1.Bone remodeling in the Masquelet-induced membrane model of rat femur by modulation of H-type vessels by total flavonoids of rhizome drynariae
Zhikui ZENG ; Wei XIONG ; Weidong LIANG ; Guowen QIAN ; Chaoyi LIANG ; Bin PAN ; Ling GUO ; Wenqiang WEI ; Xunxiang QIU ; Wenfang DENG ; Lingmei YUAN
Chinese Journal of Tissue Engineering Research 2024;28(32):5130-5135
BACKGROUND:Several studies have found that the total flavonoids of rhizome drynariae can promote neovascularization in the induced membrane,improve the biological properties of the induced membrane,and accelerate bone remodeling in the induced membrane,but the related molecular mechanisms still need to be further explored. OBJECTIVE:To explore the effect of total flavonoids of rhizome drynariae on bone remodeling in rat femoral Masquelet-induced membrane model by regulating H-type blood vessels. METHODS:Thirty-six male Sprague-Dawley rats were stratified by body mass and then randomly divided into blank group,model group and traditional Chinese medicine group,with 12 rats in each group.A 4-mm femoral bone defect model was established in all the rats.Bone defects in the model group and traditional Chinese medicine group were filled with polymethylmethacrylate bone cement.At 6 weeks after modeling,the tail bone of the rats was implanted in the blank group,as well as in the other two groups after removal of bone cement.The traditional Chinese medicine group was given 157.5 mg/kg per day of total flavonoids of rhizome drynariae at 3 days after bone implantation,while the model and blank groups were given the same amount of saline by gavage until the 8th week after bone implantation.Bone graft samples were taken for relevant testing at 8 weeks after implantation. RESULTS AND CONCLUSION:X-ray films showed that in the blank group,the fracture line in the defect area was clear,and only a small amount of bone callus formed;in the model group,the bone defect area still existed,where discontinuous cortical bone was visible;in the traditional Chinese medicine group,the defect area was filled with newborn bone tissues,the bone marrow cavity and part of the cortical bone formed,and the fracture line disappeared.Micro-CT scans showed that the amount of new bone in the defect area was low in the blank group,the number of bone trabeculae in the defect area was significantly increased in the model group,and a large amount of new bone tissue was filled in the bone defect area in the traditional Chinese medicine group.Hematoxylin-eosin staining results showed that in the blank group,only a small amount of new bone formed in the defect area and the quality of osteogenesis was poor;in the model group,there was more new bone tissue in the defect area,but some fibrous connective tissues were interspersed within the bone tissue;and in the traditional Chinese medicine group,a large amount of new bone formed in the defect area and the quality of osteogenesis was the best.CD31/Emcn immunofluorescence double-labeling staining results showed that the number of H-type blood vessels in the newborn bone tissue in the bone defect area of the blank group was sparse and sparsely distributed;compared with the blank group,there were more H-type blood vessels in the bone tissue in the bone defect area of the model group,and the blood vessels were distributed in relatively regular strips;the number of H-type blood vessels in the bone defect area of the traditional Chinese medicine group was the highest and the blood vessels were densely distributed.To conclude,the total flavonoids of rhizoma drynariae can upregulate the expression of H-type blood vessels to enhance the angiogenic-osteogenic effect,improve the osteogenic efficiency of the rat femoral Masquelet induced membrane model,and promote bone remodeling.
2.Preparation of berberine-naringin dual drug-loaded composite microspheres and evaluation of their antibacterial-osteogenic properties.
Wei XIONG ; Lingmei YUAN ; Liangxia WANG ; Guowen QIAN ; Chaoyi LIANG ; Bin PAN ; Ling GUO ; Wenqiang WEI ; Xunxiang QIU ; Wenfang DENG ; Zhikui ZENG
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(12):1505-1513
OBJECTIVE:
To develop a drug-loaded composite microsphere that can simultaneously release the berberine (BBR) and naringin (NG) to repair infectious bone defects.
METHODS:
The NG was loaded on mesoporous microspheres (MBG) to obtain the drug-loaded microspheres (NG-MBG). Then the dual drug-loaded compound microspheres (NG-MBG@PDA-BBR) were obtained by wrapping NG-MBG with polydopamine (PDA) and modifying the coated PDA with BBR. The composite microspheres were characterized by scanning electron microscopy, X-ray diffraction, specific surface area and pore volume analyzer, and Fourier transform infrared spectroscopy; the drug loading rate and release of NG and BBR were measured; the colony number was counted and the bacterial inhibition rate was calculated after co-culture with Staphylococcus aureus and Escherichia coli for 12 hours to observe the antibacterial effect; the biocompatibility was evaluated by live/dead cell fluorescence staining and cell counting kit 8 assay after co-culture with rat's BMSCs for 24 and 72 hours, respectively, and the osteogenic property was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining after 7 and 14 days, respectively.
RESULTS:
NG-MBG@PDA-BBR and three control microspheres (MBG, MBG@PDA, and NG-MBG@PDA) were successfully constructed. Scanning electron microscopy showed that NG-MBG@PDA-BBR had a rough lamellar structure, while MBG had a smooth surface, and MBG@PDA and NG-MBG@PDA had a wrapped agglomeration structure. Specific surface area analysis showed that MBG had a mesoporous structure and had drug-loading potential. Low angle X-ray diffraction showed that NG was successfully loaded on MBG. The X-ray diffraction pattern contrast showed that all groups of microspheres were amorphous. Fourier transform infrared spectroscopy showed that NG and BBR peaks existed in NG-MBG@PDA-BBR. NG-MBG@PDA-BBR had good sustained drug release ability, and NG and BBR had early burst release and late sustained release. NG-MBG@PDA-BBR could inhibit the growth of Staphylococcus aureus and Escherichia coli, and the antibacterial ability was significantly higher than that of MBG, MBG@PDA, and NG-MBG@PDA ( P<0.05). But there was a significant difference in biocompatibility at 72 hours among microspheres ( P<0.05). ALP and alizarin red staining showed that the ALP positive area and the number of calcium nodules in NG-MBG@PDA-BBR were significantly higher than those of MBG and NG-MBG ( P<0.05), and there was no significant difference between NG-MBG@PDA and NG-MBG@PDA ( P>0.05).
CONCLUSION
NG-MBG@PDA-BBR have sustained release effects on NG and BBR, indicating that it has ideal dual performance of osteogenesis and antibacterial property.
Rats
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Animals
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Osteogenesis
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Delayed-Action Preparations/pharmacology*
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Microspheres
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Berberine/pharmacology*
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Anti-Bacterial Agents/pharmacology*
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Escherichia coli
3.Effects of lifestyle changes on body weight and metabolic parameters during the early stage of COVID-19 epidemic and regular epidemic prevention period in physical examination population
Xin ZHAO ; Wen GUO ; Pei QIN ; Xiuru LIANG ; Wenfang ZHU ; Xiaona LI ; Qingqing DIAO ; Nianzhen XU ; Qun ZHANG
Chinese Journal of Health Management 2022;16(11):751-757
Objective:To analyze the influence of lifestyle changes on body weight and metabolic parameters during the early stage of corona virus disease 2019 (COVID-19) epidemic and regular epidemic prevention period in physical examination population.Methods:A total of 801 subjects from Nanjing enterprises and institutions who underwent physical examination in the Department of Health Promotion Center, the First Affiliated Hospital of Nanjing Medical University in May of 2019 to 2021 were included in this study. The basic information and data of body mass index, fasting blood glucose, total cholesterol, low density lipoprotein cholesterol (LDL-C), triglyceride and uric acid were collected. Information about dietary and exercise habits was obtained through pre-examination questionnaires. The data of 2019 was set as baseline data, data of 2020 represented information during the early stage of COVID-19 epidemic and data of 2021 represented information in regular epidemic prevention period. The subjects were divided into underweight group, normal weight group, overweight group, and obese group according to the body mass index at baseline. The Friedman test was applied to compare changes in body mass index and metabolic parameters across the population. Chi-square test was used to compare changes in dietary and exercise habits. Multivariate logistic regression analysis was adopted to explore the influencing factors of body mass index changes.Results:During the early stage of COVID-19 epidemic, the proportion of weight gain was the highest in the underweight group (42.9%), the proportion of weight loss was the highest in the overweight group (24.2%), and the obese group has the most stable body weight (70.6%) ( P=0.004). Men ( OR=0.56, 95% CI: 0.36-0.87) were less likely to gain weight than women ( P=0.010). There was no significant differences in weight change among all groups in the regular epidemic prevention period ( P=0.380). During the early stage of COVID-19 epidemic, the levels of fasting blood glucose, total cholesterol and LDL-C were significantly lower than those of baseline [4.98 (4.66, 5.42) vs 5.23 (4.91, 5.66) mmol/L, 4.98 (4.36, 5.67) vs 5.11 (4.54, 5.77) mmol/L, 2.90 (2.45, 3.33) vs 3.23 (2.77, 3.74) mmol/L], and the uric acid level was higher [333.0 (275.5, 397.0) vs 311.0 (257.5, 368.0) μmol/L] (all P<0.001). In regular epidemic prevention period, the levels of body mass index, fasting blood glucose, total cholesterol and LDL-C were significantly higher than those in the early stage of the epidemic [24.0 (21.9, 26.3) vs 23.8 (21.7, 26.1) kg/m 2, 5.18 (4.85, 5.62) vs 4.98 (4.66, 5.42) mmol/L, 5.12 (4.42, 5.76) vs 4.98 (4.36, 5.67) mmol/L, 3.06 (2.59, 3.57) vs 2.90 (2.45, 3.33) mmol/L], while the uric acid was significantly lower [319.0 (265.0, 377.0) vs 333.0 (275.5, 397.0) μmol/L] (all P<0.001). During the early stage of the epidemic, the reduction proportion of unhealthy diet in the home group was significantly higher than that in the outing group (19.5% vs 11.4%), and the increment proportion of exercise in the outing group was significantly higher than that in the home group (5.1% vs 1.6%) (both P<0.05). In regular epidemic prevention period, the increase rate of unhealthy diet in the home group was significantly higher than that in the outing group (26.8% vs 13.0%) ( P<0.001), and there was no significant difference in exercise between the two groups ( P=0.325). During the early stage of COVID-19 epidemic and in the regular epidemic prevention period, unhealthy diet>3 times per week ( OR=3.85, 3.01, 95% CI: 1.74-8.51, 1.41-6.39) was positively correlated with weight gain, and regular exercise ( OR=4.35, 2.61, 95% CI: 2.05-9.23, 1.15-5.91) was positively correlated with weight loss (all P<0.05). Conclusions:During the early stage of COVID-19 epidemic and in the regular epidemic prevention period, the lifestyle in the physical examination population has an impact on body weight and metabolic indicators. In the early stage of the epidemic, unhealthy diet and exercise decreased, and metabolic indicators such as blood glucose and lipids decreased. People with low body weight tend to gain weight. In the regular epidemic prevention period, the subjects′ exercise increased but unhealthy diet also increased, and blood glucose, lipid and body weight elevated significantly.
4.Multi-omics Analyses Provide Insight into the Biosynthesis Pathways of Fucoxanthin in Isochrysis galbana
Chen DUO ; Yuan XUE ; Zheng XUEHAI ; Fang JINGPING ; Lin GANG ; Li RONGMAO ; Chen JIANNAN ; He WENJIN ; Huang ZHEN ; Fan WENFANG ; Liang LIMIN ; Lin CHENTAO ; Zhu JINMAO ; Chen YOUQIANG ; Xue TING
Genomics, Proteomics & Bioinformatics 2022;(6):1138-1153
Isochrysis galbana is considered an ideal bait for functional foods and nutraceuticals of humans because of its high fucoxanthin(Fx)content.However,multi-omics analysis of the regula-tory networks for Fx biosynthesis in I.galbana has not been reported.In this study,we report a high-quality genome assembly of I.galbana LG007,which has a genome size of 92.73 Mb,with a contig N50 of 6.99 Mb and 14,900 protein-coding genes.Phylogenetic analysis confirmed the monophyly of Haptophyta,with I.galbana sister to Emiliania huxleyi and Chrysochromulina tobinii.Evolutionary analysis revealed an estimated divergence time between I.galbana and E.huxleyi of~133 million years ago.Gene family analysis indicated that lipid metabolism-related genes exhibited significant expansion,including IgPLMT,IgOAR1,and IgDEGS1.Metabolome analysis showed that the content of carotenoids in I.galbana cultured under green light for 7 days was higher than that under white light,and β-carotene was the main carotenoid,accounting for 79.09%of the total carotenoids.Comprehensive multi-omics analysis revealed that the content of β-carotene,antheraxanthin,zeaxanthin,and Fx was increased by green light induction,which was significantly correlated with the expression of IgMYB98,IgZDS,IgPDS,IgLHCX2,IgZEP,IgLCYb,and IgNSY.These findings contribute to the understanding of Fx biosynthesis and its regulation,pro-viding a valuable reference for food and pharmaceutical applications.
5.Study on Chemical Constituents of Ethanol Extract from Yao Medicine Cissampelopsis spelaeicola
Shuang LIANG ; Wenfang MA ; Kewen LIANG ; Lichun ZHAO ; Gang DENG ; Dongyang XIE
China Pharmacy 2019;30(20):2835-2839
OBJECTIVE: To study the chemical constituents of ethanol extract from Yao medicine Cissampelopsis spelaeicola. METHODS: The petroleum ether, ethyl acetate and n-butanol fraction from 75% ethanol extract of C. spelaeicola were isolated and purified by silica gel, SephadexLH-20 gel column and AB-8 macroporous resin column, etc. The structures of the compounds were analyzed and identified by physicochemical properties and spectral data (mass spectrometry, hydrogen spectrum, carbon spectrum). RESULTS & CONCLUSIONS: Twelve compounds were isolated and identified from 75% ethanol extract of C. spelaeicola. β-sitosterol(Ⅰ) and Stigmasterol(Ⅱ) were isolated from petroleum ether fraction; p-hydroxybenzoic acid(Ⅲ), β-daucossterol(Ⅳ), protocatechuic acid(Ⅴ), 6β-hydroxyeremophi-7(11)-en-12,8β-olide(Ⅵ), 10β-hydroxyeremophil-7(11)-en-8,12-olide(Ⅶ), 10β-hydroxyeremophi-7(11),8(9)-dien-8,12-olide(Ⅷ), Quercetin(Ⅸ), Hyperin(Ⅹ) and 4α-hydroxy- eudesman-11-ene(Ⅺ) were isolated from ethyl acetate fraction; quercetin-3-O-robinobioside(Ⅻ) was isolated from n-butanol fraction. Compounds Ⅰ-Ⅻ are isolated from C. spelaeicola for the first time. The study can lay material foundation for activity stady of C. spelaeicola.
6.Role of NO signal in ABA-induced phenolic acids accumulation in Salvia miltiorrhiza hairy roots.
Lihong SHEN ; Jiahui REN ; Wenfang JIN ; Ruijie WANG ; Chunhong NI ; Mengjiao TONG ; Zongsuo LIANG ; Dongfeng YANG
Chinese Journal of Biotechnology 2016;32(2):222-230
To investigate roles of nitric oxide (NO) signal in accumulations of phenolic acids in abscisic.acid (ABA)-induced Salvia miltiorrhiza hairy roots, S. miltiorrhiza hairy roots were treated with different concentrations of sodium nitroprusside (SNP)-an exogenous NO donor, for 6 days, and contents of phenolic acids in the hairy roots are determined. Then with treatment of ABA and NO scavenger (2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide, c-PTIO) or NO synthase inhibitor (NG-nitro-L-arginine methyl ester, L-NAME), contents of phenolic acids and expression levels of three key genes involved in phenolic acids biosynthesis were detected. Phenolic acids production in S. miltiorrhiza hairy roots was most significantly improved by 100 µmoL/L SNP. Contents of RA and salvianolic acid B increased by 3 and 4 folds. ABA significantly improved transcript levels of PAL (phenylalanine ammonia lyase), TAT (tyrosine aminotransferase) and RAS (rosmarinic acid synthase), and increased phenolic acids accumulations. However, with treatments of ABA+c-PTIO or ABA+L-NAME, accumulations of phenolic acids and expression levels of the three key genes were significantly inhibited. Both NO and ABA can increase accumulations of phenolic acids in S. miltiorrhiza hairy roots. NO signal probably mediates the ABA-induced phenolic acids production.
Abscisic Acid
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pharmacology
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Benzofurans
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metabolism
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Free Radical Scavengers
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pharmacology
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Hydroxybenzoates
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metabolism
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Nitric Oxide
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metabolism
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Phenylalanine Ammonia-Lyase
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metabolism
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Plant Roots
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metabolism
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Salvia miltiorrhiza
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metabolism
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Tyrosine Transaminase
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metabolism
7.Status of post-traumatic growth of severe burn patients and its influencing factors
Ying LIANG ; Wenfang XIAO ; Dandan JIAO
Modern Clinical Nursing 2016;15(1):58-61
Objective To understand the status of post-traumatic growth of severe burn patients and analyze its influencing factors. Methods General information questionnaire and post-traumatic growth inventory (PTGI) were used to investigate thirty patients with severe burn. Multi-linear regression method was used to analyze the post-traumatic influencing factors. Results The average score of PTGI was (68.30 ± 10.90), which was in media level. The main influencing factors included burn area, active exercise, time and expense of treatment, marital status and sear proliferation. Conclusion In order to improve the post-traumatic growth,nurses should enhance psychological intervention, teach effective coping strategies and increase the social level to the burn patients.
8.Diversity of SCCmec type diversity in clinically isolated methicillin-resistant Staphylococcus epidermidis
Yongchang YANG ; Liang CHEN ; Daiwen XIAO ; Hua YU ; Hua LIU ; Wenfang HUANG
International Journal of Laboratory Medicine 2016;37(16):2273-2274,2277
Objective To investigate the SCCmec types of clinically isolated methicillin‐resistant Staphylococcus epidermidis (M RSE) .Methods Eighty‐four strains of clinically isolated Staphylococcus epidermidis identified by the fully automatic microbio‐logical identification system were collected and performed the MRSE identification by PCR for amplifying esp and mecA genes and SCCmec typing .Its distribution characteristics were analyzed .Results Esp gene was amplified in 84 strains and the detection rate of mecA was 76 .19% (64/84) ,in which the MRSE detection rates in blood ,sputum ,urine and wound secretion were 76 .8% , 68 .8% ,100% and 71 .4% respectively .The multiple PCR amplification displayed that among 64 strains of MRSE ,19 strains were SCCmec simple type ,in which 19 strains were SCCmec type Ⅰ and 3 strains were SCCmec type Ⅲ ;42 strains were SCCmec mixed type ,in which 2 strains were SCCmec mixed type Ⅰ and Ⅱ ,14 strains were SCCmec mixed type Ⅰ and Ⅲ ,12 strains were SCCmec mixed type Ⅰ ,Ⅱ and Ⅲ ,5 strains were SCCmec mixed type Ⅱ and Ⅲ ,a strains were and SCCmec mixed type Ⅲ and Ⅳ .Conclu‐sion The SCCmec type in clinically isolated MRSE shows obvious diversity and its majority is SCCmec mixed type .
9.Clinical Characteristics and Current Treating Status in Atrial Fibrillation Patients With Different Gender From 20 Emergency Departments in China
Xinjie WANG ; Yan LIANG ; Jun ZHU ; Yanmin YANG ; Wenfang MA ; Mingming LIU
Chinese Circulation Journal 2015;(8):744-748
Objective: To analyze the clinical characteristics and current treating status from atrial ifbrillation (AF) patients with different gender in 20 emergency departments.
Methods: A total of 2015 consecutive AF patients from 20 emergency departments nationwide from 2008-11 to 2011-10 were retrospectively investigated. The patients were divided into 2 groups: Female group,n=1104 and Male group,n=911. The baseline clinical characteristics and current treating status were compared between groups, the risk of stroke in non-valvular atrial ifbrillation (NVAF) patients was evaluated by CHADS2 score and the factors affecting walfarin application were studied by Logistic regression analysis.
Results: Compared with Male group, Female group had the elder age (69.11 ± 12.96) years vs (67.67 ± 13.63) years,P=0.015, lower body mass index (BMI) (23.24 ± 3.73) kg/m2 vs (23.89 ± 3.47) kg/m2,P=0.000, more patients combining with heart failure (39.7% vs 34.6%,P=0.019), more patients with valvular heart disease (26.6% vs 12.4%,P=0.000). In contrast, Male group had more smokers (41.4% vs 5.1%,P=0.000), more patients combining with coronary artery disease (45.1% vs 39.1%,P=0.007) and more patients with previous history of myocardial infarction (9.5% vs 5.5%,P=0.001). The average CHADS2 score was higher in Female group than Male group (2.0 ± 1.4) vs (1.8 ± 1.4),P=0.008 and the proportion of patients with CHADS2 score ≥2 was higher in Female group than Male group (58.0% vs 51%,P=0.005). There were 407 patients of valvular heart disease with AF and 167 (41%) of them received walfarin treatment including 119 female and 48 male,P=0.713; 1608 NVAF patients with CHADS2 score≥ 2 and 105 of them received anticoagulation therapy including 54 female and 51 male,P=0.636.
Conclusion: The clinical characteristics and current treating status are different by gender from AF patients in 20 emergency departments in China.
10.Construction of mutant strains of methicillin resistant Staphylococcus epidermidis with psm-mec gene deletion
Yongchang YANG ; Honghua HU ; Liang CHEN ; Hua LIU ; Hua YU ; Wenfang HUANG
Chinese Journal of Microbiology and Immunology 2015;(9):672-677
Objective To construct mutant strains of methicillin resistant Staphylococcus epidermi-dis (MRSE) with psm-mec gene deletion and to investigate the function of psm-mec gene.Methods The drug sensitivity test and DNA sequence analysis were performed to screen out the tetracycline and chloram -phenicol sensitive clinical strains of MRSE , whose upstream and downstream sequences of psm-mec gene were identical to those of the Staphylococcus epidermidis reference strain RP62A.The recombinant plasmid pBT2-Δpsm-mec was constructed by using the fusion PCR and a temperature sensitive shuttle plasmid .After being identified , the plasmid was transformed into the Staphylococcus aureus RN4220 strain by electropora-tion, and then transformed into the selected clinical isolates of MRSE .The mutant strains of MRSE with psm-mec deletion were screened out and identified after homologous recombination .The differences in biofilm formation between the mutant and wild-type strains were analyzed for further elucidation the relationships be-tween the psm-mec gene and biofilm formation in MRSE strains .Results Three clinical MRSE isolates for the construction of mutant strains with psm-mec gene deletion were screened out and identified by using drug sensitivity test and sequence alignment analysis .The mutants constructed via homogenous recombination were screened out and identified .Compared with the corresponding wild-type strains, the three mutants with psm-mec gene deletion showed significantly decreased ability of biofilm formation , demonstrating that the psm-mec genes strains induced the biofilm formation of MRSE .Conclusion The Δpsm-mec mutant strains were successfully constructed .The psm-mec gene played an important role in the biofilm formation of Staphy-lococcus epidermdis.

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