Fungal keratitis represents a significant cause of blindness, with current therapeutic approaches yielding limited success. The disease's onset and progression are primarily driven by fungal virulence factors and the host's immune response. The innate immune system is the first to respond, with neutrophils playing a pivotal role in the antifungal defense. Although neutrophils are critical for pathogen clearance, their excessive or abnormal activation can lead to tissue damage, exacerbating the disease. Thus, elucidating the mechanisms underlying neutrophil activity in fungal keratitis is crucial for refining treatment strategies. This article aims to systematically review the principal antimicrobial mechanisms employed by neutrophils, including phagocytosis, degranulation, and the formation of neutrophil extracellular traps(NETs). Furthermore, it explores the crosstalk between neutrophils and macrophages, alongside their collective impact and underlying mechanisms in the context of fungal keratitis. Exploration of the mechanisms of fungal keratitis facilitates precise intervention and enhances the efficacy of treatment.

Yunnan Province borders with Myanmar, Vietnam, and Laos, the China-Myanmar border area is the key area for prevention of re-establishment from imported malaria after the disease was eliminated in China. Since the malaria elimination action plan was launched in Yunnan Province in 2011, 129 counties (cities, districts) were classified into three categories according to malaria incidence and transmission risk, and different technical strategies and measures were implemented with adaptations to local circumstances. A total of 68 malaria consultation service stations were established on the Chinese side of the China-Myanmar border and 80 malaria prevention and control stations were established on the Myanmar side by Yunnan Province in 2014. Then, the “Three Lines of Defense” strategy was implemented for malaria elimination in the China-Myanmar border area in Yunnan Province during the period from 2015 to 2018, and this strategy was further refined and adjusted to the “3 + 1” strategy for prevention of re-establishment from imported malaria in 2019. Through decades of multifaceted efforts, the malaria elimination goal was achieved in Yunnan Province in June 2021. However, the number of imported malaria cases appeared a tendency towards a rise in Yunnan Province in 2023 and 2024, due to changes in the situation in Myanmar and the gradual resumption of international travel and border crossings following the adjustment of the COVID-19 prevention and control policy in China. The joint malaria prevention and control cooperation between China and Myanmar was initiated with the pilot project for joint malaria prevention and control in the China-Myanmar border area in 2005, and this project was progressed into the joint malaria and dengue fever prevention and control project in parts of the Greater Mekong Subregion border areas in 2010. The threat of overseas malaria epidemics to border areas in Yunnan Province was effectively reduced through implementation of coordination meetings with Myanmar health departments, establishment of efficient information exchange mechanisms, establishment of overseas surveillance sentinel sites, technical training, provision of material supports, joint propagation activities and joint responses to malaria epidemics. This project was incorporated into the Five-Year Plan of Action on Lancang-Mekong Cooperation (2018—2022) in China in 2018, with 5 liaison offices and 20 liaison workstations established in Myanmar, Laos, Vietnam, Cambodia, and Thailand, and 21 cross-border malaria surveillance sites assigned in border areas of Myanmar, Laos and Vietnam, and a long-term malaria prevention and control cooperation mechanisms was established through meetings, training, propagation, and joint investigations. Currently, Yunnan Province is poised to engage in more extensive and in-depth cooperation with neighboring countries, including malaria diagnosis and treatment techniques, drug and vaccine research and development, talent cultivation, information sharing, cross-border human health services, and health promotion, under the guidance of the Five-Year Plan of Action on Lancang-Mekong Cooperation (2023—2027).

Fungal keratitis is a serious blinding eye disease. The development of fungal infections depends primarily on the interaction of fungal virulence with host immune defense factors. The cornea is considered an immune-privileged organ, and resident macrophages are the main immune cells that respond to the heterogeneity exhibited by the microenvironment with their polarization. In the early stage of infection, macrophages polarize towards M1, which promotes inflammation and facilitates fungal clearance but produces a cellular storm that exacerbates immune damage; in the late stage of infection, macrophages polarize towards M2, which suppresses the inflammatory response and facilitates tissue repair, but may be immunosuppressed or even immune escape to the detriment of pathogen clearance. The balance between pro-inflammatory and anti-inflammatory responses is key to maintaining the functional integrity of the cornea. Current antifungal drug therapy is limited, so it is particularly important to find a therapeutic target for the inflammatory response triggered by the immune response in addition to antifungal therapy. In this review, the functional and phenotypic characterization of macrophage subsets associated with fungal keratitis was reviewed, more in-depth research is needed to explore the specific mechanisms by which macrophage polarization and their impact on fungal keratitis. Targeted regulation of macrophage differentiation based on their phenotype and function could be an effective approach to treat and manage fungal keratitis in the future.

Objective:To observe the transparency and tissue structure changes of human corneal stromal lenticules after long-term cryopreservation and explore a simple and feasible method for long-term effective preservation of corneal stromal lenticules.Methods:Two hundred samples of intact human corneal stromal lenticules from 200 eyes were obtained during femtosecond laser small-incision lenticule extraction (SMILE) in Hainan Eye Hospital, Zhongshan Ophthalmic Center from 2013 to 2020.The samples were divided into 1-month, 24-month, 60-month and 80-month group and were stored in an ultra-low temperature freezer for 1, 24, 60 and 84 months respectively at -80 ℃ according to grouping, with 50 samples in each group.Transmittance of the corneal lenticules at wavelength of 300-800 nm was measured with an ultra-micro spectrophotometer and every lenticule was measured for 10 times with a 50 nm interval.The histomorphology and collagen fiber structure of the corneal lenticules were examined by hematoxylin-eosin staining and Masson staining, respectively.The arrangement of collagen fibers and ultrastructure changes of keratocytes in the samples were inspected with a transmission electron microscope.The apoptosis rate of keratocytes was determined by TUNEL staining.The study protocol was approved by an Ethics Committee of Hainan Eye Hospital at Zhongshan Ophthalmic Center (No.2013-003). This study complied with the Declaration of Helsinki.Written informed consent was obtained from each subject before surgery.Results:The corneal lenticules were clear and intact in all groups and no significant difference in the transmittance within 450-800 nm wavelength was seen among the 4 groups (all at P>0.05). Masson staining revealed that the collagen fibers in the lenticules were neatly arranged and tightly packed in the 1-month group.In the 24-month group, interfibrous vacuoles were found in some collagen fibers.The arrangement of the collagen fibers was loose and more vacuoles were displayed in the 60-month group, and the loss of some collagen fibers appeared and the lenticules were thinned in the 84-month group.It was found through hematoxylin-eosin staining that the morphological changes of corneal stromal lenticules corresponded to the alterations of collagen fibers.Transmission electron microscopy showed that in the 1-month group, the collagen fibers of the corneal stroma lenticules were neatly arranged and regular, and the corneal stromal cells were elongated and spindle-shaped, and the nuclear membrane was intact and the cytoplasm was abundant.In the 24-month group, the collagen fibers showed slightly loose arrangement, and the corneal stromal cells were deformed with incomplete nuclear membrane.In the 60-month group, the collagen fibers were in loose and irregular arrangement, and the nuclei were atrophied and deformed.The 84-month group showed disorganized arrangement of collagen fibers, wrinkled and atrophied corneal stromal cells, discontinuous nucleus membrane and nucleoplasmic lysis.TUNEL staining showed that the percentage of apoptotic corneal cells in lenticules was (87.80±1.17)%, (89.50±1.05)%, (89.30±1.51)% and (90.20±1.47)% in the 1-month, 24-month, 60-month and 84-month groups, respectively, with no statistically significant difference found in overall comparison ( F=4.525, P=0.053). Conclusions:The disorder of collagen fibers and apoptosis of keratocytes occur in the human corneal stromal lenticules till 84 months after cryopreservation, however, the transparency and integrity remain excellent.The ultra-low temperature preservation technique provides an effective and simple solution for long-term storage of human corneal stromal lenticule.

Objective To compare the clinical efficacy of one-stage laparoscopic cholecystectomy (LC) plus laparoscopic common bile duct exploration (LCBDE) with primary suture (PS) versus two-stage endoscopic retrograde cholangiopancreatography (ERCP)/endoscopic sphincterotomy (EST) followed by LC in treatment of cholecystolithiasis complicated with choledocholithiasis.Methods The clinical data of 171 patients with cholecystolithiasis complicated with choledocholithiasis,who underwent minimally invasive surgical treatment from January 2016 to December 2017,were retrospectively analyzed.Of these patients,90 underwent one-stage LC + LCBDE + PS (the one-stage group),and 81 underwent two-stage ERCP/EST followed by LC (the two-stage group).The main clinical variables between the two groups of patients were compared.Results The surgical success rates (94.4％ vs.95.1％),residual stone rates (3.3％ vs.4.9％) and incidences of postoperative complications (6.7％ vs.8.6％) showed no significant difference (all P ＞ 0.05) between the one-stage and two-stage groups.Compared with the two-stage group,the operative time was shorter (110.4 vs.135.7 min),the length of postoperative hospital stay was shorter (3.3 vs.7.1 d) and the total hospitalization cost was reduced (22 756.2 vs.31 429.3 yuan) in the one-stage group.The incidence of long-term complications (2.2％ vs.9.9％) in the one-stage group was also lower than that in the two-stage group (both P ＜ 0.05).Condusions Both one-stage LC + LCBDE + PS and two-stage ERCP/EST + LC are safe and effective in the treatment of cholecystolithiasis complicated with choledocholithiasis.One-stage LC + LCBDE + PS shows obvious advantages in hospitalization stay,hospitalization cost and in the preservation of function of the Oddi sphincter,and therefore should be the first choice in most cases.

Objective To compare the clinical efficacy of one-stage laparoscopic cholecystectomy (LC) plus laparoscopic common bile duct exploration (LCBDE) with primary suture (PS) and two-staged endoscopic retrograde cholangiopancreatography (ERCP)/endoscopic sphincterotomy (EST) followed by LC in treatment of cholecystolithiasis complicated with choledocholithiasis.Methods Of these patients,58 received one-staged LC + LCBDE + PS (the one-stage group),and 71 underwent two-staged ERCP/EST followed by LC (the two-stage group).Results The surgical success rate,residual stone rate,incidence of postoperative complications and operative time showed no significant difference (x2 =0.344,0.344,0.108,t =-0.240,all P ＞ 0.05) in both the one-staged and two-stage groups.Compared with the two-staged group,the hospital stay was shorter (4.1d vs.6.9d) and the total hospitalization cost was lower (23 126 yuan vs.32 982 yuan) in the one-staged group.Conclusion Both one-staged LC + LCBDE + PS and two-staged ERCP/EST + LC are safe and effective in the treatment of cholecystolithiasis complicated with choledocholithiasis on base for base basis.

ObjectiveTo construct a protein interaction network based on high-throughput sequencing data of liver cancer-related non-coding RNAs, to perform a functional enrichment analysis, and to screen out circular RNAs (circRNAs) participating in the development and progression of liver cancer via the mechanism of competitive endogenous RNA (ceRNA). MethodsThe circRNA-miRNA-mRNA network was constructed using gene expression omnibus (GEO) data based on the ceRNA theory. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) analyses were performed to identify circRNAs with potential ceRNA function and explore their functions. ResultsA total of 9 co-expressed circRNAs, 20 co-expressed miRNAs, and 153 co-expressed mRNAs were screened out from the GEO database, and the liver cancer-related circRNA-miRNA-mRNA network was successfully constructed. The GO analysis revealed 90 biological processes, which mainly involved 12 functional clusters including hepatocyte differentiation, phase-change regulation of cell cycle, and negative regulation of transcription factor activity. The KEGG analysis showed that the co-expressed circRNAs were also involved in the p53 and PI3K-Akt signaling pathways. ConclusionThis study provides new insights for circRNAs mediating the development and progression of liver cancer through the mechanism of ceRNA.

Objective To study the feasibility and efficacy of treatment of extrahepatic cholangiolithiasis using laparoscopic self-releasing J-tube drainage combined with primary suturing of common bile duct.Methods The clinical data of 172 patients with extrahepatic cholangiolithiasis who underwent operations from March 2013 to January 2015 were retrospectively studied.The patients were divided into two groups:the J-tube drainage group (n =82) and the T-tube drainage group (n =90).Surgical duration,intraoperative bleeding,postoperative hospital stay,treatment cycles,hospital costs,incidences of postoperative bile leakage and residual extrahepatic cholangiolithiasis were compared between the two groups.Results The length of postoperative hospital stay in the J-tube drainage group was (4.9 ± 1.2) days and in the T-tube drainage group was (8.0 ± 2.0) days.The treatment cycles in the J-tube drainage group were (4.9 ± 1.2) days while in the T-tube drainage group were (24.1 ± 3.2) days.The hospital costs in the J-tube drainage group were (12 817.1 ±3 167.1) yuan and the costs in the T-tube drainage group were (15 012.5 ±2 354.8) yuan.There were significant differences in hospital stay,treatment cycles and hospital costs between the two groups (all P ＜ 0.05).The surgical duration in the J-tube drainage group was (108.2 ± 10.2) minutes and the duration in the T-tube drainage group was (110.1 ± 13.1) minutes.The amount of intraoperative bleeding in the J-tube drainage group was (35.0 ± 20.0) ml and the amount in the T-tube drainage group was (42.0 ±30.0) ml.There were no significant differences in intraoperative bleeding and surgical duration between the two groups (all P ＞ 0.05).No significant differences were observed in the incidences of post-operative bile leakage and in residual extrahepatic cholangiolithiasis between the two groups (P ＞ 0.05).Conclusions Self-releasing J-tube drainage combined with laparoscopic primary suturing of common bile duct was safe and efficacious.It was minimally invasive in treating patients with extrahepatic cholangiolithiasis.

To understand the genetic origin and variation of porcine epidemic diarrhea virus (PEDV) prevailing in Henan province,a total of 22 PEDV positive samples from suckling piglets with severe diarrhea were collected from 16 pig farms and amplified by RT-PCR.ORF3 genes were then cloned into pMD18-T vector and sequenced.The result of sequencing showed that the length of ORF3 genes all were 675 bp which encoded 224 amino acids.The strains in this study had eight amino acid substitutions when compared with CV777 strain.The nucleotide homologies of the 22 strains were 95.9％-100％ and amino acid homologies were 96.4％-100％.By comparing with those of European strain CV777 and vaccine strain truncated CV777,the nucleotide homologies were 97.1％-97.9％ and 94.8％-95.4％,respectively.Based on the phylogenetic relationship of ORF3 genes,the PEDV field strains and PEDV reference strains could be divided into two groups,and all the field strains identified in this study belong to group 2.It suggested that prevalent PEDV strains in this study seem to be closely related to Henan isolates in previous years,domestic strains,Janpanese strains,American strains as well as Korean strains but differ genetically from European strains or vaccine strains used in China.This experiment analyzed the ORF3 gene sequence features in prevailing of Henan region in recent years,which provides a new support of epidemiology study and protein function research of PEDV ORF3 gene.

Objective To introduce the application of the network training on clinicians'knowledge of malaria diagnosis and treatment in Yunnan Province,and evaluate its effect. Methods Through the platform Yiboshi(www.yiboshi.com),the medi-cal and health personnel at the units of provincial,prefectural,county levels and 25 townships of 25 border counties were trained on the knowledge of malaria diagnosis,treatment,prevention and control,and the effects were evaluated by examina-tions,questionnaires and interviews. Results Totally 7152 participants were trained,the average participation,completion and pass rates of the training were 95.26%,98.55% and 97.30%,respectively. The trainees mainly learned malaria control knowledge from 3 aspects,namely policy of malaria elimination,malaria epidemiology,malaria diagnosis and treatment. The questionnaires showed that 95.94%of the participants considered that their theoretical and technical levels improved,97.30%were interested in the training content,93.24% recognized the arrangement of the training time was reasonable,and 91.89%were satisfied with the service of the platform. Conclusions The network training on knowledge of malaria diagnosis and treat-ment in Yunnan Province has achieved good effect. The network training meets the need of training a large number of clinicians in the malaria elimination and post-elimination stage.