Adolescent ; Adult ; Catheterization, Central Venous ; instrumentation ; Device Removal ; Humans ; Male ; Vena Cava, Inferior ; Vena Cava, Superior

Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.

Objective To explore the relationship between the expression levels of excision repair cross complementation group 1(ERCC1), breast cancer susceptibility gene 1(BRCA1), thymidylate synthase (TS) mRNA and clinicopathological features, prognosis in advanced colorectal cancer, and the correlation between the expression levels of ERCC1 and BRCA1. Methods The expression levels of ERCC1, BRCA1 and TS mRNA of postoperative paraffin embedded tissue were tested by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) in 49 advanced colorectal cancer cases. The results were analyzed by χ2 test of the correlation between the expression levels and clinicopathological characteristics. Patients were followed up by clinic or telephone. The prognosis was analyzed by small sample Kaplan-Meier survival analysis and Log-rank time series analysis, and P<0.05 was statistically significant. Results The expression level of ERCC1 mRNA in patients with colorectal carcinoma had no obvious correlation with the clinical and pathological characteristics such as gender, age, tumor location, lymph node metastasis, distant metastasis, CEA and differentiation degree (P>0.05). The expression level of BRCA1 mRNA had no significant correlation with the above clinical and pathological features (P>0.05) except distant metastasis (P=0.030) and differentiation degree (P=0.002). The expression level of TS mRNA had no significant correlation with the above clinical and pathological features (P>0.05) except distant metastasis (P=0.003). The expression level of ERCC1 and BRCA1 mRNA obviously correlated (P=0.002). The 1 year overall survival rate was 95.92%(47/49);the 2 year overall survival rate was 83.67%(41/49);and the 3 year overall survival rate was 73.47%(36/49). Overall survival and progression-free survival time in ERCC1 mRNA low expression group (47.8, 41.0 months) was higher than that in ERCC1 mRNA low expression group (27.3, 20.0 months) respectively (P=0.001, P=0.001). Overall survival and progression-free survival time in BRCA1 mRNA low expression group (43.7, 42.7 months) was higher than that in BRCA1 mRNA high expression group (29.3, 25.1 months) respectively (P=0.009, 0.006). Overall survival time in TS mRNA low expression group (39.8 months) was higher than that in BRCA1 mRNA high expression group (25.2 months). Conclusions The expression level of ERCC1 mRNA is not correlated with its clinical and pathological characteristics, but with its biological characteristics. BRCA1 and TS levels are correlated with invasion and metastasis. Low levels of ERCC1 and BRCA1 expression have a better prognostic effect on platinum based first-line chemotherapy for advanced colorectal cancer, and they are correlated. Low level of TS also has longer disease-free survival. Three joint detection could be used as a prognostic factor for colorectal cancer chemotherapy.

Objective To explore the correlation of posttraumatic growth with social support in patients with gastrointestinal neoplasms at the end of life.Methods Cluster random sampling was used to select 280 patients with gastrointestinal neoplasms who were predicted to survive no longer than 6 months in 10 first-class hospitals in Beijing and Tangshan,and they were investigated by Posttraumatic Growth Inventor (PTGI) and Social Support Rate Scale (SSRS).Results The total score of PTGI (61.13 ± 16.78)and the scores of appreciation of life (23.71±7.34),personal strength(10.67±3.55),new possibility(10.07±2.11),relating to others(12.54±5.14),spititual change(10.89±4.35)were higher than median value.The total score of SSRS(41.21±8.48) and objective support(10.27±2.75) were higer than the normal level((39.05±6.35),(8.42±2.27)) (P＜0.05).The appreciation of life was positively correlated with subjective support and the total score of SSRS (r=0.25,r=0.26,P＜0.05),personal strength was positively correlated with objective support,subjective support and the total score of SSRS(r=0.24,r=0.29,r=0.23,P＜0.05),new possibility was positively correlated with subjective support(r=0.21,P＜0.05),relating to others was positively correlated with objective support and the total score of SSRS (r=0.26,r=0.26,P＜0.05),spititual change was positively correlated with objective support and subjective support (r=0.28,r=0.31,P＜0.05).The total score of PTGI was positively correlated with subjective support and the total score of SSRS (r=0.29,r=0.29,P＜0.05).Conclusion Patients with gastrointestinal neoplasms at the end of life experience a certain extent of posttraumatic growth and high level of social support.Posttraumatic growth is positively correlated with social support.

Objective To discussion the value of palate standard section in prenatal ultrasound diagnosis of isolate cleft palate,and summarize the two-dimensional ultrasound characteristics of isolate cleft palate.Methods Two-dimensional ultrasound was performed in 1 8 073 fetuses during 1 8 to 40 gestational weeks of pregnancy.Fetal palate structure were scanned through the oral cleft,the cheek,the neck part,and the standard sagittal section,transverse section and coronal section were obtained to diagnosis cleft palate. The results were compared with labour or postpartum findings.Results In 1 8 073 cases,20 cases of isolate cleft palate,1 9 cases were diagnosed as fetal isolate cleft palate by prenatal ultrasonography,of which 1 8 cases were correctly diagnosed,1 case was misdiagnosed,1 case was missed diagnosed.The sensitivity, specificity,accuracy of prenatal ultrasound detection were 95%,100%,95%,respectively.The ultrasonic characteristics of isolate cleft palate:lack of crack were located in the midline,soft cleft palate was shown as the midline of soft palate interrupted,short of crack was“><”,“‖”,“/\”shape,hard cleft palate was shown as the midline of hard palate interrupted,short of crack was open back“V”or“U”shape.Conclusions The palate standard section has high value in prenatal ultrasound diagnosis of cleft palate,which can assess fracture morphology,length,width,direction and involving range.

OBJECTIVETo investigate the relationship between the protein expression of calpain-2 and calcineurin (CaN) and atrial fibrillation (AF) in patient with valvular heart disease (VHD).

METHODSA total of 40 patients who underwent valve replacement surgery in our hospital from March 2013 to March 2014, right atrial appendages were excised during operation and patients were divided into sinus rhythm (SR) group (n = 17) and AF group (n = 23). The protein expression of calpain-2 and the α-isoform of CaN catalytic subunit (CnA) in the right atrial appendages were determined by Western blot.

RESULTSThe protein levels of the full-length CnAa (60,000), the 45,000 fragment of CnAa without autoinhibitory domain, and calpain-2 were significantly upregulated in the AF group compared to the SR group (1.25 ± 0.51 vs. 0.76 ± 0.37, 1.08 ± 0.37 vs. 0.76 ± 0.25, and 0.82 ± 0.44 vs. 0.51 ± 0.19, respectively, all P < 0.05).

CONCLUSIONActivated calpain-2-CaN signal pathway might be involved in the pathogenesis of AF.

Objective To evaluate the performance of the ABX Micro C-reactive protein(CRP)in determination of CRP.Methods The analytic characteristics including precision,carry-over,linearity, stability,interference and comparability were examined.Results The coefficient of variation(CV)was less than 5.1%,10% and d.3% for within-run,between-run and between-day,respectively.Carryover was less than 1.2%.Whole blood samples held at either room temperature or 4℃ were stable for 48 hours with relative deviation less than 6.0% relatively.Linear range was 1.0-70.0 mg/L using undiluted samples.The comparison between the ABX Micro CRP and Behring Nephelometer Ⅱ was well correlated Both serum:Y=0.996 7X-0.398 5,r~2=0.965 9;serum for BN Ⅱ,whole-blood samples for the ABX Micro CRP:Y=0.908 8X-0.138 2,r~2=0.959 4;both serum and whole-blood samples for the ABX Micro CRP: Y=1.001 7X-0.898 2,r~2=0.952 7.No obvious interference was observed by hyperhemoglobinemia and hyperlipidemia.Conclusion The determination of CRP test with ABX Micro is accurate and reliable.

Objective To determine the effect of interferon-gamma (IFN-γ) on the phagocytosis and killing activity of a murine macrophage cell line RAW264.7 against T.asahii,and to estimate the possibility of treating T.asahii infection with IFN-γγ.Methods T.asahii was cultured with or without the presence of different concentrations (10,100,1000 U/ml) of IFN-γfor 18 hours followed by the incubation with RAW264.7 cells for different durations.After additional culture for 45 minutes,the number of T.asahii cells phagocytosed by RAW264.7 cells was counted under an inverted microscope,and the rate of phagocytosis was calculated.The number of colony forming units of T.asahii per milliliter (cfu/ml) was counted after 4-hour additional culture and the growth inhibition rate was determined.Data were processed by the SPSS 16.0 software,and comparisons of parameters between these groups were done by Bonferroni method and analysis of variance after homogeneity test of variance.Results The number of phagocytosed T.asahiicells was 25.12 ± 1.81,35.88 ± 3.56,52.12 ± 3.23,with the phagocytosis rate being 25.12％,35.88％ and 52.12％,respectively in RAW264.7 cells incubated with IFN-γ of 10,100,1000 U/ml,significantly different from that in untreated RAW264.7 cells (13.62 ± 2.39,13.62％,all P ＜ 0.01).The colony forming units of T.asahii per ml after incubation with untreated RAW264.7 cells differed significantly from those after incubation with IFN-γ (10,100,1000 U/ml)-treated RAW264.7 cells ((68.12 ± 3.39) × 500 vs.(58.62 ± 4.89) × 500,(45.50 ± 3.02) × 500 and (34.62 ± 4.24) × 500,all P＜0.01),with the growth inhibition rate being 25.21％,41.95％ and 55.83％ respectively for RAW264.7 cells incubated with IFN-γ of 10,100,1000 U/ml.Statistical differences were also observed in the killing activity between RAW264.7 cells incubated with different concentrations of IFN-γ (all P ＜ 0.01).Conclusion IFN-γ (10-1000 U/ml) may enhance the phagocytosis and killing activity of RAW264.7 cells against T.asahii in a concentration-dependent manner.

Flashing light effect on microalgae could significantly improve the light efficiency and biomass productivity of microalgae. In this paper, the baffles were introduced into the traditional flat plate photobioreactor so as to enhance the flashing light effect of microalgae. Making Chlorella sp. as the model microalgae, the effect of light intensity and inlet velocity on the biomass concentration of Chlorella sp. and light efficiency were evaluated. The results showed that, when the inlet velocity was 0.16 m/s, with the increase of light intensity, the cell dry weight of Chlorella sp. increased and light efficiency decreased. With increasing the inlet velocity, the cell dry weight of Chlorella sp. and light efficiency both increased under the condition of 500 μmol/(m2 x s) light intensity. The cell dry weight of Chlorella sp. cultivated in the novel flat plate photobioreactor was 39.23% higher than that of the traditional one, which showed that the flashing light effect of microalgae could be improved in the flat plate photobioreactor with inclined baffles built-in.
Biomass ; Chlorella ; growth & development ; Culture Techniques ; instrumentation ; Light ; Microalgae ; growth & development ; Photobioreactors

Objective To evaluate the changes in refractive state in high-myopic patients with rhegmatogenous retinal detachment after vitrectomy combined with cataract surgery.Methods Clinical data of 30 patients of rhegmatogenous retinal detachment in the Affiliated Hospital of Qingdao University from January 2016 to March 2017 was retrospectively analyzed,and these patients were randomly dividedly into two groups:high myopia (15 patients of 15 eyes) and non high myopia group (15 patients of 15 eyes),in which patients firstly underwent vitrectomy combined with phacoemulsification and silicone oil tamponade,followed by intraocular lens (IOL) implantation after the removal of silicone oil 3 months later.The two groups were continuously followedup for 6 months after IOL implantation.Before vitrectomy,the axial length,corneal curvature were measured for calculating the predictive spherical equivalent (SE).Then axial length and corneal curvature at 1 month,3,6 months after IOL implantation were measured for calculating the actual SE.The best corrected visual acuity (BCVA) was recorded before and after operation and the LogMAR was calculated in each groups.The SE,axial length,corneal curvature before vitrectomy and 1 month,3,6 months after IOL implantation were respectively compared in each group.The relationship between the refractive prediction errors and the preoperative axial length in the high myopia group was evaluated.Results The LogMAR visual acuity was improved in both groups,and the difference between before and after treatment was statistically significant (both P ＜0.05).The SE at 1 month,3,6 months after surgery showed significantly myopic shift in the high myopic eyes,and the myopic shift gradually increased with time,with statistically significant at each time point (all P ＜ 0.05),but there was no significant difference in myopic shift between before and after surgery as well as among each time point after surgery in the non-high myopic eyes (P ＞ 0.05).Axial length and corneal curvature at 1 month,3,6 month after surgery in the high myopia group were significantly increased (all P ＜ 0.05),whereas those in the non high myopia group did not change significantly (all P ＞ 0.05).By simple linear regression analysis,the refractive prediction errors had a negative correlation with the preoperative axial length in the high myopia group (R2 =0.580,P =0.001).Conclusion The LogMAR visual acuity after vitrectomy combined with cataract surgery is improved in patients with rhegmatogenous retinal detachment.Postoperative axial length growth and corneal curvature increase are presented in patients with high myopia,and the longer the preoperative axial length is,the more myopic shift the eye shows.