To study the effects of puerarin on the aromatase P450 (P450(arom)) mRNA expression and the effects of low-dose puerarin on transcription factors of the P450(arom) gene (P II) 5'-flanking region.

Objective To produce and purify recombinant adeno-associated virus(AAV) loaded with anti-amyloid ? peptide single-chain antibody gene for gene therapy of Alzheimer's disease.Methods The plasmid pSNAV2.0-Abeta-scFv was used to transform BHK-21 cell by Lipofectamine 2000 for stable package cell line establishment.The helper virus HSV1-rc/?UL2 was used to transfect package cell for anti-amyloid ? peptide single-chain antibody gene loaded recombinant adeno-associated virus production.The chloroform-PEG/NaCl-chloroform extraction and ion-exchange chromatography were employed for recombinant AAV purification.SDS-PAGE and PCR amplification were adopted for purified virus identification.The final virus physical titer was determined by digoxin-labeled DNA probes.The effect of gene therapy was tested with transgenic mice by water maze test.Results The purity of recombinant adeno-associated virus with our target gene reach up to 98% after stable cell package and serial purification.The physical titer of the final virus was 1?1012vg/mL.The latency of treated mice in water maze test were reduced significantly.Conclusion The adeno-associated virus carrying anti-amyloid? peptide single-chain antibody gene was produced by HSV1system and purified.The animal behavior test demonstrated the recombinant AAV waseffective in the gene therapy of Alzheimer's disease.

Objective To screen the specific antigen mimotopes of influenza A(H1N1)virus by phage display technology,in order to pave a way to develop novel influenza vaccine.Methods Using human convalescent serum of pandemic influenza A(H1N1)in 2009 as solid-phase selective molecule,an artificial synthesized phage randomly displaying cyclic 7-mer peptide was screened with three rounds of "absorption-elution-amplification" selection.At the end of the third round selection,32 clones were randomly chosen from the top-agar phage plaques and placed onto E.coli ER2378 in logarithmic growth phase.After culturing for 5 hours,the positive clones were identified by enzyme linked immunosorbent assay(ELISA),cross reaction test and competitive inhibition assay.The identified positive clones were analyzed by DNA sequencing.The decoded amino acids sequences,which displayed on the surface of phage,were aligned with the hemagglutinin(HA)gene of influenza virus by homology comparison for definition of the mimotopes of influenza A(H1N1)virus.Results After enriching the specific antibody-binding phages from phage displaying peptide library,12 positive clones were chosen from 32 randomly selected clones.DNA sequencing and homology comparison showed that one epitope PLHARLP was confirmed as mimotope of swine influenza A(H1N1)virus antigen,which was composed of the 52nd,53rd,59th,60th,61st,83rd and 271st amino acid.Conclusions Swine influenza A(H1N1)mimotope has been obtained by cyclic 7-mer peptide phage library screening.This result provides a basis for developing new influenza virus vaccine from virus antigen mimotopes.

Objective To evaluate CT imaging and pathological features of maxillofacial malignant myoepithelioma (MME). Methods A total of eight patients with pathologically confirmed MME of the maxillofacial region were underwent non-enhanced and contrast-enhanced CT scans.CT features of the lesions were assessed as follows:location,number,shape,size,enhancement pattern and CT number on non-enhanced and contrast-enhanced scan.CT features were compared with the pathological results.Results All the lesions were unilateral,in which mixed type with clear cells and spindle cells in 5 cases,mixed type with epithelioid cells and plas-ma cells in 2 cases,and clear cell type in one case.8 cases were manifested as lobulated mass.On non-enhanced scan,the density of all the lesions were hetergeneous with cystic area,1 case with calcification and 2 cases with bone destruction.On the contrast-en-hanced CT,all the lesions were showed hetergeneously moderate or obvious enhancement,and cystic,line and crack areas without the enhancement were observed.Minor vessel and spiculate protuberance of marginal zone on the arterial phase were observed in most le-sions.Conclusion MME has different pathological subtypes and characteristic CT features.CT is an effective method to diagnose MME.

Objective: To evaluate the value of computed tomography (CT) multi-planner reconstruction (MPR)in the treatment of lung cancer with percutaneous argon-helium cryoablation.Method:A total of 66 patients with stage T2 or T3 non-small cell lung can-cer who had complete follow-up data were treated with percutaneous argon-helium cryoablation with conventional axial CT(conven-tional group)or MPR guidance(MPR group)between January 2013 and 2016.There were 31 patients in the conventional group and 35 in the MPR group.The total number of punctures,the total time of operation,and the incidence of complications,tumor residual rates,and local control rates were compared between the two groups.Results:When the number of needles was the same,the aver-age number of punctures and puncture errors in the two groups were significantly lower in the MPR group than in the conventional group(P<0.05).Comparing the average operation time,there was no statistical difference between the MPR group and the conven-tional scan group(P>0.05).The incidence of postoperative complications was significantly lower in the MPR group than in the conven-tional scan group.Among them,the incidence of puncture combined hemorrhage was 1.52% vs.13.64%,and pneumothorax second-ary to puncture was 3.03% vs.19.70%(P<0.05).During the follow-up period to 12 months after surgery,the therapeutic benefit indica-tors such as local residual rate and total effective rate in the MPR group were also significantly better than those in the conventional group,which were 1.52% vs.10.61 and 51.52% vs.36.36%(P<0.05),respectively.Conclusions:Using MPR technology to guide percuta-neous argon-helium cryoablation treatment for lung cancer confers a small puncture error,low incidence of complications,and signifi-cant treatment benefits.This method is worthy of clinical promotion.

ObjectiveTo analyze the relationship of the static single-leg stance balance with ankle muscles strength and position sense in individuals with functional ankle instability (FAI). MethodsFrom March to May, 2023, 44 FAI individuals were recruited in Hebei Normal University, and the maximum isokinetic muscle strength (60°/s and 180°/s) and position sense as ankle flexion, extension, inversion and eversion were measured using isokinetic dynamometer; while the center of pressure (COP) sway distance and envelope area were measured as static single-leg stance with eyes open or closed using three-dimensional force plate. ResultsThe maximum isokinetic torque as ankle eversion at 180°/s correlated with the COP anterior-posterior sway distance with eyes closed (r = -0.378, P < 0.05). ConclusionExplosive ankle eversion strength is related with the balance in static single-leg stance, which may help to evaluate and intervent the ankle function of FAI individuals.

Objective To understand the status of new coronavirus contamination in the biosafety laboratory environment, identify key areas prone to contamination, and provide evidence for disinfection of central objects. Methods surfaces of high-frequency contact environment and protective equipment were sampled with moistened sterile cotton swabs after experiment and before disinfection, the results of the one-step real-time reverse transcription polymerase chain reaction (RT-PCR) of open reading frame 1ab and N fragment were used to evaluate the pollution status. Results Environmental surveys found 4 of 217 samples of environmental objects to be positive for new coronavirus RNA, that positive rate was 1.84%. Among them, BSL-3, BSL-2, and BSL-1 were sampled 23, 184, and 10 respectively. The 3 positive samples were from surfaces of nucleic acid extraction room of BSL-2 and from the handles of pass-through box, laboratory door handles and the outer surface of the alcohol watering pot respectively. The 1 positive sample was from the forearm of the protective clothing in BSL-2 laboratory. Conclusion There was a certain degree of virus pollution in key areas of the new coronavirus laboratory. The BSL-2 laboratory has a higher risk of environmental pollution than the BSL-3 and BSL-1 laboratories.

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*