BACKGROUND: AdiponecUn (apM1) has been a new target for gene therapy of ischemic diseases; apM1 gene cloning is the key of successful apM1 gene therapy. There are two main gene cloning methods: directional cloning and T-A cloning method. Directional cloning method is complicated, while the T-A cloning method is relatively simpler, with higher successful rate. OBJECTIVE: T-A cloning of apM1 gene coding region was applied to verify the sequence in comparison with GenBank. DESIGN, TIME AND SETTING: The gene verification experiment was performed at the laboratories of Fujian Hypertension Research Institute which is in The First Affiliated Hospital of Fujian Medical University and Academy of Integrative Medicine which is in Fujian University of Traditional Chinese Medicine, from June 2006 to December 2008. MATERIALS: Adipose tissue sample of omental fat pad was obtained from a surgical patient in the First Affiliated Hospital of Fujian Medical University. Trizol produced by Invitrogen company; M-,-MLV, Gel Extract Kit produced by PROMEGA company; Taq enzyme produced by TIANGEN company; Restriction Endonucleases BamH Ⅰ and Sal Ⅰ, pMD18-T Vector produced by TAKARAcompany were used in this study.METHODS: Total mRNA was extracted from human greater omentum adipose tissue. The coding region of human apM1 (hapM1) gene was amplified by RT-PCR. The coding region of hapM1 gene was cloned into pMD18-T vector .The recombinant plasmid was identified with restriction enzyme digestion analysis and nucleotide sequencing. MAIN OUTCOME MEASURES: The electrophorasis verification of the recombinant plasmids coding target gene using double enzyme digestion, comparison of the similarity between the sequences of the plasmids and hapM1 in GenBank. RESULTS: Sense and antisense coding region of hapM1 gene were cloned. The sequencing results showed that the sequences of the cloned DNA were completely identical to that of hapM1 in GenBank.CONCLUSION: The coding region of hapM1 gene was successfully cloned.

Abstract Human adenovirus (HAdV), which is characterized by infectivity, complex pathogenesis and multiple target organs, causes multiple organ infections in the respiratory system, gastrointestinal system and eyes, which seriously endangers human health. Various subspecies of HAdV has different tissue tropism, which presents diverse clinical symptoms and epidemiological characteristics. Based on the molecule biological characteristics of HAdV, this review summarizes the clinical symptoms and epidemiological characteristics of HAdV infections depending on tissue tropism, and describes the trends in HAdV epidemiology, so as to provide insights into prevention and control of HAdV infections.

In this article, the authors introduced his experiences of treating insomnia by dispersing the depressed liver Qi, clearing heat, resolving phlegm, reinforcing spleen Qi, nourishing Yin, repressing fire, soothing the nerves, and activating blood to remove blood stasis Stasis-Removing.

Kawasaki disease has been the major etiology of aquired heart problems in children. Mecha-nisms of Kawasaki disease are still unclear in the past years. Some Kawasaki disease patients were not responsive to intravenous immunoglobulin therapy. And corticosteroid therapy remains controversial in the treatment of Ka-wasaki disease. A few patients with Kawasaki disease presented severe systemic symptoms,and even died for it. It is difficult to evaluate the severity of coronary artery lesions and prognosis of Kawasaki disease. In this article, we reviewed some new concepts on diagnosis and treatments on Kawasaki disease.

Objective To explore the effect of different sleeping state and body position nursing on the surface temperature, heart and lung function, and development of premature infants. Methods A total of 160 cases of premature infants were selected as research objects, divided into active-prone group, active-supine group, quiet-prone group, quiet-supine groupwith random number tables. Each group on the basis of conventional nursing care was given the above position. Respiratory frequency (RR), heart rate (HR), oxygen saturation (SpO2), RR deviation, HR deviation and surface temperature were observed before and after nursing. After continuous nursing for 7 days, the differences of development indexes of each group were also observed. Results Surface temperature of forehead, right flank, right forearm, right leg were (36.4 ± 0.2), (36.4 ± 0.3), (35.6 ± 0.3),(34.4 ± 0.5)℃in group C, which were significantly higher than those in the other groups (F=2.89-3.31, P<0.05). HR, RR, HR-deviation, RR-deviation and its change value in group C were (128.5 ± 1.8) beats/min, (34.6 ± 2.2) beats/min, (9.1 ± 1.2)%, (212.5 ± 31.5)%, which were significantly lower than those in the other groups (F=4.61-4.96,P < 0.05), and SpO2 change value was 0.027±0.001, which was significantly higher than those in the other groups (F=3.65,P<0.05). Weight gain, the length growth, head circumference growth and upper arm around growth in group C and A in preterm infants after 7 days of continuous nursing were (118.6±6.3) g, (11.3±1.2) mm, (6.4±0.4) mm, (4.4± 0.2) mm and (110.3 ± 5.7) g, (11.0 ± 1.1) mm, (6.0 ± 0.5) mm, (4.2 ± 0.3) mm, which were significantly better than (74.5 ± 4.4) g, (6.9 ± 0.8) mm, (4.5 ± 0.7) mm, (3.2 ± 0.2) mm of active-supine group and (78.6 ± 4.9) g, (7.6±0.7) mm, (4.8±0.6) mm, (3.5±0.3) mm of quiet-supine group (F=3.57, 4.98,P<0.05). And the above indicators of quiet-prone group were higher than that of active-prone group (P<0.05). Conclusions The quiet-prone position can improve the heart and lung function and development of premature infants.

Aim To investigate the expression of the lentiviral vector carrying a human apM1(hapM1) gene in 293 T cells and its bioactivity.Methods The CDS region of hapM1 gene was subcloned into the lentiviral vector from a cloning vector,and the transfer plasmid(PNL-hapM1-IRES2-EGFP)of the lentiviral vector was constructed.The three plasmids of the lentiviral vector: PNL-hapM1-IRES2-EGFP,HELPER,and VSVG were cotransfected to 293 T cells to produce the recombinant lentivirus;293 T cells were infected by the recombinant lentivirus and were identified with fluorescent microscope,RT-PCR and western blot.Inhibitory effect of the recombinant hapM1 on proliferation of vascular smooth muscle cells(VSMCs) was identified.Results The sequencing results showed that the sequence of the subcloned hapM1 gene was completely identical to that reported in GeneBank.The virus titre of the recombinant hapM1 lentivirus was 2.0?105 TU?L-1.After the recombinant lentivirus infected 293 T cells,RT-PCR and Western blot results showed that the expression of hapM1 in the hapM1-293 T group could be measured.The recombinant hapM1 inhibited the proliferation of VSMCs.Conclusions The recombinant lentivirus carrying a human apM1 gene was constructed successfully.The recombinant hapM1 could be expressed in 293 T cells and had its bioactivity.

Objective To investigate the utility of plasma D-dimer and fibrinogen (FIB) for the severity assessments and predicting the prognosis of patients with community-acquired pneumonia (CAP).Methods The clinical data of patients with CAP admitted to First Affiliated Hospital of Fujian Medical University were retrospectively analyzed.The patients were divided into Ⅰ-Ⅴ level groups according to pneumonia severity index (PSI),and they were divided into non-survivors and survivors according to 30-day prognosis.The data including gender,age,PSI score,platelets count (PLT),white blood cell count (WBC),D-dimer,FIB,and C-reactive protein (CRP) were compared among groups.The correlations between PSI score and D-dimer,CRP as well as FIB were analyzed by Spearman or Pearson correlation analysis.Receiver operating characteristic curve (ROC) was used to assess the prognostic value of these indicators.Results A total of 499 patients with CAP were enrolled with 298 male and 201 female,the average age was (63.4 ± 17.8) years old,and the 30-day mortality was 6.4％ (32/499).There were 77,80,104,162 and 76 patients in PSI Ⅰ-Ⅴ level groups,and there were more male patients in PSI Ⅵ and Ⅴ level groups.There were no significant differences in PLT and FIB among the groups of different PSI levels,but the levels of WBC,D-dimer and CRP were significantly increased as PSI level increased from Ⅰ to Ⅴ (F1 =3.810,x 22 =102.361,F3 =7.070,all P ＜ 0.01).Compared with survivors,the non-survivors were elder (t =-4.773,P ＜ 0.001) with lower PLT (t =3.026,P =0.003)and higher WBC,PSI score,D-dimer and CRP levels (t1 =-2.545,t2 =-8.421,Z3 =-6.947,t4 =-3.770,all P ＜ 0.05).Plasma D-dimer levels in elderly patients (≥ 65 years old) were statistically higher than those in younger patients (＜ 65 years old;Z =-5.338,P ＜ 0.01).It was shown by correlation analysis that PSI score was positively correlated with D-dimer and CRP (r values were 0.475 and 0.260,both P ＜ 0.001),and no correlation was found between PSI score and FIB (r =-0.062,P =0.170).The area under the ROC curve (AUC) for predicting 30-day death of PSI score,D-dimer and CRP was 0.858 [95％ confidence interval (95％CI) =0.802-0.914],0.867 (95％CI =0.812-0.922) and 0.732 (95％CI =0.641-0.823).The combination of D-dimer and PSI score was better than any single indicator for predicting the prognosis with higher AUC up to 0.905 (95％CI =0.867-0.944),all P ＜ 0.001.The sensitivity and specificity for PSI in predicting 30-day death respectively were 78.1％ and 82.4％ with the cut-off of greater than 122,and those for D-dimer were 75.0％ and 82.9％ with the cut-off of greater than 2.10 mg/L,50.0％ and 84.4％ for CRP with the cut-off of greater than 100.50 mg/L.Conclusions D-dimer could well reflect the severity of CAP and be a good indicator for predicting the prognosis.The combination of D-dimer and PSI might improve the accuracy in predicting prognosis.

Cardiomyopathy, Dilated ; surgery ; therapy ; Humans ; Stem Cell Transplantation