Isolation and Culture of Human Meibomian Gland Ductal Cells.

Purpose: Hyperkeratinization of meibomian gland (MG) ducts is currently recognized as the primary pathologic mechanism of meibomian gland dysfunction (MGD). This research figured out a method to isolate the MG ducts and established a novel system to culture the human meibomian gland ductal cells (HMGDCs) for investigating the process of MGD. Methods: The MG ducts were obtained from the eyelids of recently deceased donors and subjected to enzymatic digestion. The acini were then removed to isolate independent ducts. These MG ducts were subsequently cultivated on Matrigel-coated wells and covered with a glass plate to obtain HMGDCs. The HMGDCs were further cultivated until passage 2, and when they reached 60% confluence, they were treated with IL-1ß and rosiglitazone for a duration of 48 hours. Immunofluorescence staining and Western blot techniques were employed to identify ductal cells and analyze the effects of IL-1ß on HMGDCs in an in vitro setting. Results: Ophthalmic micro-forceps and insulin needles can be employed for the purpose of isolating ducts. Within this particular culture system, the rapid expansion of HMGDCs occurred in close proximity to the duct tissue. MG ducts specifically expressed keratin 6 (Krt6) and hardly synthesized lipids. Furthermore, the expression of Krt6 was significantly higher (P < 0.0001) in HMGDCs compared to human meibomian gland cells. Upon treatment with IL-1ß, HMGDCs exhibited an overexpression of keratin 1, which was effectively blocked by the administration of rosiglitazone. Conclusions: The present study successfully isolated human MG ducts and cultured HMGDCs, providing a valuable in vitro model for investigating the mechanism of MGD. Additionally, the potential therapeutic efficacy of rosiglitazone in treating hyperkeratinization of ducts in patients with MGD was identified.

Amniotic membrane transplantation combined with conjunctival flap covering surgery for the treatment of corneal perforations in fungal keratitis.

Purpose: To investigate the efficiency of amniotic membrane transplantation (AMT) combined with conjunctival flap covering surgery (CFCS) for patients with corneal perforations in fungal keratitis (FK). Methods: In this non-comparative, retrospective case series, 16 participants of corneal perforation in FK were successfully treated by a combination of multilayer AMT and bipedicle conjunctival flap with partial tenon's capsule. Corneal healing, recurrence of FK, visual acuity, and relevant complications were reported as outcome measures. Results: Sixteen patients (13 male, 3 female) had a mean age of 58.8 ± 10.3 (range 29-72) years. The mean diameter of corneal perforation was 1.9 ± 0.7 (range 0.5-2.8) mm. Corneal perforations healed and all the patients preserved their eyeballs. During the 11.0 ± 4.4 (range 6-18) months of follow-up, there was no recurrence of FK in any of these cases. Visual acuity improved in 15 eyes (93.8 %) and remained unchanged in 1 patient (6.3 %) who had no light perception when first admitted. All 6 patients who accepted secondary keratoplasty showed improved best corrected visual acuity of more than 4 lines. The most frequently found fungi were Aspergillus species (6 of 16, 37.5 %) and Fusarium species (4 of 16, 25.0 %), followed by 1 Scedosporium apiospermum (1 of 16, 6.3 %). Conclusions: Combination AMT with CFCS is a safe and effective surgery for patients with corneal perforations in FK, particularly where eye banks and fresh corneas are not available. This surgery could preserve the integrity of the eyeball and avoid the recurrence of FK. Besides, it provides a greater opportunity for further optical keratoplasty.

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.

Conjunctival Limbal Autograft Combined with Amnion-Assisted Conjunctival Epithelial Redirection for Unilateral Total Limbal Stem Cell Deficiency after Severe Chemical Burn.

(1) Background: To evaluate the efficacy of conjunctival limbal autograft (CLAU) combined with the amnion-assisted conjunctival epithelial redirection (ACER) procedure for patients with unilateral total limbal stem cell deficiency (LSCD) caused by severe chemical burn. (2) Methods: A retrospective interventional case series of unilateral total LSCD after chemical burn who underwent CLAU combined with ACER surgery between September 2021 and July 2023 was collected. Outcome measures included epithelialization of the cornea with donor limbus-derived epithelium, best corrected visual acuity (BCVA), and complications. (3) Results: Nine males and one female were included in this study. The mean age was 40.9 ± 9.63 (range, 26 to 55) years. The average duration between injury and CLAU combined with the ACER procedure was 7.67 ± 3.97 (range, 4 to 18) months. All patients achieved corneal epithelialization and improved BCVA. Postoperative complications occurred in four cases, including delayed corneal epithelial healing in one case, delayed amniotic membrane dissolution and detachment in two cases, and recurrence of symblepharon in one case. No complications were noted in the healthy donor eyes. (4) Conclusions: CLAU combined with ACER is a safe and effective treatment for unilateral total LSCD caused by severe chemical burn. This combined surgery restores visual function for patients with corneal blindness caused by chemical burn, reducing the burden on the families and society.

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±77.1 µm, ranging from 350 to 535 µm. The mean corneal thickness was 597.4±117.3 µm, ranging from 458 to 851 µm. 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.

The inhibition of p38 MAPK blocked inflammation to restore the functions of rat meibomian gland epithelial cells.

Meibomian glands (MGs) are vital for ocular surface health. However, the roles of inflammation in the progression of meibomian gland dysfunction (MGD) are largely unknown. In this study, the roles of the inflammation factor interleukin-1ß (IL-1ß) via the p38 mitogen-activated protein kinases (MAPK) signaling pathway on rat meibomian gland epithelial cells (RMGECs) were explored. Eyelids from adult rat mice at 2 months and 2 years of age were stained with specific antibodies against IL-1ß to identify inflammation levels. RMGECs were exposed to IL-1ß and/or SB203580, a specific inhibitor of p38 MAPK signaling pathway, for 3 days. Cell proliferation, keratinization, lipid accumulation, and matrix metalloproteinases 9 (MMP9) expression were evaluated by MTT assay, polymerase chain reaction (PCR), immunofluorescence staining, apoptosis assay, lipid staining, and Western blot analyses. We found that IL-1ß was significantly higher in the terminal ducts of MGs in rats with age-related MGD than in young rats. IL-1ß inhibited cell proliferation, suppressed lipid accumulation and peroxisome proliferator activator receptor γ (PPARγ) expression, and promoted apoptosis while activating the p38 MAPK signaling pathway. Cytokeratin 1 (CK1), a marker for complete keratinization, and MMP9 in RMGECs were also up-regulated by IL-1ß. SB203580 effectively diminished the effects of IL-1ß on differentiation, keratinization, and MMP9 expression by blocking IL-1ß-induced p38 MAPK activation, although it also inhibited cell proliferation. The inhibition of the p38 MAPK signaling pathway blocked IL-1ß-induced differentiation reduction, hyperkeratinization, and MMP9 overexpression of RMGECs, which provides a potential therapy for MGD.

Ductal Hyperkeratinization and Acinar Renewal Abnormality: New Concepts on Pathogenesis of Meibomian Gland Dysfunction.

Meibomian gland dysfunction (MGD) is a functional and morphological disorder of the meibomian glands which results in qualitative or quantitative alteration in meibum secretion and is the major cause of evaporative dry eye (EDE). EDE is often characterized by tear film instability, increased evaporation, hyperosmolarity, inflammation, and ocular surface disorder. The precise pathogenesis of MGD remains elusive. It has been widely considered that MGD develops as a result of ductal epithelial hyperkeratinization, which obstructs the meibomian orifice, halts meibum secretion, and causes secondary acinar atrophy and gland dropout. Abnormal self-renewal and differentiation of the acinar cells also play a significant role in MGD. This review summarizes the latest research findings regarding the possible pathogenesis of MGD and provides further treatment strategies for MGD-EDE patients.

Characteristics of In Vitro Culture and In Vivo Confocal Microscopy in Patients with Fungal Keratitis in a Tertiary Referral Hospital in Central China.

PURPOSE: To investigate the characteristics of in vitro culture and in vivo confocal microscopy (IVCM) in patients with fungal keratitis (FK) presented in a tertiary referral hospital in central China. METHODS: In this noncomparative retrospective study, patients with the diagnosis of FK between October 2021 and November 2022 were reviewed. An IVCM and fungal culture (corneal scraping specimens) were performed, and the characteristics were analyzed. RESULTS: During October 2021 and November 2022, 85 patients were diagnosed with FK. From 63 culture-positive cases, 8 species of fungus were identified. The proportions of isolated fungal species were Fusarium and Aspergillus equally accounting for 33.3% (21 of 63), Alternaria 9.5% (6 of 63), Curvularia 6.3% (4 of 63), Scedosporium apiospermum 6.3% (4 of 63), Paecilomyces lilacinus 3.2% (2 of 63), Exserohilum 3.2% (2 of 63), and Candida 4.8% (3 of 63), respectively. In positive culture cases, IVCM was found to be positive for hyphae or spores in 61 of 63 patients (96.8%). Different fungal species had a variety of cultural characteristics and IVCM manifestations. CONCLUSIONS: In a tertiary referral hospital in central China, Fusarium species, Aspergillus species, and Alternaria species were the 3 most common isolated fungal pathogens, and the proportion of Aspergillus species was significantly higher than that in other regions of China. Careful lesion depth examination by IVCM and OCT should be taken before lamellar keratoplasty to avoid postoperative recurrence. Identifying the IVCM image and culture characteristics will facilitate rapid diagnosis and proper treatment, but IVCM cannot yet replace fungal cultures to distinguish between different fungal species.

Botulinum Neurotoxin Type a Injection Combined with Absorbable Punctal Plug Insertion: An Effective Therapy for Blepharospasm Patients with Dry Eye.

Blepharospasm patients often have dry eye manifestations. Botulinum neurotoxin type A (BoNT-A) injection has been the main management for blepharospasm and absorbable punctal plug (APP) insertion is shown to be effective in the treatment of dry eye. However, there have been no studies investigating the combined treatment of BoNT-A and APP in blepharospasm patients with dry eye. In this retrospective study, 17 blepharospasm patients with dry eye treated by BoNT-A injection and 12 receiving BoNT-A plus APP treatment were enrolled. The efficacy was evaluated according to the Jankovic rating scale, Ocular Surface Disease Index (OSDI), fluorescein staining (FL), fluorescein tear break-up time (FBUT) and Schirmer I test (SIT). Both BoNT-A and BoNT-A+APP treatment effectively reduced the functional impairment of blepharospasm. At baseline, all the patients had high OSDI scores (BoNT-A group: 82.48 ± 7.37, BoNT-A+APP group: 78.82 ± 4.60, p = 0.112), but relatively low degrees of FL (BoNT-A group: 3.18 ± 1.01, BoNT-A+APP group: 3.50 ± 1.24, p = 0.466), FBUT (BoNT-A group: 1.71 ± 0.77, BoNT-A+APP group: 2.17 ± 0.58, p = 0.077) and SIT (BoNT-A group: 2.53 ± 0.99, BoNT-A+APP group: 3.17 ± 1.23, p = 0.153). After treatment, OSDI, FL, FBUT and SIT were all obviously restored in the two groups. When comparing the changing rates, only OSDI (BoNT-A group: -52.23% ± 15.57%, BoNT-A+APP group: -61.84% ± 9.10%, p = 0.047) and FL (BoNT-A group: -22.55% ± 25.98%, BoNT-A+APP group: -41.94% ± 14.46%, p = 0.016) showed significant differences between the two groups. This study suggests that OSDI is not applicable in the diagnosis of dry eye among blepharospasm patients. For blepharospasm patients with severe dry eye symptoms, especially those with fluorescein staining in the cornea, the combined treatment of BoNT-A and APP is more effective than using BoNT-A alone.

Sandwich (Amnion/Conjunctival-Limbal Autograft/Amnion) Transplantation for Recurrent Pterygium with Restrictive Strabismus.

(1) Background: This study aimed to evaluate the clinical outcome of Sandwich (Amnion/Conjunctival-Limbal Autograft/Amnion) transplantation for recurrent pterygium with restrictive strabismus. (2) Methods: This retrospective study included 11 eyes in 11 patients diagnosed with recurrent pterygium with restrictive strabismus who received sandwich transplantation. The outcomes were measured by pterygium recurrence, best-corrected visual acuity, esotropia (prism diopters), and treatment complications. (3) Results: Eleven patients (six males, five females) had a mean age of 60.5 (range 36-80) years. The previously received pterygium excision surgery number was 1.8 ± 1.02 (range 1-4). The mean follow-up period was 19.9 ± 8.41 (range 12-36) months. All patients had a restriction of abduction in the previously operated eye, causing esotropia in the primary position. Pre-operative esotropia was 17.2 (range 10-30) prims diopter (PD). Five eyes (45.5%) had symblepharon before surgery. All patients were orthotropic until the last follow-up. Symblepharon was released in all eyes. Free ocular motility was present in all eyes. No donor site scar formation, scleral melt, or corneal ulcer was noted. (4) Conclusions: Sandwich transplantation for recurrent pterygium with restrictive strabismus is safe and effective.

Topical Chemotherapy for Treating Ocular Surface Squamous Neoplasia with a Combination of Interferon α-2b and 5-Fluorouracil.

INTRODUCTION: This study evaluated the efficacy of combined interferon α-2b (IFNα2b) and 5-fluorouracil (5-FU) as primary treatment for ocular surface squamous neoplasia (OSSN). METHODS: In this retrospective study, 27 eyes with OSSN followed by topical application of combined IFNα2b and 5-FU were examined. Reported outcome measures were tumor response, visual acuity, time to complete resolution, recurrence and treatment complications. RESULTS: Twenty-six patients (17 male, 9 female) had a mean age of 63.9 (median, 67; range 22-83) years. Complete tumor response was observed in 24 eyes (88.9%). Three eyes (11.1%) showed partial response to the chemotherapy agents and later underwent surgical tumor removal. The median time to complete resolution was 6 (mean, 6.1; range, 3-11) weeks. Of these, the patients received between one to three cycles of 5-FU therapy (median, 2; mean, 1.8). Complications noted were transient irritation at 5-FU cycle (11 eyes, 40.7%). There was no tumor recurrence at mean follow-up of 16.1 (median, 12; range 6-38) months. CONCLUSIONS: Combination therapy of IFNα2b and 5-FU was a safe and effective treatment, inducing a short duration of administration and low recurrence rate for OSSN. TRIAL REGISTRATION: Retrospectively registered, UHCT22048.

Amniotic membrane transplantation with topical interferon alfa-2b after excision of ocular surface squamous neoplasia.

To evaluate the outcome of amniotic membrane transplantation (AMT) after tumor excision followed by topical interferon alfa-2b (IFNα2b) drops for primary ocular surface squamous neoplasia (OSSN). Twelve eyes of 12 patients with a mean age of 66±10y were included. The average follow-up was 23±10mo. All 12 patients had limbal involvement. Smooth ocular surface and transparent cornea were achieved in all cases. No sign of inflammation, neovascularization, symblepharon or recurrence was noted at the last follow-up. We conclude that AMT with topical IFNα2b drops restores a healthy ocular surface in OSSN without recurrence.

Characterization of Ex Vivo Expanded Oral Mucosal Epithelium Cells on Acellular Porcine Corneal Stroma for Ocular Surface Reconstruction.

PURPOSE: To ex vivo expand oral mucosal epithelium cells (OMECs) on acellular porcine corneal stroma (APCS) without using feeder cells and serum and to compare the morphologic and phenotypic characteristics of cultured oral cells on APCS to those of cells on deluded human amniotic membrane (HAM). METHODS: SD rat oral mucosal biopsies were cultured on APCS and HAM. Reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to analyze the characterization of stem cells and epithelial differentiation of the outgrowth products. RESULTS: Stratified and optimal transplantable OMECs were obtained after being cultured three to four weeks. Both RT-PCR and immunohistochemistry showed that cultured OMECs expressed markers of epithelial differentiation cytokeratin K3 and epithelial stem cell markers of p63 and ABCG2. CONCLUSIONS: OMECs can be successfully cultured on APCS without using xenobiotic feeder cells and serum. Characterization showed that these sheets retain the morphologic and phenotypic characteristics of OMECs within differentiated cells and stem cells. The optimal transplantable sheets can prove to be particularly beneficial to both bilateral limbal stem cell deficiency and deep corneal lesions.

Small-incision lenticule extraction versus femtosecond lenticule extraction for myopic: a systematic review and Meta-analysis.

AIM: To examine differences in efficacy, accuracy, safety, aberrations and corneal biomechanical between Small incision lenticule extraction (SMILE) and femtosecond lenticule extraction (FLEx) for myopia. METHODS: Comprehensive studies were conducted on the PubMed, MEDLINE, EMBASE, and Cochrane Controlled Trials Register before 31 July, 2015. Meta-analyses were performed on the primary outcomes [loss of ≥2 lines of corrected distance visual acuity (CDVA), uncorrected distance visual acuity (UDVA) ≥20/20, spherical equivalent (SE) within ±0.50 diopters (D), final refractive SE], secondary outcomes were high-order aberrations (HOAs) and corneal biomechanical [central corneal thickness (CCT), corneal hysteresis (CH) and corneal resistance factor (CRF)]. RESULTS: Seven trials describing a total of 320 eyes with myopia were included in this Meta-analysis. No significant differences were found in the efficacy [UDVA weighted mean difference (WMD) -0.01; 95%CI: -0.04 to 0.01; P=0.37, UDVA ≥20/20, OR 1.49; 95%CI: 0.78 to 2.86; P=0.23], accuracy (SE WMD -0.03; 95%CI: -0.12 to 0.07; P=0.58, SE within ±0.5 D OR 1.25; 95%CI: 0.34 to 4.65; P=0.74), HOAs (WMD -0.04; 95%CI: -0.09 to 0.01; P=0.14) and CCT WMD 1.83; 95%CI: -7.07 to 10.72; P=0.69, CH WMD -0.01; 95%CI: -0.42 to 0.40; P=0.97, CRF WMD 0.17; 95%CI: -0.33 to 0.67; P=0.50) in the last fellow-up. But for safety, FLEx may achieve fewer CDVA lost two or more two lines (OR 11.11; 95%CI: 1.27 to 96.86; P=0.03) than SMILE, however CDVA (WMD 0.00; 95%CI: -0.03 to 0.02; P=0.77) is similar. CONCLUSION: SMILE and FLEx are comparable in terms of both efficacy, accuracy, aberrations and corneal biomechanical measures in the follow-up, but FLEx seems to be better in safety measures. The results should be interpreted cautiously since relevant evidence is still limited, although it is accumulating. Further large-scale, well-designed randomized controlled trials are urgently needed.

Retinal nerve fiber layer thickness changes in obstructive sleep apnea syndrome: a systematic review and Meta-analysis.

AIM: To evaluate the retinal nerve fiber layer (RNFL) thickness changes in patients with obstructive sleep apnoea syndrome (OSAS), and detect possible prevalence of glaucoma in this population. METHODS: Comprehensive studies were conducted on the Cochrane Library, PubMed and Embase through March, 2015. Only studies that fit the selection criteria about RNFL and OSAS would be included. For the measures, we calculated the 95% confidence interval (CI) and weighted mean differences (WMD). The systematic review and Meta-analysis was performed by RevMan 5.2 software. RESULTS: Nine case-control studies were analyzed containing a total of 1086 cases and 580 controls. Average RNFL thickness in OSAS was reduced significantly compared with healthy controls in random effects model (WMD=-2.56, 95% CI: -4.82 to -0.31, P =0.003, I2=57%). A significant RNFL thickness reduction were found between the two groups in inferior quadrant (WMD=-3.11, 95% CI: -5.53 to -0.69, P=0.01), superior quadrant (WMD=-2.37, 95%CI: -4.7 to 0.04, P=0.05). In nasal quadrant (WMD=-2.54, 95% CI: -6.53 to 1.45, P=0.21) and temporal quadrant (WMD=-1.26, 95% CI: -2.19 to 0.47, P=0.15) there was no difference of RNFL thickness between the two groups. CONCLUSION: The results show that RNFL thickness is lower in patients with moderate or severe OSAS than in normal subjects or patients with mild OSAS according to the nine homogeneity studies.

[Development and application of Jingcen DY-1 spraying tanker for Oncomelania hupensis snail control].

OBJECTIVE: To develop a Jingcen DY-1 type spraying tanker for Oncomelania hupensis snail control and evaluate its effect of field application as well as the cost. METHODS: The currently available tractor was used as a vector, and the mechanical and electrical equipments and containers were integrated with shafts, pipelines and electric lines to produce a spraying tanker for snail control, with the functions of carrying people and molluscicides, generating electric power and getting water, mixing stocking solutions, adjusting molluscicide solutions evenly, and spraying drugs. The volume of the molluscicide solution, flow rate of water injection, and the flow rate, range and advance speed of the spray gun were tested, and the solution concentrations of molluscicide in the tanker and at the muzzle of the spray gun at different time were detected. Meanwhile, the molluscicidal effect and cost of the spraying tanker were analyzed by the field test. RESULTS: The volume of the liquid storage pot of the Jingcen DY-1 type spraying tanker was 1 800 L, the flow rate of water injection was 400 L/min, the flow rate and the spray range of the standard spray gun were 110-200 L/min and 19.70-23.50 m, respectively, the efficiency of drug spraying of the spraying tanker was 6 000 m/h, and the ratio of spray width (m) to march speed (m/min) was 1:200. When 5 min post mother liquid recirculating, the average concentration of the molluscicide at the upper-, middle- and lower-layers of the liquid storage pot was (1030.39 ± 43.00) mg/L, with a variation coefficient of 4.17%. The average concentration of the molluscicide in the spraying process (spraying for 2, 4, 6, 8, 9 min) was (953.00 ± 68.87)mg/L, with a variation coefficient of 7.22%. The concentration of the residual drug in the liquid storage pot post spraying was 1 000.43 mg/L, which reached the effect concentration for snail control. After spraying for 7 days in the field, the average density of living snails reduced by 88.20% as compared to that before spraying, and the adjusted mortality of snails was 87.65%. The unit cost of Jingcen DY-1 spraying tanker was 0.086 7 Yuan/m². which reduced by 58.20% as compared to that of the conventional spraying tanker. CONCLUSIONS: Jingcen DY-1 type spraying tanker for snail control which integrates various equipments together can effectively control the concentration and dose of the molluscicide, and the machine is labor-saving, efficient, economic and well adapted, and is worthy to be widely applied.

[Application effect of Oncomelania hupensis snail crushers in batch detection].

OBJECTIVE: To evaluate the application effect of Oncomelania hupensis snail crusher in batch detection. METHODS: The O. hupensis snail crushers and triangular flasks for cercariae shedding were made. The uninfected snails were divided into 4 groups with the snail number of 50, 100, 200, 300, respectively, and then they were put in 40 triangular flasks, 10 flasks each group. Three flasks in each group were randomly selected, and 12 schistosome infected O. hupensis snails were put into the flasks (1 snail per flask). The 12 flasks with infected O. hupensis snails were put back to each group and were numbered uniformly. The O. hupensis snails in each group were crushed by the O. hupensis snail crushers, and the crushing rate of snails, and average crushing time and duration were recorded. When all the O. hupensis snails were crushed, water was added into the flasks, and water films from each flask were taken and observed under microscopes, the number of cercariae was recorded and the detection rates of each group were calculated. Meanwhile, the O. hupensis snails in the field were collected and detected by the crushing method and crushing and shedding method with O. hupensis snail crushers, and the coincidence rate of the two methods was calculated. RESULTS: The crushing rate of snails in all the 4 groups were 100% , the average crushing times of groups with 50, 100, 200, 300 snails were 15.70, 23.20, 32.20, 39.20, respectively, and the average duration was 1.01, 1.70, 2.00, 3.00 min, respectively. Both the crushing time and duration were positively correlated with the number of snails detected (r = 0.68, 0.73, both P <0.01). The detection rates of cercariae in all the 4 groups were 100%. In the field application, 109 snails were detected by the crushing method, 620 ones were detected by the crushing and shedding method with O. hupensis snail crushers, and no infected snails were found, the coincidence rate of the two methods were 100%. CONCLUSION: The O. hupensis snail crusher with the advances of easy operation and labor-saving is valuable of field promotion.

[Effect of schistosomiasis control in Jingzhou City from 2009 to 2013].

OBJECTIVE: To evaluate the effect of schistosomiasis control in Jingzhou City of Hubei Province from 2009 to 2013, so as to provide the reference for the establishment of control strategy in the future. METHODS: The annual reports on schistosomiasis control from 2009 to 2013 in nine counties (cities, districts) in Jingzhou City were collected and analyzed sta- tistically. RESULTS: The estimated number of schistosomiasis patients in Jingzhou City dropped from 78 903 in 2009 to 28 034 in 2013, with a reduction rate of 63.37%. The schistosome infection rate of residents dropped from 2.21% in 2009 to 0.65% in 2013, which showed an exponential decline trend (y = 3.196e(-0.317x)). The number of cattle with schistosomiasis decreased from 2 132 in 2009 to 0 in 2013, and the infection rate of cattle dropped from 1.87% in 2009 to 0% in 2013. Only 2 cases with acute infection of schistosome were found in 2009, and no schistosome infected snails were found out in 2 consecutive years, 2012 and 2013. CONCLUSIONS: The endemic level of schistosomiasis in Jingzhou City has decreased significantly. In order to consolidate the achievement of control, the comprehensive strategy including infection source control, snail control with mol- luscicides and snail surveillance should be implemented.

[Investigation on field feces in schistosomiasis endemic areas in Jingzhou City].

OBJECTIVE: To understand the status of field feces in Jingzhou City, so as to provide the evidence for improving the control measures to interrupt the transmission routes of schistosomiasis. METHODS: The distribution of field feces was investigated in 27 schistosomiasis endemic villages in Gong' an, Jianli, Jiangling, Honghu and Shishou counties (cities) from 2010 to 2012. The schistosome positive status of the field feces was surveyed with the hatching method. RESULTS: There were 1366 field feces and the average density was 0.0892 feces per 100 square meters in this survey. The cattle feces, human feces, dog feces and elk feces respectively accounted for 99.71%, 0.07%, 0.15% and 0.07% in the survey. The infection rates of the field feces were 1.46% and 2.42% in the channels and bottomlands, respectively (P > 0.05). The average rate of infected field feces was 3.21% in 2010, 0.36% in 2011, and 1.60% in 2012, and the difference between 2010 and 2012 was not statistically significant (P > 0.05). CONCLUSIONS: The main field feces come from cattle, and the main distribution of infected field feces is in channels and bottom-lands. Therefore, the management of cattle and treatment of field feces should be strengthened.

[Classification and grading of environments with Oncomelania hupensis snails inside embankment of marshland and lake areas].

OBJECTIVE: To classify and grade the environments with Oncomelania hupensis snails inside embankment in marshland and lake areas, so as to improve the work efficiency and realize the systematic management. METHODS: The schistosomiasis endemic area Liaodi and Xiongdi villages in Jiangling County, Hubei Province were selected as the experiment villages, and the environments with snails in the two villages were divided into sections with a length of 500 m, then the snail situation were surveyed by the systematic sampling method with 10 m and 50 m a frame respectively. The environments were classified according to the discovery of infected snails and wild fecal contamination, and the numbers of sites with snails and their areas in different classes were recorded. Meanwhile, the data of sites with infected snails in Jingzhou City during the recent 5 years were collected and graded according to the average density of snails, so as to discuss the correlativity between the grades of environments with snails and the numbers of sites with infected snails and the occurrence rate of frames with snails. RESULTS: There were 1 967 sites with infected snails in Jingzhou City during the recent 5 years, and there was a positive correlation between the grades of environments with snails and the occurrence rate of frames with snails (r(s) = 0.77, P < 0.01), when the grade was 5, the constituent ratio of numbers of sites with infected snails was 3.1%, and when the grade was 2-3, the ratio raised to 56.3%. There were 39 sites with snails in the two villages, the Class 1, 2 and 3 environments included 1, 18 and 20 sites, with the areas of 1080, 51,640 m2 and 41,220 m2, respectively. There were no statistically significant difference between the grades divided by snail survey with 10 m and 50 m a frame ( chi2 = 4.667, P > 0.05), but the time-consuming of the former was 3 times of the latter. CONCLUSIONS: The classification and grading of the environments with snails inside embankment in marshland and lake areas can master the key point of the snail survey. The subsection and setting frames at a suitable distance can save time and manpower, improve work efficiency, as well as understand the distribution of snail status of the environment with snails, which can realize the sort management of the snail environments inside embankment.