ObjectiveTo investigate the role and mechanism of Go-Ichi-Ni-San complex subunit 1 （GINS1） in the progression of hepatocellular carcinoma （HCC） and the development of chemotherapy resistance. MethodsThe tumor database GEPIA2 was used to analyze the differential expression of GINS1 between HCC patients and healthy individuals， and pathological tissue samples were collected from 40 HCC patients who were admitted to The Affiliated Tumor Hospital of Xinjiang Medical University and the First Affiliated Hospital of Xinjiang Medical University from May 2017 to January 2021. Immunohistochemical staining was used to measure the difference in the expression of GINS1 between HCC tissue and corresponding adjacent tissue， and the correlation between the expression level of GINS1 and the clinical TNM stage of HCC was analyzed. Western blot was also used to measure the difference in the expression of GINS1 between HCC Huh7/Hep3B/Li-7/MHCC97H cell lines and normal human QSG7701 hepatocytes. The method of lentivirus transfection was used to establish the MHCC97H cell line with stable GINS1 knockdown and its negative control cell line. CCK-8 assay and colony formation assay were used to measure cell proliferative capacity； scratch assay was used to measure cell migration ability； Transwell assay was used to measure cell invasion ability； cells were treated with oxaliplatin to measure their sensitivity to chemotherapy drugs. Nude mice were used to establish a tumor-bearing model and observe the effect of GINS1 knockdown on the growth of HCC in vivo. Western Blot was used to measure the expression levels of the proteins associated with the Notch pathway and the JAK/STAT pathway. The cells were treated with the Notch receptor agonist Jagged-1 to analyze the association between GINS1 and the Notch/JAK/STAT pathway. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsThe expression of GINS1 was upregulated in HCC patients， HCC tissue， and HCC cell lines （all P<0.05）， and the expression level of GINS1 was positively correlated with the clinical TNM stage of HCC （r=0.822， P=0.011）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in proliferation， migration， and invasion activities （all P<0.01） and a significantly enhanced sensitivity to oxaliplatin （P<0.01）. Compared with the nude mice in the control group， GINS1 knockdown caused significant inhibition of tumor weight and volume in vivo in nude mice （all P<0.001）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in the expression levels of Notch1， Notch3， p-JAK2， and p-STAT3 （all P<0.05）， while there were no significant differences in the overall expression levels of JAK2 and STAT3 （P>0.05）. After Jagged-1 treatment， the GINS1-knockdown MHCC97H cells showed significant increases in proliferation， migration， and invasion activities and a significant reduction in sensitivity to oxaliplatin， as well as significant increases in the levels of p-JAK2 and p-STAT3 （all P<0.05）. ConclusionGINS1 is upregulated in HCC and can promote HCC progression and chemotherapy resistance through the Notch/JAK2/STAT3 pathway.

Objective:To study the risk factors of thyroid nodules in patients with type 2 diabetes.Methods:Data of patients with type 2 diabetes with normal thyroid function admitted to the Department of Endocrinology of Heze Municipal Hospital from January to June 2024 were collected. Binary logistic regression was used to analyze the influencing factors of thyroid nodules in patients with type 2 diabetes, and the receiver operating characteristic curve (ROC curve) was used to evaluate the diagnostic efficacy of each influencing factor.Results:Among 162 patients with type 2 diabetes, 96 had thyroid nodules, accounting for 59.3%. The incidence of thyroid nodules in women was significantly higher than that in men (χ 2 = 4.56, P = 0.034). Multivariate logistic regression analysis showed that age (≥50 years old), overweight and obesity [body mass index (BMI)≥24.0 kg/m 2], high glycated hemoglobin (≥10%), and high total cholesterol ( > 6.5 mmol/L) were independent risk factors for thyroid nodules in patients with type 2 diabetes ( OR = 1.83, 1.67, 1.08, 3.65, P < 0.05), and men was an independent protective factor ( OR = 0.63, P = 0.039). The ROC curve results showed that total cholesterol and total cholesterol combined with glycated hemoglobin could distinguish patients with thyroid nodules from those without thyroid nodules, with AUC = 0.64 and 0.68, respectively, and the differences were statistically significant ( P < 0.05). Conclusions:The incidence of thyroid nodules in patients with type 2 diabetes is relatively high. Age, overweight and obesity, high glycated hemoglobin, and high total cholesterol are independent risk factors for thyroid nodules in patients with type 2 diabetes, and total cholesterol has the ability to distinguish patients with thyroid nodules.

Objective:To explore the effect of β-galactoside α-2-6sialyltransferase1(ST6GAL1)on glycolysis,migration and invasion of colorectal cancer(CRC)HCT116 cells and its possible molecular mechanisms.Methods:The difference in the expression of ST6GAL1 in CRC patients and healthy people was analyzed using the GEPIA2 database.WB was performed to detect the differences in the expressions of ST6GAL1 in CRC cell lines HCT116,SW480,Caco-2,HT29,LoVo and human normal colon epithelial cell line NCM460.The difference in the expressions of ST6GAL1 in CRC tissues and corresponding adjacent tissues was analyzed by immunohistochemistry.HCT116 cell lines with stably knocked down or overexpressed ST6GAL1 were constructed by lentivirus transfection.Cell migration ability was detected by scratch test.Cell invasion ability was detected by Transwell test.WB assay was performed to detect the expression levels of cell glycolysis-related proteins and Notch1 intracellular domain(Notch1 ICD)as well as the phosphorylation level of PI3K/AKT/mTOR pathway.The expression level of Notch1 ICD and its entry into nucleus were observed by immunofluorescence assay.The Notch1 receptor agonist Jagged1 was added to HCT116 cells,and the expression levels of glycolysis-related proteins and Notch1 ICD and PI3K/AKT/mTOR pathway phosphorylation level were detected by WB.Results:The expression of ST6GAL1 was up-regulated in CRC tissues and cells(all P<0.05).Compared with the control and overexpression groups,knockdown of ST6GAL1 resulted in significantly lower levels of Notch1 ICD expression and PI3K/AKT/mTORC1 phosphorylation in HCT116 cells,lower levels of cellular glycolysis-related protein expressions and weaker cell migration and invasion abilities(all P<0.05).Overexpression of ST6GAL1 increased Notch1 ICD expression levels within HCT116 cells and promoted their entry into the nucleus.Cell glycolysis-related protein expression levels were elevated(all P<0.05).Cell migration and invasion abilities were enhanced(all P<0.05).Conclusion:ST6GAL1 activates the PI3K/AKT/mTORC1 pathway through activation of Notch1 receptor and phosphorylation,thus enhancing the glycolytic level and migration and invasion abilities of CRC cells.

Objective:To study the risk factors of thyroid nodules in patients with type 2 diabetes.Methods:Data of patients with type 2 diabetes with normal thyroid function admitted to the Department of Endocrinology of Heze Municipal Hospital from January to June 2024 were collected. Binary logistic regression was used to analyze the influencing factors of thyroid nodules in patients with type 2 diabetes, and the receiver operating characteristic curve (ROC curve) was used to evaluate the diagnostic efficacy of each influencing factor.Results:Among 162 patients with type 2 diabetes, 96 had thyroid nodules, accounting for 59.3%. The incidence of thyroid nodules in women was significantly higher than that in men (χ 2 = 4.56, P = 0.034). Multivariate logistic regression analysis showed that age (≥50 years old), overweight and obesity [body mass index (BMI)≥24.0 kg/m 2], high glycated hemoglobin (≥10%), and high total cholesterol ( > 6.5 mmol/L) were independent risk factors for thyroid nodules in patients with type 2 diabetes ( OR = 1.83, 1.67, 1.08, 3.65, P < 0.05), and men was an independent protective factor ( OR = 0.63, P = 0.039). The ROC curve results showed that total cholesterol and total cholesterol combined with glycated hemoglobin could distinguish patients with thyroid nodules from those without thyroid nodules, with AUC = 0.64 and 0.68, respectively, and the differences were statistically significant ( P < 0.05). Conclusions:The incidence of thyroid nodules in patients with type 2 diabetes is relatively high. Age, overweight and obesity, high glycated hemoglobin, and high total cholesterol are independent risk factors for thyroid nodules in patients with type 2 diabetes, and total cholesterol has the ability to distinguish patients with thyroid nodules.

Pain is one of the five vital signs,and the leading complication of war trauma,so analgesia is critical to combat casualty care.The U.S.Tactical Combat Casualty Care guidelines have defined the battlefield graded analgesic strategies.This paper reviews the assessment of pain grade of war wounds,involving the preliminary evaluation according to categories and conditions of trauma,and the quantitative evaluation according to subjective feeling and objective index monitoring.The analgesic strategies are analyzed,involving such as analgesic drugs local anesthetics,non-steroidal analgesics,opioids,N-methyl-D-aspartate receptor antagonists,and such analgesic techniques as regional nerve block,nerve radiofrequency,nerve ablation and spinal cord electrical stimulation technology.This review is expected to provide a useful reference for improving pain management and war injury treatment in China's army.

Background and objective Lung cancer is the most common type of cancer,accounting for more than half of all cancer cases,with lung adenocarcinoma(LUAD)representing over half of lung cancer patients.Currently,the 5-year survival rate for metastatic LUAD patients remains low and there is an urgent need for new biomarkers as targets for targeted therapy.Go-Ichi-Ni-San 1(GINS1),an important member of the GINS family,is closely related to the occurrence and devel-opment of human malignant tumors.This study aims to explore the role of GINS1 in glycolysis,proliferation,and metastasis of LUAD cells and the related molecular mechanisms.Methods The expression of GINS1 was analysed using bioinformatics between LUAD patients and healthy controls.The expression levels of GINS1 in LUAD and adjacent tissues were detected by immunohistochemistry and Western blot.Western blot and real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)were used to detect the expression of GINS1 in LUAD cell lines A549,SK-LU-1,Calu-3,H1299 and BEAS-2B.Stably knockdown GINS1 in A549 cells and its negative control cell line,as well as stably overexpress GINS1 in H1299 cells and its negative control cell line,were constructed by lentiviral transduction.Colony formation test was used to detect cell proliferation.Scratch test was used to detect cell migration.Transwell test was used to detect cell invasion,and the test kits were used to detect glucose consumption and lactate production.The expression levels of glycolysis-related proteins,Notch signaling pathway proteins and phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)signaling pathway proteins were detected by Western blot.The Notch receptor agonist Jagged1 was added to cells from the shGINS1-A549 group and the Notch receptor inhibitor LY3039478 was added to cells from the GINS1-OE-H1299 group for the regression assay.Results The expression of GINS1 was up-regulated in LUAD patients,tissues and cell lines,and corre-lated with overall survival(P＜0.05).Knockdown of GINS1 significantly inhibited the proliferation,migration and invasion of A549 cells(P＜0.05),while overexpression of GINS1 significantly enhanced the proliferation,migration and invasion of H1299 cells(P＜0.05).Furthermore,knockdown of GINS1 resulted in reduced glucose consumption,reduced lactate production,and reduced expression levels of glycolytic-related proteins in A549 cells(P＜0.05);overexpression of GINS1 enhanced glycolytic level in H1299 cells(P＜0.05).The expression levels of Notch1,Notch3,phosphorylated-PI3K(p-PI3K),phosphorylated-AKT(p-AKT)and phosphorylated-mTORC1(Ser2448)[p-mTORC1(Ser2448)]in A549 cells were significantly decreased by GINS1 knockdown(P＜0.05),while the expression levels of P13K,AKT,mTOR and p-mTORC2(Ser2481)were not significantly changed(P＞0.05).Overexpression of GINS1 increased the levels of Notch1,Notch3 and PI3K/AKT/mTORC1 pathway phosphorylated proteins in H1299 cells(P＜0.05).Jagged1 significantly reversed the inhibition of glycolysis,prolif-eration and metastasis induced by GINS1 knockdown in A549 cells(P＜0.05),and LY3039478 significantly inhibited the en-hancement of glycolysis,proliferation and metastasis induced by GINS1 overexpression in H1299 cells(P＜0.05).Conclusion The expression of GINS1 enhances the expression of Notch1 and Notch3 receptors,and then phosphorylates and activates the downstream PI3K/AKT/mTORC1 signaling pathway to enhance the glycolysis,proliferation and metastasis of LUAD cells.

Objective:To study the protective effects of various doses of Glycyrrhizin on hippocampus of young rats with status epilepticus (SE).Methods:Lithium chloride and pilocarpine were injected intraperitoneally into male Sprague-Dawley (SD) rats (with a postnatal age of 18-21 days), so as to induce SE in rats.The rats were divided into 5 groups according to the random number table method: control group, SE group, SE+ low dose Glycyrrhizin group, SE+ medium dose Glycyrrhizin group and SE+ high dose Glycyrrhizin group.Three different doses of Glycyrrhizin (20 mg/kg, 40 mg/kg and 60 mg/kg) were injected intraperitoneally into the rats.The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in serum of SE rats were determined by enzyme linked immunosorbent assay.Quantitative real-time PCR (qRT-PCR) was used to detect the mRNA expression levels of TNF- α, IL-1β and IL-6 in hippocampus of SE rats.The expression levels of Bax, Bcl-2 and Caspase-3 in hippocampus were detected by Western blot.The damage of neurons was measured by hematoxylin and eosin (HE) staining and Nissl staining.Neurons apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The mitochondrial changes were observed under transmission electron microscopy.One-way ANOVA followed by Tukey post-hoc test was used for statistical analysis. Results:Compared to the control group, TNF-α[(369.69±58.07) ng/L vs. (75.46±14.64) ng/L], IL-1β[(242.27±25.23) ng/L vs. (45.29±5.90) ng/L] and IL-6[(288.15±24.60) ng/L vs. (46.59±8.80) ng/L] in the serum of SE rats were significantly up-regulated(all P<0.05). Compared to SE group, low, medium and high doses Glycyrrhizin could effectively reduce the levels of TNF-α[(216.67±8.31) ng/L, (158.81±5.03) ng/L and (113.69±12.54) ng/L vs. (369.69±58.07) ng/L], IL-1β[(131.21±5.50) ng/L, (86.60±7.79) ng/L and (65.06±4.39) ng/L vs. (242.27±25.23) ng/L] and IL-6[(150.24±9.48) ng/L, (101.70±5.85) ng/L and (91.60±2.81) ng/L vs. (288.15±24.60) ng/L] released in serum after SE occurred (all P<0.05). The neuronal damage, loss, apoptosis and mitochondrial damage were found in the hippocampus of SE rats.Glycyrrhizin could ameliorate these symptoms.Compared to the control group, Bax levels(0.57±0.01 vs. 0.14±0.01)and Caspase-3 levels(0.54±0.00 vs. 0.11±0.01)in the hippocampus of SE rats were markedly increased, while Bcl-2 levels(0.27±0.01 vs. 0.57±0.02)were decreased(all P<0.05). Compared to the SE group, low, medium and high doses Glycyrrhizin could effectively reduce the levels of Bax(0.51±0.02, 0.45±0.03 and 0.40±0.02 vs. 0.57±0.01)and Caspase-3(0.47±0.02, 0.42±0.02 and 0.37±0.01 vs. 0.54±0.00), and increase the levels of Bcl-2(0.41±0.02, 0.45±0.02 and 0.51±0.01 vs. 0.27±0.01)(all P<0.05). Conclusions:Glycyrrhizin can effectively protect the hippocampus of young rats with SE.

Objective To investigate the association of energy metabolic markers with the risk of spontaneous bacterial peritonitis (SBP) in patients with decompensated hepatitis B virus-related liver cirrhosis (HBV-LC). Methods A retrospective analysis was performed for the clinical data of the patients with decompensated HBV-LC who were admitted to Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2019, and baseline clinical parameters and energy metabolic markers were compared between the patients with SBP and those without SBP within 2 weeks after admission. A multivariate logistic regression analysis was performed to investigate the risk factors for SBP. The t -test was used for comparison of normally distributed continuous data between two groups, and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between two groups; the Fisher's exact test was used for comparison of categorical data between two groups. The receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency of the newly established logistic regression model, and with the corresponding point of Youden index as the cut-off value, the DeLong test was used to compare the area under the ROC curve (AUC). Results A total of 50 patients with decompensated HBV-LC were included, among whom 23 (46%) developed SBP within 2 weeks after admission and 27 (54%) had no SBP during hospitalization. Compared with the non-SBP patients, the SBP patients had significantly lower triglyceride, prealbumin, and prothrombin time activity (PTA) and significantly higher international normalization ratio, C-reactive protein (CRP), and Model for End-Stage Liver Disease score (all P < 0.05). Comparison of baseline energy metabolic markers showed that compared with the non-SBP patients, the SBP patients had significantly lower respiratory quotient (RQ) [0.79(0.76-0.86) vs 0.85(0.79-0.91), P =0.041] and carbohydrate oxidation (CHO) rate [20.50%(15.25%-41.05%) vs 41.6%(22.25%-68.05%), P =0.041]. The multivariate logistic regression analysis showed that PTA was an independent risk factor for SBP in the patients with decompensated HBV-LC during hospitalization (odd ratio=0.004, P =0.008), and the regression model established based on the variables including PTA, CRP, RQ, and CHO had an AUC of 85.0% and a cut-off value of 0.60 at the maximum Youden index, with a specificity of 85.19% and a sensitivity of 73.91%, suggesting that this model had a better discriminatory ability than CRP (AUC=74.5%, P =0.049) and procalcitonin (AUC=56.4%, P < 0.01). Conclusion There are significant reductions in the energy metabolic markers RQ and CHO in the patients with decompensated HBV-LC who develop SBP within a short term, and their combination with PTA, CRP, and CHO/RQ ratio can help clinicians identify the patients at a high risk of SBP in the early stage and enhance nutrition support for such patients.

Objective:To investigate and analyze the energy metabolism characteristics and the correlation between energy metabolism and the risk of secondary bacterial infection in patients with hepatitis B virus-related chronic liver disease (HBV-CLD).Methods:Data of 183 cases admitted to the Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2020 were retrospectively analyzed. 79 cases of chronic hepatitis B, 51 cases of hepatitis B-related liver cirrhosis, and 53 cases of hepatitis B-related liver failure were collected. Among them patients with liver failure and decompensated liver cirrhosis were defined as severe liver disease group. The Quark RMR indirect calorimetry (COSMED Corporation, Italy) was used to exam the patients' energy metabolism condition, and the incidences of secondary bacterial infection of the patients during hospitalization were recorded. Shapiro-Wilk test and normal QQ plot were used to analyze the normal distribution of continuous variable data, which was consistent with the normal distribution and was described by mean ± standard deviation. In addition, if it did not conform to the normal distribution, the median and interquartile distance were used to describe it. Levene’s test was used to test the homogeneity of variance of the data, which was consistent with the normal distribution. The t-test was used to compare the means of the two groups of samples. One-way analysis of variance was used to compare the mean values of the three groups of samples, and then the Tukey's test was used to compare the two groups. If the variance was uneven or did not conform to the normal distribution, the Wilcoxon rank sum test was used to compare the differences between the two groups. Kruskal-Wallis test (H test) was used to compare the differences between the three groups of samples, and then the Dunnett’s test (Z test) was used for comparison between the two groups. Categorical variable data were analyzed using chi-square test. Logistic regression analysis was used to screen independent risk factors, and the criteria for variable inclusion ( P < 0.05). Results:The respiratory entropy (RQ) and non-protein respiratory entropy (npRQ) of the three groups had statistically significant difference ( P < 0.05). Among them, the RQ and npRQ of the chronic hepatitis B group were higher than hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group. There were statistically significant differences in fat oxidation rate (FAT%) and carbohydrate oxidation rate (CHO%) between the three groups ( P < 0.05). Compared with hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group, chronic hepatitis B group ( P < 0.05) had lower FAT% and higher CHO%. There were no statistically significant differences in the measured and predicted resting energy expenditure and protein oxidation rate (PRO%) between the three groups. The incidence of secondary bacterial infection in patients with severe liver disease was 48.39% (45/93). Compared with the non-infected group, the RQ and npRQ values ??of the infected group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Logistic regression analysis showed that glutamyltransferase, cholesterol, and npRQ were independent risk factors for secondary bacterial infections in patients with severe liver disease. Glutamyltransferase elevation, and cholesterol and npRQ depletion had suggested an increased risk of secondary bacterial infection. Subgroup analysis of patients with hepatitis B-related liver failure also showed that compared with non-infected group, RQ value and npRQ value of secondary bacterial infection group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Conclusion:Patients with hepatitis B virus-related chronic liver disease generally have abnormal energy metabolism. Low RQ, npRQ, CHO% and high FAT% are related to the severity of the disease; while npRQ reduction is related to the risk of secondary bacterial infection in patients with severe liver disease, and thus can be used as a clinical prognostic indicator.

Objective:To explore the application of weighted adjustments of dropout rates in sensitivity analysis of medically repeated measurements data and the implementation with SAS 9.4 software.Methods:By compiling SAS codes, mixed-effects models for repeated measures were used to conduct the covariance analysis of multivariable repeated measurements data. Meanwhile, the overall dropout rate and the dropout rates of each group were used to make weighted adjustments by applying pattern-mixture models, which was considered to be a sensitivity analysis to validate the stability of results.Results:The dropout rates of placebo group, low-dose and high-dose groups were 8.77%, 11.79% and 16.15%, respectively, the differences were significant ( P=0.025). The results of mixed-effects models for repeated measures showed the differences of curative effect indicators changes from baselines of between high-dose, low-dose groups and placebo group were significant ( P=0.008 and P=0.002). The results of pattern-mixture models considering weighted adjustments of the respective groups' dropout rates were consistent with those of mixed-effects models for repeated measures. Conclusions:The pattern-mixture models considering weighted adjustments of dropout rates can be used in the sensitivity analysis of repeated measurements data. The SAS codes can provide a practical basis for the popularization and application of weighted adjustments of dropout rates in the sensitivity analysis of repeated measurements data.