PCR-reverse dot blot human papillomavirus genotyping as a primary screening test for cervical cancer in a hospital-based cohort

Yafang Kang,Pengming Sun,Xiaodan Mao,Binhua Dong,Guanyu Ruan,Lihua Chen 대한부인종양학회 2019 Journal of Gynecologic Oncology Vol.30 No.3

Objective: To evaluate the polymerase chain reaction (PCR)-reverse-dot-blot (RDB) human papillomavirus (HPV) genotyping test as a feasible assay for the cervical cancer primary screening. Methods: In a hospital-based cohort, a total of 21,568 women were voluntarily enrolled from March 2009 to November 2016 for evaluating the 3 current cervical cancer screening strategies: co-test, cytology primary and high-risk HPV (HR-HPV) primary by using PCRRDB HPV genotyping and liquid-based cytology (thinprep cytologic test [TCT]). Women with HR-HPV infection and/or abnormal cytology were referred for colposcopy, and the biopsy or conization was performed according to the American Society for Colposcopy and Cervical Pathology (ASCCP) guidelines. Results: Overall, 18.20% (3,935/21,568) of the women were detected as HR-HPVpositive, 5.04% (1,088/21,568) were diagnosed with cervical intraepithelial neoplasia 2 or higher (CIN2+), and 3.43% (739/21,568) with CIN3+. The cumulative incidence rates for CIN2+/CIN3+ in patients with HPV-16/18-positive were 48.28%/37.20%, while they were 0.86%/0.38%, 0.30%/0.15% and 0.18%/0.09% in cytology-negative, HR-HPV-negative and co-test-negative population, respectively. Using CIN2+ and CIN3+ as the observed endpoints, the sensitivity and negative predictive value (NPV) of HR-HPV genotyping as a primary screening tool were 90.99%/99.49% and 91.57%/99.80%. Moreover, using HR-HPV genotyping primary screening could detect the same more CIN2+/CIN3+ cases in baselinedetection as co-testing (990/700 vs. 991/701) and far more than cytology primary screening (903/656, p<0.05). It also achieved the lowest misdiagnosis rate (8.01%/5.02%). Although HPV genotyping primary screening required an increased number of colposcopies (2.75/3.89 per CIN2+/CIN3+ case), it yielded an acceptable rate. Conclusions: The PCR-RDB HPV genotyping test is a cost-effective and beneficial cervical cancer primary screening for hospital-based opportunistic screening.

Improving electrochemical performance of LiMnPO_4 by Zn doping using a facile solid state method

Yourong Wang,Yafang Chen,Siqing Cheng,Liangnian He 한국화학공학회 2011 Korean Journal of Chemical Engineering Vol.28 No.3

Olivine structure LiMnPO_4/C as cathode materials for Li-ion batteries were synthesized via a simple solidstate reaction. Improvement of the electrochemical performance of LiMnPO_4/C cathode material was realized significantly by the method of doping Zn. The obtained LiMn_(0.95)Zn_(0.05)PO_4/C electrode material was studied by the measurements of X-ray diffraction pattern, scanning electronic microscopy, electrochemical impedance spectroscopy and electrochemical performance. The results indicate that the LiMn_(0.95)Zn_(0.05)PO_4/C materials exhibit discharge specific capacity of 140.2 mA h g^(−1) at 0.02 C rate and better rate capability. These excellent results are elucidated by EIS test, which showed that there was the decrease of charge transfer resistance and faster lithium-ion diffusion in LiMnPO_4/C cathode materials after Zn doping.

Construction, Investigation and Application of TEV Protease Variants with Improved Oxidative Stability

( Enkhtuya Bayar ),( Yuanyuan Ren ),( Yinghua Chen ),( Yafang Hu ),( Shuncheng Zhang ),( Xuelian Yu ),( Jun Fan ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.12

Tobacco etch virus protease (TEVp) is a useful tool for removing fusion tags, but wild-type TEVp is less stable under oxidized redox state. In this work, we introduced and combined C19S, C110S and C130S into TEVp variants containing T17S, L56V, N68D, I77V and S135G to improve protein solubility, and S219V to inhibit self-proteolysis. The solubility and cleavage activity of the constructed variants in Escherichia coli strains including BL21(DE3), BL21(DE3)pLys, Rossetta(DE3) and Origami(DE3) under the same induction conditions were analyzed and compared. The desirable soluble amounts, activity, and oxidative stability were identified to be reluctantly favored in the TEVp. Unlike C19S, C110S and C130S hardly impacted on decreasing protein solubility in the BL21(DE3), but they contributed to improved tolerance to the oxidative redox state in vivo and in vitro. After two fusion proteins were cleaved by purified TEVp protein containing double mutations under the oxidized redox state, the refolded disulfide-rich bovine enterokinase catalytic domain or maize peroxidase with enhanced yields were released from the regenerated amorphous cellulose via affinity absorption of the cellulose-binding module as the affinity tag.

Prediction of Cellulose Crystallinity in Liquid Phase Using CBM-GFP Probe

Xiaoyu Guo,Fan Yang,Huixue Liu,Yingmin Hou,Yafang Wang,Jie Sun,Xiaoyi Chen,Yanan Liu,Xianzhen Li 한국고분자학회 2019 Macromolecular Research Vol.27 No.4

Carbohydrate-binding modules (CBMs) have been developed to investigate the presence of crystalline and amorphous regions of cellulose. However, systematic and quantitative assessment of cellulose crystallinity using such non-hydrolytic fusion proteins in liquid phase has not been reported. In this work, cellulose directed CBM probes containing a green fluorescent protein (GFP) were constructed and named CG17, CG28, and CG2a. The probe binding condition was determined as incubating 30 μg/mL probes in 10 mM phosphate buffer at 30oC for 60 min. Under the optimized condition, the linear correlations between CBM probe binding capability and X-ray diffraction (XRD) crystallinity were well established. Using linear regression equations, the crystallinity of several cellulosic materials was well calculated. Amorphous component and cellulosic surface area probably had a less effect on binding capability of CG2a than that of CG17 and CG28. Therefore, crystalline-region specific probe CG2a should be an efficient tool for interpreting the crystallinity of cellulosic materials.

Backbone Cyclization of Flavin Mononucleotide-Based Fluorescent Protein Increases Fluorescence and Stability

Lin Tingting,Ge Yuanyuan,Gao Qing,Zhang Di,Chen Xiaofeng,Hu Yafang,Fan Jun 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.12

Flavin mononucleotide-binding proteins or domains emit cyan-green fluorescence under aerobic and anaerobic conditions, but relatively low fluorescence and less thermostability limit their application as reporters. In this work, we incorporated the codon-optimized fluorescent protein from Chlamydomonas reinhardtii with two different linkers independently into the redox-responsive split intein construct, overexpressed the precursors in hyperoxic Escherichia coli SHuffle T7 strain, and cyclized the target proteins in vitro in the presence of the reducing agent. Compared with the purified linear protein, the cyclic protein with the short linker displayed enhanced fluorescence. In contrast, cyclized protein with incorporation of the long linker including the myc-tag and human rhinovirus 3C protease cleavable sequence emitted slightly increased fluorescence compared with the protein linearized with the protease cleavage. The cyclic protein with the short linker also exhibited increased thermal stability and exopeptidase resistance. Moreover, induction of the target proteins in an oxygen-deficient culture rendered fluorescent E. coli BL21 (DE3) cells brighter than those overexpressing the linear construct. Thus, the cyclic reporter can hopefully be used in certain thermophilic anaerobes.

A FKBP5 mutation is associated with Paget’s disease of bone and enhances osteoclastogenesis

Bingru Lu,Yulian Jiao,Yinchang Wang,Jing Dong,Muyun Wei,Bin Cui,Yafang Sun,Laicheng Wang,Bingchang Zhang,Zijiang Chen,Yueran Zhao 생화학분자생물학회 2017 Experimental and molecular medicine Vol.49 No.-

Paget’s disease of bone (PDB) is a common metabolic bone disease that is characterized by aberrant focal bone remodeling, which is caused by excessive osteoclastic bone resorption followed by disorganized osteoblastic bone formation. Genetic factors are a critical determinant of PDB pathogenesis, and several susceptibility genes and loci have been reported, including SQSTM1, TNFSF11A, TNFRSF11B, VCP, OPTN, CSF1 and DCSTAMP. Herein, we report a case of Chinese familial PDB without mutations in known genes and identify a novel c.163G4C (p.Val55Leu) mutation in FKBP5 (encodes FK506-binding protein 51, FKBP51) associated with PDB using whole-exome sequencing. Mutant FKBP51 enhanced the Akt phosphorylation and kinase activity in cells. A study of osteoclast function using FKBP51V55L KI transgenic mice proved that osteoclast precursors from FKBP51V55L mice were hyperresponsive to RANKL, and osteoclasts derived from FKBP51V55L mice displayed more intensive bone resorbing activity than did FKBP51WT controls. The osteoclast-specific molecules tartrate-resistant acid phosphatase, osteoclast-associated receptor and transcription factor NFATC1 were increased in bone marrow-derived monocyte/ macrophage cells (BMMs) from FKBP51V55L mice during osteoclast differentiation. However, c-fos expression showed no significant difference in the wild-type and mutant groups. Akt phosphorylation in FKBP51V55L BMMs was elevated in response to RANKL. In contrast, IκB degradation, ERK phosphorylation and LC3II expression showed no difference in wild-type and mutant BMMs. Micro-CT analysis revealed an intensive trabecular bone resorption pattern in FKBP51V55L mice, and suspicious osteolytic bone lesions were noted in three-dimensional reconstruction of distal femurs from mutant mice. These results demonstrate that the mutant FKBP51V55L promotes osteoclastogenesis and function, which could subsequently participate in PDB development.