‍ ObjectiveTo investigate the value of preoperative fibrosis 4 score （FIB-4） combined with prognostic nutritional index （PNI） in predicting recurrence after radiofrequency ablation （RFA） for early-stage hepatocellular carcinoma （HCC）. MethodsA retrospective analysis was performed for the clinical data of 365 patients with the initial diagnosis of early-stage HCC who underwent RFA at Tianjin Third Central Hospital from January 2013 to December 2017， and a statistical analysis was performed for recurrence and survival. The receiver operating characteristic （ROC） curve was plotted for FIB-4 and PNI with postoperative tumor recurrence as the positive event， and their optimal cut-off values were selected. FIB-4 and PNI were graded and combined as FIB-4-PNI score， based on which the patients were divided into 0-point group with 207 patients， 1-point group with 93 patients， and 2-point group with 65 patients. The chi-square test was used for comparison of categorical data between groups. The Kaplan-Meier survival analysis and the log-rank test were used to compare the recurrence-free survival （RFS） and overall survival （OS） between groups， and the Cox regression model was used to investigate the influencing factors for RFS and OS. ResultsThe 1-， 3-， and 5-year RFS rates of all patients were 79.2%， 49.8%， and 34.3%， respectively， with a median RFS of 35 months， while the 1-， 3-， and 5-year OS rates of all patients were 98.9%， 86.9%， and 77.3%， respectively. There were significant differences in cumulative RFS and OS rates between the patients with different levels of FIB-4， PNI， and FIB-4-PNI （RFS rate： χ²=17.890， 29.826， and 32.397， all P<0.001； OS rate： χ²=16.896， 21.070， and 26.121， all P<0.001）. The multivariate Cox regression analysis showed that history of diabetes （hazard ratio ［HR］=1.418， 95% confidence interval ［CI］： 1.046‍ ‍—‍ ‍1.922， P=0.024）， two tumors （HR=1.516， 95%CI： 1.094‍ ‍—‍ ‍2.101， P=0.012）， three tumors （HR=2.146， 95%CI： 1.278‍ ‍—‍ ‍3.604， P=0.004）， FIB-4-PNI 1 point （HR=1.875， 95%CI： 1.385‍ ‍—‍ ‍2.539， P<0.001）， and FIB-4-PNI 2 points （HR=2.35， 95%CI： 1.706‍ ‍—‍ ‍3.236， P<0.001） were independent risk factors for RFS， while two tumors （HR=1.732， 95%CI： 1.005‍ ‍—‍ ‍2.983， P=0.048）， three tumors （HR=3.511， 95%CI： 1.658‍ ‍—‍ ‍7.433， P=0.001）， FIB-4-PNI 1 point （HR=2.094， 95%CI： 1.230‍ ‍—‍ ‍3.565， P=0.006）， and FIB-4-PNI 2 points （HR=3.908， 95%CI： 2.306‍ ‍—‍ ‍6.624， P<0.001） were independent risk factors for OS. ConclusionFIB-4-PNI score can be used as an independent predictive factor for recurrence and overall survival time after RFA for early-stage HCC， and it can be combined with tumor features to predict postoperative recurrence and survival.

ObjectiveTo investigate the expression of lysophosphatidic acid （LPA） in patients with liver cancer， as well as its influence on malignant biological behavior of liver cancer and related regulatory mechanism. MethodsFrom January 2016 to December 2022， 26 patients with liver cancer， 28 patients with liver cirrhosis， and 28 individuals undergoing physical examination were enrolled. ELISIA was used to measure the content of LPA in plasma and peritoneal effusion of the patients with liver cancer or liver cirrhosis accompanied by peritoneal effusion， and the content of LPA was measured in plasma of the normal population at the same time， so as to clarify the difference in the expression of LPA in different populations， such as the patients with liver cancer and those with liver cirrhosis. MTT cell proliferation assay and cell migration assay were used to observe the influence of LPA and its inhibitor pertussis toxin （PTX） on the proliferation， migration， and invasion of SMMC7721 cells. In order to investigate the effect of LPA on the expression of RhoA and its upstream and downstream molecules FAK and P53 after binding to its receptor， qPCR and Western blot were used to observe the effect of LPA on the mRNA and protein expression levels of P53， FAK， and RhoA in SMMC7721 cells. A one-way analysis of variance was used for comparison of the means of continuous data between multiple groups， and the SNK-q test was used for comparison between two groups. ResultsCompared with the patients with liver cirrhosis， the patients with liver cancer had a significantly higher concentration of LPA in plasma （4.99±0.55 μmol/L vs 2.63±0.43 μmol/L， P<0.05） and peritoneal effusion （5.19±0.63 μmol/L vs 2.91±0.46 μmol/L， P<0.05）， and the patients with liver cancer also had a significantly higher level of plasma LPA than the normal population （4.99±0.55 μmol/L vs 1.61±0.39 μmol/L， P<0.05）. The cell proliferation assay showed that LPA significantly promoted the proliferation of SMMC7721 cells， and cell proliferation rate increased with the increase in dose and time； in particular， the middle-and high-dose groups had a significantly higher proliferation rate than the control group （P<0.05）. PTX inhibited the proliferative capacity of SMMC7721 cells in a time-dependent manner， and there was a significant difference between the groups （P<0.05）. The proliferation rate of the 72-hour high-dose LPA group was 3.6 times that of the control group， while the proliferation rate of the PTX group was 0.6 times that of the control group； the proliferation rate of the 72-hour high-dose LPA+PTX group was 1.2 times that of the control group. In addition， LPA increased the migration ability of hepatoma cells， while PTX inhibited their migration， in a time-dependent manner， and there was a significant difference between the groups （P<0.05）. The migration rate of the 72-hour high-dose LPA group was 3.09 times that of the control group， while the migration rate of the PTX group was 0.4 times that of the control group； the migration rate of the 72-hour high-dose LPA+PTX group was 0.99 times that of the control group. qPCR and Western blot showed that there were significant reductions in the mRNA and protein expression levels of P53 in SMMC7721 cells after LPA treatment， while there were significant increases in the mRNA and protein expression levels of FAK and RhoA； there was a significant difference between the LPA group and the control group （P<0.05）. ConclusionThere is an abnormal increase in the expression of LPA in patients with liver cancer， and LPA can promote the proliferation of liver cancer cells and increase their migration ability. At the same time， LPA changes the expression levels of P53， FAK， and RhoA， which may be associated with the promotion of tumor development and progression by LPA.

Objective To explore the effects of ginsenoside Rg1 on blood-brain barrier, neuroinflammation and behavioral function of traumatic brain injury (TBI) mouse model.MethodsThe experiment was divided into two parts. In the first part, 27 SPF male BALB/c mice were randomly divided into blank group, sham operation group and TBI model group, with 9 mice in each group. TBI model group was made by controlled cortical impact (CCI) after craniotomy, while sham operation group was only performed craniotomy without any treatment, and the blank group was not treated at all. The effect of modeling was evaluated after operation. In the second part, 50 male BALB/c mice were randomly divided into sham operation group, three different drug dosage groups and solvent (DMSO) control group, with 8 mice in each group. The drug treatment groups were injected with ginsenoside Rg1 at the doses of 10, 20 and 40 mg/kg respectively 6 hours after TBI model had been successfully established, while the DMSO control group was given the same amount of 1% DMSO for one week, twice a day. Modified neurological severity scores (mNSS) were performed on the 1st, 3rd, 7th and 14th day after modeling, and the blood-brain barrier leakage was detected by Western blotting on the 3rd day after modeling. On the 14th and 16th day, the elevated cross maze test and water maze test were used to detect the neurobehavioral function. On the 28th day after anesthesia and perfusion, the brains were taken out, and the neuroinflammation such as activation of microglia and astrocytes was observed by immunofluorescence staining.ResultsThe expression level of MMP-9, a marker of blood-brain barrier, decreased in ginsenoside Rg1 treatment group (P<0.01). The number of microglia （Iba-1 positive) and astrocyte (GFAP positive) cells decreased significantly (P<0.05), which indicated that neuroinflammation was inhibited, and the best effect was achieved at the dosage of 20 mg/kg (P<0.01). The mNSS of mice in ginsenoside Rg1 treatment group were significantly lower than those in DMSO control group (P < 0.01), and the proportion of times they entered the open arm was significantly higher than that in DMSO control group (P < 0.05). The time ratio in the quadrant where the water maze experimental platform was located and the times of crossing the platform were significantly higher than those in control group (P < 0.05), and the dosage of 20 mg/kg had the best effect.ConclusionThe TBI mouse model was successfully constructed and applied to the study of ginsenoside Rg1 repair of mouse traumatic brain injury. Ginsenoside Rg1 can significantly improve blood-brain barrier, alleviate neuroinflammation and improve neurobehavioral function in TBI model mice, and the effect is the most significant at the dose of 20 mg/kg.

Objective To investigate the level of glycosylated albumin (GA) in liver cirrhosis patients with different Child-Pugh classes and its application value in predicting liver function. Methods A total of 486 patients with liver cirrhosis who were hospitalized in Tianjin Third Central Hospital from January 1 to December 31, 2019, were enrolled, among whom 227 patients had liver cirrhosis without diabetes and 259 patients had liver cirrhosis with diabetes. The patients were divided into groups according to Child-Turcotte-Pugh (CTP) score, and fasting blood glucose, glycosylated hemoglobin, and percentage of GA (GA%) were measured. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between three groups, and the Dwass-Steel-Critchlow-Fligner test was used for further comparison between two groups. Scatter plots and fitting curves were plotted for CTP score and GA% to evaluate the association between them and calculate the cut-off value. Results For the cirrhosis patients without diabetes, there were significant differences between the patients with different Child-Pugh classes in GA% ( χ 2 =24.809, P < 0.001), fasting blood glucose ( χ 2 =11.899, P =0.003), and glycosylated hemoglobin ( χ 2 =13.607, P =0.001); further pairwise comparison showed that there was a significant difference in GA% between Child-Pugh class A/B liver cirrhosis patients without diabetes and Child-Pugh class C liver cirrhosis patients ( P < 0.05), Child-Pugh class A patients had a significantly higher level of fasting blood glucose than Child-Pugh class B patients ( P < 0.05), and Child-Pugh class A patients had a significantly higher level of glycosylated hemoglobin than Child-Pugh class B/C patients ( P < 0.05). For the patients with liver cirrhosis and diabetes, there were significant differences between the patients with different Child-Pugh classes in GA% ( χ 2 =10.734, P =0.005) and fasting blood glucose ( χ 2 =16.295, P < 0.001); further pairwise comparison showed that Child-Pugh class C liver cirrhosis patients with diabetes had a significantly lower GA% than Child-Pugh class A/B patients ( P < 0.05) and Child-Pugh class A patients had a significantly lower fasting blood glucose level than Child-Pugh class B patients ( P < 0.05). The fitting curve showed that GA% increased with the increase in CTP score in the liver cirrhosis patients without diabetes, reached the highest value at the CTP score of 6.5, and then started to decrease, with the lower value at the CTP score of 11.5, which showed a curvilinear relationship; in the liver cirrhosis patients with diabetes, GA% first increased and then decreased with the increase in CTP score, with a cut-off value of 8. Conclusion GA% first increases and then decreases along with the progression of liver cirrhosis. There is a significant difference in GA between liver cirrhosis patients with different Child-Pugh classes, suggesting that the reduction in GA is closely associated with liver function decompensation in end-stage liver cirrhosis.

Intestinal flora imbalance plays a certain role in the development and progression of liver cancer, while probiotics have a certain impact on liver cancer, both of which are the focus of clinical research. This article introduces the mechanism of action of intestinal flora imbalance in the pathogenesis of liver cancer and the preventive effect of probiotics against liver cancer. Intestinal flora imbalance can participate in the pathological process of liver cancer by activating Toll-like receptor 4, regulating the level of metabolites, producing endotoxin, and inducing bacterial translocation and intestinal bacterial overgrowth, while probiotics can effectively prevent liver cancer by maintaining enterohepatic circulation, enhancing immune function, promoting the reproduction of intestinal probiotics, and reducing the toxicity of carcinogens, which can be further studied as the focus of subsequent liver cancer prevention in clinical practice.

Microglia are involved in the inflammatory response and retinal ganglion cell damage in glaucoma. Here, we investigated how microglia proliferate and migrate in a mouse model of chronic ocular hypertension (COH). In COH retinas, the microglial proliferation that occurred was inhibited by the P2X7 receptor (P2X7R) blocker BBG or P2X7R knockout, but not by the P2X4R blocker 5-BDBD. Treatment of primary cultured microglia with BzATP, a P2X7R agonist, mimicked the effects of cell proliferation and migration in COH retinas through the intracellular MEK/ERK signaling pathway. Transwell migration assays showed that the P2X4R agonist CTP induced microglial migration, which was completely blocked by 5-BDBD. In vivo and in vitro experiments demonstrated that ATP, released from activated Müller cells through connexin43 hemichannels, acted on P2X7R to induce microglial proliferation, and acted on P2X4R/P2X7R (mainly P2X4R) to induce microglial migration. Our results suggest that inhibiting the interaction of Müller cells and microglia may attenuate microglial proliferation and migration in glaucoma.
Adenosine Triphosphate/pharmacology* ; Animals ; Cell Proliferation ; Glaucoma/metabolism* ; Mice ; Microglia/metabolism* ; Receptors, Purinergic P2X4/metabolism* ; Receptors, Purinergic P2X7/metabolism* ; Retinal Ganglion Cells/metabolism*

Objective:To analyze the risk factors of death within two years of the patients with liver cirrhosis after transjugular intrahepatic portosystemic shunt(TIPS), and to explore the predictive value of 6 common clinical evaluation systems on the risk of death after TIPS.Methods:TIPS clinical data from 132 patients with liver cirrhosis were analyzed retrospectively. According to the 2-year clinical outcome after TIPS, the patients were divided into the death group and the survival group. Logistic regression was used to analyze the risk factors of death within 2 years after TIPS. According to the scores of CTP, MELD, MELD Na, BioCliM, FIB-4, and ALBI evaluation systems, the prediction efficiency of death risk of the six evaluation systems was evaluated by using the receiver operating characteristic (ROC) curves and the area under the curve (AUC).Results:During the 2-year follow-up period after TIPS, the age, urea nitrogen level, platelet count, and proportion of hepatic encephalopathy in the death group were higher than those in the survival group one month after TIPS, and the serum sodium level was lower than those in the survival group (all P<0.05). Multivariate analysis showed that the elderly and hepatic encephalopathy one month after operation were independent risk factors for death (all P<0.05). At 1 week after the surgery, there were significant differences in CTP, MELD, and MELD-Na scores between the survival group and the death group (all P<0.05). One week after operation, the AUC of ROC of CTP, MELD, MELD-Na, and ALBI scores were 0.685, 0.721, 0.805, and 0.658 respectively, and the optimal critical values were 8.5, 12.99, 14.51 and -1.52 respectively. Conclusions:The elderly and the occurrence of hepatic encephalopathy one month after TIPS are independent risk factors for the death of liver cirrhosis patients after TIPS. The evaluation of CTP, MELD, MELD-Na, and ALBI one week after TIPS can predict the death risk of decompensated liver cirrhosis patients within 2 years after TIPS, and MELD-Na has the best predictive effect.

Objective:To explore the effects of miR-181a-5p on the occurrence and development of gastrointestinal stromal tumors (GIST) through targeting CTDSPL mediating TGF-β signaling pathway.Methods:Surgical treatment of GIST patients in the First People’s Hospital of Shangqiu City from Jan. 2016 to Dec. 2019 were selected as research objects, and tumor tissue and adjacent normal tissue were collected intraoperatively. The clinicopathological data of the patients were analyzed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression of CTDSPL gene and miR-181a-5p expression. Western blot was used to detect the protein level of CTDSPL and TGF-β signaling pathway related factors. Human gastrointestinal stromal tumor cell lines (GIST-T1) were transfected with miR-181a-5p mimic, miR-181a-5p inhibitor, or CTDSPL overexpression vector. MTT was used to detect cell proliferation activity, Transwell assay was utilized to detect cell invasion, flow cytometry was used to determine cell apoptosis in each group.Results:Compared with adjacent tissues, expression of miR-181a-5p and TGF-β signaling pathway related factors was activated while CTDSPL expression was inhibited. Tumor size, invasion depth and modified NIH grading were related to the mRNA expression level of CTDSPL gene in GIST tumor tissues (All P<0.05) . Compared with Blank group, inhibition of miR-181a-5p or CTDSPL overexpression had the ability to inhibit the cell viability and invasion, induce apoptosis. The effects of miR-181a-5p mimic on GIST-T1 can be saved by CTDSPL overexpression. Conclusion:miR-181a-5p can promote the occurrence and development of GIST by down-regulating the CTDSPL gene level and activating TGF-β signaling pathway.

Objective: To learn the prevalence and influencing factors of dyslipidemia among the residents aged 35-75 years in Zhejiang Province, so as to provide basis for prevention and control of dyslipidemia. Methods: Based on the national high-risk early screening and comprehensive intervention program for cardiovascular disease, multi-stage stratified cluster sampling method was adopted to select 35-75 year-old permanent residents from six cities （counties or districts） in Zhejiang Province. A questionnaire survey was used to collect demographic data;physical examination and laboratory test were carried out. A multivariate logistic regression model was used to analyze the influencing factors for dyslipidemia. Results : A total of 44 440 residents were investigated and 40 751 residents responded,accounting for 91.70%. The crude prevalence rate of dyslipidemia was 36.88% （15 027 cases） and the age-standardized one was 34.58%. The crude prevalence rates of hypercholesterolemia, hypertriglyceridemia and high low density lipoprotein-cholesterol were 18.10%, 14.05%, 10.57% and 15.78%, respectively,the age-standardized ones were 16.11%, 13.76% and 14.53%, respectively.The results of multivariate logistic regression analysis showed that age （OR=1.006, 95%CI:1.004-1.008）,urban residence （OR=1.139, 95%CI: 1.087-1.194）,smoking （OR=1.099, 95%CI: 1.033-1.170）,drinking （OR=0.915,95%CI: 0.863-0.970）,underweight （OR=0.623, 95%CI: 0.528-0.735）, overweight （OR=1.624,95%CI: 1.552-1.699）, obesity （OR=2.128, 95%CI: 1.985-2.281）, diabetes （OR=1.600, 95%CI: 1.493-1.715）, hypertension （OR=1.218, 95%CI: 1.165-1.273） and hyperurcemia （OR=1.789, 95%CI: 1.679-1.905） were associated with dyslipidemia. Conclusions The age-standardized prevalence rate of dyslipidemia among the residents aged 35-75 years in Zhejiang Province was lower than that in the whole country,but the prevalence rates of hypercholesterolemia,hypertriglyceridemia and high low density lipoprotein-cholesterol were relatively high.

OBJECTIVES: To investigate the distribution of serum calcium and the relationship between serum calcium and serum metabolic parameters in endometrial carcinoma (EC) patients. METHODS: Retrospective assessment of patients diagnosed with endometrial cancer from Peking University People's Hospital from 2004 to 2009. Clinical characteristics as well as pretreatment serum calcium, albumin, fasting plasma glucose (FPG), serum triglycerides (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and total cholesterol (TC) value were extracted from patient records. Serum calcium was corrected for albumin. Unpaired t test and analysis of covariance were used to compare serum calcium among categorical variables. Simple correlation analyses and partial correlation analyses were used to assess the associations between serum calcium and continuous variables. RESULTS: Two-hundred twenty patients were included in this study. After adjusting for confounders, postmenopausal patients had higher total serum calcium (p=0.002) and albumin-corrected serum calcium (p=0.012) than premenopausal patients, endometrioid endometrial carcinoma (EEC) patients had higher total serum calcium than non-endometrioid endometrial carcinoma (NEEC) patients (p=0.037). Significant positive correlations were found between total serum calcium and FPG (p=0.017), TG (p=0.043), HDL (p=0.042), LDL (p < 0.001), and TC (p < 0.001) after adjusting for multiple variables, and the corrected serum calcium showed no significant correlation with metabolic parameters. CONCLUSION: Total serum calcium might be a more sensitive parameter for metabolic syndrome in endometrioid endometrial cancer patients than lipids.
Beijing ; Blood Glucose ; Calcium* ; Cholesterol ; Endometrial Neoplasms* ; Fasting ; Female ; Humans ; Lipoproteins ; Retrospective Studies ; Triglycerides