Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.
Animals ; Animals, Genetically Modified ; Antithrombin III ; biosynthesis ; Chromatography, Affinity ; Female ; Goats ; Heparin ; Humans ; Mammary Glands, Animal ; metabolism ; Milk ; chemistry ; Recombinant Proteins ; biosynthesis

Objective To explore the change of cerebral microcirculation before and after clipping of intracranial aneurysm using CT perfusion (CTP)imaging.Methods CTP images of 70 patients diagnosed intracranial aneurysm by CTA were retrospectively an-alysed.They were divided into four groups according to have or not intracranial arterial spasm:A group:Preoperative CVS,postop-erative CVS (4 cases),B group:Preoperative CVS,postoperative nCVS (13 cases),C group:Preoperative nCVS,postoperative CVS (20 cases),D group:Preoperative nCVS,postoperative nCVS (33 cases).CBV/CBF/MTT were measured in four groups be-fore and after surgery,and analyzed statistically.Results (1)The postoperative CBV/CBF/MTT was compared with the preopera-tive data.There were statistically significant in CBVA/B/C/D/CBFA/B/D/MTTA/B during the surgery (P <0.05 ).There were no statistically significant in the CBFC and MTTC/D during the surgery (P >0.05).(2)There were statistically significant in the△CBVBC/BD/△CBFAB/BC/BD/△MTTAB/BD (P < 0.05 ).△CBVAB/AC/AD/CD/△CBFAC/AD/CD/△MTTAC/AD/BC/CD were no statistically significant (P >0.05).Conclusion CTP can early and accurately predict the change of cerebral microcircu-lation after surgery.

Objective To explore the association of catechol-O-methyhransferase(COMT) gene polymorphisms with schizophrenia susceptibility and its symptoms assessed by Positive and Negative Syndrome Scale (PANSS)in southern Chinese population.Methods COMT gene rs4633,rs4680 and rs8185002 polymorphisms were genotyped using Sequenom genotyping technology in 700 schizophrenia patients (300 Zhuang and 400 Han) and 700 healthy controls (300 Zhuang and 400 Han),and Positive and Negative Syndrome Scale (PANSS) was used for clinical symptoms assessment of patients.Statistical analysis was performed using PLINK software.Results rs4633,rs4680 and rs8185002 polymorphisms were not significantly associated with schizophrenia susceptibility in Zhuang or Han population respectively(P＞0.05).After merging Zhuang and Han samples,rs4633(I 2 =0.000,Pmeta =0.040) and rs4680 (I2=0.000,Pmeta =0.014) were significantly associated with the susceptibility to schizophrenia.In addition,haplotype T-A-T was significantly associated with schizophrenia susceptibility (P=0.049).However,these three polymorphisms were not significantly associated with total score,positive scale score,negative scale score and general psychopathology scale score assessed by PANSS(P＞0.05).Conclusion COMT gene rs4633 and rs4680 polymorphisms are involved in the susceptibility to schizophrenia in southern Chinese population.

Objective To detect the bacteria and drug sensitivity in new borus with ventilator-associated pneumonia. Methods The study included 128 new horns. We detected the bacteria and drug sensitivity of trachea secretion in neonatal VAP. Results Klebsiella pneumonia, acinetobacter baumannii, enterobacter cloacae, pseudomonas aerubinosa and escherichia coli were the most common bacteria in VAP. They were widely resistance to main antibioties. Conclusion Gram-negative bacilli are the most bacteria. Mixed infection and muhidrug resistance are important clinical characteristic in VAP.

In recent years, multi-walled carbon nanotubes (MWCTs) are very favorable to the adsorption of middle molecular substances in the hemoperfusion because of their multiporous structure, large surface area and high reactivity, which are beneficial to the excellent absorption properties. The purpose of this study was to study the MWCTs on the adsorption capacity of the middle molecular substances. Vitamin B12 (VB12) was selected as a model of the middle molecular substances. The morphologies of MWCTs and activated carbon from commercial "carbon kidney" were observed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The adsorption behavior of VB12 was compared to each other with UV-visible absorption spectra. The MWCTs formed a sophistaicate gap structure, and compared to the activated carbon, MWCTs had a larger surface area. By Langmuir equation and Freundlich equation fitting analysis, VB12 adsorption on MWCTs is fit for multi-molecular layer adsorption, and the adsorption type of activated carbon is more inclined to the model corresponding to Langmuir monolayer adsorption. The adsorption rate of MWCTs is faster than that of the activated carbon and the adsorption capacity is greater, which could be expected to become the new adsorbent in the hemoperfusion.
Adsorption ; Charcoal ; chemistry ; Nanotubes, Carbon ; chemistry ; Porosity ; Toxins, Biological ; chemistry ; Vitamin B 12 ; chemistry

Objective To investigate the feasibility of 80 kV and low tube current on brain CT perfusion(CTP).Methods Sixty patients underwent 80 kV CTP examination,and were randomly divided into five groups according to different tube currents,A group (40 mAs),B group (60 mAs),C group (80 mAs),D group (100 mAs),E group (150 mAs).Twelve patients in each group were selected to analyze the parameters of the caudate nucleus and white matter frontals and temporal,including CBV,CBF and MTT.Subjective image quality,CT value,SD,SNR,CNR and radiation dose were measured and calculated in the 5 groups respectively.Results Differences of CBV and CBF in the caudate nucleus and white matter frontals were existed among A group,B group,and E group (P＜0.05).There were no significant differences of CT value among the 5 groups (P＞0.05).For the SD and SNR,E group was better than A-D groups (P＜0.05).The CNR was higher in E group than that in A group (P＜0.05).The subjective image quality score evaluation in C,D groups were not lower than that in E group(P ＞0.05).The ED of A-D groups were all lower than that of E group (P＜0.05).Conclusion 80 kV/80 mAs brain CTP can obtain accurate perfusion parameter and satisfied image quality,and the radiation dose can be reduced by 46.22％ (2.42 mSv).

Objective To study the structure and function changes of heart in elderly patients with hypertension. Methods Two hundred old patients with hypertension in our hospital were enrolled in this study. Which were divided into elderly aged group (≥ 80 years old, 76 cases) and non-elderly aged group (<80 years, 124 cases). The SBP, DBP, LVDd, VPWd, IVST, LVDs, LAD, EF and FS were compared between the two groups. Results There were no significant differences of SBP and DBP between the two groups(P>0.05). The LVDd, LVDs, and LAD in elderly aged group were higher than that in non-elderly aged group (P<0.05). The LVPWd and IVST in elderly aged group were lower than that in non-elderly aged group (P<0.05). There were no significant differences of EF and FS between the two groups (P>0.05). Conclusion The cardiac structural changes are more early than function changes in elderly hy-pertensive patients.

Animals ; Caenorhabditis elegans ; metabolism ; radiation effects ; Caenorhabditis elegans Proteins ; genetics ; metabolism ; Light ; Lysine ; analogs & derivatives ; genetics ; metabolism ; Promoter Regions, Genetic ; Protein Transport ; RNA, Transfer ; genetics ; metabolism ; Tumor Necrosis Factor Ligand Superfamily Member 14 ; metabolism

Objective To evaluate the risk factors and sonographic findings of pregnancies complicated by placenta increta or placenta percreta. Methods Totally, 2219 cases were retrospectively analyzed from 20 tertiary hospitals in China from January 2011 to December 2015. The data were collected based on the original case records. All cases were divided into two groups, the placenta increta (PI) group (79.1％, 1755/2219) and the placenta percreta (PP) group (20.9％, 464/2219), according to the degree of placental implantation. The risk factors and sonographic findings of placenta increta or percreta were analyzed by uni-factor and logistic regression statistic methods. Results The risk factors associated with the degree of placental implantation were age, gravida, previous abortion or miscarriage, previous cesarean sections, and placenta previa (all P<0.05), especially, previous cesarean sections (χ2=157.961) and placenta previa (χ2=91.759). Sonographic findings could be used to predict the degree of placental invasion especially the boundaries between placenta and uterine serosa, the boundary between placenta and myometrium, the disruption of the placental-uterine wall interface and loss of the normal retroplacental hypoechoic zone(all P<0.01). Conclusions Previous cesarean sections and placenta previa are the main independent risk factors associated with the degree of placenta implantation. Ultrasound could be used to make a prenatal suggestive diagnosis of placenta accreta spectrum disorders.

Antibody-drug conjugates(ADCs)are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies.In this study,we report the generation of a monoclonal antibody against monomethyl auristatin E(MMAE)and the development,validation,and application of sensitive and high-throughput enzyme-linked immunosor-bent assays(ELISA)to measure the concentrations of MMAE-conjugated ADCs and total antibodies(tAb,antibodies in ADC plus unconjugated antibodies)in cynomolgus monkey sera.These assays were suc-cessfully applied to in vitro plasma stability and pharmacokinetic(PK)studies of SMADC001,an MMAE-conjugated ADC against trophoblast cell surface antigen 2(TROP-2).The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit,Lys(m-dPEG24)-Cit,and Val-Cit linkers.The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation be-tween serum concentrations and the OD450 values,with R2 at 1.000,and the dynamic range was 0.3-35.0 ng/mL and 0.2-22.0 ng/mL,respectively;the intra-and inter-assay accuracy bias％ranged from-12.2％to-5.2％,precision ranged from-12.4％to-1.4％,and the relative standard deviation(RSD)was less than 6.6％and 8.7％,respectively.The total error was less than 20.4％.The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters,which suggested that these can be applied to quantify MMAE-conjugated ADCs,as well as in PK studies.Furthermore,these assays can be easily adopted for development of other similar immunoassays.