| Objective: To analyze the distribution characteristics of conjunctival cyst flora in patients with chronic dacryocystitis who used antibiotics before and after DCR(Dacryocystorhinostomy)surgery and those who did not use antibiotics before and after surgery.And analyze the difference of the flora and the dominant flora among the three groups.Methods: Prospective control study.Part I: Healthy control group: A total of 26 cases of conjunctival cyst bacteria from orbital tumor patients,cataract patients and healthy volunteers without eye surface diseases in Tianjin Medical University Eye Hospital from November 2018 to August 2019 were collected.Groups older than 50 years old and groups younger than 50 years old were analyzed based on 16 Sr RNA highthroughput sequencing technology,and QIIME software,R software,Lef SE and other software were used for biological analysis of conjunctival cyst bacteria.Part II: The experimental group: Collecting dacryocystitis patients who underwent DCR surgery in the Department of Orbital Diseases and Ophthalmology,Tianjin Medical University Eye Hospital from November 2018 to August 2019,divided into the antibiotic group and the non-antibiotic group,according to the collection of dacryocystitis patients Age composition The first part of the age is greater than 50 years old as a healthy control group.Collect the conjunctival sac flora 1 day before surgery,1 week after surgery,2 weeks after surgery and 4 weeks after surgery,and analyze the flora using the same test method as above to compare the effect of antibiotics on the structure of conjunctival bursa.Result: I.Clinical data analysis According to the inclusion criteria,the healthy control group finally selected 26 eligible subjects,including 11 males and 15 females.A total of 14 cases in the group over 50 years old,6 males and 8 females,age(56-84 years old),average age(72.4 ± 9.2);a total of 12 cases in the group younger than 50 years old,5 males and 7 females,Age(24-49 years old),average age(35.8 ± 9.5).A total of 19 cases were collected in the experimental group,and 7 cases of antibiotics were used according to the experimentaldesign method(all antibiotics were levofloxacin eye drops).Among them,6 were female and 1 was male.Twelve patients did not use antibiotics before admission and after surgery,including 9 females and 3 males,aged(58-84 years),average age(70.2 ± 8.0).According to the enrollment criteria,15 subjects with nasal bacterial flora were selected,including 9 females and 6 males,aged(58-83),and average age(69.3 ± 7.26).II.Structure of conjunctival cysts in normal people 1.A total of 462,936 valid sequences were obtained from 26 samples.A total of 4521 type OUT were obtained at the door level under 97% sequence similarity.The average age of OTU is 2214 for those over 50 years old(C1),and the average number of 2307 OUT is under 50 years old(C2).It shows that the species number of C2 group is more than that of C1 group.2.The Chao1,Simpson and Shannon indexes of the C1 group were lower than those of the C2 group,and Chao1 and Simpson had statistical significance(P <0.05).3.PCA analysis showed that the difference between group C1 and group C2 was obvious.C1 aggregated well,group C2 was more dispersed and partially mixed with group C1.The results showed that the richness and diversity of bacteria in C2 group were greater than that in C1 group.4.The statistical analysis of the top 20 abundances of the genus level abundance.The core bacteria of the conjunctival sac of group C1 mainly include Cupriavidus(24.3%),Sphingobium(5.3%),and Thermus(2.3%),Acinetobacter(2.2%),Sphingobium(1.4%),Ochrobactrum(0.6%);the core flora of group C2 mainly includes: Thermus(15.3%),Cupriavidus(10.3%),Bifidobacterium(7.7%),Acinetobacter(5.6%),Sphingomonas(3.4%),Agrobacterium(3.0%),Ralstonia(2.5%),seudomonas(1.8%),Ochrobactrum(1.5%),Mycoplana(1.5%).5.Lef SE analysis was used to compare the differences between the two groups.The predominant bacterial groups in group C1 mainly include: Tenericutes、Proteobacteria、Spirochaetaceae、Sphingomonadales、Cytophagales;the C2 are mainly include: Fusobacteria 、 Armatimonadetes 、 Deinococci 、 Gammaproteobacteria 、Enterobacteriales、Bifidobacterium.Ⅲ.Structural changes of conjunctival sac flora after dacryocystorhinostomy for chronic dacryocystitis.1.A total of 3934982 effective sequences were obtained from 114 samples.The number of OTU at the portal level was compared with the control group 1 day before surgery,1 week after surgery,2 weeks after surgery,and 4 weeks after surgery were: 2353,2450,2927,and 2215,the antibiotic groups were: 2329,2498,2435,2451,2215.The OUT Numbers before and after nasal surgery were 3164 and 3225,respectively.2.The diversity index of the unused antibiotic group is significantly higher than the healthy control group except the Simpson index,but only the Shannon index is statistically significant(P <0.05);the Chao1,Ace,Shannon index of the antibiotic group is greater than the control group,Chao1,Ace is statistically significant(P <0.05),Simpson index is less than the control group;preoperative,postoperative 1 week,postoperative 2 weeks diversity index group using antibiotics group is higher than the group without antibiotics,using 4 weeks after surgery The Simpson and Shannon indexes of the antibiotic group were higher than those without antibiotics,and the rest were lower than those without antibiotics,but they were not statistically significant(P> 0.05).3.In the PCA analysis,there is a clear separation distance between the antibiotic group and the unused antibiotic group and the control group.The samples are scattered and mixed with each other.The sample distribution in the unused antibiotic group gradually approaches the healthy control group at 4 weeks.Antibiotic group compared with nonantibiotic group: The samples before antibiotics,1 week after surgery,and 2 weeks after surgery were distributed in the lower area.The samples were scattered in the 4th week after surgery.The distribution is scattered,and some are mixed with the unused antibiotic group.The distribution of samples before and after the 4th week of operation was scattered and mixed between the two groups.4.Count the top 20 flora of the genus level abundance.The composition of the core flora of the group with antibiotics and the group without antibiotics is similar.The main groups are Cupriavidus,Sphingomonas,Thermus,and Acinetobacter and Lactobacillus,but there is a big difference in composition ratio.The core bacterial groups of the nasal cavity are mainly Copper Cupri,Sphingomonas,Thermophilus,Streptococcus,Acinetobacter,Paleobacillus,Monascus,Lactobacillus,Cilia,Bacillus Bacillus,Bacteroides,etc.Among them,Lactobacillus mainly exists before surgery,Streptococcus mainly exists in the nasal cavity at the 4th week after operation,and Stenotrophomonas only exists in the nasal cavity after operation.5.Lef SE analysis of the differences between groups(LDA Score> 2)found that the predominant bacterial group in the antibiotic group compared with the control group was mainly Bacteroides,Bacillus,and Lactobacillus before surgery;the first week after surgery was mainly actinomycetes Phylum,acidobacteria,cyanobacteria,and green chlamydophyta;thick wall bacteria in the 2nd week after operation;Wps-2 and armor bacteria in the 4th week after operation.No significant differences were found between the antibiotic-free group and the control group.Comparison between the antibiotic group and the non-antibiotic group: the increased flora before surgery is Lactobacillus,Streptococcus,Trichosporaceae,Flavobacteriaceae,Treponema,Treponema;the reduced flora is mainly red ring Mycobacteriaceae,Halomonasceae,Achromobacter,Enterococcus;Clostridia,Rumenococci,Rhodobacteraceae increased 1 week after surgery;the reduced flora includes Proteobacteria,Tussock Cephalosporaceae,ciliates,bacillus,ML635J-21,cellulomycetes,campylobacter,marine spirilles,enterococci;increased thick wall bacteria,RF39,soft membranes 2 weeks after surgery Mycobacteria,Clostridiaceae,Soft-walled Phylum,Fecalococcus,Erysipelotidae;Reduced flora include Enterobacteriaceae,Stenobacteria,Azospirillum;Increased flora 4 weeks after surgery include Ruminococcus Genus,Roseateles,Nocardiopsis,Ralstonia,Turicibacter;the reduced bacterial flora are Nocardia,Bacillus,and Abnormal Coccus.The relative abundance of bacteria in the nasal cavity after the operation is mainly increased: Halomonasceae,Marine Spirulina,Staphylococcus,Acidophilus,Mycobacterium;the relative abundance of bacteria after surgery is mainly: Lactobacillus,oleobacteria.Conclusion: 1.Conjunctival sac flora in normal people: the abundance and diversity of conjunctival sac flora are older than 50 years old to a certain extent than those less than 50 years old;the core flora of the two age groups is mostly similar,mainly at the genus level Are: Corynebacterium,Sphingomonas,Thermophilus,Acinetobacter,Paleobacillus,but there are differences in the composition ratio,while Bifidobacterium,Agrobacterium,Ralstonia Genus and Mycobacterium are only present in the conjunctival sac less than50 years old.And the type and number of pathogenic bacteria in the group less than 50 years old is higher than that in the group more than 50 years old.2.The proportion of some flora in the unused antibiotic group gradually decreased after surgery,and by the 4th week after surgery,some of the flora reached the same proportion as the normal control group,but the number of most flora did not change significantly.3.Long-term use of antibiotics increases the abundance and diversity of the flora,making the structure of the flora more complicated,and samples.There was no significant change in the number of various bacterial groups from before operation to the fourth week after operation,and they were significantly higher than that of the control group,and the use of antibiotics may prolong the time of normalization of the bacterial group.4.Patients with chronic dacryocystitis use antibiotics and no antibiotics before and after the operation.The core flora composition is similar.The genus level is mainly: Cupriavidus,Sphingomonas,Thermophilus,No Acinetobacter and Lactobacillus,but there is a big difference in composition ratio.The use of antibiotics will increase the abundance and diversity of conjunctival cyst bacteria to a certain extent.At the same time,long-term use of antibiotics may increase the number of drug-resistant pathogenic bacteria in the conjunctival sac.5.In the patients with chronic dacryocystitis,the composition of the nasal cavity in the 4th week after En-DCR has been basically the same as that before the operation. |
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