A surgical alternative of fusiform penetrating keratoplasty for the management of severe infectious keratitis

AIM:To describe the surgical procedure of fusiform penetrating keratoplasty(FPK)using multiple trephines of different sizes for treating patients with severe infectious keratitis.METHODS:Fourteen eyes underwent FPK,and 15 eyes received conventional penetrating keratoplasty(PK)were included in the study.The best-corrected visual acuity(BCVA),refractive outcomes,endothelial cell density,and postoperative complications were recorded.RESULTS:The FPK group was followed for an average of 15.3±2.1mo,whereas the PK group was followed for 16.1±1.9mo.The corneal ulcers were elliptical-shaped in all 14 eyes in the FPK group.The mean BCVA(logMAR,0.26±0.13)showed no statistically significant differences from that in the PK group(logMAR,0.21±0.12,P>0.05)at 1y after surgery.But the mean curvature,mean astigmatism,and mean spherical equivalent in the FPK group were lower than those in the PK group(P<0.05).Peripheral anterior synechia was observed in one patient in the FPK group,whereas 6 patients in the PK group.Suture loosening and neovascularization were observed in 4 and 5 eyes in the PK group,respectively.No graft immune rejection or elevation of intraocular pressure was observed in the two groups.CONCLUSION:For patients with elliptical-shaped corneas or corneal ulcers,FPK can avoid disrupting of corneal limbus,reduce the risk of postoperative complications,and can result in satisfactory visual quality.

Hydrogel dressings on neurotrophic keratitis in an experimental animal model

AIM:To investigate the therapeutic effects of hydrogel dressings on neurotrophic keratitis in rats.METHODS:Male Wistar rats,aged 42–56d,were randomly divided into control,experimental,and treatment groups,each consisting of five rats.The experimental and treatment groups underwent neurotrophic keratitis modeling in both eyes.After successful modeling,biomedical hydrogels formed with polyvinyl alcohol and polyvinyl pyrrolidone were used in treatment group for 7d.Ocular irritation response and keratitis index scores,Schirmer’s test,tear film break-up time(BUT),sodium fluorescein staining,and hematoxylin and eosin(HE)staining were used to evaluate the effectiveness of the treatment.RESULTS:The neurotrophic keratitis model was successfully established in rats with severe ophthalmic nerve injury,characterized by keratitis,ocular irritation,reduced tear secretion measured by decreased BUT and Schirmer test values,corneal epithelial loss,and disorganized collagen fibers in the stromal layer.Following treatment with hydrogel dressings,significant improvements were observed in keratitis scores and ocular irritation symptoms in model eyes.Although the recovery of tear secretion,as measured by the Schirmer’s test,did not show statistical differences,BUT was significantly prolonged.Fluorescein staining confirmed a reduction in the extent of corneal epithelial loss after treatment.HE staining revealed the restoration of the structural disorder in both the epithelial and stromal layers to a certain extent.CONCLUSION:Hydrogel dressing reduces ocular surface irritation,improves tear film stability,and promotes the repair and restoration of damaged epithelial cells by maintaining a moist and clean environment on the ocular surface in the rat model.

CD3ε of a pan T cell marker involved in mouse Aspergillus fumigatus keratitis

AIM:To explore whether CD3ε is involved in the adaptive immunity of Aspergillus fumigatus(A.fumigatus)keratitis in mice and the role of innate and adaptive immunity in it.METHODS:Mice models of A.fumigatus keratitis were established by intra-stromal injection and corneal epithelial scratching.Subconjunctival injections of natamycin,wedelolactone,LOX-1 inhibitor(poly I)or Dectin-1 inhibitor(laminarin)were used to treat mice with A.fumigatus keratitis.Mice were pretreated by intraperitoneal injection of anti-mouse CD3ε.We observed the corneal infection of mice under the slit lamp microscope and made a clinical score.The protein expression of CD3ε and interleukin-10(IL-10)was determined by Western blotting.RESULTS:With the disease progresses,the degree of corneal opacity and edema augmented.In the intrastromal injection models,CD3εprotein expression began to increase significantly on the 2^(nd) day.However,in the scraping epithelial method models,CD3ε only began to increase on the 3^(rd) day.After natamycin treatment,the degree of corneal inflammation in mice was significantly attenuated on the 3^(rd) day.After wedelolactone treatment,the severity of keratitis worsened.And the amount of CD3ε protein was also reduced,compared with the control group.By inhibiting LOX-1 and Dectin-1,there was no significant difference in CD3ε production compared with the control group.After inhibiting CD3ε,corneal ulcer area and clinical score increased,and IL-10 expression was downregulated.CONCLUSION:As a pan T cell marker,CD3ε participate in the adaptive immunity of A.fumigatus keratitis in mice.In our mice models,the corneas will enter the adaptive immune stage faster.By regulating IL-10,CD3ε exerts antiinflammatory and repairs effects in the adaptive immune stage.

Advances in Drug Treatment of Fungal Keratitis

Fungal keratitis is an important cause of corneal blindness in China, accounting for 45% of infectious keratitis. The main pathogenic bacteria include yeast, filamentous bacteria and nearly 100 kinds of fungi, which are difficult to diagnose, difficult to treat and poor prognosis. When the infected fungal strains have strong virulence and poor drug sensitivity, it is easy to prolong the disease. Once the fungal infection involves the whole limbus and reaches the whole layer of the cornea, it will be followed by intraocular tissue infection such as anterior chamber, lens and vitreous body. When the infection is difficult to control and the visual function is seriously damaged, the enucleation of eye contents has to be performed, which causes irreversible harm to the patient’s appearance and physical and mental health. Therefore, in order to gain greater hope for the vision of patients with fungal keratitis, In recent years, with the continuous progress of clinical medicine and microbiological diagnostics, the treatment methods of fungal keratitis have been constantly updated. This article will briefly review the new progress in drug and surgical treatment of fungal keratitis in recent years to provide patients with better visual prognosis. .

Herpes simplex keratitis:A brief clinical overview

The aim of our minireview is to provide a brief overview of the diagnosis,clinical aspects,treatment options,management,and current literature available regarding herpes simplex keratitis(HSK).This type of corneal viral infection is caused by the herpes simplex virus(HSV),which can affect several tissues,including the cornea.One significant aspect of HSK is its potential to cause recurrent episodes of inflammation and damage to the cornea.After the initial infection,the HSV can establish a latent infection in the trigeminal ganglion,a nerve cluster near the eye.The virus may remain dormant for extended periods.Periodic reactivation of the virus can occur,leading to recurrent episodes of HSK.Factors triggering reactivation include stress,illness,immunosuppression,or trauma.Recurrent episodes can manifest in different clinical patterns,ranging from mild epithelial involvement to more severe stromal or endothelial disease.The severity and frequency of recurrences vary among individuals.Severe cases of HSK,especially those involving the stroma and leading to scarring,can result in vision impairment or even blindness in extreme cases.The cornea's clarity is crucial for good vision,and scarring can compromise this,potentially leading to visual impairment.The management of HSK involves not only treating acute episodes but also implementing long-term strategies to prevent recurrences and attempt repairs of corneal nerve endings via neurotization.Antiviral medications,such as oral Acyclovir or topical Ganciclovir,may be prescribed for prophylaxis.The immune response to the virus can contribute to corneal damage.Inflammation,caused by the body's attempt to control the infection,may inadvertently harm the corneal tissues.Clinicians should be informed about triggers and advised on measures to minimize the risk of reactivation.In summary,the recurrent nature of HSK underscores the importance of both acute and long-term management strategies to preserve corneal health and maintain optimal visual function.

Food Microbiology and Safety 实验课的教改探索

该文作者所在大学与新西兰梅西大学合作开展了食品科学与工程专业国际化培养本科教育项目,全英文课程Food Microbiology and Safety是该项目的核心课程之一。该实验课以应用性为前提,以课程思政为引领,强调食品微生物学与食品安全学的融合,中外师资合作开展了系列教学探索与实践。课程设计以食品产品和食品生产为情境,以微生物安全为话题,模拟真实生产场景,丰富内涵和拓展外延;自主设计实验激发主观能动性、自主规划进程锻炼统筹能力、线上资源平台支撑自主学习与评价、随机分组培养团队合作能力。实践表明,课程有效实现了食品微生物学与食品安全学的有机融合,目标驱动型实验教学过程更利于激发学生的主动性,大量综合性和设计性实验提高了课程的两性一度。与同期对照班级相比,该项目所培养学生具有更强的专业素养和实践能力,为食品专业其他实验课程的教学改革提供了参考。

Two-step strategy—conjunctival flap covering surgery combined with secondary deep anterior lamellar keratoplasty for the treatment of high-risk fungal keratitis

AIM:To investigate whether the two-step strategy[conjunctival flap covering surgery(CFCS)combined with secondary deep anterior lamellar keratoplasty(DALK)]is effective for patients with high-risk fungal keratitis(FK).METHODS:In this noncomparative,retrospective case series,10 subjects(6 males,4 females)with a mean age of 56.5±7.1(range 47-72)y with high-risk FK undergone the two-step strategy were included.Reported outcome measures were healing of the corneal ulcer,recurrence of FK,reject reaction,improvement in best corrected visual acuity(BCVA)and relevant complications.RESULTS:The average diameter of corneal infiltrates was 7.50±0.39 mm,ranging from 6.94 to 8.13 mm.The mean depth of corneal infiltrates was 422.4±77.1μm,ranging from 350 to 535μm.The mean corneal thickness was 597.4±117.3μm,ranging from 458 to 851μm.Hypopyon and endothelial plaques were presented in all patients.The period between the two steps was 3.65±0.9(ranging from 3 to 5)mo.The graft diameter was 7.75±0.39 mm.At the last follow-up(average 9.25±3.39,ranging from 5.5 to 17mo),no fungal recurrence or graft rejection appeared,and all patients showed improvement of BCVA.One patient suffered from liver function impairment due to oral voriconazole for 4wk and recovered spontaneously after 1wk of drug withdrawal.CONCLUSION:The two-step strategy is safe and effective in the treatment of high-risk FK by transforming intentional therapeutic penetrating keratoplasty during acute infection to later optical DALK.It is a practical strategy,especially in areas lacking fresh donor corneas and eye bank services.

Clinical observation of recombinant human nerve growth factor in the treatment of neurotrophic keratitis

AIM:To characterize changes of corneal nerve morphology and tear indices in patients with neurotrophic keratitis(NK)treated with recombinant human nerve growth factor(rhNGF).METHODS:In a prospective observational study,six patients(nine eyes)were locally treated with rhNGF.Visual acuity,corneal fluorescein staining score,the heights of the tear river,lipid layer thickness(LLT),tear ferning(TF)test,conjunctival impression cytology(CIC)examination,the densities of cornea subbasal nerve fibers were determined before and after treatment.RESULTS:Compared with baseline,there was a significant difference in corneal fluorescence staining scores(P<0.01);all patient corneal epithelial defects recovered completely within 8wk,but there was no significant improvement in the height of the tear river(P=0.202).LLT was significantly increased when compared with baseline(P=0.042);however,the function of conjunctival goblet cells and mucin content did not significantly improve using the TF test and CIC examination(P=0.557,P=0.539).After 8wk of treatment,the average corneal subbasal nerve fiber density increased significantly(P<0.01),as did the number of corneal nerve fiber branches(P=0.001).CONCLUSION:RhNGF can increase the density of corneal subbasal nerve fibers,promote the healing of persistent corneal epithelial defects and corneal ulcers in patients with NK,also improving tear function partially.

Construction Method of Ideological and Political Theories Teaching in the Course of Environmental Microbiology for Environmental and Ecological Engineering Major

Environmental Microbiology is a professional basic course in the curriculum system of Environmental and Ecological Engineering major,which follows the requirements of ideological and political theories teaching in the course in higher education,and should be implemented in every course.Therefore,the construction of ideological and political theories teaching in the course should be an important task in the course construction of Environmental Microbiology.In view of the curriculum characteristics of Environmental Microbiology,such as multi-disciplinary feature,wide coverage and rapid development,it is an effective method for the construction of ideological and political theories teaching in the course of Environmental Microbiology by fully tapping the ideological and political elements of the curriculum,such as patriotism,scientific spirit and practical innovation,and integrating the ideological and political theories teaching in the course into the whole curriculum design and classroom teaching construction.

Should ocular Demodex be checked and treated in refractory keratitis patients without blepharitis?

AIM:To evaluate the correlation between Demodex infestation and keratitis,and to assess demodicosis using a simple approach.METHODS:A modified slit lamp illumination(at 40×magnification)was used to observe Demodex tails in 40patients with refractory keratitis and 80 healthy controls.Bacterial smear and culture of the conjunctival sac and corneal lesion were performed to identify the pathogen.Tea tree oil ointment(TTOO)was added as a Demodex killing agent for lid scrubs to the treatment when Demodex infestation was confirmed.RESULTS:Demodex tails were found in all patients compared to 42/80 of the controls(P<0.01).Seventeen patients presented blepharitis,while 23 were free of scales and inflammation at the lid margin.The demodicosis was mild,moderate,and severe in 8,19,and 13 patients,respectively,compared to mild in 42 controls(P<0.01).The keratitis was mild,moderate,and severe in 13,19,and 8patients,respectively.The severity of Demodex infestation was not correlated to the severity of keratitis(P=0.126).The growth of Staphylococcus was revealed in nine patients who did not react to antibiotic eye drops prior to the TTOO treatment.Patients’signs and symptoms got resolved after the lid scrub with TTOO.CONCLUSION:Ocular Demodex needs to be checked and treated in refractory keratitis patients with or without blepharitis.A slit-lamp illumination under high magnification favors the judgment of the severity of Demodex infestation.

Differential expression of antimicrobial peptides in human fungal keratitis

AIM:To analyze a series of antimicrobial peptides(AMPs)in corneal tissue from individuals with fungal keratitis(FK)during the active phase of the fungus infection and after healing.METHODS:Patients undergone lamellar keratoplasty for the treatment of severe FK or corneal scar had their corneal buttons sampled.Quantitative real-time polymerase chain reaction(PCR)was used to ascertain the gene expression of human beta-defensin(HBD)-1,-2,-3,-9,S100A7,8,9,and LL-37.RESULTS:All AMPs’messenger ribonucleic acid(mRNA)expression was considerably elevated in all samples(n=12).In contrast to controls,where HBD-2,-3,and S100A7 mRNAs were expressed at very low levels,it was discovered that HBD-1,-9,S100A8,S100A9,and LL-37 were constitutively expressed in all healed samples(n=4).HBD-1,-2-3,S100A7,and LL-37 mRNAs were significantly increased in all active FK samples(n=8).The levels of HBD-9,S100A8,and S100A9 mRNAs were moderately upregulated in all active FK samples.Subgroup comparison showed that HBD-2 was significantly increased in Fusarium keratitis samples(n=5),and LL-37 mRNAs were significantly enhanced in Aspergillus keratitis samples(n=3).Whereas there was not significantly increased of HBD-1,-3,-9,S100A7,8,9 mRNA in Aspergillus keratitis samples compared with Fusarium keratitis samples.CONCLUSION:AMPs expression is increased in active FK,but not all AMPs are equally expressed.HBD-2 and LL-37 expression levels are the highest,showing some specificity of AMP expression related to FK.Human AMPs,particularly HBD-2 may play a significant role in Fusarium keratitis and LL-37 might be the key player in Aspergillus keratitis.

Clinical Analysis of 137 Cases of Fungal Keratitis

Objective: To analyze the epidemiology, clinical diagnosis and treatment of fungal keratitis (FK) in hospitalized patients at the First Affiliated Hospital of Yangtze University in recent years. Methods: A retrospective investigation was conducted on the data of 137 cases of FK in our hospital from January 2019 to December 2022. The epidemiological characteristics, identification results of fungal strains, clinical treatment, and prognosis of the patients were analyzed. Results: Among the 137 FK patients, 89 were males and 48 were females, and the ratio of male to female was 1.85:1, The age of onset was the largest number of patients in 50-59 years old and 60-69 years old. The disease occurred most in autumn, winter and summer farming season (from September to December, January, May, June). 72 cases (52.6%) had a clear history of corneal injury, and 43 cases (31.4%) had a history of plant injury. Other risk factors include eye surface diseases, ophthalmic surgery, and wearing corneal contact lenses. The top three pathogens were fusarium (38.7%), aspergillus (23.3%), and alternaria (17.5%). 101 eyes showed improvement or cure after treatment with medication, 9 eyes underwent corneal stromal injection, 11 eyes were covered with conjunctival flap covering or amniotic membrane transplantation, 12 eyes were covered with corneal transplantation, and enucleation of ocular contents was performed in 4 eyes;Visual acuity was improved or maintained in 123 patients (about 89.8%). Conclusions: The incidence of FK in our hospital was mostly middle-aged and elderly men, mostly caused by corneal injury in the process of agricultural labor. The pathogens were mainly fusarium and aspergillus. The preferred treatment was medication, with severe cases requiring combined surgical treatment. Most patients can maintain or improve their vision after treatment.

Study of the Effect of Bacterial Resistance Monitoring in Clinical Microbiology Testing

Objective:To observe and analyze the effect of bacterial resistance monitoring in clinical microbiology testing.Methods:600 microbial specimens collected in our hospital in the past year(April 2021 to April 2022)were used as the test subjects of this study.The specimens were divided into Group A(control group)and Group B(research group),with 300 cases in each group.Group A consisted of blood culture specimens,while Group B consisted of sputum specimens.After the tests were completed,the rates of unfavorable and favorable results,bacterial species distribution,and bacterial drug resistance of the specimens in both groups were compared.Results:Among group A specimens,29 cases were positive(9.67%)and 271 cases were negative(90.33%);among group B specimens,99 cases were positive(33.00%)and 201 cases were negative(66.00%);the difference between the two groups of data was statistically significant(P<0.05).As for the distribution of the types of bacteria,there were 472 cases of Gram-negative bacteria and 128 cases of Gram-positive bacteria.Conclusion:Bacterial resistance monitoring is helpful in clinical microbiology testing.Through proper monitoring,bacterial resistance can be well understood.In this way,patients get to receive appropriate treatment measures and suitable antibacterial prescriptions,thereby improving the patient outcome.

Microbiological Quality of “Rabilé”, a Yeast Used for Fermentation of Dolo, a Local Beer in Dédougou, Burkina Faso

Sorghum beer or dolo is part of the eating habits of part of the population of Dédougou because of its low price compared with industrial beers. Its production is an ancestral tradition that uses traditional equipment and gives dolo organoleptic properties that are not found in industrial beers. The production process involves several stages, including fermentation, which itself comprises natural lactic fermentation followed by alcoholic fermentation using traditional yeasts, which are not controlled in any way. The general aim of this study is to assess the microbiological quality of these fermentative yeasts in the town of Dédougou, in order to contribute to the health safety of the population and the promotion of these local beers. Twenty samples of fermenting yeast were analyzed according to ISO standards, to isolate enterobacteria, total and faecal coliforms according to standard procedures for isolating these micro-organisms. The isolated strains were identified using the API20E gallery. Microbiological analyses revealed the presence of 51.17% enterobacteria, 45.38% total coliforms and 3.45% thermotolerant coliforms. We counted 40% Escherichia coli, 20% Enterobacter cloacae, 20% Klebsiella pneumoniae and 20% Klebsiella spp. All the strains detected are capable of surviving in hostile conditions and could harm the quality of the dolo, consumer health and cause real collective food poisoning in the town of Dédougou. This enabled us to assess the microbial quality of these yeasts and to propose more suitable measures for producing and preserving dolo under hygienic conditions to protect consumer health.

Microbiological Quality of Cyperus esculentus L. Products Sold in Ouagadougou and Bobo Dioulasso (Burkina Faso)

This study aimed to assess the microbiological quality of tiger nut (Cyperus esculentus L.) products sold in Ouagadougou and Bobo Dioulasso. The microbial load of tubers and tiger nut milk was determined using ISO 4833-1 (2003) standard (total mesophilic aerobic flora), NF ISO 7954 (1998) standard (yeasts and molds), and ISO 4832 (2006) standard (total coliforms, thermotolerant coliforms). The results showed that tiger nut tubers had an average microbial load between 4.86 ± 7.03 × 106 UFC/g and 1.82 ± 2.30 × 107 UFC/g for total mesophilic aerobic flora, from 4.34 ± 10.6 × 105 UFC/g to 3.52 ± 4.54 × 106 UFC/g for yeasts and molds, from 2.23 ± 2.25 × 105 UFC/g to 1.41 ± 2.16 × 105 UFC/g for total coliforms and from 1.83 ± 2.03 × 105 UFC/g to 7.0 ± 10.8 × 104 CFU/g for thermotolerant coliforms. For tiger nut milk samples, the average microbial load varied from 3.48 ± 2.98 × 106 CFU/g to 2.80 ± 5.69 × 107 CFU/g for total mesophilic aerobic flora, from 5.00 ± 7.21 × 104 CFU/g to 1.88 ± 3.31 × 105 CFU/g for yeasts and molds, from 4.58 ± 10.4 × 104 CFU/g to 6.31 ± 9.17 × 105 CFU/g for total coliforms and 7.00 ± 7.00 × 103 CFU/g to 2.87 ± 5.86 × 105 CFU/g for thermotolerant coliforms. This study revealed that the tubers and tiger nut milk sold in Ouagadougou and Bobo Dioulasso had a high microbial load which could lead to the degradation of these products and food poisoning for consumers.

Antifungal efficacy of natamycin in experimental fusarium solani keratitis

AIM: To evaluate the efficacy of topical administration Natamycin, which is produced by China, in an experimental rabbit model of Fusarium solani keratitis, to provide experimental basis for the application of clinical safety. METHODS: Fusarium solani was induced in the right eye of 30 New Zealand rabbits. Forty-eight hours after inoculation, the animals were divided into 3 different treatment groups, 10 rabbit eyes of each group: Group 1 (Natamycin) treated with topical Natamycin, group 2 (Natacyn) treated with topical Natacyn, group 3 (control) treated with topical saline solution. The eyes of each group was examined clinically with slit lamp using ulcer scoring system on day 4, 10, 15, and 21 for status of healing, corneal vascularisation, iritis, hypopyon and macular nebula. The findings were recorded on day 10 and day 21. RESULTS: Ulcer score on day 10, day 15, day 21: The score of Natamycin group are 1.45 +/- 0.16, 1.08 +/- 0.11, 0.70 +/- 0.40. The score of Natacyn group are 1.35 +/- 0.12, 1.10 +/- 0.12, 0.65 +/- 0.35. the score of control group are 1.30 +/- 0.08, 3.63 +/- 0.28, 3.80 +/- 0.16. Natamycin group and Natacyn group were different from control group (P <0.01). There is no difference between Natamycin group and Natacyn group. Status of healing on day 10 and day 21: The cure rate of the Natamycin group is 90% on day 10, and 100% on day 21. The cure rate of the Natacyn group is 80% on day 10, and 100% on day 21.Natamycin group and Natacyn group were different from control group (P<0.01). There is no difference between Natamycin group and Natacyn group. Corneal vascularisation, iritis, hypopyon and macular nebula on day 10 and day 21: in Natamycin group, the number of the eyes which have Corner vascularisation, iritis, hypopyon and macular nebula are 2,0,0,2. In Natacyn group, the number of the eyes which have Corner vascularisation, iritis, hypopyon and macular nebula are 1,0,0,2. In control group, the number of the eyes which have Corner vascularisation, iritis, hypopyon and macular nebula are 9,9,8,9.Natamycin group and Natacyn group were different from control group (P<0.01). There is no difference between Natamycin group and Natacyn group. CONCLUSION: Natamycin was found to be effective in fungal keratitis, similar to Natacyn, and it can stop the corner vascularisation, iritis, hypopyon and macular nebula to happen. Natamyin manufactured in China is effective against fungal keratitis, with esay availability and low toxicity in its use.

Inhibition of LOX-1 alleviates the proinflammatory effects of high-mobility group box 1 in Aspergillus fumigatus keratitis

AIM: To investigate the inflammatory amplification effect of high-mobility group box 1(HMGB1) in Aspergillus fumigatus(A. fumigatus) keratitis and the relationship between lectin-like oxidized low-density lipoprotein receptor 1(LOX-1) and HMGB1 in keratitis immune responses.METHODS: Phosphate buffer saline(PBS), and Boxb were injected into BALB/c mice subconjunctivally before the corneas were infected with A. fumigatus. RAW264.7 macrophages and neutrophils were pretreated with PBS and Boxb to determine the HMGB1 inflammatory amplification effects. Abdominal cavity extracted macrophages were pretreated with Boxb and Poly(I)(a LOX-1 inhibitor) before A. fumigatus hyphae stimulation to prove the the relationship between the two molecules. LOX-1, interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), macrophage inflammatory protein-2(MIP-2) and IL-10 were assessed by polymerase chain reaction and Western blot.RESULTS: Pretreatment with Boxb exacerbated corneal inflammation. In macrophages and neutrophils, A. fumigatus induced LOX-1, IL-1β, TNF-α and MIP-2 expression in Boxb group was higher than those in PBS group. Poly(I) treatments before infection alleviated the proinflammatory effects of Boxb in abdominal cavity extracted macrophages. Pretreatment with Boxb did not influence Dectin-1 mRNA levels in macrophages and neutrophils.CONCLUSION: In fungal keratitis, HMGB1 is a proinflammatory factor in the first line of immune response. HMGB1 mainly stimulates neutrophils and macrophages to produce inflammatory cytokines and chemokines during the immune response. LOX-1 participates in HMGB1 induced inflammatory exacerbation in A. fumigatus keratitis.

Conjunctival microbiome changes associated with fungal keratitis: metagenomic analysis

AIM: To investigate the ocular surface microbiome profile of patients with fungal keratitis(FK) through bacterial 16 S r DNA sequencing. METHODS: The swab samples were collected from 8 patients with FK(Group 1 from the corneal ulcer, Group 2 from the conjunctival sac of the infected eyes, and Group 3 from the conjunctival sac of the fellow eyes) and 10 healthy eyes(Group 4 from the conjunctival sac). Bacterial 16 S rDNA V4-V5 region sequencing was performed to characterize the bacterial communities on the ocular surfaces of the patients with FK. RESULTS: Our metagenomic data showed that 97% of the sequence reads were categorized into 245 distinct bacterial genera, with 67.75±7.79 genera detected in Group 1, 73.80±13.44 in Group 2, 74.57±14.14 in Group 3, and 89.60±27.49 in Group 4. Compared with the healthy eyes(Group 4), both infected(Groups 1 and 2) and fellow eyes(Group 3) of the patients with FK showed reduced bacterial diversity and altered ocular surface microbiota compositions, with lower abundance of Corynebacterium and Staphylo coccus and higher abundances of Pseudomonas, Achromobacter, Caulobacter and Psychrobacter. CONCLUSION: Our report depicts the altered ocular surface bacterial community structures both in the affected and fellow eyes of patients with FK. These changes may contribute to the pathogenesis of FK or the increased risk for FK.

Effects of COX-2 inhibitor NS-398 on IL-10 expression in rat fungal keratitis

AIM: To investigate the expression of interleukin-10 (IL-10) and the effect of NS-398 (COX-2 inhibitor) on the expression of IL-10 in fungal keratitis in rats, and analyze its effects on anti-fungus immunity. METHODS: Ninety Wister rats were randomly divided into 3 groups. Group A was blank control group (10 eyes). Group B was fungal keratitis group (40 eyes). Group C was fungal keratitis group treated with NS-398 (40 eyes). PAS staining, 100g/L potassium hydroxide (KOH) smear and fungal culture confirmed the successful establishment of fungal keratitis model. After the central epithelium was scraped, Fusarium solani colonies were applied and contact lens was put on the right cornea of group B and C, and plane contact lens was put on the left cornea of control eyes. Phosphate buffered saline (PBS) eyedrops were given for group B and NS-398 eyedrops for group C. The expression of IL-10 on corneas of group B and C on the 1(st) day, 3(rd) days, 75(th) days, and 14(th) days were detected by immunohistochemistry and semi- quantitative reverse transcription- polymerase chain reaction (RT-PCR). RESULTS: Histopathologic examination showed neutrophil infiltration and severe tissue necrosis in ulcer cornea. PAS staining confirmed the existence of hyphae and spores in the superficial layer of stroma. In the blank and control groups almost no expression of IL-10 was detected at any observing points. In group B the expression of IL-10 increased at first and decreased thereafter. Its expression also showed significant difference at any observing points (P < 0.01). Compared with group B, the expression of IL-10 in group C showed no difference on the 1(st)day, decrease on the 3(rd) day, but a significant increase on the 7(th) day and 14(th) day. CONCLUSION: IL-10 takes part in the occurrence and development of fungal keratitis. NS-398 can upgrade the expression of IL-10 in fungal keratitis in the later period of the ulcer. Meanwhile, pathologic observation showed a slightly corneal opacity. IL-10 may play an important role in the process of cornea anti-damage repair.

Oral Microbiology:Past,Present and Future

Since the initial observations of oral bacteria within dental plaque by van Leeuwenhoek using his primitive microscopes in 1680, an event that is generally recognized as the advent of oral microbiological investigation, oral microbiology has gone through phases of ＂reductionism＂ and ＂holism＂. From the small beginnings of the Miller and Black period, in which microbiologists followed Koch＇s postulates, took the reductionist approach to try to study the complex oral microbial community by analyzing individual species; to the modem era when oral researchers embrace ＂holism＂ or ＂system thinking＂, adopt new concepts such as interspecies interaction, microbial community, biofilms, poly-microbial diseases, oral microbiological knowledge has burgeoned and our ability to identify the resident organisms in dental plaque and decipher the interactions between key components has rapidly increased, such knowledge has greatly changed our view of the oral microbial flora, provided invaluable insight into the etiology of dental and periodontal diseases, opened the door to new approaches and techniques for developing new therapeutic and preventive tools for combating oral polymicrobial diseases.