.Glottis exposure was evaluated using Cormack-Lehane grade.Epistaxis during intubation, successful intubation, time and degree of glottis expo-sure, intubation time and development of tachycardia and hypertension and requirement for assisted ventila-tion with face mask during intubation were recorded.The patients were followed up postoperatively, and the development of intubation-related complications was also recorded.Results Compared with group FOB, glottis exposure time and incubation time were significantly shortened(P<0.05), and no significant change was found in Cormack-Lehane grade, rate of successful incubation, rate of successful intubation at first attempt or intubation-related complications in group DE(P>0.05). Hypertension and tachycardia were not found and no patients required assisted ventilation with face mask during intubation in the two groups.Conclusion DE-guided nasotracheal intubation provides similar efficacy to that of FOB with shorter time and is an optimal selection for the patients with difficult airway.

Objective:To observe the body fluid and cellular immune function of children with cerebral palsy (CP) in the plateau area, as well as the exchanges of these factors during the comprehensive rehabilitation treatment.Methods:A total number of 144 children admitted to Xining Hospital of Traditional Chinese Medicine from June 2018 to October 2019 were selected as the CP group for comprehensive rehabilitation treatment (consecutive courses). The peripheral blood immunoglobulin/complement (IgA, IgG, lgM, C3, C4) level, T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and neuron-specific enolase (NSE) content was examined in the clinical specimens before and after treatment by using the immunoturbidimetry, flow cytometry, electrochemiluminescence analysis according to the Gross Motor Function Classification System (GMFCS) and Gross Motor Function Test Scale (GMFM-88). Children were divided as the different degrees to evaluate the rehabilitation efficacy. A total number of 50 healthy children taken a health check/physical examinations during the same period were considered as the control group. For statistical Analysis, the χ2 test and independent sample t test were performed. Results:The levels of humoral immune IgG, IgA, IgM, C3 and C4 in CP Group [(6.42±1.05), (0.64±0.13), (0.89±0.13), (0.80±0.08), (0.17±0.03) g/L, respectively] in CP groups′ children were lower than those in the control group [(10.25±0.62), (1.04±0.06), (1.06±0.17), (1.04±0.04), (0.27±0.04) g/L, respectively]. The humoral immune IgG and IgA levels [severe (5.40±0.69) and (0.55±0.09)g/L, moderate (6.63±0.30) and (0.66±0.14)g/L, mild (7.57±0.63) and (0.74±0.09)g/L, P<0.05] were also related to the children with CP of different GMFCS grades. Moreover, the level of T lymphocyte subsets (CD3+,CD4+,CD8+,CD4+/CD8+) in the CP group were not statistically different to that in the control groups children. Receiving the rehabilitation treatment, the levels of serum humoral IgG and IgA in CP Group (7.69±1.14) and (0.79±0.17) g/L were significantly enhanced; whereas the serum NSE (12.82±2.49) μg/L was lower than that before treatment (18.57±3.08) μg/L, and the total score of GMFM-88 (121.35±26.51) was higher than that before treatment (101.04±27.62). The differences were statistically significant ( P<0.05). IgM, C3, C4 and T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) had no significant difference compared with those before treatment ( P>0.05). Conclusions:Children with CP at high altitude have abnormal humoral immune function. IgG and IgA may be related to the severity of CP and neuronal damage. Comprehensive rehabilitation can not only improve the motor function of children with CP, relieve neuronal damage, but also enhance their humoral immunity status.

Objective To evaluate the effect of propofol on high-mobility group box 1 protein (HMGB1)/Toll-like receptor 4 (TLR4) signaling pathway during hepatic ischemia-reperfusion (I/R) injury in rats.Methods Thirty-six clean-grade healthy male Sprague-Dawley rats,aged 3 months,weighing 250 -300 g,were divided into 3 groups (n=12 each) using a random number table method:sham operation group (group S),hepatic I/R group (group I/R) and propofol group (group P).Hepatic I/R injury was induced by occluding the portal vein and hepatic artery supplying the left and middle lobes of the liver for 1 h followed by 6-h reperfusion in anesthetized rats.Propofol was infused via the tail vein at a rate of 12 mg ·kg-1 · h-1 starting from 20 min before ischemia until 6 h of reperfusion in group P.The rats were sacrificed at 6 h of reperfusion,and the left lobe of the liver was removed for microscopic examination of the pathological changes which were scored and for determination of the expression of HMGB1,TLR4,tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-6) in liver tissues (by Western blot).Results Compared with group S,pathological scores of liver tissues were significantly increased,and the expression of HMGB1,TLR4,TNF-α and IL-6 was up-regulated in I/R and P groups (P＜0.05).Compared with group I/R,pathological scores of liver tissues were significantly decreased,and the expression of HMGB1,TLR4,TNF-α and IL-6 was down-regulated in group P (P＜ 0.05).Conclusion The mechanism by which propofol reduces liver I/R injury is associated with blocking HMGB-1/TLR4 signaling pathway and inhibiting inflammatory responses in rats.

Objective To examine the anticancer effect of a novel histone deacetylase inhibitor (HDACi), JZ004, on colon cancer cells HCT-8 and HT-29, and to investigate the molecular mechanisms of proliferation inhibition and apoptosis induction of cancer cells treated by JZ 004.Methods Colon cancer cells were treated with a series of concentrations of JZ004 .MTT assay was used to detect the proliferation of cancer cells .The cell cycle distribution and apoptosis were deter-mined by flow cytometry .Rhodamine 123 and DCFH-DA were applied to detect the mitochondrial membrane potential (ΔΨm) and reactive oxygen species ( ROS) production.The protein expressions of acetyl-histone H3, p21, cyclin-dependent kinase(CDK)4, Bcl-2, Mcl-1 and Bax were assayed by Western blotting .Results JZ004 was found to inhibit proliferation and induce apoptosis of colon cancer cells in a time-and dose-dependent manner , accompanied by a dose-dependent hyperacetylation of histone H3.JZ004 induced the cancer cell arrest in G 0/G1 phase by increasing the expres-sion level of p21 while CDK4 was downregulated .JZ004 also increased cellular ROS production and reduced ΔΨm by regu-lating the expressions of Bcl-2 family proteins .Conclusion As a novel HDACi , JZ004 effectively inhibits proliferation and increases ROS production to induce apoptosis of colon cancer cells .The results indicate that JZ004 is a potential compound to be developed as an anti-colon cancer agent for clinic application .