Objective To evaluate the biological activity of a eukaryotic expressed anti-H5N1-M1 cell entry single molecule antibody(TAT-ScFv-mFc). Methods The immune binding activity and affinity of TAT-ScFv-mFc to H5N1-M1 protein were detected by Western blot and localized surface plasmon resonance(LSPR)respectively;The inhibitory effect of TAT-ScFv-mFc on influenza virus H1N1 was detected by CCK-8 assay;The membrane penetration ability of TAT-ScFv-mFc to MDCK cells was verified by immunofluorescence assay. A total of 30 female BALB/c mice were injected with TAT-ScFv-mFc via tail vein,200 μL per mouse. Blood samples were collected at 5,60,120,240 and 360 min after injection. Serum samples were separated and detected for the titers by ELISA,and the half-life of TAT-ScFv-mFc was calculated according to the half-life curve drawn by Origin 2021 software. Results TAT-ScFv-mFc showed specific binding to H5N1-M1 protein with a binding rate constant of 6. 67 × 10~4[1/（M*s）]. The survival rate of MDCK cells infected by H1N1 increased gradually with the increase of TAT-ScFv-mFc concentration in a dose-dependent manner,which obviously inhibited the replication of H1N1. TAT-ScFv-mFc penetrated the cell membrane of MDCK cells in a short time,entered the cell and bound to virus M1protein,thus inhibiting virus replication and assembly. The half-life of TAT-ScFv-mFc in mice was 212 min. Conclusion TAT-ScFv-mFc has good immune binding activity and affinity with H5N1-M1,can effectively inhibit the replication of H1N1,has good penetration ability to MDCK cell membrane,and has a long half-life in mice,which lays a foundation of the drug treatment,vaccine research and preventive treatment of H5N1 infection.

Objective To compare the therapeutic effects of laparoscopic and open surgery treatment for choledocholithiasis.Methods Forty-six cases with choledocholithiasis were randomly divided into laparoscopic surgery group(n = 21)and open surgery group(n = 25).Comparison was conducted between the two groups in terms of operative time,hospitalization day,anorechlnl exhaust time,cases of postoperative pain,the amount of intraoperative bleeding and postoperative complications.Results Hospitalization day(10.5 ±2.4d),anorechlnl exhaust time(27.2 ±9.4h),cases of postoperative pain(28.6%)in the laparoscopic surgery group were significantly better than those in open surgery group(14.3 ±2.5d,54.6 ±12.5h,88.0%)(P ＜0.05).The operative time and the amount of intraoperative bleeding was not significantly different between two groups(P ＞ 0.05).The rate of infection of incisional wound in the laparoscopic surgery group(0%)was lower than that of open surgery group 24.0%(P ＜ 0.05).No residual calculi and bile leakage were found in both groups.The other postoperative complications were not significantly different between the two groups(P ＞ 0.05).Conclusion Laparoscopic choledocholithotomy is safe,effective and feasible.Compare with open surgery,it is a less invasive proceduce for the treatment of choledocholithiasis with the virtue of little trauma,fast recovery,short hospitalization days and few complication.

Objective To explore the fast diagnostics of activated infection of human herpes virus type 6 (HHV-6) with IEA/ex3( an antibody specific for an immediate early protein encoded by variant B HHV-6 U90 gene)in infants with fever and rash . Methods To collect the 55 samples of peripheral blood mononuclear cells (PBMCs) from clinical infants whose having a fever and rash, age are below 2 years old.The immediate early antigen (IEA) of HHV-6 were analyzed with IEA/ex3 in PBMCs by indirect immunofluorescence then using confocal microscope to observe, and using fluoromicroscope to observe after the PBMCs of 44 samples were incubated with cord blood cell at 37℃ in a humidified 5% CO2 atmosphere for 48 hours. The virus was isolated from the PBMCs. Results The conformation rate of the HHV-6 IEA using confocal microscope to observe and virus isolation was 92.73%, sensitivity was 90.32%, specificity was 95.83% and it was respectively 87.50%, 77.27%, 100% using fluoromicroscope to observe after the PBMCs were incubated for 48 hours. Conclusion The study suggested that detection of HHV-6B IEA in PBMCs using confocal microscope to observe after the PBMCs were incubated for 48 hours can fast diagnose HHV-6 activated infection of infant with fever and rash than technique of virus isolation.

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OBJECTIVE:To establish a method for the simultaneous determination of psoralen,isopsoralen,epimedin B,epi-medin C and icariin in Chanlong dingchuan mixture. METHODS:HPLC was performed on the column of Welch Materials C18 with mobile phase of acetonitrile- methanol(1∶1,V/V)-water(gradient elution)at a flow rate of 0.9 ml/min,the detection wavelength was 246 nm(psoralen,isopsoralen)and 270 nm(epimedin B,epimedin C and icariin),column temperature was 30 ℃,injection volume was 20 μl. RESULTS:The linear range was 8.24-164.80 μg/ml for psoralen(r=0.999 7),5.15-103.00 μg/ml for isopso-ralen(r=0.999 3),4.06-81.20 μg/ml for epimedin B(r=0.999 6),5.88-117.60 μg/ml for epimedin C(r=0.999 5)and 4.90-98.00μg/ml for icariin(r=0.999 8);the limits of quantitation were 0.385 μg/ml,0.179 μg/ml ,0.124 μg/ml,0.218 μg/ml and 0.348 μg/ml, limits of detection were 0.127μg/ml,0.059μg/ml ,0.041μg/ml,0.072μg/ml and 0.115μg/ml;RSDs of precision,stability and re-producibility tests were lower than 2.0%;recoveries were 97.93%-100.06%(RSD=0.80%,n=6),96.91%-100.16%(RSD=1.37%,n=6),96.95%-99.63%(RSD=0.98%,n=6),96.69%-99.33%(RSD=1.03%,n=6) and 96.76%-98.53%(RSD=0.70%,n=6),respectively. CONCLUSIONS:The method is simple and reliable,and suitable for the simultaneous determination of psoralen,isopsoralen,epimedin B,epimedin C and icariin in Chanlong dingchuan mixture.

Objective To investigate the risk factors of sepsis associated encephalopathy( SAE) in children.Methods We collected the clinical data of 152 cases of children with sepsis admitted in PICU of Shenzhen Children′s Hospital from June 2013 to April 2015.All 152 cases were divided into the SAE group (n=46) and non SAE group(n=106).Single factor analysis and multi-fatc or Logsi ticr egression analysis were used to study the risk fatc ors of the occurrence and mro tality of SAE in children.Re sults The inci-dence of SAE was 30.3%amongs epsis.The mortality rate of children with SAE was obviously higher than that of children without SAE(17.4%,8/46 vs.1.9%,2/106; χ2 =13.234,P<0.001).Lo gistic regression analysi showed that coagulation disorder,hepatic insfu ficiency and peid atir c cliin cal illness score≤80 were indpe endent risk factro s of SAE.C oagulationd isorder was independent riks facot rs of SAE death.Conclusion The incidence and mortality of SAE are high.Ch ildren with coa gulta ion dsi order,hepatic insufficiecn y and pediatric clinical illness score≤80 shouldb e observed lc osely.

Critically ill children suffer from pain and anxiety additionally in the face of severe organ dysfunction and often require analgesia and sedation.Different medications and combinations of medications can be used to achieve the desired effect.It is also important to keep in mind the possible adverse reactions and side effects associated with each medication.The side effect of the medications often used in children and prevention and therapy of common complications associated with analgesia and sedation is reviewed.

Alzheimer's disease (AD) is a most common neurodegenerative disease. The mechanisms underlying AD, especially late-onset AD, remain elusive. In the past few years, results from genome-wide association studies (GWAS) and systems approaches indicated that innate immune responses mediated by microglia played critical roles in AD. Functional analysis on animal models also showed that immune receptors or proteins expressed in microglia mediated Abeta-induced inflammation, or Abeta phagocytosis by microglia. Microglia plays double sword roles in AD. More work is warranted to elucidate the exact roles of microglia in AD, which will facilitate our better understanding of the mechanisms underlying AD.

OBJECTIVE:To study the direct powder compression technology of Angelica micropowder. METHODS: The formula of Angelica micropowder was optimized by mixture uniform design with tablet weight variation and friability as restrictive conditions, and a regression equation was established based on the comprehensive grading on the hardness and disintegration to figure out the optimal solution. DPS software was applied in the calculation. RESULTS: Prepared by the direct powder compression technology, the optimal formula of Angelica micropowder was obtained as follows: the hardness of the optimized tablet could reach as high as 75 N and the disintegrating time was about 10.24 min, with smooth and intact surface. CONCLUSION:The technology is reasonable in formulation and satisfactory in molding, and it can serve as theoretical basis for the production of Angelica tablets.