【Objective】 To determine the optimal process conditions for efficiently extracting human prothrombin complex concentrates from human plasma. 【Methods】 Using human plasma as the materials and the yield of prothrombin complex concentrates as the evaluation standard, the preparation process parameters were studied and optimized through design of exporement(DOE), orthogonal experiments, and single factor experiments. 【Results】 The optimal process conditions were as follows: DEAE Sephadex A50 gel was selected, which balanced to pH 7.6, and then the amount of 1.7-2.5 g/L of plasma weight is added into the cryoprecipitate supernatant for adsorption for 40 minutes; Washing solution (0.15-0.175 mol/L sodium chloride) with 3 times the volume of gel was washed 3 times, and eluent (0.5-2.0 mol/L sodium chloride) was washed 3 to 5 times; Add stabilizer (heparin 35 IU, sodium chloride 0.1 mol/L) for ultrafiltration dialysis. 【Conclusion】 By using the optimized process mentioned above, the yield(measured by human coagulation factor IX)can reach 620 000 to 630 000 IU/ton of plasma, which is suitable for large-scale production.

【Objective】 To compare the performance of five commonly used variable selection methods in high-dimensional biomedical data variable screening so as to explore the effects of sample size and association among candidate variables on screening results and provide evidence for the development of variable selection strategy in high-dimensional biomedical data analysis. 【Methods】 Variable selection algorithms were implemented based on R-programming language. Monte Carlo method was used to simulate high-dimensional biomedical data under different conditions to evaluate and compare the performance of different variable selection methods. Variable selection performance was evaluated based on the true positive rate and true negative rate in screening. 【Results】 For specified high-dimensional data, the variable selection performance was improved for all the methods when sample size was increased, and the association between candidate variables did affect variable screening results. Simulation results indicated that the elastic network algorithm yielded the best screening performance, LASSO algorithm took the second place, and ridge algorithm did not work at all. 【Conclusion】 Elastic network algorithm is an ideal variable screening method for high-dimensional data variable screening.

It’s still controversial whether there is specificity of acupoints, because numerous studies showed that stimulation to acupoints and sham acupoints produced parallel clinical efficacies. For example, acupoint specificity was not detected when Visual Analogue Scale was adopted to evaluate pain, while hemodynamic and neuroimaging studies suggested the existence of acupoint specificity. This article reviewed and summarized relevant studies that supported or didn’t support acupoint specificity. Further researches are required to prove the existence of specificity of acupoints.

Objective To investigate the ability of invasion and migration of breast cancer MDA-MB-231 cells under serum starvation and hypoxia, and the effect of antiangiogenic drugs, rh-endostatin and bevacizumab, on the ability of invasion and migration of breast cancer cells under serum starvation and/or hypoxia, in order to explore the potential risk of antiangiogenic therapy in clinics.Mte hods The cells were randomized into 4 groups, i.e., group A:10%fetal bovine serum ( FBS) group;group B:hypoxia +10%FBS group;group C:serum starvation group;group D: hypoxia +serum starvation group; each group was further divided into three subgroups as blank control, treated with rh-endostatin and bevacizumab, respectively.Cell counting kit-8( CCK-8) was used to assess the inhibition rate of cell growth induced by endostatin and bevacizumab, in order to determine the proper working concentration and time of the two drugs.Transwell assay was conducted to detect the cell invasion and migration in vitro.The expressions of c-Met and MMP-9 were detected by Western blot.The cells treated with rh-endostatin or bevacizumab under serum starvation were tested by hybridization using Exiqon miBase 18.0 microarray.The miRNAs which exibited significant differences ( P<0.05) in miRNA hybridization were verified by real-time PCR assay. Results CCK-8 assay showed that the inhibition rates of MDA-MB-231 cells cultured with 800 mg/L rh-endostatin for 48 h and 24 h were (32.2±2.5)%and (27.0±1.3)%, respectively, showing a significant difference ( P=0.023) .The inhibition rates of MDA-MB-231 cells cultured with 80 mg/L bevacizumab for 48 h and 24 h were (30.5±1.4)%and (26.1±2.4)%, respectively , showing also a significant difference (P=0.015).The Transwell assay showed that in the starvation blank group, the number of invaded and penetrated cells were 28.8 ±2.2 and 31.4 ±1.5, respectively, significantly different from that in the rh-endostatin and bevacizumab groups (P<0.05).The relative expressions of c-Met and MMP-9 were 0.213± 0.017 and 0.542±0.048, respectively, with a significant difference from those of the groups treated with each drug ( P<0.05 for both) .The numbers of penetrated cells in the Transwell assay treated with rh-endostatin in hypoxia were 17.5±2.1 and 16.5 ±2.8, respectively, and the numbers of penetrated cells in the Transwell assay treated with bevacizumab were 16.3±3.5 and 17.5±2.4, respectively, showing no significant difference among them ( P>0.05 for both ) .The ability of migration and invasion of MDA-MB-231 cells and the expression of c-Met and MMP-9 were not impacted by hypoxia ( P>0.05) .Real-time PCR assay showed that only the levels of miR-2355 and miR375 were significantly and stably decreased in the cells which had increased ability of invasion and migration.The relative expression levels of miR375 and miR-2355 in the serum starvation blank group were 0.550±0.036 and 0.852±0.121, respectively, significantly lower than that in the groups treated with rh-endostatin or bevacizumab (P<0.05).In the serum starvation group, the expression levels of miR375 and miR-2355of cells treated with rh-endostatin were 02.95±0 .012 and 0.253±0.011, and the expression levels of cells treated with bevacizumab were 0.234±0.020 and 0.309±0.02,2 respectively, ( P>0.05 for all).Compared with the serum starvation blank group, the expression levels of miR2355 and miR375 were significantly decreased when cells were treated with rh-endostatin/bevacizumab under serum starvation, but no significant difference was found between the two drugs ( P>00.5 ).However, hypoxia did not affect the expressions of miR2355 and miR375 ( P>0.05).Conclusions The results of this study suggest that serum starvation can increase the ability of invasion and migration of breast cancer cells.Furthermore, both rh-endostatin and bevacizumab may enhance their invasion and penetration ability under serum starvation condition.

Objective To investigate the ability of invasion and migration of breast cancer MDA-MB-231 cells under serum starvation and hypoxia, and the effect of antiangiogenic drugs, rh-endostatin and bevacizumab, on the ability of invasion and migration of breast cancer cells under serum starvation and/or hypoxia, in order to explore the potential risk of antiangiogenic therapy in clinics.Mte hods The cells were randomized into 4 groups, i.e., group A:10%fetal bovine serum ( FBS) group;group B:hypoxia +10%FBS group;group C:serum starvation group;group D: hypoxia +serum starvation group; each group was further divided into three subgroups as blank control, treated with rh-endostatin and bevacizumab, respectively.Cell counting kit-8( CCK-8) was used to assess the inhibition rate of cell growth induced by endostatin and bevacizumab, in order to determine the proper working concentration and time of the two drugs.Transwell assay was conducted to detect the cell invasion and migration in vitro.The expressions of c-Met and MMP-9 were detected by Western blot.The cells treated with rh-endostatin or bevacizumab under serum starvation were tested by hybridization using Exiqon miBase 18.0 microarray.The miRNAs which exibited significant differences ( P<0.05) in miRNA hybridization were verified by real-time PCR assay. Results CCK-8 assay showed that the inhibition rates of MDA-MB-231 cells cultured with 800 mg/L rh-endostatin for 48 h and 24 h were (32.2±2.5)%and (27.0±1.3)%, respectively, showing a significant difference ( P=0.023) .The inhibition rates of MDA-MB-231 cells cultured with 80 mg/L bevacizumab for 48 h and 24 h were (30.5±1.4)%and (26.1±2.4)%, respectively , showing also a significant difference (P=0.015).The Transwell assay showed that in the starvation blank group, the number of invaded and penetrated cells were 28.8 ±2.2 and 31.4 ±1.5, respectively, significantly different from that in the rh-endostatin and bevacizumab groups (P<0.05).The relative expressions of c-Met and MMP-9 were 0.213± 0.017 and 0.542±0.048, respectively, with a significant difference from those of the groups treated with each drug ( P<0.05 for both) .The numbers of penetrated cells in the Transwell assay treated with rh-endostatin in hypoxia were 17.5±2.1 and 16.5 ±2.8, respectively, and the numbers of penetrated cells in the Transwell assay treated with bevacizumab were 16.3±3.5 and 17.5±2.4, respectively, showing no significant difference among them ( P>0.05 for both ) .The ability of migration and invasion of MDA-MB-231 cells and the expression of c-Met and MMP-9 were not impacted by hypoxia ( P>0.05) .Real-time PCR assay showed that only the levels of miR-2355 and miR375 were significantly and stably decreased in the cells which had increased ability of invasion and migration.The relative expression levels of miR375 and miR-2355 in the serum starvation blank group were 0.550±0.036 and 0.852±0.121, respectively, significantly lower than that in the groups treated with rh-endostatin or bevacizumab (P<0.05).In the serum starvation group, the expression levels of miR375 and miR-2355of cells treated with rh-endostatin were 02.95±0 .012 and 0.253±0.011, and the expression levels of cells treated with bevacizumab were 0.234±0.020 and 0.309±0.02,2 respectively, ( P>0.05 for all).Compared with the serum starvation blank group, the expression levels of miR2355 and miR375 were significantly decreased when cells were treated with rh-endostatin/bevacizumab under serum starvation, but no significant difference was found between the two drugs ( P>00.5 ).However, hypoxia did not affect the expressions of miR2355 and miR375 ( P>0.05).Conclusions The results of this study suggest that serum starvation can increase the ability of invasion and migration of breast cancer cells.Furthermore, both rh-endostatin and bevacizumab may enhance their invasion and penetration ability under serum starvation condition.

OBJECTIVETo investigate the incidence, pathogens, and clinical features of infection in consecutive cases from 2010 to 2012 in Peking Union Medical College Hospital.

METHODThe incidence, pathogen, treatment, and outcomes of patients with hematological diseases who had positive findings of bacterium in their samples from 2010 to 2012 were retrospectively analyzed.

RESULTSThere were 449 positive samples (5.8%) from 4 890 patients during this period, among which 388 were proved to be with pathogenic bacteria. Samples separated from patients with community-aquired infections accounted for 8.4% of all positive samples. Most community-aquired infections were caused by Gram-negative bacteria (75%), although no multidrug-resistant bacteria was observed. Samples separated from patients with nosocomial infections accounted for 91.6% of all positive samples. Respiratory tract (49.4%) and peripheral blood (32.6%) were the most common samples with positive results. Skin soft tissues (10.4%), and urine (3.7%) were less common samples. Most of the pathogenic bacteria of the nosocomial infections were Gram-negative (66.9%). The most common Gram-negative bacteria included Escherichia coli (13.8%), Pseudomonas aeruginosa (12.1%), and Klebsiella pneumonia (12.1%), while Staphylococcus aureus (10.4%), Enterococcus faecium (7.0%), and Staphylococcus epidermidis (5.1%) were the most common Gram-positive bacteria. Gram-negative bacteria consisted of most of sputum samples and peripheral blood samples. Samples from the surface of skin wound and anal swab were composed largely by Gram-positive bacteria (63.8%). The detection rates of extended-spectrum beta-lactamase-producing Klebsiella pneumonia/Klebsiella oxytoca, Escherichia coli, and Proteus mirabilis were 24.0%, 87.9% and 38.4%, respectively. The resistance to Acinetobacter baumannii was serious. Multidrug-resistant, extensive drug resistant and pan drug resistant A. baumannii acountted for 74% of all A. Baumannii infections. Stenotrophomonas maltophilia showed low resistance to sulfamethoxazole/trimethoprim, levofloxacin and minocycline. Also, 22 methicillin-resistant Staphylococcus aureus and 9 methicillin-resistant Staphylococcus Epidermidis were detected, which were only sensitive to vancomycin, teicoplanin, and linezolid. All patients were treated in the haematology wards and most of them were under agranulocytosis or immunosuppression. Finally, 22 patients reached clinical recovery through anti-infective therapy, whereas 49 patients died. Among those deaths, 42 patients attributed to severe infections and infection-associated complications. Fourteen of all the deaths might be infected with drug-resistance bacteria. There were 61 samples proved to be bacteria colonization. Nonfermenters such as Acinetobacter baumannii and Stenotrophomonas maltophilia made up for a large amount of bacteria colonization.

CONCLUSIONSThe pathogens of nosocomial infections in the hematology ward are mainly Gram-negative bacteria. The incidences and pathogens vary from different infection sites. Nosocomial infection still has a higher mortality rate. Once nonfermenters are detected positive, the pathogenic or colonial bacteria should be distinguished.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; isolation & purification ; Bone Marrow Transplantation ; Cross Infection ; microbiology ; Female ; Hematologic Diseases ; complications ; microbiology ; Hematology ; Hospital Departments ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To investigate the correlation between the distribution of lesions on MRI and the findings of computerized tomography angiography (CTA) in patients with posterior circulation infarction.Methods Patients with acute posterior circulation infarction were enrolled in the study.The sites of the infarcts were divided into proximal,middle and distal infarctions according to the results of MRI.All the patients received head and neck CTA.The correlation between the distribution of posterior circulation infarcts and the sites of vertebrobasilar system lesions was analyzed.Results A total of 203 patients with acute posterior circulation infarction were enrolled.Their primary clinical symptoms and signs were unilateral limb weakness (n =77,37.93％),dizziness (n =129,63.55％),dysarthria (n =31,15.27％),nausea and vomiting (n =61,30.05％),headache (n =79,38.92％),gait abnormal (n =50,24.63％),nystagmus (n=34,16.75％),and ataxia (n=21,10.34％).Proximal infarction (n=35,17.24％):medullary infarction (n =28,13.79％),posterior inferior cerebellar artery infarction (n =7,3.45％); middle infarction (n =95,46.79％):pontine infarction (n =80,39.4％),anterior inferior cerebellar infarction (n =15,7.39％); distal infarction (n=73,35.96％):middle cerebral infarction (n=6,2.96％),superior cerebellar infarction (n =16,7.88％),thalamic infarction (n =34,(16.75％),occipital lobe infarction (n =10,4.93％),temporal lobe infarction (n =7,3.44％).Extracranial vertebral artery lesions were most common in the distal infarction group.It reached 53.42％,and was significantly higher than 22.86％ in the proximal infarction group (P =0.003) and 33.68％ in the middle infarction group (P =0.010).Intracranlal vertebral artery lesions were most common in the proximal infarction group.It reached 57.14％,and then followed by the middle infarction (41.05％).They were all significantly higher than 15.07％ in the distal infarction group (all P =0.000).Basilar artery lesions were most common in the middle infarction group.It reached 20.00％ and was significantly higher than 4.11％ in the distal infarction group (P=0.002).Posterior cerebral artery lesions were most common in the distal infarction group.It reached 27.40％ and was significantly higher than 5.71％ in the proximal infarction group (P =0.009) and 5.26％ in the middle infarction group (P=0.000).Conclusions The range of vascular lesions of the distribution of lesions shown on MRI and the findings of CTA on vertebrobasilar artery system in patients with posterior circulation infarction had some connection.During the proximal and middle infarctions,the possibility of having intracranial vertebral artery lesions was greater; during the distal infarction,the possibility of having extracranial vertebral artery and posterior cerebral artery lesions was greater.

Objective: To explore the feasibility and safety of three dimensional (3D) electro-anatomical mapping system (Carto3) in treating the patients with paroxysmal supra-ventricular tachycardia (PSVT) by radiofrequency catheter ablation (RCFA).
Methods: A total of 180 PSVT patients were divided into 2 groups, n=90 in each group. 3-D group, the patients received RCFA with 3-D reconstructed valve ring model under Carto3 guidance. 2-D group, the patients received RCFA under conventional X-ray guidance. The procedural and X-ray exposure times, rates of success and complications, tachycardia recurrence at 6 months after procedure and the cost were observed and compared between 2 group.
Results: The procedural time was similar between 2 groups, P=0.1403. The patients in 3-D group had the lower X-ray exposure time (2.1 ± 0.7 vs 7.8 ± 3.6) min, particularly in those with right-sided accessory pathway (3.4 ± 0.7 vs 20.2 ± 7.1) min, and dual atrio-ventricular (A-V) nodal pathways (1.1 ± 0.3 vs 5.5 ±1.7) min, all P<0.0001. There was 1 patient in 3-D group without RCFA and all the others were successes. 2-D group had 3 patients with failed RFCA including 2 of right-sided accessory pathway, 1 of dual A-V nodal pathways and received 2nd RCFA under Carto3 guidance. 3-D group had no complication, no recurrence. In 2-D group, 1 patient suffered from complete A-V block (AVB) during ablation and 1-year later, the Holter showed II° to III° AVB;2 patients with recurrence including 1 of dual A-V nodal pathways and had successful 2nd ablation. The cost was higher in 3-D treatment.
Conclusion: RFCA was feasible for treating PSVT patients under Carto3 guidance, which had the higher success rate with lower X-ray exposure and complication.

Our previous work has shown that mesenchymal stem cells (MSC) have little therapeutic effect on rat arthritis induced by collagen. This study was aimed to further investigate whether the MSC lysates exhibit beneficial effects on rheumatoid arthritis. Aliquots of cell lysates from 1×10(7) human bone marrow MSC were intraperitoneally injected into collagen-induced arthritis (CIA) Wistar rats weekly for 4 consecutive weeks. Methotrexate at a dose of 1 mg/kg or normal saline was served as positive and negative controls respectively. On week 4 the symptom scores were recorded and the hind joints of the rats were pathologically examined and X-ray examination was performed. The results showed that on week 4, the symptom scores of the rats that received MSC lysates (6.87 ± 0.83) and MTX (6.44 ± 1.13) were significantly lower than that of control rats (7.33 ± 0.77, P < 0.01). Meanwhile, pathological examination on the involved ankle showed that the synovitis and arthritis scores of MSC lysates and control groups were 2.28 ± 0.48 and 2.28 ± 0.55 respectively, significantly higher than that of MTX treatment rats (0.71 ± 0.48, P < 0.05). However, X-ray examination on the ankle joints showed that the injury score of control rats was 4 ± 0.57, greatly higher than those from MSC lysates (2.71 ± 0.75) and MTX treatment groups (2.57 ± 0.78, P < 0.05 for both groups). It is concluded that MSC lysate infusion has beneficial effects on CIA rat, but the effectiveness seems inferior to MTX.
Animals ; Arthritis, Experimental ; chemically induced ; therapy ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Collagen ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; Methotrexate ; pharmacology ; Rats ; Rats, Wistar

Objective To construct a lentiviral vector carrying angiopoietin-1 (Ang-1) and DsRed gene, and to package a virus particles. Methods The Ang-1 lentiviral vector with DsRed (PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2) was constructed by Gateway technology, and identiifed by PCR and gene sequencing. The lentiviral vector was mixed with helper vector pLV/helper-SL3, pLV/helper-SL4 and pLV/helper-SL5 by Lipofectamine 2000 to prepare DNA-Lipofectamine?2000 complexes. The complexes were then added to transfect 293FT cells and package virus. The virus titers and infection ef-ifciency were determined by lfuorescence expression. Results Ang-1 lentiviral vector PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2 was constructed successfully as identified by PCR and gene sequencing. Lentivirus with high-efficiency infection was produced by transfection to 293FT cells and the virus titer was 5×108 TU/ml. Conclusions The recombinant len-tiviral vector for Ang-1 was successfully constructed by Gateway technology and the lentivirus with high-efifciency infection packaging can be used for further experiment of Ang-1 gene.