Animals ; Biocompatible Materials ; therapeutic use ; Bone Regeneration ; Bone Substitutes ; therapeutic use ; Bone Transplantation ; Collagen ; therapeutic use ; Guided Tissue Regeneration, Periodontal ; methods ; Humans ; Membranes, Artificial ; Periodontal Diseases ; surgery ; Polytetrafluoroethylene ; therapeutic use

Alveolar Process ; surgery ; Dental Implantation, Endosseous ; methods ; Esthetics, Dental ; Gingiva ; surgery ; Humans

The major challenge in the development of recombinant biologics lies in generating and isolating rare high-producing stable single clone in a short period of time. The selection marker is an essential component of the plasmid vector, it plays an important part in the generation and screening of producing cell lines. Engineering the selection marker to enhance the stringency of selection for high producing cells is one of the most effective approaches to improve the cell line development process. Here, using Chinese hamaster overy (CHO) cells as an example, we introduce the application of selection marker for generation of recombinant biologics producing mammalian cell lines, methods of engineering the selection markers to enhance the selection stringency, and propose considerations on cell substrate stability and selection marker safety, in order to provide references for high-efficiency development of recombinant biologics.

Chlorine ; Computer Simulation ; Computers, Analog ; Diffusion ; Software

Objective This paper aims at establishing a inductively coupled plasma mass spectrometry ( ICP-MS) method for quantification and evaluation of iodine in human urine and serum in routine clinical laboratory .Methods This study was methodology validation research on iodine evaluation using ICP-MS.Ammonia, isopropanol and ultrapure water were mixed at certain ratio to dilute samples in the ratio of 1:10, and then the diluted samples were analyzed by ICP -MS.Re was used as the internal standard.And linearity, lower limit of detection, recovery, precision, accuracy, carryover and stability was evaluated thoroughly .Results of iodine of pregnant women who required iodine tests were retrospectively analyzed to evaluate the status of iodine .Results The method only needs 30s for analysis of one sample .It was sensitive with a lower limit detection of 0.87μg/L, the correlation coefficient was higher than 0.999 9 in ten measurements.The recovery in both serum and urine was approximately 100% (95.3% -109.9%). Based on the NIST standard reference material 3668 comparison, the bias was less than 4%( -0.9% -3.9%).The inter-coefficient variation (CV) for serum iodine and urine iodine was 1.2%-3.0%, 2. 0%-2.9%, respectively;and total CV for serum iodine and urine iodine were 3.0%-3.8%, 4.1%-4.9%, respectively.The mean carryover of this method was 0.03% and iodine was stable for at least one month at -20℃ and 4℃.The urine and serum iodine for pregnant women was (154.8 ±89.7) μg/L (mean ±SD),(75.8 ±21.4) μg/L, respectively.The correlation between urine and serum iodine was 0.21. Conclusion Establishe a rapid and simple ICP -MS method for urine and serum iodine measurement with high accurate and precise in routine clinical laboratory .

Objective To study the effect of a Chinese medicine, Huanglian Jiedu Decoction ( HLJDD) , on ath-erosclerotic plaque, inflammatory factors and regulatory T cells in ApoE-/- mice. Methods High fat diet was given to ApoE-/- mice to establish an atherosclerosis model. 40 male ApoE-/- mice were randomly divided into five groups:model group, simvastatin group, low, moderate and high dose HLJDD groups (n =8). HLJDD was intragastrically administered in a dose of 3. 5, 7. 0, or 14. 0 g/( kg?bw) once dailg for 16 weeks. The dose of simvastatin was 5. 0 g/( kg?bw). An-other 8 male C57BL/6J mice were taken as control group. At the end of the 29-week experiment, all of the mice were sacri-ficed. The aortic plaques, level of blood lipids, inflammatory factors, Tregs number and the level of Foxp3 mRNA were de-tected and analyzed by ELISA, flow cytometry and RT-PCR, respectively. Results Compared with the control group, the aortic plaques were much larger in the model group, and the levels of TC, TG, LDL-C, IL-6, hs-CRP, and TNF-α were significantly higher than those in the control group (P<0. 01 for all). Meanwhile, the levels of HDL-C, IL-10, TGF-βand Foxp3 mRNA were much lower than those in the control group ( P<0. 01 for all) , and the Tregs numbers were less than that in the control group (P<0. 01). HLJDD regulated the blood lipids in ApoE-/ - mice and decreased the levels of IL-6, hs-CRP, andTNF-α, however increased the levels of IL-10, TGF-β and Foxp3 mRNA. At the same time, it in-creased Tregs number in the ApoE-/ - mice. Compared with the model group, the difference was statistically significancet (P<0. 01). Conclusions HLJDD can significantly alleviate the aortic plaque damages in ApoE-/ - mice. It may be re-lated to the up-regulation of Tregs, which can lead to decrease the expression of serum pro-inflammatory factors such like IL-10, hs-CRP and TNF-α.

Duck circovirus(DuCV),a potential immunosuppressive virus,was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction(PCR)based method. In this study,a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of～35%. It was found that ducks between the ages of 40～60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders,growth retardation or lower-than-average weight. The complete genomes of 9. strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes,compared with others genomes downloaded from GenBank,ranged in size from 1988 to 1996 base pairs,with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes,Group I(the Euro-USA lineage)and Group II(the Taiwan lineage),with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species,including Duck,Muscovy duck,Mule duck,Cheery duck,Mulard duck and Pekin duck.

To look for a more stable and convenient way of constructing short hairpin RNA expression vectors targeting the latent membrane protein-1(LMP-1) encoded by Epstein-Barr virus(pshLMP1), and to study the inhibition function of pshLMP1 expression vectors in HNE1 cells, we designed the pshLMP1 expression cassette and pshLMP1 expression vectors by both the annealing method and PCR method and then co-transfected with pEGFP-N1-1158 into HNE1 cells to observe the mRNA and protein levels of LMP-1 genes by green fluorescence analysis, RT-PCR and western blot. pshLMP1 expression vectors were successfully obtained by both methods but better cloning efficiency was achieved and fewer deletions and mutations of nucleotides were achieved with the PCR method. Furthermore, the mRNA and protein levels of LMP-1 genes were down-regulated by pshLMP1 expression vectors. According to our research, we found that the PCR method provides a more efficient way to construct pshLMP1 expression vectors which have the ability to inhibit the function of LMP-1 genes expressed in HNE1 cells, and also provides a novel application of RNA interference technology against-EBV.

Biological Factors ; analysis ; Drug Monitoring ; Drugs, Chinese Herbal ; analysis ; Humans ; Pharmaceutical Preparations ; analysis ; Technology, Pharmaceutical

Background and purpose:Cervical cancer is one of the most common cancer in Xinjiang, especially for Uygur from southern Xinjiang and its pathogenesis is not clear. MicroRNA (miRNA) is a class of small non-coding RNA playing an important regulatory role. Its expression and dysfunction is closely related to the development of tumors. In this study, we screen and preliminary analyse expression of miRNA in cervical squamous cell carcinoma samples with human papillomavirus (HPV) 16 positive of Uygur patients. The target genes of miRNA were predicted.Methods:miRNAs were pre-screened by using miRNA microarray technology in 5 cases of HPV16 positivity Uygur patients from southern Xinjiang with cervical squamous cell carcinoma. Fifteen cases specimens were examined by qRT-PCR for preliminary veriifcation, and 83 cases of cervical cancer were detected and analysed the expression of miRNA; Targeted genes were predicted by using four softwares of target scan, miRwalk, miRanda and Pictar.Results:Eighteen differentially expressed miRNAs were selected by SAM software in 5 cases of HPV16 positivity southern Xinjiang Uygur cases with cervical squamous cell carcinoma.miRNA-138 and miRNA-720 were found expressed signiifcantly different by initial veriifcation. Contrasted with 40 normal cases, miR-138 and miR-720 were down-regulated in 83 Uygur patients from southern Xinjiang with cervical squamous cell carcinoma (P<0.05),and correlated with lymph node matastasis and vascular invasion (P<0.05), no correlation with age and the range of cervical wall involvement and HPV16 (P>0.05). miRNA-720 was correlated with clinical stage and tumor size (P<0.05); And the commonly targeted gene between miRNA-138 and miRNA-720 was EZH2.Conclusion:miRNA-138 and miRNA-720 were downregulated in Uygur patients from southern Xinjiang with cervical squamous cell carcinoma, and the common target gene was EZH2.The expression of miR-720 and miR-138 were correlated with relevant risk factors of invasion and metastasis.