[Efficacy of a recombinant bovine basic fibroblast growth factor gel for the treatment of moderate dry eye: a multicenter randomized double-blind parallel controlled clinical trial].

Objective: To compare the clinical efficacy of a recombinant bovine basic fibroblast growth factor (rb-bFGF) gel and a gel matrix in the treatment of moderate dry eye. Methods: It was a prospective random double-blind controlled study. One hundred patients diagnosed as moderate dry eye in Eye Institute and Affiliated Xiamen Eye Center of Xiamen University, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology, Beijing Tongren Hospital, Capital Medical University, Eye & ENT Hospital of Fudan University and Zhongshan Ophthalmic Center from August 2015 to April 2019 were divided into two groups: experimental group and control group. Two groups of patients were allocated to receive either a rb-bFGF gel or a gel matrix 4 times per day for 4 weeks. Subjective symptoms, break-up time of the tear film (BUT), Schirmer Ⅰ test (SⅠt) and corneal fluorescein sodium staining were assessed at baseline, 2 and 4 weeks after treatment. Bulbar impression cytology was evaluated at baseline and 4 weeks after treatment. Irritation of the rb-bFGF gel and the gel matrix was estimated after treatment. T test, Wilcoxon signed-rank test or Mann-Whitney U test was used for quantitative data, and Chi-square test was used for enumerative data. Results: Eighty-four subjects were included for statistical analyses after the exclusion of 16 subjects who were lost for followup, with an age of 43±14 years. There were 42 cases in the experimental group and the control group, respectively. There was no statistically significant difference between the two groups in demographic baseline characteristics before treatment (P>0.05). The total score of subjective symptoms was 7.17±3.60 and 5.95±3.25 at 2 and 4 weeks after therapy in the experimental group, which were lower than 9.48±3.88 before treatment (t=6.226, 6.563; both P<0.05); in the control group, it was 7.01±3.25 and 6.32±3.85 at 2 and 4 weeks after treatment, with a significant reduction in comparison with that before treatment (9.15±3.58; t=4.693, 4.726; both P<0.05). The median (lower quartile, upper quartile) BUT was 4.00 (2.40, 5.00) s and 4.64 (3.00, 5.00) s at 2 and 4 weeks after therapy in the experimental group, which were longer than 3.72 (2.00, 4.39) s before treatment (Z=-2.485, -3.152; both P<0.05). The BUT was 4.41 (2.79, 5.12) s at 2 weeks after therapy in the control group, which was of no statistical difference compared with 3.89 (2.09, 4.25) s before treatment (Z=-1.953, P>0.05). The BUT was 5.21 (3.00, 5.02) s at 4 weeks after therapy in the control group, which was longer than that before treatment (Z=-2.485, P<0.05). The SⅠt score was 7.31 (3.75, 10.00) mm and 8.50 (4.00, 11.00) mm at 2 and 4 weeks after therapy in the experimental group, which were significantly higher than 6.69 (2.00, 8.13) mm before treatment (Z=-2.031, -2.236; both P<0.05); in the control group, it was 6.82 (2.00, 8.25) mm and 6.86 (3.00, 9.25) mm at 2 and 4 weeks after therapy, which were not significantly increased compared with 6.50 (2.00, 7.75) mm before treatment (Z=-0.179, -1.161; both P>0.05). The corneal fluorescein sodium staining points were 5.00 (2.00, 5.00) and 3.71 (0.00, 5.00) at 2 and 4 weeks after therapy in the experimental group, which were significantly lower than 7.10 (5.00, 7.00) before treatment (t=-2.895, -4.639; both P<0.05); those in the control group were 5.52 (0.00, 7.00) and 6.19 (0.75, 6.25) at 2 and 4 weeks after treatment, with a significant reduction in comparison with 8.90 (5.00, 10.50) before treatment (t=-2.776, -1.991; both P<0.05). The differences in the average total score of subjective symptoms, BUT, SIt, and corneal fluorescein sodium staining points between both groups were not statistically significant at each time point. The impression cytology grade was decreased from 1.72 (1.00, 2.00) before treatment to 0.94 (0.00, 2.00) at 4 weeks after therapy in the experimental group (Z=-2.803, P<0.05). The staining grade of conjunctival imprinted cells in the control group was 1.42 (1.00, 2.00) at 4 weeks, which showed no statistical significance compared with 1.56 (1.00, 2.00) before treatment (Z=1.195, P>0.05). The impression cytology grade was significantly reduced in the experimental group compared with the control group at 4 weeks after treatment (Z=-3.308, P<0.05). The number of goblet cells was 10.90 (5.00, 20.00) at 4 weeks after therapy in the experimental group, which was significantly higher than 6.30 (5.00, 8.00) before treatment (Z=-2.383, P<0.05); in the control group, it was 8.36 (4.00, 12.00) at 4 weeks after treatment, with no significant increase in comparison with that before treatment [7.55 (5.00, 11.00)] (Z=-0.095, P>0.05). The number of goblet cells was not significantly increased in the experimental group compared with the control group at 4 weeks after treatment (Z=-1.162, P>0.05). Most patients indicated that the drug was non-irritating, and no patient had intolerable irritation affecting daily lives at 4 weeks after therapy; there was no difference between the two groups (Z=-0.290, P>0.05). Conclusions: Both the rb-bFGF gel and the gel matrix can effectively improve the symptoms and signs of moderate dry eye. However, compared with the gel matrix, the rb-bFGF gel shows obvious advantages in promoting conjunctival epithelial cell repair and increasing the number of goblet cells. (Chin J Ophthalmol, 2021, 57: 930-938).

[Effect of binocular accommodation and vergence function examinations and interventions on subjective discomfort of dry eye].

Objective: To investigate the accommodation and vergence (AV) function of patients with mild to moderate refractory dry eye symptoms, and determine the impact of relevant interventions on subjective symptoms. Methods: A total of 103 patients with dry eye disease (DED) in Zhongshan Ophthalmic Center between December 2017 and June 2019 were included. After 3-month conventional treatment, the patients entered the treatment-responsive group if ocular surface disease index (OSDI) decreased ≥12.5, and others were recruited into the refractory symptoms group. Binocular AV function, OSDI, tear break-up time (TBUT), Schirmer's test (ST) and fluorescein staining (FL) were determined in all the patients. Corrective therapy on the AV dysfunction was added in the refractory symptoms group besides the conventional therapy. The above-mentioned indexes were reexamined 8 weeks later. The incidence of AV dysfunction was compared between the refractory symptoms group and the treatment-responsive group. Moreover, the differences of OSDI and tear film stability were compared before and after the corrective therapy in the refractory symptoms group. Results: Sixty of 103 DED patients were classified into the refractory symptoms group [mean age: (27±6) years; 18 males and 25 females] and 43 into the treatment-responsive group [mean age: (30±6) years; 32 males and 28 females]. The incidence of AV dysfunction in the refractory symptom group (100%) was higher than that of the treatment responsive group (72.1%) (P<0.001). Forty patients with refractory symptoms accomplished the 8-week corrective therapy, and the OSDI score was significant improved (23.4±16.0 vs 40.6±15.7, P<0.001). However, further changes in ST, TBUT and FL were not detected in these patients (all P>0.05). Conclusions: There is a high prevalence of AV dysfunction in patients with refractory symptomatic DED. The corrective therapy on AV may improve the subjective symptoms in these patients.

[Focusing on the functional dry eye].

Dry eye is one of the most common eye diseases characterized by disturbance of tear film homeostasis and a series of ocular discomforts. Discrepancy of ocular subjective symptoms and objective signs, especially unexplained severe symptoms without visible signs has been a long-time confusion for ophthalmologists. Functional disorder is a medical condition that impairs normal functioning of bodily processes due to autonomic nervous dysfunction caused by cerebral cortical dysfunction, but at the exterior, there is no appearance of abnormality. Clinical manifestations are the disease symptoms with a lack of exact physical signs and examination results to confirm the diagnosis. According to the definition and characteristics of functional disorder, we put forward the concept of functional dry eye. We assume that those patients with dry eye symptoms but without detectable signs could be classified into such a functional dry eye subtype. The possible mechanism, clinical manifestations, and treatment strategies are further discussed. (Chin J Ophthalmol, 2019, 55: 885-890).

[Diagnostic criteria of demodex blepharitis].

Demodex infestation is a common cause of blepharitis and now is drawing more and more attention from clinicians. Demodex mite is the most common age-related ectoparasite of human being and may be detected from asymptomatic normal population. Furthermore, the pathogenesis of demodicosis remains unclear. Therefore, the commonly accepted diagnostic criteria of demodex infestation related blepharitis have not been established. Herein, we summarize the related studies about ocular demodicosis and comment the debates about the diagnosis of the disease. We also propose diagnostic criteria of demodex blepharitis. (Chin J Ophthalmol, 2017, 53: 648-652).

[Analysis of the influence of iron overload in glucose metabolism in thalassemia major patients].

Objective: This study aimed at determining the characteristics of the glucose homeostasis and its relationship with iron overload of the patients with ß-thalassemia major (ß-TM). Method: From Sun Yat-sen Memorial Hospital between January 2014 and December 2015, a total of 57 transfusion-dependent ß-TM patients with 5-18 years old were enrolled in this study and fasting blood glucose(FBG) and insulin level, serum ferritin (SF), serum iron, transferrin, total iron binding capacity, unsaturated iron binding capacity were determined.Insulin resistance index (IRI), insulin sensitivity index and ß-cell function index (BFI) were also estimated. Besides, in 36 patients cardiac T2* and liver T2* were estimated. Result: (1) Four patients(7%) with ß-TM were diagnosed diabetes mellitus, and 14(24%) had impaired fasting glucose. (2) The incidence of abnormal glucose metabolism was significantly different according to levels of SF and degrees of the cardiac iron overload(χ(2)=9.737, P<0.05; χ(2)=17.027, P<0.05). It rose while the level of SF increased and the degree of cardiac iron overload aggravated. (3) The incidence of abnormal glucose level was not significantly different in cases with different degree of liver iron overload.The severe group of liver iron overload had significantly higher levels of INS, HOMA-ßFI, HOMA-ISI, HOMA-ßFI than the non-severe group (Z=-2.434, -2.515, F=8.658, all P<0.05), while no differences were found in the level of FBG, HOMA-ßFI between two groups. (4) The result of logistic regression analysis indicated that the cardiac T2* was a significant predictor for the incidence of abnormal glucose metabolism in TM patients (P=0.035, OR=1.182%, 95%CI=1.048 to 1.332). Conclusion: The high prevalence of abnormal glucose metabolism in ß-TM patients was mainly closely related with the internal iron overload, especially in organs.The cardiac T2* was an independent risk factor for the incidence of abnormal glucose metabolism in TM patients.

The impact of daycare attendance on outdoor free play in young children.

Background: Outdoor free play is important for healthy growth and development in early childhood. Recent studies suggest that the majority of time spent in daycare is sedentary. The objective of this study was to determine whether there was an association between daycare attendance and parent-reported outdoor free play. Methods: Healthy children aged 1-5 years recruited to The Applied Research Group for Kids! (TARGet Kids!), a primary care research network, were included. Parents reported daycare use, outdoor free play and potential confounding variables. Multivariable linear regression was used to determine the association between daycare attendance and outdoor free play, adjusted for age, sex, maternal ethnicity, maternal education, neighborhood income and season. Results: There were 2810 children included in this study. Children aged 1 to <3 years (n = 1388) and ≥3 to 5 years (n = 1284) who attended daycare had 14.70 min less (95% CI -20.52, -8.87; P < 0.01) and 9.44 min less (95% CI -13.67, -5.20; P < 0.01) per day of outdoor free play compared with children who did not attend daycare, respectively. Conclusions: Children who spend more time in daycare have less parent-reported outdoor free play. Parents may be relying on daycare to provide opportunity for outdoor free play and interventions to promote increased active play opportunities outside of daycare are needed.

Conservation and population genetic diversity of Curcuma wenyujin (Zingiberaceae), a multifunctional medicinal herb.

Curcuma wenyujin is an important multifunctional medicinal herb in China. Currently, populations of C. wenyujin are decreasing, and wild individuals have almost disappeared from their natural habitats. Moreover, little is known regarding the molecular characteristics of this plant. In this study, we investigated the genetic diversity and variation of five populations of C. wenyujin, using ran-dom amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. We found that the percentages of polymorphic loci (PPL) at the species level (98.25% by RAPD and 100% by ISSR) were significantly higher than those at the population level (66.32% by RAPD and 67.14% by ISSR). The highest values of PPL, expected heterozygosity, and Shannon's information index were in Pop1, while the lowest values were in Pop2. Both DNA markers revealed a short genetic distance between Pop1 and Pop2 (0.1424 by RAPD and 0.1904 by ISSR). Phylogenetic trees produced similar results, with Pop1, Pop2, and Pop5 in one group and Pop3 and Pop4 in another. There were no significant correlations between their genetic distances and their geographical distances. The highest genetic diversity was in Pop1 and the lowest was in Pop2, and genetic diversity at the species level was relatively low, but much higher than that at the population level. We recommended the establishment of a germplasm bank, in situ con-servation, and propagation of wild individuals. The present study will improve the evaluation, protection, and utilization of the population resources of C. wenyujin.

Increasing testicular temperature by exposure to elevated ambient temperatures restores spermatogenesis in adult Utp14b (jsd) mutant (jsd) mice.

Because mutations in the human UTP14C gene are associated with male infertility, we sought to develop a method for fertility restoration in azoospermic mice with a mutation in the orthologous Utp14b(jsd) (jsd) gene that have spermatogonial arrest. The method is based on our observation that elevation of testicular temperatures restores spermatogonial differentiation in jsd mutant mice. To non-surgically raise intrascrotal temperatures we placed these mice in incubators at different elevated ambient temperatures. Exposure of jsd/jsd mice to ambient temperatures of 34.5 °C or 35.5 °C for 24 days increased the proportion of tubules with spermatocytes from 0% in untreated controls to over 80%. As those higher temperatures interfere with spermatid differentiation, the mice were then transferred to incubators at 32-32.5 °C for the next 24 days. These environments allowed differentiation to progress, resulting in up to 42% of tubules having late spermatids and about half of the mutant mice having spermatozoa in testicular suspensions. When these spermatozoa were used in intracytoplasmic sperm injection, all gave rise to viable healthy offspring with normal weight gain and fertility. The successful restoration of fertility in Utp14b mutant mice suggests that transient testicular warming might also be useful for spermatogenesis recovery in infertile men with UTP14C gene mutations.

Spironolactone prevents dietary-induced metabolic syndrome by inhibiting PI3-K/Akt and p38MAPK signaling pathways.

OBJECTIVES: Aim of the study is to evaluate the impact of spironolactone (SPL) on indexes of metabolic syndrome (MS) and further investigate the mechanisms underlying its protective effects. METHODS: A rat model of MS was established by administering a fat- and salt-enriched diet (FS diet). The occurrence of MS was examined by measurement of blood pressure (BP), aldosterone (ALD) content, blood lipid (BL), glucose and insulin levels. Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Pancreatic gland tissue injury was assessed by ß-cell apoptosis. Mineralocorticoid receptor (MR) activity, phosphatidylinositol 3- kinase/Akt (PI3-K/Akt), and phosphorylation of p38MAPK (Pp38MAPK) in pancreatic gland tissue were evaluated by western blot analysis. RESULTS: SPL prevented hypertension, and dyslipidemia during MS induced by the intake of FS diet, but had no effect on K+ and Na+ disturbances. Furthermore, SPL significantly attenuated ALD and MR expression levels after FS diet. Finally, SPL inhibited phosphorylation protein kinase B (p- PKB) activation in the pancreatic gland tissue, a downstream target of PI3-K, and phosphorylation of p38MAPK pathway, critical for cellular apoptosis. CONCLUSIONS: This study demonstrates that SPL exerts a protective effect on hypertension and dyslipidemia. This protective effect may depend, at least in part, on MAPK and PI3-K pathways.

Anti-inflammatory activities of fractions from Geranium nepalense and related polyphenols.

Geranium nepalense Sweet is a common Chinese herbal medicine and has been used as influenza, dysentery, antiphlogistic and analgesic tonic, hemostatic, stomachic, and antidiabetic drugs. The anti-inflammatory effects of G. nepalense on tetradecanoyl phorbol acetate (TPA)-induced mouse ear edema were studied in this work. The results showed that ethyl acetate fraction of the water extract of G. nepalense possessed significant activity at 2.5 g/kg (p < 0.01) with aspirin as a positive control (0.6 g/kg). Six polyphenolic compounds, including three flavonoids, i.e. kaempferol, kaempferol-7-O-ß-D-glucopyranoside, and quercetin-7-O-α-rhamnopyranoside, and two tannins, i.e. pyrogallol and gallic acid, and one lignin, i.e. epipinoresinol, were isolated and characterized from ethyl acetate fraction. The isolation of polyphenols provides a clue for beneficial effects of G. nepalense in the demonstrated anti-inflammatory activity.

MUC19 expression in human ocular surface and lacrimal gland and its alteration in Sjögren syndrome patients.

This study investigated the expression of MUC19, a newly discovered gel-forming mucin gene, in normal human lacrimal functional unit components and its alteration in Sjögren syndrome patients. Real-time PCR and immunohistochemistry were performed to determine the expression of MUC19 and MUC5AC in human cornea, conjunctiva, and lacrimal gland tissues. Conjunctival impression cytology specimens were collected from normal control subjects and Sjögren syndrome patients for Real-time PCR, PAS staining, and immunohistochemistry assays. In addition, conjunctiva biopsy specimens from both groups were examined for the expression differences of MUC19 and MUC5AC at both mRNA and protein level. The MUC19 mRNA was found to be present in cornea, conjunctiva and lacrimal gland tissues. The immunohistochemical staining of mucins showed that MUC19 was expressed in epithelial cells from corneal, conjunctival, and lacrimal gland tissues. In contrast, MUC5AC mRNA was only present in conjunctiva and lacrimal gland tissues, but not in cornea. Immunostaining demonstrates the co-staining of MUC19 and MUC5AC in conjunctival goblet cells. Consistent with the significant decrease of mucous secretion, both MUC19 and MUC5AC were decreased in conjunctiva of Sjögren syndrome patients compared to normal subjects. Considering the contribution of gel-forming mucins to the homeostasis of the ocular surface, the decreased expression of MUC19 and MUC5AC in Sjögren syndrome patients suggested that these mucins may be involved in the disruption of the ocular surface homeostasis in this disease.

Two variants of zebrafish p100 are expressed during embryogenesis and regulated by Nodal signaling.

Human p100 protein was first identified as a transcriptional coactivator of Epstein-Barr virus nuclear antigen 2, and has been shown to be a coactivator of other cellular transactivators. Its roles in development of vertebrate embryos, however, have not been reported. We have identified a zebrafish ortholog of the human p100 coactivator. The zebrafish p100 transcript is processed to two alternative variants, long and short forms, referred to as p100L and p100S, respectively. Both GFP-p100L and GFP-p100S fusion proteins are located in the cytoplasm of transfected culture cells and microinjected embryonic cells. Analysis of transcripts with Northern blots revealed the presence of p100L and lower amounts of p100S mRNAs from the one-cell stage throughout the life cycle. Whole-mount in situ hybridization shows that p100L and p100S share the same spatiotemporal expression pattern. Their zygotic expression is initially restricted to axial mesoderm precursors during gastrulation, and then spreads over other tissues during segmentation, and finally is constrained to some internal organs at day 5. We also find that Nodal signaling is essential for the zygotic expression of p100. These studies pave the way to understanding in depth the role of p100 during vertebrate embryogenesis.