Background Retinal ischemia-reperfusion (RIR) injury is a common pathologic change.Its mechanism has not been identified.Objective This study was to investigate the relationship of microRNA-181a (miR-181a) ,tumor necrosis factor-α (TNF-α) and retinal ganglial cells (RGCs) in RIR injury.Methods RIR models were induced in 68 rats,then the rats were randomly divided into control group and RIR groups,including 0hour group,24-hour group and 72-hour group by random number table.Predicted target gene TNF-α was chosen,according to M iRanda,Targetscan and miRBase databases.Immunofluorescent labeling, Western blot and quantitative real-time PCR were used to identify the expression levels of miR-181a,TNF-α and RGCs.Immunofluorescent labeling of RGCs in retinal flat mounts was analyzed for RGCs counts.Results Compared with the control group, RGCs densitiy was obviously decreased in 24-hour and 72-hour RIR groups (P＜0.001).The expression level of mir-181a significantly decreased with reperfusion time in the RIR groups (P＜0.05).Futhermore, the expression level of miR181a was positively correlated with RGCs numbers (r=0.995 ,P=0.005).TNF-α and miR-181a were mainly located in inner layers of retina.As opposed to the changes in RGCs numbers and miR-181a expression,TNF-α in 24-hour group was obviously higher than that of the 0-hour group, though there was no statistical significance in overall correlation analysis.Conclusions In RIR,miR-181a may be involved in regulating RGCs apoptosis.TNF-α may be a target gene of miR-181 a.Interventions within 24 hours after reperfusion might be critical.Further study of miR181 a may help to explore new molecular targets for neuroprotection treatment.

Insulin is the most common medicine used for diabetic patients, unfortunately, its effective time is short, even the long-acting insulin cannot obtain a satisfactory effect. Fibroblast growth factor (FGF)-21 is a recently discovered glucose mediator and expected to be a potential anti-diabetic drug that does not rely on insulin. In this study, db/db mice were used as the type 2 diabetic model to examine whether mFGF-21 has the long-term blood lowering effect on the animal model. The results showed that mFGF-21 could stably maintain the blood glucose at normal level for a long-term in a dose-dependent manner. Administration of mFGF-21 once a day with three doses (0.125, 0.25 and 0.5 mg x kg(-1)) could maintain blood glucose of the model animals at normal level for at least 24 h. Administration of mFGF-21 every two days with the same doses could maintain blood glucose of the model animals at normal level for at least 48 h, although it took longer time for blood glucose to reach to normal level depending on doses used (twenty injections for 0.125 mg x kg(-1) and 0.25 mg x kg(-1) doses, ten injections for 0.5 mg x kg(-1) dose). Surprisingly, the blood glucose of the treated model animals still maintained at normal level for 24 h after the experiment terminated. Glycosylated hemoglobin level of the animals treated with mFGF-21, which represented long-term glucose status, decreased significantly compared to the control group and the insulin group. The results suggest that FGF-21 has potential to become a long-acting and potent anti-diabetic drug.

Objective To investigate the effects of L-carnitine on myocardial enzymes in infants with congenital heart disease undergoing open cardiac operation with cardio pulmonary bypass (CPB).Methods From Mar.2011 to Nov.2012,there were 60 infants with ventricular septal defects,whom were divided randomly into the test (n =30) and control (n =30) groups.L-carnitine was put in the cold crystal cardiac arresting liquid in the test group (6 g/L),other experimental conditions were the same between two groups.Before CPB,at 1 h,24 h,7 d after the pump-off,venous blood was drawn to test the level of serum creatine kinase (CK),CK-MB isozyme,cardiac Troponin I (cTnI).Blood samples were taken at different time points for the analysis of tlymphocyte and its subtype by flow cytometry.Observing the dopamine and other vasoactive drug 8 hours and 24 hours after the pump-off,using data obtained from the t-test done statistically.Results The levels of CK and CK-MB and cTnI had no difference between two groups before operation.From the end of CPB,the levels of CK and CK-MB and cTnI were significantly lower in the test group than that in the control group (P ＜0.05).The automatic jump higher rate (95％ vs 70％,P ＜ 0.05) and the amount of dopamine were significantly less than the control group (P ＜ 0.05).Conclusions L-carnitine improved obviously microcirculation of ischemic cardiac reperfusion injury undergoing open cardiac operation under CPB and heart function.It had a good protective effect on myocardium in infants.

Objective To investigate the effect of dual-specificity phosphatase-2 (DUSP2) on the cell proliferation and apoptosis in the gastric cancer and its mechanisms. Methods Firstly, the effects of different expressions of DUSP2 on the overall survival of 876 gastric cancer patients were analyzed by online analysis tool KM plotter, and the expressions of DUSP2 in various gastric cancer cell lines (MKN-45, SGC-7901, HGC-27 and N-87) were verified. Secondly, DUSP2 overexpressed lentiviral vector was constructed, and MKN-45 was transfected by packaged virus. DUSP2-overexpression gastric cancer cell line was gained by drug screening. Meanwhile, gastric cancer cells infected with empty vector virus were used as control. Then the effect of DUSP2 upregulation on the proliferation ability of gastric cancer cells was evaluated by MTS cell proliferation assay, and the apoptosis was determined by Annexin V-FITC / PI double staining. The protein expressions of DUSP2, ERK, p-ERK (Thr202/Tyr204), P38 and p-P38 were tested by the Western blot analysis. Results Gastric cancer patients with high DUSP2 expression showed a significant survival advantage compared with those with low DUSP2 expression, and DUSP2 levels were decreased in several gastric cancer cell lines. The Western blot analysis revealed that the expression of DUSP2 markedly increased in overexpressed DUSP2 group (experimental group) compared with that of control group. The MTS experiment showed that the cell viability was significantly decreased in experimental group than that of the control group. Correspondingly, the cell apoptosis test showed that the cell apoptosis rate was obviously higher in the experimental group than that of the control group. The results of Western blot assay indicated that p-ERK (Thr202/Tyr204) and p-38 were significantly down-regulated in the experimental group compared with those of control group. Conclusion The over-expression of DUSP2 can efficiently inhibit cell proliferation and promote its apoptosis in gastric cancer cells, and the mechanism is related to DUSP2 inhibiting the phosphorylation levels of ERK and P38.

Objective:To explore whether BHLHE40 can affect the sensitivity of thyroid cancer (TC) cells to cisplatin by activating oxidative phosphorylation (OXPHOS) pathway by targeting high mobility group A2 (HMGA2) .Methods:The mRNA expression of HMGA2 and its upstream transcription factor BHLHE40 in TC tissues was analyzed by TCGA-THCA and hTFtarget online databases. The si-HMGA2, oe-HMGA2, oe-BHLHE40, negative control si-NC and oe-NC were transfected into TC cells (K1 and SW579) by liposome transfection method. The mRNA expression levels of BHLHE40 and HMGA2 in TC cells (SW579, FTC-133, and K1) and normal thyroid cells (Nthy ori3-1) were detected by real-time quantitative PCR (qRT-PCR). The cell viability was detected by MTT assay, the half inhibitory concentration (IC 50) value of cisplatin was calculated by CCK-8 assay, the apoptosis level was detected by flow cytometry, and the expression of OXPHOS complex was detected by Western blotting. Seahorse XFe 96 was used to analyze the oxygen consumption rate of the TC cells. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay were used to analyze the binding relationship between BHLHE40 and HMGA2. Results:TCGA database results showed that the mRNA expression levels of HMGA2 and BHLHE40 in TC tissues (10.57±2.58, 13.89±1.13) were higher than those in normal thyroid tissues (4.82±1.69, 12.28±1.01), with statistically significant differences ( t=16.69, P<0.001; t=10.43, P<0.001). The results of qRT-PCR showed that the relative mRNA expression levels of HMGA2 in normal thyroid cells (Nthy ori3-1) and TC cells (SW579, FTC-133, and K1) were 1.00±0.13, 2.94±0.23, 4.71±0.41 and 6.29±0.49, while those of BHLHE40 were 1.00±0.12, 2.60±0.23, 3.39±0.35 and 6.18±0.51 respectively, both with statistically significant differences ( F=130.50, P<0.001; F=125.20, P<0.001). Further pairwise comparison showed that mRNA expression levels of HMGA2 and BHLHE40 in TC cells were significantly higher than those in normal thyroid cells (all P<0.001). According to MTT experimental results, si-HMGA2 treatment significantly reduced the cell viability of K1 cells compared to the si-NC group (all P<0.05). In addition, compared to the oe-NC group, oe-HMGA2 treatment significantly increased the cell viability of SW579 cells (all P<0.05). Compared to the oe-NC+DMSO group, the oe-HMGA2+DMSO group showed enhanced cell viability of SW579 cells, while the OXPHOS pathway inhibitor Gboxin was able to reverse the effect of overexpressing HMGA2 on cell viability (all P<0.05). The results of flow cytometry and CCK-8 experiments showed that compared to the si-NC group (apoptosis level: 6.19%±0.28%; cisplatin IC 50 value: 17.47 μmol/L), knocking down HMGA2 could increase the apoptosis level (11.96%±0.32%; t=19.17, P<0.001) and cisplatin sensitivity (IC 50 value: 1.49 μmol/L) of K1 cells. In addition, compared to the oe-NC group (apoptosis level: 9.98%±0.32%; cisplatin IC 50 value: 8.17 μmol/L), overexpressing HMGA2 significantly decreased the apoptosis level (4.32%±0.25%; t=19.65, P<0.001) and cisplatin sensitivity (IC 50 value: 34.95 μmol/L) of SW579 cells. The results of dual-luciferase assay showed that compared with the si-NC group, knocking down the expression of BHLHE40 in human kidney epithelial 293T cells significantly reduced the luciferase activity of wild-type HMGA2 (0.31±0.02 vs. 1.00±0.11; t=10.69, P=0.004). However, there was no significant effect on the luciferase activity of mutant-type HMGA2 (1.06±0.11 vs. 1.00±0.07; t=0.80, P=0.470). ChIP results showed that the mRNA expression level of HMGA2 in K1 cells was significantly increased in the anti-BHLHE40 group (6.57±0.62) compared with the IgG group (1.00±0.10; t=15.36, P<0.001). Compared to the oe-NC+DMSO group, the oe-HMGA2+DMSO group showed decreased apoptosis level ( P<0.05) and cisplatin sensitivity of SW579 cells, with a significant increase in the expression of OXPHOS complexes Ⅰ-Ⅴ and cellular oxygen consumption rates (all P<0.05). The effect of overexpressing HMGA2 was reversed by treatment with oe-HMGA2+Gboxin (all P<0.05). The recovery experiment showed that compared to the oe-NC+si-NC group, overexpression of BHLHE40 in SW579 cells increased cell viability and the expression of OXPHOS complexes Ⅰ-Ⅴ, while decreasing apoptosis levels and increasing cellular oxygen consumption rates and cisplatin IC 50 values (all P<0.05). However, simultaneous knockdown of HMGA2 reversed the effect of overexpressing BHLHE40 (all P<0.05) . Conclusion:BHLHE40 can activate the OXPHOS pathway by targeting and regulating the expression of HMGA2, thereby affecting the sensitivity of TC cells to cisplatin.

Objective To evaluate the clinical value of free glycoprotein non-metastatic melanoma protein B(GPNMB)as a drug resis-tance and prognostic marker for non-small cell lung cancer(NSCLC)patients with epidermal growth factor receptor(EGFR)amplifica-tion accompanied by mutations.Methods Fifty-five cases of NSCLC patients with EGFR amplification associated with mutations who received treatment from March 2018 to September 2019 were included as the observation group.All patients received an EGFR-tyrosine kinase inhibitor(EGFR-TKI)as the first-line treatment;67 blood samples from the physical examination center during the same period were randomly included as healthy control.We compared the expression levels of free GPNMB between the two groups,explored the correlation between GPNMB expression and the clinicopathological information in the observation group;and combined the clinical efficacy to evaluate its value as a drug resistance marker.Through follow-up,the progress free survival(PFS)of patients was statistically analyzed,and through multivariate Cox regression analysis,independent risk factors affecting the survival in the observation group were explored.Results Compared with that in the control group,the expression level of free GPNMB in the observation group was signi-ficantly up-regulated.The expression level of free GPNMB in the observation group is significantly related to the clinical efficacy of EGFR-TKI(P = 0.016).Patients with high GPNMB expression have significantly stronger drug resistance,and patients with high GPNMB expression have significantly shorter PFS duration(P = 0.032).A high free GPNMB expression(HR = 4.029,95%CI:1.942-8.358,P＜0.001)is also an independent risk factor affecting patient survival.Conclusion The expression level of free GPNMB in patients with EGFR amplification accompanied by mutant NSCLC is significantly up-regulated,and its high expression is significantly related to the enhancement of the patient's drug resistance.High GPNMB expression is significantly related to the poor prognosis of patients and is an independent risk factor affecting patient survival.

Objective:To investigate the correlation between mental health and symptom distress in breast cancer patients.Methods:The clinical data of 110 female breast cancer patients at Shanxi Bethune Hospital from June 2017 to March 2018 were collected. The patients were assessed for symptomatic distress with depression self-rating scale (SDS). Functional assessment of cancer therapy-breast (FACT-B) and the functional assessment of chronic illness therapy-spiritual well-being (FACIT-SP) were used to make the mental assessment. Multiple linear regression analysis was used to analyze the effect of symptom distress on the mental health of the patients.Results:The scores of symptom distress, mental health, FACT-B and FACIT-SP were (19.94±5.78), (50.68±10.64), (16.85±4.75), (33.83±8.33), respectively. Multivariate analysis showed that mental health score of the patients with symptom distress > 18 scores was reduced by 5.15 points ( P=0.01) compared with the patients with symptom distress≤18 scores. Compared with the patients with annual household income < 50 000 yuan, the mental health score of patients with annual household income of 50 000-79 000 yuan was increased by 9.46 points ( P < 0.01), and the mental health score of patients with annual family income ≥ 80 000 yuan was increased by 5.92 points ( P < 0.01); compared with the patients in phase I, the mental health score of the patients in phaseⅡwas decreased by 2.62 points ( P=0.02), and the mental health score of the patients in phase Ⅲ was decreased by 4.98 points ( P < 0.01). Conclusions:Symptom distress is an independent risk factor for affecting mental health of breast cancer patients. Solving symptom distress of patients can improve mental health status of the patients.

Objective:This study examined the associations between the levels of bile acids in early pregnancy and the occurrence of overweight.Methods:From 2010 to 2012, 22 302 pregnant women were recruited by Tianjin Women and Children′s Health Center to investigate gestational diabetes. Two hundred and forty-three women with gestational diabetes mellitus provided overnight fasting blood samples in the first trimester, and 243 counterparts without gestational diabetes mellitus matched on age were selected randomly to establish a nested case-control study. The association between bile acids and overweight were evaluated by binary logistic regression with data from 166 overweight pregnant women (body mass index≥24.0 kg/m 2) and 320 normal weight subjects (body mass index <24.0 kg/m 2). Results:Compared to non-overweight group, the level of primary unconjugated bile acids in overweight group was significantly higher. After adjustment of confounding factors, the OR of cholic acid (CA)>0.086 nmol/mL for overweight was 2.09 (95% CI 1.14-3.80, adjusted P=0.040), and OR of chenodeoxycholic acid (CDCA)>0.043 nmol/mL was 2.15 (95% CI 1.22-3.78, adjusted P=0.040) compared with the lower groups. However, the significant associations between the other bile acids and overweight were not detected. Stepwise selection was used to identify significant bile acid species in logistic regression. We found that only CA was independently associated with overweight, and the OR of CA>0.086 nmol/mL vs≤0.086 nmol/mL was 2.03 (95% CI 1.11-3.74, P=0.022). Conclusion:CA and CDCA in early pregnancy maybe associated with overweight, and CA might be independently associated with overweight.

Objective:To explore the changes in acoustic features of 9-10-year-old primary school students with depressive symptoms, and based on these features and the random forest (RF) algorithm, construct a model for identifying depressive symptoms in primary school students, so as to provide an intelligent psychological health screening tool for schools and education departments.Methods:This was a case-control study.A total of 1 186 primary school students aged 9-10 from three primary schools in three regions of Jiangsu Province were selected as research subjects for psychological health screening from October 26, 2022 to February 13, 2023.Their demographic data, Depression-Anxiety-Stress Scale (DASS-21) scores, Insomnia Severity Index scores, and voice recordings were collected.Based on the DASS-21 scores, the participants were divided into a control group ( n=1 086) and a depression group ( n=100).Voice recordings were made using the neutral text " The North Wind and the Sun". openSMILE was used to extract 523 acoustic features from the pre-processed voice recordings.Group differences were assessed using independent-samples t-tests or chi-square tests.Pearson correlation analysis was conducted to examine the relationship between acoustic features and depression scores.Depressive symptoms were set as the dependent variable, and the correlated acoustic features were set as the independent variable to construct a classification model using the RF algorithm.The model performance was assessed using the receiver operating characteristic (ROC) curve, the area under the curve (AUC), precision, accuracy, recall, and F1 score. Results:Compared with the control group, the depression group showed significant differences in 105 acoustic features (44 spectral features, 49 source features, and 12 prosodic features) (all P<0.05).Correlation analysis showed that 12 acoustic features (7 spectral features, 4 source features, and 1 prosodic feature) were significantly correlated with the depression score (all P<0.05).Among the RF algorithm-based classification models, the spectral features demonstrated superior performance compared to source features and prosodic features (AUC=0.793), and the performance of the model based on the combination of these features was the best (AUC=0.818). Conclusions:Acoustic features may be an objective indicator to identify the depression of 9-10-year-old primary school students, and the classification model established based on acoustic features can identify the depressed primary school students.

Objective: To establish a rat model for laryngeal precancerous lesions histologically and pathologically comparable to the human counterpart. Methods: Thirty-six Wistar rats were randomly divided into experimental group and control group, with 18 rats in each group, and 1% 4-nitroquinoline-1-oxide (4NQO) solution and saline were respectively applied to the laryngeal mucosas of rats in two groups. During subsequent 20 weeks, the changes of laryngeal mucosas were regularly observed with naked eyes and endoscope and lesions were determined by histology. SPSS 22.0 software was used for statistical analysis. Results: The food intake, water intake and body weight of the rats in the experimental group were lower than those in the control group, with statistically significant differences (all P<0.05). White plaque, superficial ulcer, erosion and miliary particles were present in the larynxes of rats in the experimental group, with histological manifestations of atypical hyperplasia or carcinoma in situ, and normal epitheliums were shown in the control group. The number of Ki67 positive cells in the laryngeal mucosas of rats in the experimental group at the 4 th, 8 th, 12 th, 16 th, and 20 th weeks were 13.5±2.4, 35.6±5.8, 53.4±8.3, 78.8±11.6, 80.6±12.4, respectively, no Ki67 positive cells were found in the control group at individual time points, and the differences were statistically significant (t=9.74, 10.63, 11.14, 11.77, 11.26, respectively, all P<0.01). Conclusion: 4NQO can credibly cause rats laryngeal precancerous lesions, which morphologically and histologically mimic laryngeal carcinnogenesis. This method is practical, easy and reliable to prepare the animal model of laryngeal precancerous lesions.
Animals ; Carcinoma in Situ ; Carcinoma, Squamous Cell/pathology* ; Humans ; Larynx/pathology* ; Precancerous Conditions/pathology* ; Rats ; Rats, Wistar