1.Differentiation of Pc and P2 variants in class 1 integron by high-resolution melting analysis
Quhao WEI ; Gang LI ; Xiaofei JIANG ; Qingfeng HU ; Huoyang LYU ; Yonglie ZHOU ; Ming GUAN ; Yuan LYU
Chinese Journal of Laboratory Medicine 2017;40(2):95-100
Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .
2.Changes in intracellular Ca2 + in the ureter smooth muscle cells of rats with neuropathic urinary tract dysfunction and its significance
Qingwei WANG ; Deshang TAO ; Xiaofei WANG ; Yan WANG ; Xinjian LIU ; Yutao LYU ; Jianguo WEN
Chinese Journal of Applied Clinical Pediatrics 2016;31(12):943-946
Objective To investigate the changes in intracellular Ca2+ in the ureter smooth muscle cells (USMC) of rats with neuropathic urinary tract dysfunction (NUTD) and their significance.Methods Forty-five rats were randomly and averagely divided into NUTD group,experimental control (EC) group and blank control (BC) group.The NUTD group was operated with a spinal cord transection at the first lumbar level and the sacral cord was destroyed;in EC group the spinal process was partly bitten at the same position,but the spinal cord was not transected;BC group was given no operations.One week later,the video-urodynamic was performed to observe the acontractile detrusor (ACD),vesicoureteral reflux (VUR) and urinary tract dysfunction in rats among the NUTD group,EC group and the BC group.Video-urodynamic assessment was performed at the sixth week after operation.Ureter smooth muscle cells (USMC) were obtained by collagenase digestion.Intracellular Ca2 + in the USMC were observed by laser scanning confocal microscope.Then the effects of Bay K8644(10-8 mol/L,10-7 mol/L,10-6 mol/L) on cytosolic Ca2+ concentrations([Ca2+] i) in NUTD group were studied by calculating the fluorescence intensity.Results ACD and no detrusor overactivity were found in all rats in NUTD group and without vesicoureteral reflux.Immunofluorescence method confirmed that the cells were USMC.Compared with BC group (31.44 ± 2.82) and EC group (32.06 ± 3.67),the fluorescence intensity (FI) of intracellular Ca2 + in USMC was much lower in the NUTD group (9.80 ± 1.11),and there was significant difference(P < 0.05).Bay K8644 (10-8 mol/L,10-7 mol/L,10-6 mol/L) increased the FI of [Ca2 +] i in a concentration-dependent manner,which were 3.80 ± 1.30,10.04 ± 2.15,19.89 ± 2.06,respectively,and there was significant difference (P < 0.05).Conclusions The decrease in Ca2 + concentration in the ureter smooth muscle cells may be one of the important factors for the primary ureteral dysfunction of NUTD.And calcium channel agonist can be meaningful for adjusting abnormal Ca2+ concentration in USMC of NUTD.
3.Establishment and practice of SCI papers management system
Yuanyuan KONG ; Yan CUI ; Jingping SU ; Xiaofei LYU ; Wei WEI ; Yun ZHANG ; Hufeng XU ; Hong YOU ; Youqing XIN
Chinese Journal of Hospital Administration 2015;(7):554-555
The quality of SCI papers is one of the objective indexes of evaluation on scientific and technological strength and research capabilities.This paper introduced a comprehensive management strategy to promote the publication of SCI papers with high impact factors,in terms of such dimensions ass research orientation,financial and technical support,personnel training,and scientific research management platform.The short and long term effects of the comprehensive management strategy system were analyzed using the SCI papers publication data and IF data from 201 1 to 2014 at the hospital,as a reference for building a scientific management system of SCI papers for the administrators.
4. Atypical epithelioid trophoblastic lesions after a cesarean section with cyst and fistula formation:a clinicopathological analysis of 4 cases
Feng ZHOU ; Haiyan SHI ; Yun LIANG ; Bingjian LYU ; Xiaofei ZHANG ; Xiaoduan CHEN
Chinese Journal of Pathology 2019;48(2):112-115
Objective:
To elucidate the clinicopathologic characteristics of atypical epithelioid trophoblastic lesions with cyst and fistula formation after cesarean section.
Methods:
The clinical and pathological data of 4 cases of post-cesarean atypical epithelioid trophoblastic lesions with cyst and fistula formation diagnosed at Women′s Hospital, School of Medicine, Zhejiang University during April 2007 to June 2018 were evaluated by hematoxylin and eosin stain and EnVision two-step immunohistochemical staining technique.
Results:
The age of the 4 patients ranged from 32 to 41 years, with a mean age of 36.5 years. Three patients recieved cystectomy and one underwent subtotal hysterectomy. Histologically, the lesions were well circumscribed and consisted of uniform cells of medium size, irregularly enlarged with hyperchromatic nuclei and 1 to 2 inconspicuous nucleoli embedded in abundant hyalinized matrix with fibrinoid material in the center. The cells exhibited immunohistochemical feature of chorionic-type intermediate trophoblastic cells (CK18+, p63+ and CD146-). All patients were alive without recurrence during follow-up of 1 to 40 months (mean
5.Application of whole-body magnetic resonance diffusion weighted imaging in evaluating the chemotherapy response for lung cancer
Yanming ZHANG ; Xiaobo LYU ; Xuehong ZHAO ; Dongfeng ZHANG ; Xin ZHANG ; Juan KE ; Wei WANG ; Min HUANG ; Xiaofei LI ; Hongjie HE ; Xiaojing ZHANG ; Yufeng ZHAI
Chinese Journal of Primary Medicine and Pharmacy 2019;26(1):6-9
Objective To study the clinical value of whole -body magnetic resonance diffusion weighted imaging(WB-DWI) in evaluating the chemotherapy response for lung cancer,thus to provide evidence for optimizing clinical imaging examination. Methods From October 2017 to May 2018,60 patients with lung cancer confirmed by histopathology in Linfen Central Hospital were selected. The patients underwent DWI examinations before chemotherapy and after two cycles of chemotherapy. The change of tumor size,distant metastasis and apparent diffusion coefficient (ADC) value were compared before and after chemotherapy. The correlation between the change rate of ADC value and the shrinkage rate of tumor size in the effective group was analyzed. Results Of 60 cases,1 case had new cerebral metastases after chemotherapy. There were statistically significant differences in ADC value [(1. 12 ± 0.33) ×10 -3mm2/svs.(1.56±0.40) ×10 -3mm2/s]andtumorsize[(4.63±2.75)cmvs.(2.28±1.45)cm] between before and after chemotherapy in the effective group(t= -3. 954,4. 711,all P<0. 01). There was correlation between the change of ADC value and tumor size(r=0. 34,P<0. 05). Conclusion WB-DWI can not only detect the change of tumor size and distant metastasis quickly and effectively,but also can observe the microscopic changes of tumor cells by measuring ADC value. So it can predict the early therapeutic response of the tumor and make effective evaluation for the staging and chemotherapy response of lung cancer.
6.Status on sexual partners among men who have sex with men among college students in Qingdao,2016
Xiaofei ZHANG ; Cuixia LYU ; Jietao ZHENG ; Lei DONG ; Zhenxia JIANG ; Jihua FU
Chinese Journal of Epidemiology 2017;38(12):1638-1641
Objective To investigate the situation of sexual partners and related factors among men who have sex with men (MSM) in college students.Methods Snowball sampling and Convenience sampling were both used to recruit MSM from colleges in Tsingtao in 2016.Questionnaire-based interviews were conducted to collect data of socio-demographic and situation of sexual partners.Sample Size was estimated based on cross-sectional study,and theoretical 267.SPSS 17.0 software was used for statistical analysis.Results A total of 300 MSM,average aged 20.7,were analyzed.Both first sex partner and the last same-sex sexual partner were mct instantly,with proportions as 58.7% (176/300) and 62.3% (187/300) respectively.Among all the MSM,88.3% (265/300) preferred selecting men as sex partners and 42.7% (128/300) enjoyed finding sex partners in college,while 86.0% (258/300) preferred finding their sex partners through intemet.Conclusions Intemet had been the major way of looking for sex partner among MSM in college students,the male sexual partner were met instantly.We should focus on men who have sex with men and their sexual health among college students to prevent and control HIV/AIDS.
7.Mechanism of action of Coptis chinensis in the treatment of dental caries based on network pharmacology and experimental validation
Xiaofei LYU ; Zhenhui LIU ; Nan JIANG ; Xiaotong CUI ; Cheng PENG
International Journal of Biomedical Engineering 2023;46(4):321-328
Objective:To explore the mechanism of action of Coptis chinensis in the treatment of dental caries using a network pharmacology approach and animal experiments. Methods:The active ingredients of C. chinensis and their targets were screened by the traditional Chinese medicine systems pharmacology (TCMSP) database and analysis platform, and the targets were searched online through the GeneCards database. The intersecting targets of C. chinensis and dental caries were screened at Venny 2.1, and the intersection targets were analyzed online for protein-protein interaction analysis and gene ontology (GO) and kyoto encyclopedia of genes and genomics (KEGG) enrichment. Then, Cytoscape was used to create a "component-target-pathway" network diagram. Rats were randomly divided into the model group and the C. chinensis group to establish a rat model of dental caries. Rats in the model group were repeatedly rubbed with a cotton ball soaked in 150 μl of 0.9% NaCl solution for 5 min, and rats in the C. chinensis group were repeatedly rubbed with a cotton ball soaked in C. chinensis (5.8 mg of C. chinensis in 150 μl of 0.9% NaCl solution) for 5 min. The two groups of rats were treated once a week for four consecutive weeks. The number of Streptococcus mutans colonies was counted, and serum serine/threonine protein kinase 1 (AKT1), JUN, interleukin-6 (IL-6), tumor necrosis factor (TNF), and B-cell lymphoma-2 (Bcl-2) were detected by enzyme immunoassay. Results:A total of 11 active ingredients in C. chinensis were found, which regulate multiple molecular pathways by intervening in 54 targets, thereby treating dental caries. Quercetin, berberine, flavodoxin, berberine infusion, and tetrahydroberberine were the core components, and AKT1, JUN, IL-6, TNF, and Bcl-2 were the core targets. GO analysis showed that BP mainly included cytokine activity, signaling receptor activator activity, signaling receptor modulator activity, cytokine receptor binding, and receptor ligand activity, etc.; and CC mainly included the response to lipopolysaccharides, the response to bacterial molecules, cellular responses to lipids, inflammatory responses, and negative regulation of cell population proliferation; MF mainly includes membrane rafts, membrane microregions, extracellular matrix, external encapsulated structures, and plasma membrane protein complexes, etc. KEGG analysis showed that advanced glycosylation end product-receptor for advanced glycosylation end products (AGE-RAGE), TNF, IL-17, Toll-like receptor, hypoxia-inducible factor-1 (HIF-1), mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB), epidermal growth factor receptor (EGFR), Janus kinase-signal transducer and activator of transcription (JAK-STAT), and phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathways have been associated with C. chinensis treatment. The results of animal experiments showed that serum Bcl-2 protein expression increased and serum AKT1, JUN, IL-6, TNF, and other proteins decreased after the C. chinensis treatment. Conclusions:C. chinensis can be involved in regulating the targets of dental caries through multiple pathways, with good therapeutic effects and a wide range of mechanisms of action, and is expected to be an important component in the development of proprietary Chinese medicines for the treatment of dental caries.
8.A rare Pk phenotype caused by a 433 C>T mutation of the β-1,3-N-acetylgalactosyltransferase gene.
Xiaofei LAN ; Xiaozhen HONG ; Xianguo XU ; Shu CHEN ; Kairong MA ; Ying LIU ; Ji HE ; Faming ZHU ; Hangjun LYU
Chinese Journal of Medical Genetics 2015;32(3):381-384
OBJECTIVETo study the serological characteristics and molecular mechanism for a rare Pk phenotype of the P1Pk blood group system.
METHODSThe blood group of the proband was identified by serological techniques. The coding region and flanking intronic sequences of the β-1,3-N-acetylgalactosyltransferase gene (B3GALANT1) associated with the Pk phenotype were analyzed using polymerase chain reaction sequence-based typing.
RESULTSThe proband was identified as having a rare Pk phenotype including anti-P in her serum. The blood group of her daughter and husband showed a P2 phenotype. The nucleotide sequences of the B3GALANT1 gene of her husband and two randomly-chosen individuals were the same as the reference sequence (GenBank AB050855). Nucleotide position 433 C>T homozygous mutation in the B3GALANT1 was found in the proband, which has resulted in a stop codon at amino acid position 145, which may produce a premature protein capable of decreasing or inhibiting the activity of the β -1,3-N-acetylgalactosyltransferase. The nucleotide position 433 C/T heterozygous in the B3GALANT1 was found in her daughter.
CONCLUSIONThe Pk phenotype resulted from 433 C>T mutation in the B3GALANT1 gene has been identified.
ABO Blood-Group System ; genetics ; Adult ; Base Sequence ; Blood Grouping and Crossmatching ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; N-Acetylgalactosaminyltransferases ; genetics ; Pedigree ; Phenotype ; Point Mutation
9.Molecular basis for an individual with rare p phenotype in P1Pk blood group system.
Kairong MA ; Xiaofei LAN ; Xianguo XU ; Xiaozhen HONG ; Shu CHEN ; Ying LIU ; Yanling YING ; Ji HE ; Faming ZHU ; Hangjun LYU
Chinese Journal of Medical Genetics 2015;32(2):250-253
OBJECTIVETo explore the molecular basis for an individual with rare p phenotype in the P1Pk blood group system.
METHODSErythrocyte blood group antigens and antibodies in serum were identified in the proband and five family members with a serological method. Coding regions and flanking untranslated regions of the α1,4-galactosyltransferase gene (A4GALT) encoding P1Pk antigens were amplified with polymerase chain reaction and directly sequenced. The haplotypes of A4GALT in the parents of the proband were also analyzed by cloning sequencing.
RESULTSThe proband was found with a rare p phenotype with anti-Tja antibody in his serum by serological method. The other family members all had a common P2 phenotype. The results of DNA sequencing showed that a cytosine was inserted at nucleotide position 1026 to 1029 (1026_1029insC) of both alleles of the A4GALT gene in the proband. The mutation has caused a reading frame shift and formed a mutant protein by extending 92 amino acid residues. The other family members were either heterozygous for the insertion or of the wild type at above position.
CONCLUSIONThe 1026_1029insC mutation of the A4GALT gene is probably responsible for the p phenotype identified for the first time in Chinese population. The individual with the p phenotype possesses anti-Tja antibody.
ABO Blood-Group System ; genetics ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Frameshift Mutation ; Galactosyltransferases ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutagenesis, Insertional ; Pedigree ; Phenotype ; Young Adult
10.Association between -1296T/C and -915A/G polymorphisms of matrix metalloproteinase inhibitor-3 gene and atherosclerotic cerebral infarction in an ethnic Han Chinese population.
Zhou ZHENG ; Xiaoping JIN ; Min ZHU ; Cai LI ; Feng ZHU ; Weiling LI ; Qing LIN ; Feng WANG ; Wanfen WANG ; Xiaofei HU ; Chenling LYU
Chinese Journal of Medical Genetics 2014;31(6):719-724
OBJECTIVETo assess the association between -1296T/C and -915A/G polymorphisms in the promoter region of matrix metalloproteinase inhibitor-3 gene (TIMP-3) and atherosclerotic cerebral infarction in an ethnic Han Chinese population.
METHODSPeripheral blood samples were collected from 485 patients with atherosclerotic cerebral infarction and 525 healthy controls. Serum levels of TIMP-3 were measured with an enzyme-linked immunosorbent assay (ELISA). The polymorphisms of the TIMP-3 gene were analyzed with DNA sequencing.
RESULTSThere were significant differences in genotype and allele frequencies in -1296T/C and -915A/G between the patients and healthy controls (chi-square: 5.227 and 5.869; P: 0.022 and 0.015, respectively). Besides, there was a strong linkage disequilibrium between -1296T/C and -915A/G (D'=1.0, r(2)=0.991). The serum levels of TIMP-3 in patients were significantly higher than the control group [(248.90 ± 97.10) pg/mL vs. (200.17 ± 79.70) pg/mL, t=2.098, P=0.039].
CONCLUSIONThe -1296T/C and -915A/G polymorphisms of the TIMP-3 gene are associated with increased risk for atherosclerotic cerebral infarction in ethnic Han Chinese and may be used as molecular markers for the disease. There is also strong linkage disequilibrium between the two loci.
Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; ethnology ; genetics ; Atherosclerosis ; blood ; epidemiology ; ethnology ; genetics ; Base Sequence ; Cerebral Infarction ; blood ; epidemiology ; ethnology ; genetics ; China ; epidemiology ; Female ; Gene Frequency ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Risk Factors ; Tissue Inhibitor of Metalloproteinase-3 ; blood ; genetics