[Topography-guided transepithelial corneal collagen cross-linking by sequential ultraviolet A irradiation in different diameters for progressive keratoconus in adults].

Objective: To compare the efficacy and safety of a novel customized topography-guided transepithelial corneal collagen cross-linking (TG-CXL) procedure by sequential ultraviolet A irradiation in different diameters and conventional transepithelial corneal collagen cross-linking (TE-CXL) in adult patients with progressive keratoconus. Methods: A prospective cohort study was conducted. Adult patients diagnosed with progressive keratoconus in the Affiliated Xiamen Eye Center of Xiamen University were continuously recruited and randomly assigned to receive the TG-CXL or TE-CXL procedure from March 2020 to March 2021. Patients in the TE-CXL group were irradiated in the central 9-mm zone of the cornea (total energy, 7.2 J/cm2; irradiance, 45 mW/cm2), while patients in the TG-CXL group were first irradiated with the protocol used in the TE-CXL group, and further irradiated in the central 6-mm zone (total energy, 3.6 J/cm2; irradiance, 9 mW/cm2). The subjective symptom of pain and corneal fluorescein sodium staining were scored within postoperative 3 days. Slit lamp examination, measurements of uncorrected visual acuity (UCVA) and best-corrected visual acuity (BCVA), corneal topography, anterior segment optical coherence tomography, in vivo corneal confocal microscopy, corneal endothelial cell count, and non-contact tonometry were performed before surgery and at 3, 6, and 12 months after surgery. Results: A total of 66 patients were enrolled (mean age, 23.0±3.3 years old), with 33 patients (33 eyes) in each group. No statistically significant differences were found in age, gender, and maximum keratometry (Kmax) between the two groups (P>0.05). On day 1 after surgery, the average pain score of the TG-CXL group (2.21±0.45) was significantly higher than that of the TE-CXL group (1.32±0.33) (P<0.05). The pain was rapidly alleviated in both groups on days 2 and 3. On days 1 and 2, the corneal fluorescein sodium staining scores in the TG-CXL group (4.15±0.83 and 2.21±0.60, respectively) were significantly higher than those in the TE-CXL group (1.76±0.56 and 0.85±0.51, respectively, P<0.001), while there was no significant difference between the two groups at day3 (P=0.184). The UCVA and BCVA of the TG-CXL group at 3, 6, and 12 months after surgery were significantly improved when compared with the baseline. At 3, 6, and 12 months, the BCVA (LogMAR) of the TG-CXL group (0.21±0.15, 0.22±0.16, and 0.22±0.16, respectively) were significantly improved when compared with those of the TE-CXL group(0.32±0.15, 0.34±0.15, and 0.36±0.16, respectively, P<0.01). However, there was no significant difference in UCVA between groups at any time point after surgery (P>0.05). The spherical and cylindrical power values of the TG-CXL group were improved when compared with the baseline (P<0.05). However, no significant difference in spherical power values was found between the two groups at any time point after surgery (P>0.05). Meanwhile, there were significant differences in cylindrical power values between the two groups at 6 and 12 months after surgery (P<0.05). The Kmax in the TG-CXL group was improved at all of the time points after surgery when compared with the baseline (P<0.001), while no significant difference in Kmax was found at any time point after surgery in the TE-CXL group when compared with the baseline (P>0.05). At 6 and 12 months after surgery, the Kmax values in the TG-CXL group were significantly lower than the TE-CXL group (P<0.05). No significant differences were found in flat keratomety, steep keratometry, the minimal thickness of the cornea, endothelial cell density, and intraocular pressure between the two groups at any time point after surgery (P>0.05). Within one month after surgery, optical coherence tomography revealed the increased density in the anterior stroma in both groups. In most patients in the TG-CXL group, a demarcation line was visible in the central and para-central corneal stroma, representing a clear and continuous, high-signal arc-shaped linear structure, which was deeper in the central cornea than the para-central cornea. In contrast, a demarcation line, fuzzy and focally discontinuous, was visible only in a few patients in the TE-CXL group, with an almost uniform depth in the central and the para-central cornea. Confocal microscopy demonstrated an apparent mesh-like cross-linked collagen structure in the superficial and intermediate corneal stroma at all time points after surgery in the TG-CXL group, with thickening stromal collagen fibers and an increased number of interconnections. In contrast, the mesh-like structure and number of interconnections in the superficial corneal stroma were significantly reduced at 12 months after surgery in the TE-CXL group, with no cross-linking structure in the intermediate corneal stroma at any time point after surgery. No serious complications such as corneal infection, sterile corneal ulcer, and persistent epithelial defect were observed in both groups during the follow-up of 12 months. Conclusions: The TG-CXL procedure by sequential irradiation in two different diameters with ultraviolet A light was effective and safe in the management of progressive keratoconus in adults, achieving significant refractive improvement. This might be a good technical alternative for refractive corneal cross-linking surgery.

[Correlation between morphological characteristics of retinal microvessels and diabetic kidney disease in patients with type 2 diabetes mellitus based on a machine learning model].

Objective: To explore the correlation between the morphological characteristics of retinal microvessels and diabetic kidney disease (DKD) in patients with type 2 diabetes mellitus (T2DM). Methods: The clinical data and fundus photography of patients with T2DM treated in the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2020 were retrospectively collected and analyzed. According to the presence of abnormal renal function, the patients were divided into DKD group and control group. The morphology and structure of fundus vessels were digitized by U-Net depth convolution neural network, and the correlation between fundus vascular morphology and DKD was analyzed by multivariate logistic regression. Results: A total of 648 patients with T2DM were enrolled, including 410 males and 238 females, and aged (53±10) years. There were 398 and 250 cases in control and DKD groups, respectively. Meanwhile, 1 296 fundus images were collected. Compared with control group, the male ratio (68.4% vs 60.1%, P=0.032), age [(54±9) vs (52±10) years, P=0.005], blood pressure [(136.8±17.3) vs (130.3±15.4) mmHg（1 mmHg=0.133 kPa）, P<0.001], total cholesterol [(4.5±1.4) vs (4.2±1.0) mmol/L, P=0.009], triglyceride [M (Q1, Q3)][1.7 (1.2, 3.0) vs 1.4 (1.0, 2.3) mmol/L, P<0.001] and Cystatin C [(0.9 (0.8, 1.0) vs 0.8 (0.7, 0.9) mg/L, P<0.001] were higher in the DKD group, while high-density lipoprotein [(1.0±0.3) vs (1.1±0.3) mmol/L, P=0.001] was lower in the DKD group. Multivariate logistic regression analysis showed that the risk of DKD in the third quartile (right eye: OR=1.825, 95%CI: 1.204-2.768, P=0.005) and fourth quartile (left eye: OR=1.929, 95%CI: 1.218-3.055, P=0.005) was higher than that in the lowest quartile of vascular curvature after adjusting for age and gender. The increase of average diameter of retinal vein was associated with the risk of DKD (left eye: OR=1.044, 95%CI: 1.013-1.075, P=0.005). The decrease of vascular fractal dimension (fourth quartile of left eye: OR=0.444, 95%CI: 0.199-0.987, P=0.046) and retinal vascular density (the second and fourth quartile of the right eye: OR=0.639, 95%CI: 0.409-0.998, P=0.049; OR=0.534, 95%CI: 0.331-0.864, P=0.010) were related to the risk of DKD. Conclusions: The abnormal morphological characteristics of retinal microvessels are related to the occurrence of DKD. The increase of retinal vein diameter and the decrease of retinal vessel density correlate with the occurrence of DKD.

[Short-term efficacy and safety of vedolizumab in patients with inflammatory bowel disease].

Objective: To evaluate the short-term efficacy and safety of vedolizumab in patients with inflammatory bowel disease (IBD). Methods: Patients with moderate and severe active IBD at the first use of vedolizumab from May 1 to October 31, 2021 were retrospectively enrolled. Then the clinical characteristics, and the efficacy and safety of vedolizumab were evaluated. Meanwhile, the clinical response rate, biological response rate and endoscopic response rate were calculated. Multivariate analysis was used to evaluate the independent influencing factors of short-term clinical efficacy and safety. Results: A total of 78 patients (44 males and 34 females) with IBD were enrolled, with a mean age of (40.5±11.9) years. The clinical remission rate, clinical response rate, biological remission rate, biological response rate and endoscopic remission rate was 60.3% (47/78), 85.9% (67/78), 70.5% (55/78), 43.6% (34/78) and 47.0% (31/66) respectively after 14 weeks of treatment. Body mass index (BMI) ≥ 18.5 kg/m2 (HR=5.04, 95%CI: 1.50-16.91, P=0.009) and biological remission at 6 weeks of treatment (HR=15.22, 95%CI: 3.16-73.38, P=0.001) were predictors of endoscopic remission at 14 weeks of treatment. Adverse reactions occurred in 57 patients, with an incidence of 73.1%. The main manifestations were liver and kidney damage (37.2%) and infection (26.9%). Conclusions: More than half of patients with moderate and severe active IBD can achieve clinical remission after 14 weeks of vedolizumab treatment. Baseline BMI level and biological remission at 6 weeks of treatment are predictors of mucosal healing at 14 weeks. The incidence of adverse reactions is not low, although serious adverse reactions are rare in short-term treatment.

A novel risk prediction model of pyroptosis-related genes for the prognosis and immunotherapy response of endometrial cancer.

OBJECTIVE: This study aims to develop a risk prediction model of pyroptosis-related genes based on its impact on immunotherapy sensitivity of uterine corpus endometrial carcinoma (UCEC), one of the most common and threatening gynecological malignancies. PATIENTS AND METHODS: Through multiple bioinformatics analysis, we obtained raw counts of RNA-sequencing data and corresponding clinical information related to UCEC from The Cancer Genome Atlas (TCGA) and Gene Expression Profiling Interactive Analysis (GEPIA) to investigate the potential mechanisms of differentially expressed pyroptosis-related genes (DEPRGs), including the correlation between DEPRGs and prognosis, tumor immune microenvironment and the immunotherapy sensitivity of UCEC patients. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment Analysis were used to figure out the functional differences. Furthermore, a mRNA-miRNA-lncRNA network was constructed to identify potential impact of pyroptosis on tumor progression. RESULTS: In this study, we achieved six DEPRGs (CASP3, GPX4, GSDMD, NOD2, PYCARD and TIRAP) and constructed a 6-gene signature which classified UCEC patients in the TCGA cohort into a low-risk group or a high-risk group. Patients in the low-risk group showed significantly longer survival time (p=0.000373). The risk score was also confirmed as an independent prognostic factor combining with the clinical characteristics. GO and KEGG functional analysis revealed the possible molecular mechanisms by which six DEPRGs influence anti-tumor immunity in UCEC patients. In addition, we found that two DEPRGs (GPX4, TIRAP) were not only significantly associated with tumor mutational burden (TMB) or microsatellite Instability (MSI), but also involved in regulating the number and function of CD8+ cells. CONCLUSIONS: Upon comprehensive bioinformatics analysis, it was concluded that pyroptosis-related genes (PRGs) could predict the prognosis of EC patients and be affected in modulating the anti-tumor immune responses for patients with EC.

[Early infantile epileptic encephalopathy caused by PACS2 gene variation: three cases report and literature review].

Objective: To explore the clinical features of three early-onset infantile epileptic encephalopathy (EIEE) patients with variations in phosphofurin acidic cluster sorting protein 2 (PACS2) gene and to review related literature. Methods: The clinical data and genetic features of three early infantile epileptic encephalopathy 66 (EIEE66) patients with a PACS2 gene variant diagnosed by the Department of Neurology, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology, from January 2019 to January 2020 were retrospectively analyzed. A literature search with "PACS2 gene" "PACS2" "epileptic encephalopathy, early infantile, 66" and"early infantile epileptic encephalopathy 66" as key words was conducted at PubMed, China National Knowledge Infrastructure (CNKI), and Wanfang Data Knowledge Service Platform (up to July 2020). Case reports of patients with PACS2 gene variants and related clinical data were chosen and reviewed. Results: Case 1, a girl aged 2 years and 2 months was hospitalized because of repetitive seizures within more than two years and 6 convulsions within 2 days due to fever. The seizures occurred at the age of 7 days, characterized by focal seizures and generalized tonic-clonic seizures. Sometimes, the frequency of seizures increased with high fever. Regular treatment had not been implemented in the early stage, later seizures were controlled by valproic acid treatment. Case 2, a female 5 months of age, was admitted due to recurrent convulsions in nearly five months. Focal seizures occured at the age of 5 days. And the brain magnetic resonance imaging (MRI) confirmed abnormal cerebellar hemispheres and cerebellar vermis, as well as cerebellar dysplasia. Several antiepileptic drugs and ketogenic diet were ineffective in the early months, and later seizures were controlled with the treatment with levetiracetam and valproic acid. Case 3, a five-month-old girl, was admitted because of recurrent convulsions for nearly five months. At the age of 3 days, she had tonic seizures, and showed good response to levetiracetam and valproic acid. All the three cases were accompanied by development delay and dysmorphic facial appearance, and got seizure-free with the treatment with valproic acid. All copy-number variant analysis and trio whole exome sequencing revealed a recurrent heterozygous missense variant (c.625G>A) in PACS2 gene. No related reports were found in Chinese journals, while 4 reports were found in English literature, describing 17 patients in total. With these 3 patients included, 20 cases had only two missense PACS2 gene variants, in whom 19 cases carried the variant c. 625G>A (p.Glu209Lys) and 1 case carried the variant c. 631G>A (p.Glu211Lys). Epilepsy was the first reported symptom in all patients, and 17 cases had seizures during the first week of life. Out of the various seizure types observed, focal seizures were the predominant types (13 cases), whereas tonic, clonic, tonic-clonic seizures and non-motor seizures (such as facial flushing) were also reported. Almost all patients showed facial dysmorphism and developmental delay to different degrees. Total of 16 patients had abnormal brain MRI recordings, and 13 cases had cerebellar hypoplasia. More specifically, 7 cases showed inferior vermian hypoplasia, and 3 cases showed hypothalamic fusion anomaly. The treatment was mainly aimed to control the symptoms. And the recommended effective treatment for epilepsy has not been reported yet. Conclusions: PACS2-related early infantile epileptic encephalopathy is an autosomal dominant disease, characterized by seizure onset within the first week of life in most cases, dysmorphic facial appearance, and various degrees of developmental retardation. Treatment with valproic acid showed good effect.

[The correlation between polycyclic aromatic hydrocarbons and idiopathic membranous nephropathy].

Objective: To explore the relationship between polycyclic aromatic hydrocarbons (PAHs), the major component of atmospheric pollutant particulate matter 2.5 (PM2.5) and idiopathic membranous nephropathy (IMN) and its possible mechanisms. Methods: Thirty-five (24 males and 11 females) patients with IMN confirmed by renal biopsy pathology in the First Affiliated Hospital of Zhengzhou University from June to September 2020 were selected as the research group, with an age of (47.3±12.9) years. Meanwhile, patients with secondary membranous nephropathy (SMN, n=10) and minimal-change disease (MCD, n=10) were selected as control group. Blood samples were collected to detect PAHs and heme oxygenase-1 (HO-1), and the relationship of PAHs with clinical indicators and oxidative stress indicator HO-1 was analyzed. Results: Compared with SMN and MCD groups [(2.6±1.7) mg/L and (2.8±1.0) mg/L], the concentration of PAHs in IMN group [(4.1±1.9) mg/L] increased (both P<0.05). The concentration of PAHs was positively correlated with the levels of ß2 microglobulin, total cholesterol and 24-hour urine total protein (r=0.509, 0.336, and 0.653, respectively, all P<0.05), but was negatively correlated with the levels of total protein and albumin (r=-0.499,-0.530, respectively, both P<0.05). Additionally, the concentration of PAHs was negatively correlated with the level of HO-1 (r=-0.358, P=0.017). Linear regression analysis showed that the concentration of HO-1 decreased by 1.737 µg/L when the concentration of PAHs increased by 1 mg/L (P=0.035). Conclusion: PAHs have a high level in IMN patients, and may be involved in the occurrence and development of IMN through oxidative stress.

[Effects and mechanisms of Xuezhikang on preventing contrast-induced acute kidney injury in diabetic rats].

Objective: To explore the effects and mechanisms of Xuezhikang on preventing contrast-induced nephropathy (CIN) in diabetic rats. Methods: Streptozotocin (65 mg/kg) was injected intraperitoneally to establish a diabetes model in 7-week-old male Sprague-Dawley (SD) rats. After 4 weeks of modeling, 24 diabetic rats were randomly divided into 4 groups: sham group, CIN group, CIN+vehicle (Veh) group and Xuezhikang group. All animals were sacrificed at 24 hours after administration of contrast. Blood and kidney tissues were collected to detect biochemical, inflammation-related, oxidative stress-related and pathological indicators. Results: After administration of contrast agent, the renal function-related indicators were decreased in Xuezhikang group compared with CIN+Veh group [serum creatinine (SCr): (59.3±3.3) µmol/L vs (73.2±4.1) µmol/L; blood urea nitrogen (BUN): (13.8±0.5) mmol/L vs (16.3±0.6) mmol/L; serum neutrophil gelatinase-associated lipocalin (sNGAL): (41.4±2.0) ng/ml vs (54.9±4.4) ng/ml; urinary kidney injury moleculer-1 (uKIM-1): (11.1±0.5) ng/ml vs (16.6±0.5) ng/ml] (all P<0.05). Histological analysis showed that the severity of renal tubule dilatation, brush border loss and renal tubular cell necrosis in Xuezhikang group was better than that of CIN+Veh group. Additionally, the oxidative stress-related indicators of Xuezhikang group improved compared with those of CIN+Veh group [malondialdehyde (MDA): (12.1±0.7) nmol/mg vs (15.5±0.8) nmol/mg, superoxide dismutase (SOD): (35.0±2.2) U/mg vs (23.7±3.4) U/mg, renal nitrite: (1.7±0.1) nmol/mg vs (1.2±0.1) nmol/mg, all P<0.05]. Meanwhile, Xuezhikang pretreatment downregulated the mRNA and protein expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (both P<0.05). Conclusion: The current study suggests that Xuezhikang protects against CIN in diabetic rats by inhibiting oxidative stress and inflammation.

RBBP6 aggravates the progression of ovarian cancer by targeting PIK3R6.

OBJECTIVE: RBBP6 is identified to be a cancer-associated gene by bioinformatics analysis. This study aims to explore the role of RBBP6 in regulating proliferation and metastasis in ovarian cancer, thus providing theoretical references for ovarian cancer treatment. PATIENTS AND METHODS: Differential expressions of RBBP6 in ovarian cancer and normal ones were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between RBBP6 and prognosis in ovarian cancer patients was analyzed. The interaction between RBBP6 and PIK3R6 was detected by bioinformatics analysis and Dual-Luciferase reporter assay. Moreover, regulatory effects of RBBP6 and PIK3R6 on proliferative and migratory potentials in A2780 and CAOV3 cells were examined by Cell Counting Kit-8 (CCK-8) and transwell assay, respectively. Finally, tumorigenicity assay was conducted in nude mice to illustrate the in vivo regulations of PBBP6 and PIK3R6 on ovarian cancer growth. RESULTS: RBBP6 was upregulated in ovarian cancer tissues than normal ones. RBBP6 was irrelevant to age, tumor size and tumor node metastasis (TNM) staging in ovarian cancer patients, but correlated to lymphatic metastasis and distant metastasis. RBBP6 was abundantly expressed in ovarian cancer cells, and among the tested cell lines, CAOV3 and A2780 expressed the highest level of RBBP6. Knockdown of RBBP6 attenuated in vitro proliferative and migratory potentials in CAOV3 and A2780 cells. PIK3R6 was the target gene binding RBBP6, which was positively regulated by RBBP6. Overexpression of PIK3R6 could abolish the inhibited proliferative and migratory potentials in ovarian cancer cells with RBBP6 knockdown. In addition, the knockdown of RBBP6 slowed the in vivo growth of ovarian cancer in nude mice, and the alleviated cancer progression was reversed by overexpression of PIK3R6. CONCLUSIONS: RBBP6 is highly expressed in ovarian cancer cases, which stimulates proliferative and migratory potentials by targeting PIK3R6. RBBP6 may be a novel therapeutic target for ovarian cancer.

[Changes of urinary monocyte chemotactic protein 1 and epidermal growth factor and their correlations with clinicopathology in idiopathic membranous nephropathy patients].

Objective: To explore the changes and clinical value of urinary monocyte chemotactic protein 1 (MCP-1), epidermal growth factor (EGF) and their ratio in patients with idiopathic membranous nephropathy (IMN). Methods: A total of 67 IMN patients were enrolled according to kidney biopsy in the Department of Nephrology of the First Affiliated Hospital of Zhengzhou University from January 2017 to December 2018. The patients included 28 males and 39 females, with an average age of (47.6±14.2) years. Eighteen age-and sex-matched healthy controls were also enrolled. Clinical and pathological data, blood and urine samples of all subjects were collected. Urinary MCP-1 and EGF level were detected by enzyme-linked immunosorbent assay (ELISA). And then the levels of urinary MCP-1, EGF and the ratio of EGF/MCP-1 and their correlations with clinicopathology were analyzed. Results: There was no statistical difference of the urine EGF levels between the two groups [8.3(6.0,12.6) vs 8.4(6.5,10.2) ng/mg Cr, P=0.575]. The urine MCP-1 levels of IMN patients were 0.37 (0.21, 0.69) ng/mg Cr, which was statistical significantly higher than those of the control group [0.09 (0.02, 0.19) ng/mg Cr] (P<0.001), while the EGF/MCP-1 ratio was lower than that of the control group [22.2(15.1,36.6) vs 87.6(71.2,132.7), P<0.001]. Urine MCP-1 was negatively correlated with eGFR (r=-0.303, P=0.012), but positively correlated with the urinary ratio of albumin to creatinin (r=0.368, P=0.002). EGF was positively correlated with eGFR (r=0.722, P<0.001), but negatively correlated with the severity of interstitial fibrosis and renal tubular atrophy (IFTA) (r=-0.312, P=0.011). EGF/MCP-1 ratio was positively correlated with eGFR (r=0.693, P<0.001), but negatively correlated with the severity of the urinary ratio of albumin to creatinin and IFTA (r=-0.261, P=0.028 and r=-0.684, P<0.001, respectively). Further multivariate logistic regression analysis showed that EGF/MCP-1 was a protective factor for moderate-to-severe IFTA (OR=0.891, 95%CI: 0.844-0.949, P=0.008). Conclusion: Patients with IMN have elevated urine MCP-1 level and decreased EGF/MCP-1 ratio, which correlate with clinical indicators. In particular, EGF/MCP-1 ratio is independently related to moderate-severe IFTA, and may be a potential clinical biomarker for diagnosis of IMN.

[Efficacy of deproteinized calf blood extract eye drops on early recovery after pterygium surgery].

Objective: To investigate the effect of deproteinized calf blood extract eye drops on early postoperative recovery in primary pterygium patients. Methods: This is a prospective randomized controlled study. Patients diagnosed with primary pterygium in single eye at affiliated Xiamen Eye Center of Xiamen University during March 2016 to May 2016 were enrolled. After Pterygium excision with autologous conjunctival transplantation, patients were randomly assigned into four groups by a random number table, treated with anti-inflammaroty drugs only (control group) or combined with the following agents: deproteinized calf blood extract eye drops (DCBE group), carboxymethylcellulose sodium eye drops (CMC group), and recombinant human epidermal growth factor eye drops (rEGF group). Short-form McGill pain questionnaire, slit lamp and corneal fluorescein sodium staining, non-contact intraocular pressure, uncorrected visual acuity (UCVA) and best corrected visual acquity (BCVA) as well as redness score of bulbar conjunctiva were performed before surgery (d0) and on day 1 (d1), day 2 (d2), day 3 (d3), day 7 (d7) and day 14 (d14) after surgery. Results: One hundred and fourteen patients including 43 males and 71 females, aged (48.9±12.5) years, were eventually included in this study. The McGill scores gradually decreased after surgery in all groups. On d2, the McGill score in DCBE group, control group, CMC group and rEGF group was (1.42±0.67), (2.21±0.88), (1.93±1.08) and (1.77±1.18), respectively; On d3, the score was (1.32±0.54), (1.93±0.72), (1.79±0.87) and (1.52±0.77), respectively. On d2 and d3, statistical difference was recorded among groups (d2, F=3.43, P=0.019; d3, F=4.047, P=0.009), and the McGill score of DCBE group was significantly lower than that of CMC group (d2, P=0.047, d3, P=0.017). On d2, the percentage of corneal epithelium defect in DCBE group, control group, CMC group and rEGF group was 8.6%±1.9%, 11.7%±1.7%, 11.5%±1.9% and 10.4%±1.8%, respectively; On d3, the percentage was 4.5%±2.2%, 9.2%±2.4%, 7.4%±2.5% and 5.9%±2.3%, respectively. On d2 and d3, statistical difference of corneal epithelium defect percentage was recorded among groups (d2, F=17.17, P<0.001; d3, F=21.4, P<0.001). On d2, the percentage of corneal epithelium defect in DCBE group was significantly lower than the other three groups (P<0.01); On d3, the percentage of corneal epithelium defect in DCBE group was significantly lower than control group and CMC group (P<0.001), while no difference was found between DCBE group and rEGF group (P>0.05). However, no statistical differences were recorded in the number of patients with vision improvement among the groups (P>0.05). The intraocular pressure remained stable. No differences in the conjunctival redness score were found among the groups after surgery (P>0.05). Conclusion: Our data demonstrated the efficacy of deproteinized calf blood extract eye drops on the postoperative management in patients with primary patients, with faster pain relief and promoted epithelium recovery. (Chin J Ophthalmol, 2019, 55:134-140).

[Accelerated transepithelial corneal collagen cross-linking for progressive keratoconus with a thin cornea: one-year results].

Objective: To evaluate the clinical results of keratoconic eyes with a thin cornea treated with accelerated transepithelial corneal collagen cross-linking (A-TE-CXL) within 1 year. Methods: Nineteen eyes of 19 patients with progressive keratoconus with a minimum corneal thickness from 380 µm to 420 µm (including the epithelium) were included in this prospective, nonrandomized clinical study and treated with A-TE-CXL. Scoring of pain and foreign body sensation, slit lamp examination, uncorrected visual acuity, best corrected distance visual acuity, corneal topography, anterior segment optical coherence tomography, in vivo corneal confocal microscopy and endothelial cell count were assessed before surgery and at 1, 3, 6 and 12 months postoperatively. Paired t test was applied for statistical analysis. Results: Mild pain and moderate foreign body sensation were reported by most patients within postoperative 24 hours, but rapidly disappeared on day 2. Extremely mild epithelial damage was observed within postoperative 24 hours, and the epithelium fully recovered on day 2. Improvement of visual acuity was recorded at 3 and 12 months. Pentacam corneal topography revealed a significant reduction of the thickness of the thinnest location from(395.2±13.8)µm preoperatively to (378.9±17.1)µm at 1 month postoperatively (t=2.982, P<0.01). Front curvature values were reduced postoperatively. K(MAX) was significantly decreased at 12 months (55.67±4.91) compared with (57.35±5.54) preoperatively, while K2 was also significantly decreased at 12 months (52.18±3.70) compared with (52.70±3.56) preoperatively (K(MAX), t=3.044, P<0.01. K2, t=2.384, P<0.05) . Within 1 month postoperatively, optical coherence tomography exhibited an increase of reflectance with a demarcation line in the anterior stroma. In vivo confocal microscopy also showed significant thickening and increased connections of collagen fibers with a maximal depth at about 90 to 120 µm. The corneal endothelial cell density remained stable (t=0.692, P>0.05). None of the patients showed postoperative complications such as corneal infection, scarring and ulceration. Conclusions: Within 1 year postoperatively, A-TE-CXL was effective and safe for the management of progressive keratoconus with a thin cornea. A-TE-CXL showed the advantages of very short time consuming in surgery, rapid recovery and very few complications, and had the potential to become a valid alternative for the treatment of keratoconus. (Chin J Ophthalmol, 2017, 53: 694-700).

An improved RT-IPCR for detection of pyrene and related polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous homogeneous chemicals which are well known by carcinogens, mutagens and endocrine disorder. Here, an improved real-time immuno-PCR (RT-IPCR) was developed for detection of pyrene and its homologs in water samples. The PAHs in sample compete with pyrene-modified DNA to bind with monoclonal antibody (McAb) coated on PCR plate. The reporter DNA was exponentially amplified by real-time PCR instrument using Fast Start universal SYBR Green Master (ROX) kit. Only two reaction steps were needed to accomplish the detection. The assay had a good linear range from 5 pmol L(-1) to 5 nmol L(-1) with a detection limit of 3.5 pmol L(-1). For application assay, the average recoveries from tap water, lake water and mineral water were 98.4%, 98.2% and 99.7%, respectively which showed a good correlation (R(2)=0.9906) with those from GC-MS. The results indicated that the improved RT-IPCR seems to be a potential method for simple and ultrasensitive detection of pyrene and some homologues in environment water samples.

Effect and Mechanism of Portal Blood Stasis Removal on Intestinal Endotoxemia and Hepatic Ischemia Reperfusion Injury.

OBJECTIVE: We used a rabbit model of hepatic ischemia reperfusion in situ to observe the change of portal venous endotoxin level before reperfusion, and the effect of portal blood stasis removal on intestinal endotoxemia and hepatic ischemia reperfusion injury. The purpose was to find an ideal method for portal blood stasis removal and provide the experimental proof for clinical application of hepatectomy. METHODS AND MATERIALS: To investigate the effect of portal blood stasis removal on intestinal endotoxemia and hepatic ischemia reperfusion injury, a rabbit hepatic ischemia reperfusion injury model was established and treated with removal of portal blood stasis before the portal blood circulation was resumed. Serum endotoxin content, alanine aminotransferase (ALT), hyaluronic acid (HA), and content of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), superoxide dismutase (SOD) and activation of nuclear factor-κB (NF-κB) in liver tissue were examined respectively. RESULTS: In portal blood stasis the level of serum endotoxin significantly decreased with each 2.5 mL blood removal (P < .01), subsequently reaching a minima at the 7.5 mL blood removal (P > .05). Removing portal blood stasis ameliorated endotoxemia and hepatic ischemia reperfusion injury as shown by ALT, HA, MDA, SOD, TNF-α, IL-6, and activation of NF-κB compared to no removal. The first 5 mL portal blood stasis contains high volume of endotoxin which may be responsible for hepatic reperfusion injury. CONCLUSION: Removal of portal blood stasis before the resume of splanchnic circulation may ameliorate intestinal endotoxemia and hepatic ischemia reperfusion injury.

Real-time immuno-PCR for ultrasensitive detection of pyrene and other homologous PAHs.

Polycyclic aromatic hydrocarbons (PAHs) are significant environmental pollutant that can lead to cancer and endocrine system disrupting. Here we developed a real-time immuno-PCR (RT-IPCR) assay based on a biotinylated reporter DNA system for ultrasensitive detection of pyrene (PYR) and homologous PAHs in water. The PAHs in sample compete with PYR-OVA coated on PCR plate to bind with monoclonal antibody (McAb). The biotinylated goat anti-mouse IgG (Bio-IgG) can be captured by the McAb bound with PYR-OVA. Then streptavidin is bound with biotin on Bio-IgG. Finally biotinylated reporter DNA is captured by the streptavidin and quantified by real-time PCR using FastStart universal SYBR Green Master (ROX) kit. The linear range of the assay was from 500 fmol L(-1) to 5 nmol L(-)) with a detection limit of 450 fmol L(-1). The average recoveries of PYR and homologous PAHs from lake water, tap water and commercial mineral water were 96.8%, 101.4% and 99.6% respectively, indicating that water samples had little interfere with the assay. The results demonstrated that the developed RT-IPCR might be a potential method for ultrasensitive detection of PYR and homologous PAHs in drinking and environment water sample.

An enzyme-linked immunosorbent assay for detection of pyrene and related polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) can form DNA-binding compounds that show genotoxicity and carcinogenicity. Pyrene, as a PAH, was covalently linked to carrier protein bovine serum albumin and ovalbumin. A monoclonal antibody (McAb) was produced that showed high cross-reactivity values with chrysene (169.73%), benzo[a]pyrene (693.34%), benzo[a]anthracene (16.36%), and indeno[1,2,3-cd]pyrene (40.96%) and showed no significant cross-reactivity values with other homologues (<0.1%). A competitive enzyme-linked immunosorbent assay (ELISA) was developed for detection of pyrene and some homologues in water samples. The detection limit of the assay was 65.08 pg ml(-1). The average recoveries of PAHs from tap water, lake water, and mineral water were 99.13, 99.74, and 99.19%, respectively, indicating that matrices of water samples do not interfere with the assay. The results demonstrated that the developed ELISA seems to be a potential method for monitoring of pyrene and some homologous PAHs in water samples.

Note: Determination of effective gas diffusion coefficients of stainless steel films with differently shaped holes using a Loschmidt diffusion cell.

In this work, an in-house made Loschmidt diffusion cell is used to measure the effective O(2)-N(2) diffusion coefficients through four porous samples of different simple pore structures. One-dimensional through-plane mass diffusion theory is applied to process the experimental data. It is found that both bulk diffusion coefficient and the effective gas diffusion coefficients of the samples can then be precisely determined, and the measured bulk one is in good agreement with the literature value. Numerical computation of three-dimensional mass diffusion through the samples is performed to calculate the effective gas diffusion coefficients. The comparison between the measured and calculated coefficient values shows that if the gas diffusion through a sample is dominated by one-dimensional diffusion, which is determined by the pore structure of the sample, these two values are consistent, and the sample can be used as a standard sample to test a gas diffusion measurement system.

Dynamic distribution of bone marrow-derived mesenchymal stromal cells and change of pathology after infusing into mdx mice.

BACKGROUND: Mesenchymal stromal cells (MSC) are attractive candidates for the treatment of Duchenne muscular dystrophy (DMD) but how the donor MSC distribute in multiple organs and whether the increased dystrophin leads to a change in the pathology of mdx mice is still uncertain. In this research we detected the distribution of MSC and the pathology of mdx mice after MSC infusion. METHODS: MSC were isolated from rat bone marrow (BM) and expanded in proliferation medium. MSC of the fifth passage were delivered intravenously into irradiated mdx mice. The distribution of MSC labeled by [3H]TdR into a recipient's organs was calculated by radioactivity. The expression of dystrophin was detected at weeks 4, 8, 12 and 16 after MSC transplantation by immunofluorescence staining, RT-PCR and Western blot. Serum creatine kinase (CK) and centrally nucleated fiber (CNF) were also detected to assess the change in pathology. RESULTS: 24-48 h after transplantation, MSC were mainly found in the BM, liver and lung. The radioactivity in these organs decreased, whereas skeletal and myocardial muscle radioactivity increased gradually over time. In accordance with the increased radioactivity in skeletal muscle, the amount of dystrophin-positive myofibers increased. Furthermore, serum CK and CNF decreased slightly, suggesting specific pathophysiologic features of the dystrophic muscle were partially restored. DISCUSSION: Upon certification of the distribution of transplanted MSC in irradiated mdx mice, we found evidence of myogenic differentiation of MSC in skeletal muscle. This research may help us understand the mechanism of therapy of MSC transplantation.

Serum cytokine levels are altered in patients with West syndrome.

The objective of this study is to explore the neuroimmunomodulator effect of interleukin (IL)-2, tumor necrosis factor (TNF)-alpha and interferon (IFN)-alpha in West syndrome (WS). Twenty-three cases of WS (13 males and 10 females, aged 4-14 months old) who first visited and consisted from 10 cryptogenic and 13 symptomatic, were enrolled in this study. Double-antibody sandwich enzyme-linked immunosorbent assay was used to measure serum IL-2, TNF-alpha and IFN-alpha levels in 23 patients with WS and the data were compared to those of 15 healthy infants who were matched with regard to age and sex. Levels of all three cytokines were significantly higher in both cryptogenic and symptomatic WS groups than the control group. Serum IL-2 levels in symptomatic WS were significantly higher than that in cryptogenic WS. There was a positive correlation between IL-2 and TNF-alpha in both cryptogenic and symptomatic WS groups. The immune systems of patients with WS are in an activated state. An imbalance in cytokine levels may be involved in the immunopathology of WS.