Comparison of postoperative recovery of primary pterygium excision combined with either limbal stem cell transplantation or amniotic membrane transplantation: a randomized controlled trial-based meta-analysis.

OBJECTIVE: To compare the postoperative recovery of primary pterygium excision combined with either limbal stem cell transplantation (LSCT) or amniotic membrane transplantation (AMT). METHODS: All relevant studies on the primary pterygium excision combined with either LSCT or AMT conducted before August 2022 were extracted from PubMed, EMBASE, Web of Science, and Cochrane Library databases. The main outcomes compared were tear film stability at 1, 3, and 6 months after surgery, postoperative corneal epithelial healing time, recurrence rate, and complications. RESULTS: Sixteen randomized controlled trials (RCTs) with 1390 eye cases were included in this meta-analysis. We found that patients of the AMT group improved significantly in the results of the tear break-up time (BUT) and Schirmer I test at 1 month after surgery (BUT: MD=-0.37, 95% CI: -0.62, -0.12, P<0.05; Schirmer I test: MD=-0.32, 95% CI: -0.57, -0.07, P<0.05) compared with those of the LSCT group, suggesting that the early stage of tear film stability after primary pterygium excision combined with AMT was superior to the LSCT combination. However, according to the Schirmer I test result, the patients in the LSCT group showed increased tear production compared to the AMT group at 3 and 6 months after surgery (3 months: MD=0.36, 95% CI: 0.08, 0.64, P<0.05; 6 months: MD=0.33, 95% CI: 0.07, 0.60, P<0.05), suggesting that the LSCT combination was superior to the AMT combination in long-term postoperative tear film stability. As for postoperative corneal epithelial healing time, the LSCT group exhibited shorter time than the AMT group (MD=-1.17, 95% CI: -2.15, -0.19, P<0.05). Furthermore, the recurrence rate was lower in the LSCT group than in the AMT group (RR=0.42, 95% CI: 0.30, 0.59, P<0.05). Lastly, there was no statistical difference in BUT and complication rate at 3 and 6 months after surgery between the LSCT and AMT groups. CONCLUSIONS: Our analysis suggests that primary pterygium excision combined with LSCT may be a better choice compared to the combination with AMT in postoperative recovery.

Population genomic, climatic and anthropogenic evidence suggest the role of human forces in endangerment of green peafowl (Pavo muticus).

Both anthropogenic impacts and historical climate change could contribute to population decline and species extinction, but their relative importance is still unclear. Emerging approaches based on genomic, climatic and anthropogenic data provide a promising analytical framework to address this question. This study applied such an integrative approach to examine potential drivers for the endangerment of the green peafowl (Pavo muticus). Several demographic reconstructions based on population genomes congruently retrieved a drastic population declination since the mid-Holocene. Furthermore, a comparison between historical and modern genomes suggested genetic diversity decrease during the last 50 years. However, climate-based ecological niche models predicted stationary general range during these periods and imply the little impact of climate change. Further analyses suggested that human disturbance intensities were negatively correlated with the green peafowl's effective population sizes and significantly associated with its survival status (extirpation or persistence). Archaeological and historical records corroborate the critical role of humans, leaving the footprint of low genomic diversity and high inbreeding in the survival populations. This study sheds light on the potential deep-time effects of human disturbance on species endangerment and offers a multi-evidential approach in examining underlying forces for population declines.

A new species of the genus Gracixalus (Amphibia: Anura: Rhacophoridae) from Mount Jinggang, southeastern China.

A new species, Gracixalus jinggangensis sp. nov., is described based on a series of specimens collected from Mount Jinggang, Jiangxi Province, southeastern China. The new species is distinguished from all other known congeners by the following combination of morphological characters: relatively small body size, SVL 27.9-33.8 mm in nine males and 31.6 mm in a single female; upper eyelid and dorsum lacking spines; skin of dorsal and lateral surface of head, body and limbs rough with sparsely scattered with tubercles; ventral skin granular; tibiotarsal projection absent; finger webbing rudimentary; toes with moderately developed webbing; brown to beige above in life, with an inverse Y-shaped dark brown marking extending from the interorbital region to the middle of dorsum; males with a single, subgular vocal sac, barely visible nuptial pads with minute granules on the dorsal surface of the bases of first and second fingers. The new species is also genetically divergent from all other Gracixalus for which comparable 16S rRNA gene sequences are available. The discovery of Gracixalus jinggangensis sp. nov. represents the twelfth known species in the genus.

A new species of the genus Leptolalax (Anura: Megophryidae) from Mt. Gaoligongshan of western Yunnan Province, China.

A new species of the genus Leptolalax is described from the Tengchong Section of Gaoligongshan National Nature Reserve, Tengchong County, Yunnan Province, China. The new species, Leptolalax tengchongensis sp. nov., can be distinguished from its congeners by a combination of the following characters: (1) small size (SVL 23.9-26.0 mm in males, 28.8-28.9 mm in females); (2) dorsal skin shagreened and scattered with fine, round reddish tubercles; (3) toes with rudimentary webbing and narrow lateral fringes; (4) tympanum distinctly discernible, almost entirely black; (5) ventrolateral glands indistinct; (6) flanks with several distinct and large dark blotches; (7) ventral surfaces white, scattered with distinct irregular dark speckling; (8) iris not bicolored, uniformly dark brown and scattered with minute, coppery reticulations throughout. To date, the new species has only been found at its type locality in evergreen broadleaf forests at elevations between 2000-2100 m.

A new species of the genus Odorrana (Amphibia: Ranidae) and the first record of Odorrana bacboensis from China.

The genus Odorrana currently contains at least 56 recognized species that inhabits montane streams in subtropical and tropical Asia. Twenty new species have been described in the last decade, indicating the potential cryptic species diversity of this genus. We collected several specimens of Odorrana species from Southern China from 2007 to 2014, and on the basis of a combined morphological characters and phylogenetic analysis, we described the new species Odorrana fengkaiensis sp. nov. herein. The new species is very similar to O. hainanensis and O. bacboensis, but can be consistently separated by morphology, and allopatric distribution. It is further reciprocally monophyletic to O. hainanensis in a mitochondrial gene trees with an average genetic divergence of 2.1% (1.9%-2.4%). The new species inhabits in lowland broad streams, rivers, pools and near the riparian areas, but its general ecology remains poorly known. The new species is characterized by its body length of adult females approximately twice as long as adult males (SVL 77.8-111.9 mm in females, 37.4-51.8 mm in males); eye large in males, eye diameter 1.01-1.16 times as long as snout length; tympanum of males large and distinct, extremely close to the eye, 0.7-1.4 mm in tympanum-eye distance; dorsolateral folds absent; dorsal skin shagreened, with several large tubercles in males; flanks with tubercles and scattered larger pustules, 8-10 of which usually arranged in a dorsolateral row; ventral skin smooth, with spines in adult males during the breeding season; the tibio-tarsal articulation stretched forward beyond the tip of snout; relative finger lengths: II < I < IV < III; dorsum brown with irregularly reticulated green markings in males and young females, uniformly brown in some old adult females; males with velvety nuptial pad on thumb, paired gular pouches; mature oocytes almost purely black in life, showed dark grey animal pole and olive vegetative pole in preservative. In addition, we found O. bacboensis, a new country record from China, indicating a range extension from north-central Vietnam to southeast Yunnan and adjacent area in Guangxi.

[Effectiveness of Meibomian Gland Tube Massage in Treating Meibomian Gland Dysfunction].

OBJECTIVE: To observe the clinical effectiveness of meibomian gland tube massage in treating meibomian gland dysfunction (MGD). METHODS: All patients were divided into medicine group (tropically administered with corticosteroid eye ointment and artificial tears)and massage group (meibomian gland tube massage in addition to these drugs) using random numbers. At different period(before treatment and after treatment 2,4 weeks), the slip-lamp microscopy and intraocular pressure measurement were performed. Ocular symptoms were evaluated by questionnaire of ocular surface disease index (OSDI), and corneal fluorescein staining scores (CFS) was used for checking the epithelial integrity,tear film breakup time (TBUT), and tear secretion (Schirmer I test,SIt). RESULTS: Before the treatment, the OSDI score,TBUT, CFS, and SIt showed no statistical significance between these two groups (all P>0.05). After the treatment, the symptoms, damage of corneal epithelium, quality of tear film,tear secretion were significantly improved in both groups(P<0.05), and were significantly superior in the massage group than in the medicine group (all P<0.01; but CFS t4w=6.60,P>0.05). CONCLUSION: The meibomian gland tube massage in combination with drug therapy can improve the treatment effectives for MGD.

Emodin ameliorates lipopolysaccharides-induced corneal inflammation in rats.

AIM: To investigate the effect of emodin on pseudomonas aeruginosa lipopolysaccharides (LPS)-induced corneal inflammation in rats. METHODS: Corneal infection was induced by pseudomonas aeruginosa LPS in Wistar rats. The inflammation induced by LPS were examined by slit lamp microscope and cytological checkup of aqueous humor. Corneal tissue structure was observed by hematoxylin and eosin (HE) staining. The activation of nuclear factor kappaB (NF-κB) was determined by Western blot. Messenger ribonucleic acid (mRNA) of tumor necrosis factor-α (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in LPS-challenged rat corneas were measured with reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Typical manifestations of acute corneal inflammation were observed in LPS-induce rat model, and the corneal inflammatory response and structure were improved in rats pretreated with emodin. Treatment with emodin could improve corneal structure, reduce corneal injure by reducing corneal inflammatory response. Emodin could inhibit the decreasing lever of inhibitor of kappaB alpha (IкBα) express, and the mRNA expression of TNF-α and ICAM-1 in corneal tissues was also inhibited by emodin. The differences were statistically significant between groups treated with emodin and those without treatment (P<0.01). CONCLUSION: Emodin could ameliorate LPS-induced corneal inflammation, which might via inhibiting the activation of NF-κB.

Evaluation of corneal graft survival in mice model.

AIM: To investigate the characteristics and criterion of graft rejection in mice model. METHODS: C57BL/6 or BALB/c mice corneal grafts were grafted onto BALB/c hosts. Each group was divided into two subgroups according to the corneal opacity scores 12d after transplantation. The characteristics of opacity and neovascularization were observed. Mice of the 12(th), 50(th) day after transplantation, the grafts biopsy of mice in allogeneic group 1, which opacity score exceed 3, were prepared for histological observation and those restore transparent were endothelial stained. RESULTS: There was no difference of corneal opacity score on the 7(th) and 12(th) day after operation; the histological results had no disparity between syngeneic group and allogeneic group. On the 12(th) day after surgery, the turbidity curve was apparent in grafts with opacity score < 2. Mononuclear cells were shown in grafts with opacity score reached 3 in allogeneic group 1. Different rejection performance was observed in tissue sections on the 50(th) day after surgery. CONCLUSION: Grafts, opacity score exceeds 3 from the 7(th) to the 12(th) day after operation could not be judged as a rejection. We should pay more attention to the variation of grafts opacity since 12d after corneal transplantation.

[Differentiation of mesenchymal stem cells induced by co-culture method].

OBJECTIVE: To investigate the potential differentiation of human mesenchymal stem cells(MSCs)into epithelium-like cells by an in vitro co-culture method. METHODS: The human conjunctival epithelium was obtained by digestion with dispase2, and the MSCs were isolated by density gradient centrifugalization. All cells were identified according to their morphologies and cell-surface antigen profiles by immunocytochemical analysis. The MSCs underwent co-culture with conjunctival epithelium in the manner without cell-to-cell contract. The morphological characterizes of cells were observed under contrast microscope, and the cytokeratin-4 expressions of the differentiated cells were identified by immunocytochemistry staining, reverse transcriptase polymerase chain reaction(RT-PCR), and Western blotting. RESULTS: Immunocytochemistry showed that positive expression of CD29 and negative expression of CD34 in the in vitro cultured MSCs. Cytokeratins4(CK4)was positively expressed in the human conjunctival epithelium. After co-cultured with conjunctival epithelial for 10 days, CK4 was detected in differentiated cells by immunocytochemistry, RT-PCR, and Western blotting. CONCLUSION: MSCs can differentiate into epithe1ium-like cells after having been co-cultured with human conjunctival epithelium.

[Effects of emodin on expression of cytokines induced by lipopolysaccharide in corneal fibroblasts].

OBJECTIVE: To investigate the effects of emodin on expression of cytokines induced by lipopolysaccharide (LPS) in cultured human corneal fibroblasts in vitro. METHODS: Primary human corneal fibroblasts of passages 4 were used in this research. Cells were treated with 10 microg/L LPS for 1, 2, 4, or 8 hours, which were pretreated with or without emodin for 30 minutes before LPS challenge. The degeneration of inhibitor of kappaB-alpha (I kappaB-alpha) and the effect of emodin on it were analyzed by Western blot analysis with a specific antibody. The cellular abundance of the mRNA of interleukin (IL)-6 and IL-8 from corneal fibroblasts under different conditions was determined by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Compared with cells without LPS treatment, I kappaB-alpha level significantly decreased in every time point after LPS challenge (P < 0.01). Emodin inhibited the LPS-induced degeneration of I kappaB-alpha by corneal fibroblasts in a dose-dependent manner (P < 0.05). Compared with cells without LPS treatment, the expressions of IL-6 and IL-8 mRNA significantly increased in every time point after LPS challenge (P < 0.01). At the same time, the expressions of the mRNA of IL-6 and IL-8 induced by LPS in corneal fibroblasts were also inhibited by emodin in a dose-dependent manner (P < 0.05). CONCLUSION: Emodin can inhibit the expressions of IL-6 and IL-8 mRNA induced by LPS in corneal fibroblasts, which maybe via inhibiting the degeneration of I kappaB-alpha and suppressing the activation of nuclear factor-kappaB.

Protective effect of emodin against lipopolysaccharides-induced corneal injury in rats.

OBJECTIVE: To investigate the effect of emodin on lipopolysaccharides (LPS)-induced corneal injury in rats. METHODS: Three parallel incisions on the central surface of corneal epithelium were made and LPS was applied on them to induce corneal injury in Wistar rats. All rats were randomly divided into emodin group (n=40) and keratitis group (n=40). Rats in the emodin group received subconjunctival injection of emodin and rats in the keratitis group received its vehicle 30 minutes before LPS exposure. At different time points--1, 3, 6, 12, and 24 hours after LPS exposure, the symptoms of all rats were observed and the severity of their ocular inflammation was examined with a slit lamp microscope, then 8 rats in each group were killed through cervical dislocation and their eyes were enucleated and prepared to observe pathological changes of corneal tissue under a light microscope. The activation of nuclear factor-kappaB (NF-kappaB) under different conditions was determined by Western blot. Immunocytochemistry staining with an antibody against intercellular adhesion molecule-1 (ICAM-1) was performed to identify positive cells in corneal tissues. RESULTS: The model of acute keratitis was successfully established in Wistar rats. LPS could induce a typical corneal inflammatory response, such as hyperemia, corneal edema and opacity, which were observed in model rats. Compared with keratitis group, both ocular behaviors and damages of the corneal structure were improved in emodin group. Furthermore, the activation of NF-kappaB and the expression of ICAM-1 induced by LPS were markedly inhibited in emodin group. CONCLUSION: Emodin can inhibit the activation of NF-kappaB and the expression of ICAM-1 induced by LPS in corneas, protect against acute corneal injury, and improve symptoms in rats.

[The effects of the inhibitor of nuclear factor on lipopolysaccharide-induced cytokine expression in cultured human corneal fibroblasts].

OBJECTIVE: To investigate the effects of pyrrolidine dithiocarbamate (PDTC) on Lipopolysaccharide (LPS)-mediated activation of nuclear factor kappa B (NF-kappaB) and cytokine expression in cultured human corneal fibroblasts. METHODS: It was a experimental study. A completely random design was employed in this research. Human corneal fibroblasts (HCFs) were obtained from human specimen. HCFs were divided into three groups: control group (group 0 h), LPS alone group and PDTC treatment group. Cells were incubated with PDTC for 30 min in the PDTC pretreatment group before LPS challenged. At different time after LPS challenged, the activities of NF-kappaB were assessed by Western Blot analysis, the secretion of IL-6 and IL-8 from cultured corneal fibroblasts was measured with enzyme-linked immunosorbent assays (ELISA); the mRNAs expression of IL-6 and IL-8 was determined by reverse transcription polymerase chain reaction (RT-PCR). The effects of PDTC on activation of NF-kappaB and the expression of IL-6 and IL-8 were also assessed in HCFs challenged with LPS. RESULTS: Compared with control group, NF-kappaB level was significantly enhanced in the nucleus in the LPS alone group, indicating that LPS mediated the activation of NF-kappaB in HCFs. The activation of NF-kappaB by LPS was markedly inhibited by PDTC (t1h = 9.3766, t2h = 15.9011, t4h = 12.5851, t8h = 10.8346, P < 0.01). Compared with control group, LPS increased IL-6 and IL-8 expression both mRNA and protein in HCFs. At the same time, PDTC partly inhibited the expression of IL-6 and IL-8 in corneal fibroblasts induced by LPS. These inhibitory effects were significant at both the mRNA and protein levels (protein of IL-6: t1h = 7.9154, t2h = 10.863, t4h = 8.2451, t8h = 13.5063. protein of IL-8: t1h = 8.5663, t2h = 20.5169, t4h = 25.1580, t8h = 34.8699. mRNA of IL-6: t1h = 12.0235, t2h = 13.2894, t4h = 24.0799, t8h = 27.2261. mRNA of IL-8: t1h = 20.9424, t2h = 24.1314, t4h = 29.8580, t8h = 47.9442. P < 0.01). CONCLUSION: These results suggest that LPS can activation of NF-kappaB in cultured HCFs and PDTC partly inhibits NF-kappaB activation.

[The expression of nuclear factor-kappa B in lipopolysaccharide-induced keratitis of rats and the effect of pyrrolidine dithiocarbamate on its expression].

OBJECTIVE: To investigate the expression of nuclear factor-kappa B (NF-kappaB) and cytokines in lipopolysaccharide (LPS)-induced keratitis of rats, and the effect of pyrrolidine dithiocarbamate (PDTC) on its expression. METHODS: A LPS-induced keratitis model was established in Wistar rats. Thirty minutes before LPS exposure, PDTC and normal saline were injected into subconjunctival tissues separately. At 0.5, 1.0, 3.0, 6.0, 12.0, 24.0 and 72.0 h after LPS exposure, the rats were examined with slit-lamp microscope. Then they were sacrificed and the corneas were excised for routine histological analysis. Immunohistochemical staining with an antibody against activated NF-kappaB was performed to detect the expression of NF-kappaB. The change of TNF-alpha mRNA expression was identified by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Histology findings demonstrated that corneas exposed to LPS showed significant changes in corneal structure, edema and pronounced inflammatory cells infiltration were observed. Both symptoms and damages of the cornea were lesser in the rats of PDTC group than that of the control keratitis group. Compared with PDTC group, the activation of NF-kappaB and the expression of TNF-alpha mRNA were significantly upregulated after LPS challenge 0.5 - 24.0 h (P < 0.01), separately; peak expression of NF-kappaB and TNF-alpha mRNA were observed at 3.0 - 12.0 h after LPS exposure. CONCLUSIONS: The expression of NF-kappaB and TNF-alpha mRNA induced by NF-kappaB plays an important role in the pathogenesis of keratitis. The inhibitor of NF-kappaB, PDTC, can relieve the cornea from damages produced by keratitis.