Animals ; Macaca mulatta ; Optic Disk ; Glaucoma ; Craniocerebral Trauma ; Intraocular Pressure ; Low Tension Glaucoma

Background: Excessive growth of keratinocytes is the critical event in the etiology of psoriasis.However, the underlying molecular mechanism of psoriatic keratinocyte hyperproliferation is still unclear. Objective: This study aimed to figure out the potential contributory role of S-phase kinase-associated protein 2 (SKP2) in promoting the hyperproliferation of keratinocytes in psoriasis. Methods: We analyzed microarray data (GSE41662) to investigate the gene expression of SKP2in psoriatic lesion skins compared with their adjacent non-lesional skin. Then, we further confirmed the mRNA and protein expression of SKP2 in human psoriatic skin tissues, imiquimod (IMQ)-induced psoriatic mice back skins and tumor necrosis factor α (TNF-α), interleukin (IL)-17A and IL-6-stimulated keratinocytes by using real-time quantitative polymerase chain reaction and western blot (WB). Furthermore, we explored the potential pathogenic role and its underlying cellular mechanism of SKP2 in promoting keratinocytes hyperproliferation through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell cycle detection, 5-ethynyl-2′-deoxyuridine staining and WB. Finally, we determined whether inhibition of SKP2 can effectively alleviate the keratinocytes hyperproliferation in vivo. Results: We identified that SKP2 is aberrantly upregulated in the psoriatic lesion skin and cytokines-stimulated keratinocytes. Moreover, upregulated SKP2 augments cytokines-induced keratinocytes hyperproliferation. Mechanistically, enhanced SKP2 increased the S phase ratio through inhibiting Cyclin-Dependent Kinase Inhibitor p27 (P27 Kip1) expression. Correspondingly, suppression of SKP2 with SMIP004 can significantly ease the epidermis hyperplasia in vivo. Conclusion Our results suggest that elevated SKP2 can empower keratinocytes proliferation and psoriasis-like epidermis hyperplasia via downregulation of P27 Kip1. Therefore, targeting SKP2-P27 Kip1 axis might be a promising therapeutic strategy for the treatment of psoriasis in future.

Objective To investigate the changes in gastrointestinal hormones during the progression of liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD), and to provide a basis for digestive function impairment. Methods A prospective analysis was performed for 326 patients with NAFLD who attended the outpatient service and were hospitalized and treated in Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine from October 2018 to June 2020, and FibroTouch was used to measure liver stiffness measurement (LSM). According to the presence or absence of liver fibrosis, they were divided into non-liver fibrosis group (group A, 161 patients with LSM < 7.3 kPa) and liver fibrosis group (group B, 165 patients with LSM ≥7.3 kPa). According to the fibrosis degree, the patients were further divided into F0-1 group (LSM < 7.3 kPa), F2 group (7.3 kPa ≤LSM < 9.7 kPa), F2-3 group (9.7 kPa ≤LSM < 12.4 kPa), F3-4 group (12.4 kPa ≤LSM < 17.5 kPa), and F4 group (LSM ≥17.5 kPa). Related data were collected, including age, sex, liver function parameters, and gastrointestinal hormones. The independent samples t -test and the one-way analysis of variance were used for comparison of normally distributed continuous data between groups, and the nonparametric Mann-Whitney U test and the Kruskal-Wallis H test were used for comparison of non-normally distributed continuous data between groups. A Spearman correlation analysis was used to investigate the correlation between LSM and liver function parameters. Results Comparison of liver function and gastrointestinal hormones showed that there were significant differences between groups A and B in alanine aminotransferase (ALT) ( Z =-3.778, P < 0.001), aspartate aminotransferase (AST) ( Z =-3.320, P =0.001), gamma-glutamyl transpeptidase (GGT) ( Z =-3.040, P =0.002), cholecystokinin (CCK) ( t =-2.944, P =0.003), and lipopolysaccharide (LPS) ( Z =-2.317, P =0.020). There were significant differences in ALT ( χ 2 =23.113, P < 0.001), AST ( χ 2 =23.415, P < 0.001), ALP ( χ 2 =15.962, P =0.003), GGT ( χ 2 =20.172, P < 0.001), and CCK ( F =2.687, P =0.031) between the F0-1 group with 161 patients, the F2 group with 89 patients, the F2-3 group with 46 patients, the F3-4 group with 16 patients, and the F4 group with 14 patients. LSM was positively correlated with direct bilirubin, ALT, AST, alkaline phosphatase, and GGT ( r =0.128, 0.266, 0.225, 0.137, and 0.213, all P < 0.05). Conclusion Liver fibrosis progression in NAFLD can affect gallbladder contraction function and gastrointestinal function, and measurement of the serum levels of CCK and LPS has an important clinical value in the early diagnosis and treatment of digestive diseases related to gallbladder contraction function and gastrointe stinal function in NAFLD patients with liver fibrosis.

Objective To investigate whether the progression of liver fibrosis affects endothelial function in patients with nonalcoholic fatty liver disease (NAFLD), and to early identify the warning of cardiovascular diseases caused by endothelial dysfunction by liver fibrosis progression. Methods A total of 280 patients who attended the outpatient service or were hospitalized in Department of Liver Disease, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, from April 2019 to October 2020 were enrolled, and they were diagnosed with fatty liver disease by ultrasound and met the diagnostic criteria for NAFLD. General information and related serological markers were collected and recorded. FibroTouch technique was performed for the NAFLD patients diagnosed by ultrasound to record their fat attenuation parameter (FAP) and liver stiffness measurement (LSM), and according to LSM, the patients were divided into non-progressive fibrosis group (239 patients with LSM < 11 kPa) and progressive fibrosis group (41 patients with LSM ≥11 kPa) to analyze the association between liver fibrosis progression and endothelin-1 (ET-1)/nitric oxide (NO) in NAFLD. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the Spearman method was used for correlation analysis. Results There were no significant differences between the non-progressive fibrosis group and the progressive fibrosis group in the expression levels of ET-1( Z =-0.190, P =0.849) and NO( Z =-1.509, P =0.131), and there were significant differences between the two groups in body mass index (BMI), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) ( Z =-3.977, -4.162, -3.471, -3.201, -3.202, and -3.311, all P < 0.05). The Spearman analysis showed that LSM was not correlated with ET-1, NO, and NO/ET-1 ( r s =-0.046, 0.086, and 0.104, all P > 0.05). Further analysis of the correlation of ET-1 and NO with each index showed that ET-1 was not correlated with age, NO, ALT, AST, GGT, total cholesterol, TG, HDL-C, low-density lipoprotein cholesterol (LDL-C), FAP, and BMI ( r s =-0.017, 0.054, -0.067, -0.016, -0.031, 0.004, 0.051, -0.084, -0.030, 0.080, and 0.044, all P > 0.05), and NO was not correlated with age, ET-1, ALT, AST, GGT, total cholesterol, TG, HDL-C, LDL-C, FAP, and BMI ( r s =0.004, 0.054, 0.011, 0.052, 0.004, -0.051, -0.052, -0.012, -0.076, -0.013, and -0.021, all P > 0.05). Conclusion This study shows that liver fibrosis progression in NAFLD has no impact on ET-1 and NO, suggesting that fibrosis progression may have no influence on endothelial function.

[Abstract] Objective: To investigate the effects of forkhead box transcription factor (FOXK2) overexpression on the proliferation, migration, invasion and adhesion of human ovarian cancer SK-OV-3 cells and its related molecular mechanism. Methods: The open reading frame (ORF) of FOXK2 was cloned into lentivirus expression vector, which was then enveloped in HEK293T cells and transfected into human ovarian cancerSK-OV-3cells.TheoverexpressionefficiencywasdetectedbyqPCRandWesternblotting.Theproliferation, migration, invasion and adhesion of SK-OV-3 cells were detected by CCK-8, Scratch-healing, Transwell and Cell adhesion assays respectively, and the expressions of epithelial-mesenchymal transition (EMT) markers were detected by qPCR. Results: The FOXK2 overexpression vector was constructed successfully and packaged into lentivirus, which was then transfected into SK-OV-3 cells. After transfection, the expression of FOXK2 was significantly increased (P<0.01); the proliferation, migration and invasion of SK-OV-3 cells were significantly reduced while the adhesion ability was significantly increased (P<0.05 or P<0.01); and the expression levels of E-cadherin and β-catenin were significantly increased while that of vimentin and fibronection were significantly decreased (all P<0.01). Conclusion: Overexpression of FOXK2 in SK-OV-3 cells leads to a significant decrease in proliferation, migration and invasion but increase in adhesion. The molecular mechanism may be related to the reversion of the EMT process in tumor cells, suggesting that FOXK2 may be a potential target for the diagnosis and treatment of ovarian cancer.

Objective: To investigate the association between the whole blood riboflavin level and the occurrence, development and prognosis of esophageal squamous cell carcinoma (ESCC) in China. Methods: From March 2014 to September 2018, ESCC patients from three hospitals (the Affiliated Hospital of Medical College of Shantou University, Shantou Central Hospital in Southern Chaoshan area and First Affiliated Hospital of Zhengzhou University in Northern Taihang Mountain) were selected as a case group; non-esophageal patients who had a physical examination were selected as a control group. The case and control group were paired by age (±5 years) and a 1:1 ration. A total of 1 528 subjects were enrolled including 764 patients in the case group and 764 patients in the control group. About 3-5 ml venous blood samples were collected, and the erythrocyte glutathione reductase activity coefficient (GRAC) was measured to assess the whole blood riboflavin level. A multivariate conditional logistic regression model was used to analyze the association between the GRAC and the risk of ESCC. The association between the GRAC and the prognosis of ESCC was analyzed by using Cox proportional risk regression model based on 288 patients with complete survival data. They were divided into two groups, the high GRAC group (GRAC≥7.87) group and the low GRAC group (GRAC<7.87) according to the strongest correlation between the total survival time, survival outcome and GRAC (GRAC=7.87). Results: Among the 1 528 patients, 958 patients were from Southern Chaoshan area, including 479 patients in the case group with an average age about (59.90±9.34) years and 479 patients in the control group with an average age about (59.55±8.77) years. Other 570 patients were from Northern Taihang Mountain area, including 285 patients in the case group with an average age (58.39±5.19) years and 285 patients in the control group with an average age about (58.74±4.57) years. The multivariate conditional logistic regression showed that the OR (95%CI) of the GRAC and the risk of ESCC was 1.009 (0.998-1.019). The Cox proportional hazard regression model analysis showed that the HR (95%CI) of the high GRAC group was 1.712 (1.034-2.824) compared with the low GRAC group in the 50-70 years group. Conclusion The whole blood riboflavin level might not be associated with the occurrence of ESCC. The high whole blood riboflavin level would be more beneficial to the prognosis of ESCC patients aged 50-70 years.

Objective To investigate the mechanism of Mi-362 targeting Six1 inhibiting proliferation and migration of cer-vical cancer cells. Methods The expression levels of microRNA-362 in cervical cancer tissues and adjacent tissues were measured in 142 patients with cervical cancer in our hospital. At the same time,Hela cancer cell group,microRNA-362 mimics group and mi-croRNA-362 inhibitor group were set up to determine the viability of cancer cells,the number of monoclonal formation of cancer cells,the apoptotic rate of cancer cells,cell cycle,the number of perforations,and the levels of microRNA-362 and Six1 in cervical cancer fluid of Hela. Results The expression level of miR-362 in cervical cancer tissue was lower than that in adjacent tissue(P<0. 05). The higher the infiltration of lymphatic vessel space,pathological stage,TNM stage,lymph node metastasis and depth of infil-tration,the lower the expression rate of miR-362(P<0. 05). The OD value and survival rate in the miR-362 mimics group were lower than those in the Hela cancer cells group(P<0. 05),while the OD value and survival rate in the mir-362 inhibitor group were higher than those in the Hela cancer cells group and the miR-362 mimics group(P<0. 05). The number of clones formed in the miR-362 mimics group was lower than that in the Hela cancer cell group(P<0. 05),and the number of clones formed in the miR-362 inhibitor group was higher than that in the Hela cancer cell group and the miR-362 mimimics group(P<0. 05). The apoptotic rate of miR-362 mimics group was higher than that of Hela cancer cell group(P<0. 05),and that of miR-362 inhibitor group was lower than that of Hela cancer cell group and miR-362 mimics group(P<0. 05). The G1 phase in the miR-362 mimics group was higher than that in the Hela cancer cell group(P<0. 05),and the G1 phase in the miR-362 inhibitor group was lower than that in the Hela cancer cell group and the miR-362 mimimics group(P<0. 05). The number of cell membrane penetration in the miR-362 mimics group was lower than that in the Hela cancer group(P < 0. 05),and that in the miR-362 inhibitor group was higher than that in the Hela cancer group and the miR-362 mimimics group(P<0. 05). The expression level of miR-362 in the miR-362 mimics group was higher than that in the Hela cancer cell group(P<0. 05),and the expression level of miR-362 in the miR-362 inhibitor group was lower than that in the Hela cancer cell group and the miR-362 mimics group(P<0. 05). The expression level of Six1 in the miR-362 mimics group was lower than that in the Hela cancer cell group(P<0. 05),and that in the miR-362 inhibitor group was higher than that in the Hela cancer cell group and the miR-362 mimics group(P<0. 05). Conclusion miR-362 plays an important inhibition in the occurrence and development of cervical cancer,and its mechanism is related to the inhibition of prolifer-ation,migration and invasion of cancer cells by microRNA-362 through negative regulation of Six1.

Objective To investigate the association between the whole blood riboflavin level and the occurrence, development and prognosis of esophageal squamous cell carcinoma (ESCC) in China. Methods From March 2014 to September 2018, ESCC patients from three hospitals (the Affiliated Hospital of Medical College of Shantou University, Shantou Central Hospital in Southern Chaoshan area and First Affiliated Hospital of Zhengzhou University in Northern Taihang Mountain) were selected as a case group; non-esophageal patients who had a physical examination were selected as a control group. The case and control group were paired by age (±5 years) and a 1:1 ration. A total of 1 528 subjects were enrolled including 764 patients in the case group and 764 patients in the control group. About 3-5 ml venous blood samples were collected, and the erythrocyte glutathione reductase activity coefficient (GRAC) was measured to assess the whole blood riboflavin level. A multivariate conditional logistic regression model was used to analyze the association between the GRAC and the risk of ESCC. The association between the GRAC and the prognosis of ESCC was analyzed by using Cox proportional risk regression model based on 288 patients with complete survival data. They were divided into two groups, the high GRAC group (GRAC≥7.87) group and the low GRAC group (GRAC<7.87) according to the strongest correlation between the total survival time, survival outcome and GRAC (GRAC=7.87). Results Among the 1 528 patients, 958 patients were from Southern Chaoshan area, including 479 patients in the case group with an average age about (59.90±9.34) years and 479 patients in the control group with an average age about (59.55 ± 8.77) years. Other 570 patients were from Northern Taihang Mountain area, including 285 patients in the case group with an average age (58.39±5.19) years and 285 patients in the control group with an average age about (58.74± 4.57) years. The multivariate conditional logistic regression showed that the OR (95%CI ) of the GRAC and the risk of ESCC was 1.009 (0.998-1.019). The Cox proportional hazard regression model analysis showed that the HR (95%CI ) of the high GRAC group was 1.712 (1.034-2.824) compared with the low GRAC group in the 50-70 years group. Conclusion The whole blood riboflavin level might not be associated with the occurrence of ESCC. The high whole blood riboflavin level would be more beneficial to the prognosis of ESCC patients aged 50-70 years.

Objective To investigate the association between the whole blood riboflavin level and the occurrence, development and prognosis of esophageal squamous cell carcinoma (ESCC) in China. Methods From March 2014 to September 2018, ESCC patients from three hospitals (the Affiliated Hospital of Medical College of Shantou University, Shantou Central Hospital in Southern Chaoshan area and First Affiliated Hospital of Zhengzhou University in Northern Taihang Mountain) were selected as a case group; non-esophageal patients who had a physical examination were selected as a control group. The case and control group were paired by age (±5 years) and a 1:1 ration. A total of 1 528 subjects were enrolled including 764 patients in the case group and 764 patients in the control group. About 3-5 ml venous blood samples were collected, and the erythrocyte glutathione reductase activity coefficient (GRAC) was measured to assess the whole blood riboflavin level. A multivariate conditional logistic regression model was used to analyze the association between the GRAC and the risk of ESCC. The association between the GRAC and the prognosis of ESCC was analyzed by using Cox proportional risk regression model based on 288 patients with complete survival data. They were divided into two groups, the high GRAC group (GRAC≥7.87) group and the low GRAC group (GRAC<7.87) according to the strongest correlation between the total survival time, survival outcome and GRAC (GRAC=7.87). Results Among the 1 528 patients, 958 patients were from Southern Chaoshan area, including 479 patients in the case group with an average age about (59.90±9.34) years and 479 patients in the control group with an average age about (59.55 ± 8.77) years. Other 570 patients were from Northern Taihang Mountain area, including 285 patients in the case group with an average age (58.39±5.19) years and 285 patients in the control group with an average age about (58.74± 4.57) years. The multivariate conditional logistic regression showed that the OR (95%CI ) of the GRAC and the risk of ESCC was 1.009 (0.998-1.019). The Cox proportional hazard regression model analysis showed that the HR (95%CI ) of the high GRAC group was 1.712 (1.034-2.824) compared with the low GRAC group in the 50-70 years group. Conclusion The whole blood riboflavin level might not be associated with the occurrence of ESCC. The high whole blood riboflavin level would be more beneficial to the prognosis of ESCC patients aged 50-70 years.

Objective To detect the expression of human epithelial growth factor receptor 2 (HER-2) in advanced lung adenocarcinoma with epithelial growth factor receptor (EGFR) mutation, and to explore the potential of HER-2 as a therapeutic target for drug resistance in patients with EGFR mutations. Methods HER-2 is commonly expressed in the advanced lung adenocarcinoma with EGFR mutations, mainly in the cell membrane. Results The overexpression rate of HER-2 protein in tissues of advanced lung adenocarcinoma with EGFR mutations was 33.3%(28/84). The overexpression rate of HER-2 protein in patients>50 years of age was 40.3%(27/67), which was significantly higher than that of patients ≤50 years of age [5.9 % (1/17)], the difference was statistically significant (χ2=7.227, P=0.007). The overexpression rate of HER-2 protein in patients with high pathological differentiation [44.4 % (8/18)] was higher than that in patients with poor pathological differentiation [30.3%(20/66)], but the difference was not statistically significant (χ2=1.273, P=0.259). The overexpression of HER-2 protein in patients with EGFR 21 exon mutation [40.5 % (17/42)] was significantly higher than that of EGFR19 exon mutation [25.0%(10/40)], but the difference was not statistical significance (χ2=2.222, P=0.136). Conclusions The overexpression rate of HER-2 protein in advanced lung adenocarcinoma patients with EGFR mutation is high, which is related to the age and tumor differentiation. HER-2 is expected to be a potential therapeutic target for drug resistance patients with EGFR mutations.