Effect of Resveratrol on MMP-2 Expression in Scleral Fibroblasts: An In Vitro Study.

PURPOSE: To investigate the effects of resveratrol (Res) on human fetal scleral fibroblasts (HFSFs) and its potential mechanism. METHODS: HFSFs were randomly divided into the Res-treated group and the control group. Following, HFSFs were treated with or without a concentration of 10 µM Res for 48 h. To detect the expression of related genes, reverse transcription quantitative PCR (RT-qPCR) and western blotting were used. The apoptosis rate of different groups was determined using flow cytometry. RESULTS: The mRNA expression of matrix metalloproteinase 2 (MMP-2), Collagen, Type I, Alpha 1 (COL1A1), Janus Kinase 2 (JAK2), and Signal Transducer and Activator of Transcription 3 (STAT3)" was downregulated in the Res-treatment group compared to the control group, according to RT-qPCR. Western blotting revealed that Res therapy reduced the expression of MMP-2, JAK2, P-JAK2, STAT3, P-STAT3, and Bcl-2 associated protein X (Bax) while increasing the expression of COL1A1 and B-cell lymphoma-2 (Bcl-2). Flow cytometry showed that the cell apoptosis rate was significantly lower in HFSFs treated with Res. CONCLUSIONS: In conclusion, these findings suggest that Res increases COL1A1 expression while inhibiting MMP-2 and cell apoptosis in HFSFs, possibly through modulation of the JAK2/STAT3 signaling pathway.

Whole genome sequence of Streptococcus pluranimalium SP21-2, a porcine strain harbouring optrA and lsa(E) with chromosomal location.

OBJECTIVES: The aim of this study was to characterise the whole genome sequence of multidrug-resistant Streptococcus pluranimalium strain SP21-2 of swine origin in China. METHODS: Illumina Miseq (200X coverage) and Nanopore PromethION platform (100X coverage) were used for genome sequencing. Rapid Annotation using Subsystem Technology (RAST) was used to annotate the genome of SP21-2. The antimicrobial resistance genes (ARGs) were identified using ResFinder-4.1. RESULTS: The assembled circular genome of S. pluranimalium SP21-2 was 1,987,058 bp in length with a GC content of 39.54%, and no plasmid sequence was detected. A total of 2086 coding sequences were predicted by RAST. Oxazolidinone-phenicol resistance gene, optrA, and pleuromutilin-lincosamide-streptogramin A resistance gene, lsa(E), are both located on chromosomes, associated with IS1216 and ISS1S, respectively. In addition, SP21-2 harbours lnu(B) (lincosamide), ant (6)-Ia and aac(6')-aph(2") (aminoglycoside), erm(B) (macrolide), and tet(O) (tetracycline). CONCLUSION: We firstly report the oxazolidinone-phenicol gene, optrA, and pleuromutilin-lincosamide-streptogramin A resistance gene, lsa(E), in S. pluranimalium. In this strain, we firstly identified ISS1S and IS1216 carrying ARGs in S. pluranimalium, which will provide a valuable reference to understanding potential transfer mechanisms of ARGs in S. pluranimalium.

MicroRNA-29a inhibits collagen expression and induces apoptosis in human fetal scleral fibroblasts by targeting the Hsp47/Smad3 signaling pathway.

Members of the microRNA-29 (miR-29) gene family have been implicated as suppressors of collagen in several human diseases. The present study aimed to explore the function of miR-29a in human fetal scleral fibroblasts (HFSFs) and to investigate potential mechanisms by which the molecule regulates cellular functioning. First, HFSFs were transfected with miR-29a mimic, miR-29a inhibitor, or their corresponding controls. Then, cell proliferation and apoptosis were assessed using a CCK-8 assay and flow cytometry, respectively. Further, using real-time PCR, western blotting, and immunofluorescence staining, levels of miR-29a, heat shock protein 47 (Hsp47), COL1A1, Smad3, P-Smad3, Bax, and Bcl-2 were investigated. Next, empty vectors and SERPINH1-overexpressing vectors were used to transfect HFSFs. Western blotting and flow cytometry were performed to assess changes in levels of HFSF protein expression and apoptosis, respectively. Results indicated that the miR-29a mimic significantly inhibited Hsp47, Smad3, P-Smad3, and COL1A1 expression. Conversely, the miR-29a inhibitor enhanced the expression of the same genes. Furthermore, miR-29a overexpression inhibited HFSFs proliferation and enhanced the rate of HFSFs apoptosis. Consistent with this finding, miR-29a overexpression led to the downregulation of Bcl-2 and upregulation of Bax. In contrast, miR-29a suppression led to the upregulation of Bcl-2 and downregulation of Bax expression and reduced the rate of apoptosis. Additional research revealed that overexpression of Hsp47 prevented HFSFs apoptosis and enhanced collagen production. Findings that miR-29a overexpression reduces collagen expression levels, slows proliferation, and promotes apoptosis in HFSFs highlight the key role of miR-29a in scleral remodeling. The effects of miR-29a on scleral remodeling might mediate by targeting Hsp47 and repressing the Smad3 pathway.

Development of the Gemini Gel-Forming Surfactant with Ultra-High Temperature Resistance to 200 °C.

In order to broaden the application of clean fracturing fluid in ultra-high temperature reservoirs, a surfactant gel for high-temperature-resistant clean fracturing fluid was developed with a gemini cationic surfactant as the main agent in this work. As the fracturing fluid, the rheological property, temperature resistance, gel-breaking property, filtration property, shear recovery performance and core damage property of surfactant gel were systematically studied and evaluated. Results showed the viscosity of the system remained at 25.2 mPa·s for 60 min under a shear rate of 170 s-1 at 200 °C. The observed core permeability damage rate was only 6.23%, indicating low formation damage after fracturing. Due to micelle self-assembly properties in surfactant gel, the fluid has remarkable shear self-repairability. The filtration and core damage experimental results meet the national industry standard for fracturing fluids. The gel system had simple formulation and excellent properties, which was expected to enrich the application of clean fracturing fluid in ultra-high temperature reservoirs.

Effect of Time Outdoors and Near-viewing Time on Myopia Progression in 9- to 11-year-old Children in Chongqing.

SIGNIFICANCE: We used an Akeso device to record the visual behavior of children with myopia in two learning modes. We found that online class mode may increase near-viewing time and reduce outdoor time compared with the traditional school mode and may be responsible for accelerating myopia progression. PURPOSE: We aimed to explore the effects of visual behavior in different learning modes on myopia progression among children 9 to 11 years old. METHODS: Forty-nine children were included and requested to use a wearable device to objectively record visual behavior in real time from November 2019 to November 2020; participants took online classes from mid-February to early May 2020 during this period. Data (including glasses-wearing time, outdoor time, and near-viewing time) were collected during two 14-day periods, which included the online class learning mode (March 2 to 15, 2020) and the traditional school mode (May 20 to June 2, 2020). Spherical equivalent refraction and axial length were obtained at baseline, at 6-month intervals, and 1 year later. RESULTS: Outdoor time during online class mode (median, 9.5 minutes; interquartile range, 0.75 to 48 minutes) was significantly lower than during the school mode (median, 29 minutes; interquartile range, 11.50 to 50 minutes; P < .001). The mean ± standard deviation of near-viewing time was significantly different between online class mode (396.58 ± 114.41 minutes) and school mode (376.52 ± 93.99 minutes; P = .007, F = 19.56). In comparison with the baseline examination (-2.33 ± 0.81 D), mean spherical equivalent refraction in oculus dexter corresponding to the 6-month examination was decreased (-2.94 ± 0.83 D, P = .001), indicating a significant increase in myopia during online class mode. CONCLUSIONS: This study provides evidence of the association of learning mode and myopia progression. Accelerated progression of myopia in online class mode may be related to increased near-viewing time and decreased time spent in outdoor activities.

Effect of Interleukin 6 on Scleral Fibroblast Proliferation, Differentiation, and Apoptosis Involved in Myopic Scleral Remodeling.

INTRODUCTION: Scleral hypoxia (HO) is present in myopic eyes, and interleukin (IL)-6 is increased in the aqueous humor of patients with high myopia. The aim of this study was to investigate the effects of IL-6 on scleral fibroblast proliferation, differentiation, and apoptosis under conditions of HO and the possible role of IL-6 in myopic scleral remodeling. METHODS: Primary human scleral fibroblasts (HSFs) were cultured using a tissue mass adherent method. First, cells were cultured under conditions of HO (2% O2) or normoxia (NO, 20% O2) for different times. A quantitative real-time polymerase chain reaction (qRT-PCR) and immunofluorescence were used to detect the expression of IL-6 in HSFs. Next, cells were divided into five groups: NO, HO, HO plus IL-6, HO plus IL-6 receptor inhibitor (IL6RI), and HO plus IL-6 and IL6RI. The groups were treated separately for 72 h. Cell counting kit-8 assay and flow cytometry were used to detect cell proliferation and apoptosis, respectively. Western blotting and qRT-PCR were used to detect the expression of various genes in the transforming growth factor-ß1/Smad2/matrix metalloproteinase-2 pathway; these methods and immunofluorescence were also used to detect transdifferentiation of HSFs. RESULTS: HO resulted in upregulation of IL-6 expression in HSFs. Compared with NO, HO resulted in diminished cell proliferation and increased apoptosis and differentiation in HSFs; the above trend was further enhanced by the addition of IL6RI. Compared with the HO group, the addition of IL-6 led to a decrease in cell proliferation and an increase in apoptosis and differentiation of HSFs; the above trends showed opposite changes after the addition of both IL-6 and IL6RI. Additionally, IL-6 and IL6RI exerted opposite regulatory effects on the transforming growth factor-ß1/Smad2/matrix metalloproteinase-2 pathway under conditions of HO. CONCLUSION: HO caused HSFs to overexpress IL-6. IL-6 has a role in scleral remodeling in myopic eyes through affecting the proliferation, differentiation, and apoptosis of HSFs.

TNFSF15 Polymorphisms are Associated with Graves' Disease and Graves' Ophthalmopathy in a Han Chinese Population.

PURPOSE: Previous studies revealed an association of tumor necrosis factor super family-15 (TNFSF15) gene polymorphisms with several autoimmune diseases, but their roles in the development of Graves' disease (GD) and Graves' ophthalmopathy (GO) remain unclear. The aim of this study was to explore the correlation between TNFSF15 single nucleotide polymorphisms (SNPs) and susceptibility to GD and GO in a Han Chinese population. MATERIALS AND METHODS: This case-control study enrolled 813 healthy controls and 315 GD patients including 229 patients with GO. Eight SNPs of TNFSF15 (rs3810936, rs6478108, rs6478106, rs4979462, rs10817669, rs4246905, rs10733612, and rs10759734) were detected by TaqMan real-time PCR assay or PCR-restriction fragment length polymorphism (PCR-RFLP) and the possible association of these eight SNPs with the risk of GD and GO was analyzed. Allele frequencies and genotype distributions were compared using χ2 test. A stratified analysis was also performed according to clinical activity score (CAS) and sex. RESULTS: In comparison with the controls, the CT genotype of rs3810936 and the T allele of rs4979462 were decreased in GD patients. GD patients with GO also showed a lower frequency of the rs3810936 CT genotype and the rs4979462 T allele than the controls. Inactive GO patients showed a lower frequency of the rs3810936 CT genotype than controls, while GD females demonstrated a lower incidence of the T allele in rs4979462 and rs6478106 than the controls. Furthermore, GO females exhibited a lower rate of the rs4979462 T allele than the controls. CONCLUSIONS: Our results suggest that TNFSF15 rs3810936 and rs4979462 polymorphisms are associated with the susceptibility of GD and GO in a Han Chinese population. Additionally, rs3810936 may affect the activity of GO, and rs4979462 may correlate with the sexual dimorphism observed in GD and GO.

Village-Scale Livelihood Change and the Response of Rural Settlement Land Use: Sihe Village of Tongwei County in Mid-Gansu Loess Hilly Region as an Example.

Rural livelihood change has great influence on the scale, structure, and morphology of rural settlement land use, thus bringing new challenges to rural revitalization and settlement reconstruction. Sihe village of Tongwei County in mid-Gansu loess hilly region (China) was taken as an example here. Based on participatory rural appraisal data, we analyzed the structure and allocation of rural households' livelihood assets as well as their livelihood diversity by using ecological asset, livelihood diversification index, and landscape pattern index models. We aimed to find a response mechanism between rural livelihood change and rural settlement land use change. The results might provide useful information for the selection of new village sites, reconstruction of rural settlements, and creation of livable rural environment. Results indicate that: (1) The total value of the average livelihood assets per household in the Sihe village increased significantly from 0.48 in 1988 to 1.288 in 2016. The four types of livelihood assets including natural, material, manpower, and financial assets changed with time. In 1988, the manpower asset was the most important type of livelihood assets, with value accounting for 76.67% of the total value of livelihood assets. With the extension of time, the proportions of the four types of assets in total livelihood assets became closer to each other. The livelihood diversification index of the Sihe village increased from 2.01 in 1988 to 3 in 2016, indicating the rural livelihoods became diverse; (2) Because of the dual influence of external environmental factors and the rural development policies of the country and the region, the livelihoods changed towards agricultural sector from 1988 to 2008, and the agricultural livelihoods tended to be diverse. The following trend of livelihood strategy change was observed: from diverse non-agricultural production group (IV) to agricultural and non-agricultural production group (III), then to diverse agricultural production group (II) and finally to agricultural production group (I). After 2008, the livelihoods changed towards non-agricultural sector, and the non-agricultural livelihoods tended to be diverse. This trend of livelihood change is opposite to that before 2008; (3) 2008 is the key year of livelihood change. Livelihood change caused changes in the scale, structure, and morphology of rural settlement land use, which eventually led to the change of rural residential land use.

Five-Year Progression of Refractive Errors and Incidence of Myopia in School-Aged Children in Western China.

BACKGROUND: To determine the change in refractive error and the incidence of myopia among school-aged children in the Yongchuan District of Chongqing City, Western China. METHODS: A population-based cross-sectional survey was initially conducted in 2006 among 3070 children aged 6 to 15 years. A longitudinal follow-up study was then conducted 5 years later between November 2011 and March 2012. Refractive error was measured under cycloplegia with autorefraction. Age, sex, and baseline refractive error were evaluated as risk factors for progression of refractive error and incidence of myopia. RESULTS: Longitudinal data were available for 1858 children (60.5%). The cumulative mean change in refractive error was -2.21 (standard deviation [SD], 1.87) diopters (D) for the entire study population, with an annual progression of refraction in a myopic direction of -0.43 D. Myopic progression of refractive error was associated with younger age, female sex, and higher myopic or hyperopic refractive error at baseline. The cumulative incidence of myopia, defined as a spherical equivalent refractive error of -0.50 D or more, among initial emmetropes and hyperopes was 54.9% (95% confidence interval [CI], 45.2%-63.5%), with an annual incidence of 10.6% (95% CI, 8.7%-13.1%). Myopia was found more likely to happen in female and older children. CONCLUSIONS: In Western China, both myopic progression and incidence of myopia were higher than those of children from most other locations in China and from the European Caucasian population. Compared with a previous study in China, there was a relative increase in annual myopia progression and annual myopia incidence, a finding which is consistent with the increasing trend on prevalence of myopia in China.

Prevalence of open-angle glaucoma in southwestern China: the Yongchuan Glaucoma study.

This study examined the prevalence of primary open-angle glaucoma (POAG) among residents aged ≥50 years living in Yongchuan district of Chongqing. Stratified cluster sampling was employed in random selection to estimate the prevalence of glaucoma from April to June, 2005. Twenty-nine villages or neighborhood communities were randomly selected in urban area (Zhongshan Road), suburban area (Shanjiao Town) and exurban area (Zhutuo Town) of this district. All the respondents underwent detailed ophthalmic examinations. The examinations included questionnaire investigation, visual acuity test, naked-eye examination, measurement of peripheral anterior chamber depth (Van Herrick's technique), detection of intraocluar pressure (IOP) with a Perkins hand-held applanation tonometer (HA-2) and examination of the optic disc by using a 78 diopters (D) lens (including the cup-disc ratio, cup/disc ratio asymmetries, horizontal and vertical diameter, notching and optic disc hemorrhages). A total of 5938 residents were actually examined, and the response rate was 85.19%. The crude prevalence of POAG was 0.86% (n=51/5938, 95% CI 0.64%-1.11%). There were 24 males and 27 females in the glaucoma group. The glaucoma prevalence was not significant different in case number between the male and female subjects (P=0.4900). Furthermore, no association between age or schooling and POAG was noted (P=0.8030, 0.0734). Out of 51 subjects with POAG, unilateral glaucoma-related blindness occurred in 38 subjects (74.5%) and bilateral glaucoma-related blindness was found in 7 subjects (13.7%). This study exhibited that the prevalence of POAG was 0.86% among residents aged ≥50 years living in Yongchuan District of Chongqing. The vision loss caused by POAG in this population was obviously higher than that previously reported in other studies. Glaucoma management, detection of affected persons and handling of the burden of glaucoma should be the priorities of the agenda of local health authorities of Western China.