Interactions of primary neuroepithelial progenitor and brain endothelial cells: distinct effect on neural progenitor maintenance and differentiation by soluble factors and direct contact

Neurovascular interactions are crucial for the normal development of the central nervous system. To study such interactions in primary cultures, we developed a procedure to simultaneously isolate neural progenitor and endothelial cell fractions from embryonic mouse brains. Depending on the culture conditions endothelial cells were found to favor maintenance of the neuroprogenitor phenotype through the production of soluble factors, or to promote neuronal differentiation of neural progenitors through direct contact. These apparently opposing effects could reflect differential cellular interactions needed for the proper development of the brain.

A Study on Clinical Management of Glandular Changes in Cells on Cervical Cytology

A cytological cervical smear abnormality of glandular origin raises a high suspicion of underlying invasive or pre invasive cancers. Objective: To look into the diagnosis and management of women presenting with glandular changes on cervical cytology and to further implement a good management plan for these women. Methods: This study is a retrospective review of all patients referred to North Cumbria Integrated Care (NCIC) NHS foundation trust, United Kingdom with glandular changes on their cervical smear result between January 2015 and December 2020. Data was collected from the hospital colposcopy data base. Results: This study comprised of a sample size of 65 women. 11 of these (17%) were referred with borderline changes in their endocervical cells and 54 (83%) referred with a ?glandular neoplasia of endocervical type. There were colposcopically significant lesions (high grade lesions or suspected adenocarcinoma) for 52 (80%) of the women. All but one (98%) had Large loop excition of transformation zone (LLETZ) after colposcopy. Histologically, 8 (12%) women were diagnosed with adenocarcinoma, 29 (45%) were diagnosed with high grade cervical glandular intraepithelial neoplasia (HGCGIN), 9 (14%) had high grade cervical intraepithelial neoplasia (CIN2/3), 2 (3%) had low grade cervical glandular intraepithelial neoplasia (LGCGIN), 4 (6%) had a normal histology and 10 (15%) had lesions of mixed origin. After their first LLETZ treatment, 24 (37.5%) needed repeat LLETZ, 8 (12%) had a hysterectomy and one woman had radical trachelectomy.

Endoscopic ultrasound-guided fine-needle aspiration cytology in pancreaticobiliary carcinomas:diagnostic efficacy of cell-block immunocyto-chemistry

BACKGROUND： Endoscopic ultrasound-guided fine-needle aspiration cytology was demonstrated to be a useful tool for the diagnosis and staging of pancreaticobiliary neoplastic le- sions. Nonetheless, the diagnostic value of this procedure may be limited by low cellularity of the specimen, contamination of intestinal cells and unfeasibility of ancillary immunocy- tochemical procedures. The present study was to evaluate its usefulness in the diagnosis of neoplastic lesions.

Comparative Study of Cell Findings by Conventional Smear and Liquid-Based Cytology for Oral Exfoliative Cytology

Background: Liquid-based cytology (LBC) is a method of manufacturing cyto-diagnostic specimens. Improved accuracy is expected from standardizing specimen production and use of this method is rapidly spreading in oral cytology. On the other hand, LBC reportedly requires training to show peculiar cell findings compared to those of conventional smear cytology (CVC). Few studies have compared detailed cell findings for oral CVC and LBC. Objectives: The aim of this study was to compare cytological findings between CVC and LBC using cytomorphological image analysis. Materials and Methods: Cytological specimens were collected from 20 patients (negative for squamous neoplasia in 10, dysplasia in 5, squamous cell carcinoma in 5) and 5 controls of the tongue between January 2017 and December 2018. Two different preparation techniques were investigated cytomorphologically for CVC and LBC (BD CytorichTM). Results: LBC showed significantly higher cell numbers than CVC for all lesions. LBC-to-CVC ratio ranged from 9.52 (hyperkeratosis) to 1.87 (deep cells in oral squamous cell carcinoma (OSCC)). Nuclear area of normal, hyperkeratosis, and inflammation were significantly higher in LBC than those of CVC. Hyperchromasia was significantly more frequent with CVC than with LBC for hyperkeratosis, inflammation, dysplasia and OSCC. There was no significant difference in circularity between CVC and LBC among all lesions. Conclusion: Only one cytomorphological disadvantage was seen with LBC, in the form of decreased hyperchromasia. Further clarification of the advantages and disadvantages of LBC is needed, including management of precision and screening practices.

Fine-needle aspiration cytology of an intrathyroidal nodule diagnosed as squamous cell carcinoma: A case report

BACKGROUND Both squamous cell carcinoma(SCC)and papillary thyroid carcinoma(PTC)are common malignant tumors in the neck.However,seldom has SCC of the thyroid been diagnosed.Further,cytological features of SCC and PTC have rarely been reported.The significance of fine-needle aspiration cytology(FNAC)in the diagnosis of neck masses has been established.Herein,we present an exceedingly rare case of an intrathyroidal SCC diagnosed using FNAC,along with its cytological features.CASE SUMMARY A 66-year-old man presented with a left-sided neck mass.Ultrasound examination showed an ill-defined nodule.The appearance was hypoechoic with a few hyperechoic spots.FNAC of the left thyroid nodule was performed.A cellular smear was obtained,and it showed a large number of neoplastic cells with rich cytoplasm and poor cell adhesion.Tumor cell nuclei showed coarse nuclear chromatin and a few enlarged prominent nucleoli.An increased nuclear/cytoplasm ratio was observed.Thus,malignancy was diagnosed without a confirmed tumor type.Percutaneous tumor biopsy was performed to make a definite diagnosis.The tumor cells showed typical squamous cell characteristics.CONCLUSION Head and neck SCC and PTC have different cytologies.Measures are needed to ensure accurate diagnosis using FNAC.

Liquid Based Cytology of Cell Remnants in Needles Used for Breast Fine Needle Aspiration

This study examined cells contained in needles used for the collection of breast fine needle aspirates for the detection of malignant cells trapped in the needles. Remnants of cells contained in 50 needles used for the collection of scanty breast fine needle aspirates were examined by the liquid based cytology technique and compared with the conventional cytological technique of specimens in the corresponding syringes. The breast specimens were collected with clean sterile needles attached to the syringes. Smears were made and stained by the conventional method. The needles were removed from the syringes and a fixative was withdrawn into the syringes and the syringes were recapped with the needles. The fixative containing the specimen was then completely discharged into a centrifuge tube through the needles and treated by the liquid based cytology technique. The study revealed that cells were found trapped in all the needles used for the collection of breast FNA. 6% of them were positive for malignancy, similar to results obtained in the conventional method. Needles used for the collection of breast FNA should be examined before malignancy is completely ruled out particularly in extremely scanty specimens with a clinical suspicion of malignancy.

Five-Year Survivors of Non-Small Cell Lung Cancer Patients with Positive Pleural Lavage Cytology

Background: The pleural lavage cytology (PLC) for non-small cell lung cancer (NSCLC) patients has been reported as a significant prognostic factor. The aim of this study is to clarify the clinicopathological characteristics of 5-year survivors of patients with positive PLC. Methods: Among 401 resected NSCLC patients, 30 (7.48%) patients with positive PLC were reviewed retrospectively. Results: Only 7 of 30 patients (23.3%) survived more than 5-year. There were no differences in age, gender, histology, pT status and pN status between 5-year survivors and non-survivors. On the other hand, the serum carcinoembryonic antigen (CEA) level was significantly higher in non-survivors. Among these 5-year survivors, 4 of 7 patients died of NSCLC recurrences more than 5 years after surgery. Remaining 3 patients survived without cancer recurrences. Interestingly these 3 survivors had not received any adjuvant therapies after surgery. Conclusion: None of the 5-year survivor with positive PLC had high serum CEA level. Furthermore 5-year survival did not equal to cure in NSCLC patients with positive PLC.

Diagnostic value of negative enrichment and immune fluorescence in situ hybridization for intraperitoneal free cancer cells of gastric cancer

Objective:To explore the intraperitoneal free cancer cell(IFCC)detection value of negative enrichment and immune fluorescence in situ hybridization(NEimFISH)on chromosomes(CEN)8/17.Methods:To verify the reliability of NEimFISH,29 gastric cancer tumors,their adjacent tissues and greater omental tissues were tested.Our study then included 105 gastric cancer patients for IFCC.We defined patients as IFCC-positive if a signal was detected,regardless of the detailed cancer cell numbers.A comparison of clinicopathological features was conducted among IFCC groups.We also compared the diagnosis value and peritoneal recurrence predictive value among different detection methods.The comparison of IFCC number was also conducted among different groups.Results:A cutoff of 2.5 positive cells could distinguish all benign tissue samples and 97%of malignant tissue samples in our study.Compared to intestinal gastric cancer,patients with diffuse gastric cancer tended to have more IFCCs(6 vs.4,P=0.002).The IFCC counts were often higher in the lymphovascular invasion positive group than negative group(3 vs.1,P=0.022).All IFCC samples that were considered positive using conventional cytology were also found to be positive using NEimFISH.When compared to conventional cytology and paraffin pathology,NEimFISH had a higher IFCC positive rate(68.9%)and higher one-year peritoneal recurrence predictive value with area under the curve(AUC)of 0.922.Conclusions:Gastric cancer could be effectively diagnosed by NEimFISH.The IFCC number found using NEimFISH on CEN8/17 is closely associated with Lauren type and vascular invasion of cancer.NEimFISH is a reliable detection modality with a higher positive detection rate,higher one-year peritoneal recurrence predictive value and quantitative features for IFCC of gastric cancer.

Detection methods and clinical significance of free peritoneal tumor cells found during colorectal cancer surgery

Peritoneal washing is now part of the standard clinical practice in several abdominal and pelvic neoplasias. However, in colorectal cancer surgery, intra-peritoneal free cancer cells(IFCC) presence is not routinely investigated and their prognostic meaning is still unclear. When peritoneal washing results are positive for the presence of IFCC a worse outcome is usually expected in these colorectal cancer operated patients, but it what is not clear is whether it is associated with an increased risk of local recurrence. It is authors' belief that one of the main reasons why IFCC are not researched as integral part of the routine staging system for colon cancer is that there still isn't a diagnostic or detection method with enough sensibility and specificity. However, the potential clinical implications of a routine research for the presence IFCC in colon neoplasias are enormous: not only to obtain a more accurate clinical staging but also to offer different therapy protocols, based on the presence of IFCC. Based on this, adjuvant chemotherapy could be offered to those patients found to be positive for IFCC; also, protocols of proactive intraperitoneal chemotherapy could be applied. Although presence of IFCC appears to have a valid prognostic significance, further studies are needed to standardize detection and examination procedures, to determine if there are and which are the stages more likely to benefit from routine search for IFCC.

Epidermal growth factor receptor mutation analysis in cytological specimens and responsiveness to gefitinib in advanced non-small cell lung cancer patients

Background： Epidermal growth factor receptor（EGFR） mutation is the key predictor of EGFR tyrosine kinase inhibitors（TKIs） efficacy in non-small cell lung cancer（NSCLC）. We conducted this study to verify the feasibility of EGFR mutation analysis in cytological specimens and investigate the responsiveness to gefitinib treatment in patients carrying EGFR mutations.Methods： A total of 210 cytological specimens were collected for EGFR mutation detection by both direct sequencing and amplification refractory mutation system（ARMS）. We analyzed EGFR mutation status by both methods and evaluated the responsiveness to gefitinib treatment in patients harboring EGFR mutations by overall response rate（ORR）, disease control rate（DCR） and progression free survival（PFS）.Results： Of all patients, EGFR mutation rate was 28.6%（60/210） by direct sequencing and 45.2%（95/210） by ARMS（P〈0.001） respectively. Among the EGFR wild type patients tested by direct sequencing, 26.7% of them were positive by ARMS. For the 72 EGFR mutation positive patients treated with gefitinib, the ORR, DCR and median PFS were 69.4%, 90.2% and 9.3 months respectively. The patients whose EGFR mutation status was negative by direct sequencing but positive by ARMS had lower ORR（48.0% vs. 80.9%, P=0.004） and shorter median PFS（7.4 vs. 10.5 months, P=0.009） as compared with that of EGFR mutation positive patients by both detection methods. Conclusions： Our study verified the feasibility of EGFR analysis in cytological specimens in advanced NSCLC. ARMS is more sensitive than direct sequencing in EGFR mutation detection. EGFR Mutation status tested on cytological samples is applicable for predicting the response to gefitinib. Abundance of EGFR mutations might have an influence on TKIs efficacy.

Effects of High Temperature Stress on Microscopic and Ultrastructural Characteristics of Mesophyll Cells in Flag Leaves of Rice

The microscopic and ultrastructural characteristics of mesophyll cells in flag leaves of two rice lines （a thermosensitive line 4628 and a thermo-resistant line 996） under high temperature stress （37℃ during 8：00-17：00 and 30℃ during 17：00-8：00） were investigated using an optical and a transmission electron microscopy. The membrane permeability and malondialdehyde content increased under the high temperature stress, and the increase of both variables was greater in the line 4628 than in the line 996. Under the high temperature stress, the line 996 showed tightly arranged mesophyll cells in flag leaves, fully developed vascular bundles and some closed stomata, whereas the line 4628 suffered from injury because of undeveloped vascular bundles, loosely arranged mesophyll cells and opened stomata. The mesophyll cells in flag leaves of the line 4628 were severely damaged under the high temperature stress, i.e. the chloroplast envelope became blurred, the grana thylakoid layer was arranged loosely and irregularly, the stroma layer disappeared, many osmiophilic granules appeared within the chloroplast, the outer membrane of mitochondria and the nucleus disintegrated and became blurred, the nucleolus disappeared, and much fibrillar-granular materials appeared within the nucleus. In contrast, the mesophyll cells in flag leaves of the line 996 maintained an intact ultrastructure under the high temperature stress. From these results, it is suggested that the ultrastructural modification of the cell membrane system is the primary plant response to high temperature stress and can be used as an index to evaluate the crop heat tolerance.

Improved Detection of Cervical Cancer and High Grade Neoplastic Lesions by a Combination of Conventional Cytology and DNA Automated Image Cytometer

OBJECTIVE: To reduce false-negative rates of population based cervical screening programs employing conventional cytology in combination with automated DNA image cytometer. METHODS: Involved cervical samples from a total of 3603 women were taken by a cervix brush and then placed into a fixative solution. The cells were separated from mucus by mechanical and chemical treatment after which they were deposited onto microscope slides by a cytospin. Two slides were prepared from each case;one slide was stained by Papanicolaou stain for conventional cytology examination, while the other slide was stained by a DNA specific and stoichiometric stain. The latter slide was used to determine the relative amount of DNA in the cell nuclei in order to assess the ploidy status of the epithelial cells. Enrolled in the study, 157 women were followed by colposcopy examination where punch biopsies were taken from the visible lesions or from suspicious areas. The results of the conventional cytology were then compared to the DNA image cytometer for all samples. RESULTS: Histopathology diagnosed 51 lesions from the 132 biopsied cases as CIN2 or higher, including 27 CIN2, 16 CIN3 and 8 invasive cancers. Conventional cytology correctly identified 29 of the 51 high grade CIN and in-vasive cancer, while DNA image cytometer correctly identified 38 high grade CIN and invasive cancer using the crite-rion that at least three cells were found on the slide that contained DNA amount in excess of 5c. 42 out of 51 high grade CIN and invasive cancer were found by conventional cytology in combination with DNA image cytometer. Sensitivities were 56.8%, 74.5% and 82.4%, while specificities were 86.2%, 81.5% and 81.5% in conventional cytology, DNA image cytometer and combination both cytology and DNA image cytometer respectively. CONCLUSION: The study demon-strated that screening for high grade neoplastic lesions and cervical cancer by DNA image cytometer or combination of conventional cytology and DNA image cytometer is more sensitive than conventional screening approach.

Pemphigus vulgaris macroscopically and cytologically resembling oral squamous cell carcinoma

We describe the clinical, macroscopic, cytological, histopathological, immunohistochemical, serodiagnostic and aspects of pemphigus vulgaris (PV) in the oral gingiva that clinically mimicked oral squamous cell carcinoma (OSCC) in a 57-year-old Japanese man. He developed slight haphalgesia of the buccal gingiva around teeth numbers 18 and 19 2 years ago. A dentist diagnosed intractable ulcer, but the patient ignored the condition for about 2 years until a sharp pain in the gingiva worsened. He consulted an otolaryngologist, who referred the patient to our hospital under a cytological diagnosis of OSCC. An oral examination revealed several extensive painful erosions/ ulcers from the buccal and lingual gingiva around teeth numbers 18 to 21 to the distal alveolar mucosa of no. 18 and the buccal and lingual gingiva around tooth number 31. A presumptive diagnosis of PV with dysplastic changes was determined from cytological smears. The cytological Nikolsky test was positive. The diagnosis of PV was confirmed from clinical and histopathological findings of a biopsy specimen obtained from the perilesional site. Although the definitive diagnosis of PV required only 2 weeks after this patient presented at our hospital, 2 years had elapsed since the onset of oral lesions.

Comparison of peritoneal free gastric cancer cells' detecting rates between laparoscopically assisted and open radical gastrectomy

Objective： To compare laparoscopic gastrectomy and conventional surgery on the dissemination and seeding of tumor cells. Methods：Intraoperative peritoneal lavage cytologic examination was performed in 65 patients with gastric cancer, during laparoscopic gastrectomy （n = 34） and conventional surgery （n = 31）. Cytology was examined twice, immediately after opening the peritoneal cavity and just before closing the abdomen. Saline was poured into the peritoneal cavity, and 100 ml fluid was retrieved after irrigation. Laparoscopic instruments were lavaged after surgery with 100 ml saline. Carbon dioxide （COz） was derived through the trocar side orifice after pneumoperitoneum during laparoscopic gastrectomy and filtered through 100 ml saline. Cytologic examination of the filtrate was performed after the filtration process. Results： The incidence of positive cytology during laparoscopic surgery was 32.26% in the preoperative lavage and 22.58% in the postoperative lavage. The incidence of positive cytology during conventional surgery was 41.18% before lavage and 26.47% after lavage. Only one positive cytology was detected in the CO2 filtrate gas. The incidence of positive cytology in the lavage of the instruments during laparoscopic surgery was 6.45 %. Conclusion： During gastric laparoscopic surgery, CO2 pneumoperitoneum does not affect tumor cell dissemination and seeding. In this study, laparoscopic techniques used in gastric cancer surgery were not associated with a higher risk for intraperitoneal dissemination of cancer cells than the conventional surgery.

Cytological studies on neurosecretory cell of the X-organ in Scylla serrata

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Renal Cell Carcinoma Associated with Xp11.2 Translocation/TFE3 Gene Fusion: A Case Report with Immunohistochemical and Cytological Features

Gene fusions involving two of the MiT subfamily factors, such as TFE3, TFEB, TFC and MiTF, have been identified in renal cell carcinoma (RCC). Xp11.2 translocation RCC is a rare pediatric neoplasm that harbors gene fusions involving TFE3, which plays an important role in cell proliferation and survival. We herein present a case of RCC associated with Xp11.2 translocation/TFE3 gene fusion in a 14-year-old Japanese boy presenting gross hematuria and body weight loss. The tumor was characterized by histopathology, cytology and TFE3-immunohistochemistry/immunocytochemistry. Knowledge of distinctive morphological and immunostaining features of this tumor can help to accurately diagnose this rare subset of translocation associated RCC in routine pathological diagnostic procedures.

DIAGNOSTIC VALUE OF DENSITY GRADIENT CENTRIFU-GATION FOR EXFOLIATIVE TUMOR CELLS IN MALIG-NANT PLEURAL EFFUSIONS

Objective To find out a specific method for diagnosis of malignant pleural effusions (MPEs)with higher sensitivity and practicality. Methods The diagnosis of MPEs were made using density gradient cen-trifugation(DGC) , smear cytologic examination( SCE) and pleural needle biopsy (PNB). Comparisons between these results and those of benign pleural effusions were also made. Results The positive rates of DGC,SCE and PNB for diagnosing MPEs were 94. 3% ,62. 9% and 44. 6% , respectively, and the positive rate of SCE combined with PNB for diagnosing MPEs was 73. 2%. The positive rate of the exfoliative tumor cells ( ETCs ) by DGC was much higher than that of SCE or/and PNB with no false-positive. Conclusion The ETCs isolated by DGC from the MPEs is quite specific for the diagnosis of malignant tumors with higher sensitivity and practicality in clinico-patho-logical practice.

Clinical significance of telomerase activity in peritoneal lavage fluid from patients with gastric cancer and its relationship with cellular proliferation

AIM： To evaluate the efficacy of telomerase activity assay and peritoneal lavage cytology （PLC） examination in peritoneal lavage fluid for the prediction of peritoneal metastasis in gastric cancer patients, and to explore the relationship between telomerase activity and proliferating cell nuclear antigen expression.METHODS： Telomeric repeated amplification protocol （TRAP）-enzyme-linked immunosorbent assay （ELISA） was performed to measure the telomerase activity in 60 patients with gastric cancer and 50 with peptic ulcer. PLC analysis of the 60 patients with gastric cancer was used for comparison. The proliferating cell nuclear antigen （PCNA） in gastric carcinoma was immunohistochemically examined.RESULTS： The telomerase activity and PLC positive rate in peritoneal lavage fluid from patients with gastric cancer was 41.7% （25/60）, and 25.0% （15/60）, respectively. The positive rate of telomerase activity was significantly higher than that Qf PLC in the group of pT, （15/16 vs 9/16, P 〈 0.05）, P1-3 （13/13 vs 9/13, P 〈 0.05） and diffuse type （22/42 vs 13/42, P 〈 0.05）. The patients with positive telomerase activity, peritoneal metastasis, and serosal invasion had significantly higher levels of average PCNA proliferation index （PI）, （55.00 ± 6.59 vs 27.43 ± 7.72, 57.26 ±10.18 vs 29.15 ±8.31, and 49.82 ± 6.74 vs 24.65 ±7.33, respectively, P 〈 0.05）.CONCLUSION： The TRAP assay for telomerase activity is a useful adjunct for cytologic method in the diagnosis of peritoneal micrometastasis and well related to higher proliferating activity of gastric cancer. The results of this study also suggest a promising future therapeutic strategy for treating peritoneal dissemination based on telomerase inhibition.

Nasal cytology with emphasis on mast cells can improve the diagnosis and treatment of chronic rhinosinusitis

Objective:Chronic rhinosinusitis (CRS) involves inflammation of the nasal and para-nasal mucosa.Due to its heterogeneous nature,unknown pathogenesis,and high recurrence rate,effective treatment is difficult.Nasal cytology is presently not a part of the routine diagnosis or treatment decision for CRS.Data sources:A literature search was performed for published papers in English between January 1990 and June 2019 using MEDLINE.Study sdection:Terms used were chronic rhinosinusids,eosinophils,etiology,immunopathology,inflammation,mast cells,nasal cytology,polyps,and treatment.Both reviews and original articles were collected and studied.Results:There is no standard nasal fluid,mucus sampling,or staining techniques for identifying inflammatory cell types.Results were divergent from different countries.Moreover,the main focus of these papers on the cells in nasal washings was eosinophils,with infrequent mentioning of other cell types that may imply different etiology and pathology.The heterogeneous cell profile of CRS and the role of mast cells have been unappreciated due to the lack of specific immunohistochemical technique or study of its unique mediators.Conclusions:Nasal cytology could help distinguish the type and the activation state of inflammatory cells.Thus it can help in providing a clearer picture of CRS pathogenesis,identifying different patient groups,and developing effective treatments.

Quantification of corneal neovascularization after ex vivo limbal epithelial stem cell therapy

AIM: To assess cultured limbal epithelial stem cell transplantation in patients with limbal stem cell deficiency by analyzing and quantifying corneal neovascularization.METHODS: This retrospective, interventional case series included eight eyes with total limbal stem cell deficiency. Ex vivo limbal epithelial stem cells were cultured on human amniotic membrane using an animalfree culture method. The clinical parameters of limbal stem cell deficiency, impression cytology, and quantification of corneal neovascularization were evaluated before and after cultured limbal stem cell transplantation. The area of corneal neovascularization,vessel caliber(VC), and invasive area(IA) were analyzed before and after stem cell transplantation by image analysis software. Best-corrected visual acuity(BCVA),epithelial transparency, and impression cytology were also measured.RESULTS: One year after surgery, successful cases showed a reduction(improvement) of all three parameters of corneal neovascularization [neovascular area(NA), VC, IA], while failed cases did not. NA decreased a mean of 32.31%(P =0.035), invasion area29.37%(P =0.018) and VC 14.29%(P =0.072). BCVA improved in all eyes(mean follow-up, 76 ±21mo).Epithelial transparency improved significantly from 2.00 ±0.93 to 0.88±1.25(P =0.014). Impression cytology showed that three cases failed after limbal epithelial stem cell therapy before 1y of follow-up.CONCLUSION: This method of analyzing andmonitoring surface vessels is useful for evaluating the epithelial status during follow-up, as successful cases showed a bigger reduction in corneal neovascularization parameters than failed cases. Using this method,successful cases could be differentiated from failed cases.