Background and purpose:Previous researches have shown that procalcitonin differentiates infec-tious from non-infectious fever and assesses the severity of infectious diseases. This study aimed to investigate the clin-ical value of procalcitonin in patients with chemotherapy-induced febrile neutropenia.Methods:A total of 147 patients with chemotherapy-induced febrile neutropenia admitted to intensive care unit from Jan. 2012 to Dec. 2014 were di-vided into infectious group and fever of unknown origin group according to clinical symptoms, signs and etiology. The infectious group was divided into sepsis, severe sepsis, and septic shock groups according to the severity of infection. The procalcitonin levels were compared between different groups.Results:A procalcitonin cut-off value>0.935 ng/mL provided a sensitivity of 90.0%, speciifcity of 90.0% and AUC=0.905. The procalcitonin level of the infectious group was signiifcantly higher than that of the fever of unknown origin group [1.805 (1.268-2.523) ng/mLvs 0.555 (0.398-0.818) ng/mL,P<0.001]. There is a signiifcant difference between the severe sepsis group and the sepsis group [13.885 (7.600-17.961) ng/mLvs 1.805 (1.268-2.563) ng/mL,P<0.001]. Compared with the severe sepsis group, the value of procalcitonin in the septic shock group was signiifcantly higher [23.800 (20.050-30.478) ng/mLvs 13.885 (4.955-19.133) ng/mL,P<0.001].Conclusion:Plasma procalcitonin is a useful marker for diagnosing neutropenia in patients with infection. Meanwhile, procalcitonin can be used to assess the severity of infection in patients with neutropenia.

Objective To investigate the clinical value of serum glycyl-proline dipeptidyl aminopeptidase(GPDA)combined with carcino-embryonic antigen (CEA),carbohydrate antigen724 (CA724),carbohydrate antigen242 (CA242) in the early diagnosis of gastric cancer.Methods Collected in Changan hospital in patients with gastric cancer and atrophic gastritis patients and healthy subjects 60 cases,by TBA-120FR biochemical analyzer glycyl-proline dipeptidyl aminopeptidase (GPDA),chemiluminescence analyzer to detect the levels of serum CEA,CA724 and CA242,analysis of single detection and joint detection and the differences between the positive rate and sensitivity.Results The detection of GPDA in gastric cancer group was significantly lower than that in atrophic gastritis group and healthy control group,the difference was statistically significant (F=69.532,P=0.000).The results of CEA,CA724 and CA242 in gastric cancer group were higher than those in atrophic gastritis group and healthy control group,the difference was statistically significant (CEA:F=59.926,P=0.001;CA724:F=51.056,P =0.001;CA242:F =72.613,P =0.000).Serum GPDA,CEA,CA724 and CA242 single detection positive rate were 70 ％,45 ％,61.7 ％ and 50 ％.Tumor markers CEA,CA724,CA242 positive rate of three joint detection was 75％.Serum GPDA and tumor markers of CEA,the positive rate of CA724 and CA242 combined detection of four was 86.7％.The positive rate of three and higher than that of single detection,the difference was statistically significant (F=49.635,P=0.003).Serum GPDA,CEA,CA724 and CA242 single detection sensitivity was 70.2 ％,50.2 ％,67.3 ％ and 53.2％.Tumor markers CEA,CA724,CA242 three joint detection sensitivity was 85.6％.Serum GPDA and tumor markers CEA,CA724 and CA242 four joint detection sensitivity was 90.3％.The sensitivity was higher than the three items and the individual tests,and the difference was statistically significant (F=52.016,P =0.001).Conclusion GPDA joint CEA,CA724 and CA242 tumor markers detection can improve the positive rate and sensitivity in early diagnosis of gastric cancer,but it will not reduce the diagnostic specificity,the clinical diagnosis of early gastric cancer has important significance and value.

Objective To investigate the relationship between promoter hypermethylation of secreted frizzled-related protein (SFRP) gene and acute leukemia (AL). Methods We examined the promoter methylation starus of SFRP1, 2, 4 and 5 in primary or relapsed AL patients, cell lines ( HL60,NB4, Molt-4 and Jurkat) and peripheral blood mononuclear cells from healthy people with methylationspecific PCR (MSP). Results None of the normal mononuclear cells showed methylation of any SFRP genes. The frequencies of aberrant methylation among the samples were 33.9% (20/59) for SFRP1,23.7%(14/59) for SFRP2, 6. 8% (4/59) for SFRP4 and 10. 2% (6/59) for SFRP5 in acute myelocytic leukemia (AML), and 39.3% (11/28) for SFRP1, 28.6% (8/28) for SFRP2, 25.0% (7/28) for SFRP4 and 32. 1% (9/28) for SFRP5 in acute lymphoblastic leukemia (ALL). Hypermethylation of SFRP1, 2, 5genes were present in the 4 AL cell lines. SFRP4 was methylated in NB4, Molt-4 and Jurkat cell lines.However, methylation and unmethylation of SFRP4 were both detected in HL60. Conclusions Hypermethylation of SFRP genes is a common event in the evolution of AL. Methylation of SFRP genes might serve as potential independent biomarkers for early detection of AL.

AIM: To evaluate the implication of CXCL10-loop3-EGF fusion protein for the activities of targeting tumor and anti-angiopoiesis. METHODS: RT-PCR was preformed to amplify CXCL10 coding sequence from PBMC activated by IFN-γ. CXCL10-loop3-EGF fusion gene, which was conducted by Over-Lap Extention PCR, was hinged up with plasmid pTG19-T, transfected to E. coli DH5α and processed positive colony selection. After ligated with plasmid pET32a(+), recombinant CXCL10-loop3-EGF fusion gene was then transfected to E. coli Origami B (DE3) and induced to express its coding fusion protein his-CXCL10-loop3-EGF. The recombinant fusion protein CXCL10-loop3 -EGF was purified by His-bind affinity chromatograph, enterokinase cleavage, ultrafiltration and dislysis. The transwell chemotatic test and HUVEC angiopoiesis inhibition test were performed to determine the anti-tumor responses and anti-angiopoiesis activity of CXCL10-loop3-EGF fusion protein. RESULTS: CXCL10-loop3-EGF fusion protein was successfully constructed and confirmed by SDS-PAGE analysis and Western blotting. Significant PBMC chematatic activity and HUVEC anti-angiopoiesis activity were observed. CONCLUSION: CXCL10-loop3-EGF fusion protein, which has perfect anti-tumor activity, is successfully constructed.

Objective The use of extracorporeal membrane oxygenation (ECMO) as a treatment for the failure of cardiopulmonary function after cardiac surgery is increasing and has been reported to be 3% to 5% in the cases with congenital heart disease. We reviewed our experience with ECMO in children who received heart surgery for congenital heart disease and complicated with severe heart failure postoperatively. Methods Eight patients received ECMO, seven was due to the failure to wean from bypass and one had fulminant myocarditis. Import membrane oxygenator,veno-arterial mode ECMO and right atriumascending aortic cannulation were used in 7 cases and peripheral cannulation via femoral veno-artery route was used in 1 case.Supportive intervention persisted from 65 to 498 hours, with flow rate maintained at 80 to 120 ml per minute per kilogram body weight. Results Five patients died, with a mortality of 62.5%, and 3 cases discharged, with a survival rate of 38%. Bleeding occurred in 5 cases, thrombosis occurred in 2 cases, hemolysis was identified in 1 case and DIC was observed in 1 case.One case had liver failure and 2 cases had malnutrition. Oxygenator plasma leakage occurred in 2 cases. Mean arterial blood pressure increased significantly after the establishment of ECMO as compared with that before the procedure [( 60.2 ± 7.8 )mmHg vs. (48. 1 ± 5.2 ) mmHg, P≤0.05]. The arterial concentration of lactate decreased significantly, from (5. 1 ± 0. 8 )mmol per liter before ECMO to ( 3.6 ±0. 5 )mmol per liter after ECMO, P ＜0.05. Conclusion For patients who survived the congenital heart surgery and no residual anatomic deformity, ECMO can be used as early as possible as a treatment for severe heart failure which resulted from coexistent of left and right ventricular and pulmonary insufficiency. An overall mortality may be decreased by ECMO technique as it plays a substitution role for gas exchange in the lung. As a result, the concentration of oxygen and the airway pressure used during ventilation, and the resultant lung injury can be reduced. Appropriate strategies involve transfusion of fresh platelet and packed red blood cells, replacement of frozen plasma and blood products, as well as rational use of vasoactive drugs and heparin, and maintaining a stable internal environment. Following strategies are also recommended: using continuous arterio-venous hemofiltration and durable heparin-coated membrne oxygenator, reducing hemorrhagic complications, monitoring pressure on both side of the film, identifying plasma leakage carefully and reducing the mechanical complications.

Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

s were 0.573±0.016, 0.707±0.008 and 0.711±0.013). Conclusions CXCL10 can express stablely in MCF-7 cell lines, which resulted in down-regulation of expression of VEGF and STAT3 gene. CXCL10 played an important role in anti-tumor effect.

Objective To explore the effects of action observation therapy on upper-extremity motor function after ischemic stroke and on the motor cortex using functional magnetic resonance imaging (fMRI).Methods Forty patients with ischemic stroke were randomly assigned to an observational group (n =20) or a control group (n =20).Both groups received conventional rehabilitation,while the observational group was additionally provided with action observation therapy for 8 weeks.Both groups were assessed using the Fugl-Meyer assessment (FMA) and the Barthel index (BI) before and after the 8 weeks of treatment and functional magnetic resonance imaging was performed before treatment.Two months after the treatment,nine patients of the experimental group and 8 of the control group who continued to receive their respective treatments after discharge were again assessed using functional magnetic resonance imaging.Results After the treatment the average FMA score and BI score of both the observational group and the control group had increased significantly.The increase in the average FMA score of the observational group was significantly greater than that of the control group.However,there was no significant difference between the two groups in the increases in BI score after 8 weeks of treatment.The fMRI results showed that there was a significantly greater rise in activity in the bilateral precentral gyrus,parietal lobe and the supplementary motor area of the patients in the observational group after the treatment compared with the control group.Conclusion Action observation therapy can improve upper extremity motor function and performance in the activities of daily living after ischemic stroke and induce changes in the excitability of the cerebral motor cortex.

Objective To establish RNase L gene knockout HEK 293 cell lines using CRISPR/Cas9 system.Methods Small guide RNA ( sgRNA) sequences of human RNase L were designed and sgRNAs were inserted into pCas-Guide and pCas-guide RNA(gRNA) vectors were obtained.The donor DNA sequences of the homologous arm were designed for RNase L knockout .In the presence of the right homologous arm , the resistance gene of hygromycin B and the left homologous arm as templates of homology-directed repair , the donor DNA template was amplified by overlopping PCR and cloned into the pBackZero-T expression vector and pBackZero-T-RNase LK vector was obtained .The pCas-gRNA vector and pBackZero-T-RNase LK vector were co-transfected into HEK293 cells to establish the stable expression cell line of RNase L gene knockout .Cells were cultured with hygromycin B , while Western blotting and DNA sequencing were used to analyze the gene of RNase L knockout from genome .Results and Conclusion The pCas-gRNA vector and pBackZero-T-RNase LK vector were successfully constructed.Five RNase L gene knockout HEK293 cell lines were generated,contributing to the study of the biological function and molecular mechanism of RNase L .

Objective To analyse the difference of effects of different tidal volume mechanical ventilation strategies in infants with severe congenital heart disease after curative.Methods Sixty-eight cases were chosen from CICU of Anhui Provincial Children's Hospital during Oct 2010 to Jan 2012.Thirty-two cases in A group were used in ventilation strategy by lower tidal volumes (6 ～ 10 ml/kg),36 cases in B group were used in ventilation strategy by larger tidal volumes (10 ～ 15 ml/kg).The time of mechanical ventilation,duration of ICU stay and postoperative complications were compared between the two groups.Results The time of mechanical ventilation of B group was shorter than that of A group [(8.6 ± 2.5) d vs (11.7 ± 3.2) d],duration of ICU stay of B group was shorter than that of A group [(11.4 ± 4.8) d vs (15.6 ± 5.7) d],there were statistical differences between two groups(P ＜ 0.0l).The incidence rate of ventilator associated pneumonia and pneumothorax were 6.3％ and 3.1％ in A group,which were 5.6％ and 5.6％ in B group,there were no statistical differences between two groups (P ＞ 0.05).Conclusion The effects of mechanical ventilation with larger tidal volumes is better than that with lower tidal volumes in infants with congenital heart disease postoperative therapy.