AIM:To explore the mechanism of PX-12 induced apoptosis of hepatocellular carcinoma cells.METHODS:Human hepatoma cell line Huh7 was se-lected as the main research object.After the cells were treated with thioredoxin inhibitor PX-12,the cell viability was detected by CCK8 method,the cell mi-gration ability was detected by cell scratch test,the cell proliferation ability was detected by cell prolif-eration kit,the levels of intracellular reactive oxygen species and apoptosis were detected by flow cy-tometry,and the expression of apoptosis-related proteins were detected by Western blot.RESULTS:Compared with the control group,the cell viability,migration ability and proliferation ability of PX-12 treatment group were significantly decreased(P＜0.05),and the level of intracellular reactive oxygen species was increased(P＜0.05)in a concentration-dependent manner.Apoptosis inhibitor Z-VAD-FMK and antioxidant NAC could restore the cell viability,and NAC could reduce the accumulation of intracel-lular reactive oxygen species induced by PX-12 and restore the apoptosis induced by PX-12(P＜0.05).CONCLUSION:PX-12 induces apoptosis of hepato-cellular carcinoma cells through oxidative stress.

Objective:To investigate the feasibility of 3.0 T glutamate chemical exchange saturation transfer (GluCEST) imaging in evaluating renal redox metabolism in renal ischemia-reperfusion injury (IRI).Methods:Rabbits in the IRI group ( n=56) underwent surgery by clamping the left renal artery for 45 min and then releasing to establish IRI. Rabbits in the sham group ( n=8) underwent the same operation without clamping the left renal artery. GluCEST MRI was performed before and at 1 h, 12 h, 1 day, 3 days, 7 days, and 14 days after the operations, with eight rabbits in the IRI group sacrificed immediately after each scanning and eight in the sham group sacrificed at 14 days after scanning. The left kidneys were removed for histopathological examination and reactive oxygen species (ROS) fluorescence staining. Differences in the magnetic resonance ratio asymmetry (MTR asym) of the renal cortex and outer medulla among different groups were compared. Correlations between the MTR asym and ROS were analyzed. Results:The MTR asym of the renal cortex in the sham and IRI subgroups were higher than that of the outer medulla ( t=8.16, P<0.001; t=4.78, P=0.002; t=4.94, P=0.002; t=5.76, P=0.001, t=6.68, P<0.001; t=6.40, P<0.001; t=5.16, P=0.001; t=3.30, P=0.013). The MTR asym of the renal cortex and outer medulla in the IRI-1h, IRI-12h, IRI-1d, IRI-3d, IRI-7d, and IRI-14d groups were lower than in the sham and IRI-pre groups (all P<0.05). The MTR asym of the renal cortex and outer medulla in the IRI-1h group were lower than in the IRI-12h, IRI-1d, IRI-3d, IRI-7d, and IRI-14d groups (all P<0.05). The MTR asym of the renal cortex in the IRI-12h group was lower than in the IRI-7d and IRI-14d groups (1.84%±0.09% vs.2.42%±0.19%, 2.41%±0.31%, all P<0.05). The MTR asym of the renal cortex in the IRI-1d group was lower than in the IRI-7d group (1.99%±0.17% vs. 2.42%±0.19%, P=0.008). The MTR asym of the outer medulla in the IRI-12h group was lower than in the IRI-3d, IRI-7d, and IRI-14d groups (1.32%±0.27% vs. 1.79%±0.31%, 1.98%±0.18%, 1.66%±0.40%, respectively, all P<0.05]. The MTR asym of the outer medulla in the IRI-7d group was higher than in the IRI-1d and IRI-14d groups (1.98%±0.18% vs. 1.52%±0.31%, 1.66%±0.40%, all P<0.05). The MTR asym of the renal cortex and outer medulla had a strong negative correlation with the mean fluorescence intensity of ROS ( ρ=-0.889, P<0.001; ρ=-0.784, P<0.001). Conclusion:3.0 T GluCEST imaging can indirectly reflect the changes of renal redox metabolism in renal IRI.

Objective To study the relationship between waist-to-height ratio(WHtR)and new-onset cerebral infarction in the elderly with hypertension.Methods A total of 9 096 elderly hypertensive patients were enrolled from the Kailuan study population.Using the first occurrence of cerebral infarction as the endpoint event,a prospective cohort study was performed.According to the WHtR value of the subjects with an interval of 0.05,the subjects were divided into G1 group(WHtR＜0.45),G2 group(0.45≤WHtR＜0.50),G3 group(0.50≤WHtR＜0.55),and G4 group(WHtR≥0.55).The cumulative incidence rate of endpoint event in each population were statistically analyzed using the life table method.Cox proportional hazards regression analysis was used to explore the role of WHtR in the occurrence of new cerebral infarction.Model 1 was a single factor Cox proportional hazards analysis model,only considering the influence of WHtR on the risk of cerebral infarction.After adjusting sex,age,body weight,serum hypersensitive C-reactive protein,systolic blood pressure,serum triglycerides,smoking history,fasting blood glucose,alcohol consumption,high-density lipoprotein cholesterol,diastolic blood pressure,exercise,low-density lipoprotein cholesterol,educational level,antihypertensive drug history,model 2 was established to observe the influence of WHtR on the occurrence of new-onset cerebral infarction.Results The follow-up lasted(9.3±2.7)years.Compared to the G2 group,multivariate Cox proportional hazards regression model showed that the hazard ratio HR(95%CI)for cerebral infarction in model 1 was 1.00(0.67～1.50),1.29(1.05～1.60)and 1.36(1.12～1.66)respectively in G1,G3 and G4;and that in model 2 was 1.09(0.72～1.65),1.24(0.99～1.54)and 1.31(1.05～1.62)respectively in G1,G3 and G4.Conclusion The risk of cerebral infarction in the elderly with hypertension is the lowest when WHtR between 0.45≤WHtR＜0.50,and WHtR greater than or equal to 0.55 is associated with an increased risk of new-onset cerebral infarction.

Ras-related C3 botulinum toxin substrate 1(RAC1)is a crucial member of the Ras GTPase super-family,exerting an overriding role in cell behaviors such as motility,proliferation and adhesion,and is involved in regu-lating vascular permeability and cellular barrier function.Recently,the significance of RAC1 in the etiology and progres-sion of various pulmonary disorders has garnered increasing attention,which sheds light on lung disease study.In this re-view,we introduce the characteristics of RAC1 systematically and recapitulate its effects and molecular mechanisms in the context of lung tumors,chronic obstructive pulmonary disease,lung injury and pulmonary arterial hypertension,hoping to provide new insights for future pulmonary disease treatment.

Objective To construct a canine model of vocal cord scar by low-termperature plasma ablation and screen the target genes closely related to the formation of vocal cord scar.Methods Four Chinese rural canines were treated with plasma ablation under the support of laryngoscope and endoscope,and the left vocal cords were injured to the muscle layer.The contralateral sides were left untreated.The gross morphology of vocal cord was observed before operation,immediately after operation,3 weeks after operation and 12 weeks after operation.The pathologi-cal structure of vocal cords was observed by HE stainning,and the ultrastructure of vocal cords was observed by transmission electron microscopy.In addition,high-throughput sequencing was used to analyze the differences in gene expression between the bilateral vocal cords,and the target genes with significantly different expression were screened out.Results In general morphology,the normal vocal cords were banded and well closed.At 3 weeks af-ter operation,the vocal cords were congested and swollen,with uneven edges and red granulation tissues were seen.At 12 weeks after operation,the vocal cord wound was localized contracture and depression,and scar was formed.HE staining showed obvious thickening of the squamous epithelium of the scarred vocal cords,thickening and disor-dered arrangement of the fiber layer,local clumping or bundle aggregation,and scattered fiber bundles were also seen in the muscle layer.Transmission electron microscopy showed interstitial thickening,uneven density,cell swelling,unclear intercellular boundary,proliferation of nuclei and mitochondria,and cells in an active state.High-throughput sequencing analysis revealed that many gene families were involved in the process of vocal cord scar re-pair,including IL family,CCL and CXCL family,MMPs family and its inhibitor TIMPs family,Wnt family,HSP family,MAPK family and TGF-β family.Conclusion We successfully constructed the canine model of vocal cord scar by low-temperature plasma ablation and screened out the target genes closely related to the formation of vocal cord scar by high-throughput sequencing,which provides certain reference value for exploring the mechanism of vo-cal cord scar.

Objective To investigate the effect of ERK inhibitor PD98059 on the proliferation and differentia-tion of rat otocyst osteoblasts.Methods SD neonatal rat osteoblasts were extracted by two-step digestion with 0.25％pancreatin and type Ⅰ collagenase,and co-cultured with ERK inhibitor PD98059 at concentrations of 0 μmol/L,10 μmol/L,25 μmol/L and 50 μmol/L,respectively.Then,the osteoblasts proliferation of the four groups were assessed by EDU method for 4 consecutive days.The proliferation trend of each group was compared and analyzed.Osteoblasts were differentiated by β-sodium glycerophosphate,L-vitamin C and dexamethasone at concentrations of 10 mmol/L,50 ug/ml and 10-7 mol/L.After 24 h,the mRNA expression levels of osteogenic fac-tors which include Ocn,Bsp,Runx2,Bmp2,OPG and RANKL in each group were detected by RT-qPCR,and the differences of the results were analyzed.Results All the concentrations of ERK inhibitor PD98059 could inhibit the proliferation of osteoblasts in SD neonatal rat,and the inhibitory effect of PD98059 at concentrations of 10 μmol/L was significantly greater than that of the other three groups(P＜0.05).In addition,all the concentrations of PD98059 could inhibit the expressions of Ocn,Bsp,Runx2,Bmp2 and OPG mRNA.The mRNA expressions of Ocn,Bsp,Runx2 and Bmp2 in 10 μmol/L PD98059 group were significantly lower than those in 0 μmol/L,25μmol/L and 50 μmol/L PD98059 groups(P＜0.05).The mRNA expressions of OPG in 10 and 25 μmol/L PD98059 groups were significantly lower than those in 0 and 50 μmol/L PD98059 groups(P＜0.05),and there was no significant difference between the first two groups(P＞0.05).The CT value of RANKL mRNA was not detec-ted in all groups.Conclusion ERK pathway inhibitor PD98059 can both inhibit the proliferation and differentiation of osteoblasts in rat otocyst.Therefore,we speculate that ERK1/2-MAPK pathway may mediate the formation of tympanosclerosis by affecting the proliferation and differentiation of rat otocyst osteoblasts.

AIM:To investigate the expression of centromere protein-H(CENP-H)in adrenocortical carcino-ma(ACC)and its relationship with disease progression and prognosis,and to explore the impact of CENP-H gene knock-down on the viability and migration of ACC cells.METHODS:The mRNA expression level of CENP-H in 76 ACC pa-tients and 128 healthy controls,and its correlations with tumor stages and prognosis were analyzed by GEPIA2 database.The mRNA expression of CENP-H in different stages of ACC and its correlation with disease prognosis were further ana-lyzed by ULCAN database.The protein expression of CENP-H was examined by immunohistochemical staining of paraffin-embedded ACC and normal adrenal gland specimens.Knockdown of CENP-H by siRNA(siCENP-H)was performed in human ACC cell line H295R.The viabilty of H295R cells transfected with siCENP-H or siNC was measured by CCK-8 as-say,the cell migration was detected by wound-healing assay,and the protein levels of CENP-H,p-ERK1/2,t-ERK1/2,p-P38,t-P38,p-JNK1/2 and t-JNK1/2 were detected by Western blot.RESULTS:The mRNA level of CENP-H was signifi-cantly higher in ACC than that in normal controls,and was correlated with tumor stages and prognosis.The protein level of CENP-H was significantly higher in ACC specimens than that in normal adrenal gland.Knockdown of CENP-H in H295R cells resulted in decreased cell viability and migration.The protein levels of p-P38 and p-JNK1/2 were decreased in si-CENP-H group.CONCLUSION:CENP-H is highly expressed in ACC,and is correlated with tumor stages and poor prognosis.Knockdown of CENP-H can inhibit the viability and migration of ACC cells,and its mechanism may related to inactivation of P38 and JNK signaling pathways.

Objective:To explore the feasibility of chemical exchange saturation transfer (CEST) imaging at 3.0 T MRI in quantifying renal redox metabolism in vitro models and experimental animals.Methods:Redox metabolites in vitro models with physiological concentrations were prepared, including reduced metabolites (glutamate, alanine, glutathione) and oxidized metabolites (2-ketoglutarate, pyruvate, glutathione disulfide, ammonium hydroxide). CEST examinations were performed at 3.0 T MRI. The imaging parameters were as follows: CEST images with different saturation pulse intensity (B 1) (1, 2, 3, 4 μT) and a fixed radio frequency (RF) duration of 2 000 ms; CEST images with different RF durations (1 500 and 2 000 ms) were acquired with a fixed B 1 value of 2 μT to obtain the optimal scanning parameters. CEST examinations with optimized parameters were performed on the left kidneys of seven healthy rabbits, and the differences in magnetic resonance ratio asymmetry (MTR asym) between rabbit renal cortex and outer medulla were measured. A paired t-test was used to compare the differences. Results:The optimal B 1 for CEST examination of redox metabolites was 2 μT, and the optimal RF duration was 2 000 ms. The MTR asym peaks of glutathione disulfide, glutathione, glutamic acid, and alanine were at 3.75, 3.5, 3, and 1.5 ppm, respectively. The MTR asym peaks of pyruvate, 2-ketoglutarate, and ammonium hydroxide were at 1 ppm. The MTR asym peak values of reduced metabolites were higher than those of oxidized metabolites. When the B 1 value was 2 μT and the RF duration was 2 000 ms, the MTR asym signal of the renal cortex was (2.60±1.10) %, (2.86±1.32) %, (3.04±1.06) %, and (2.98±0.91) % at 1, 3, 3.5, and 3.75 ppm, respectively. The MTR asym signal of the outer medulla was (1.00±0.56) %, (2.43±0.94) %, (2.29±0.88) % and (1.98±0.58) %, respectively. The MTR asym signal of the renal cortex was higher than that of the outer medulla, and the differences were statistically significant ( t=3.04, P=0.023; t=2.56, P=0.043; t=3.50, P=0.013; t=3.45, P=0.014). Conclusion:CEST imaging at 3.0 T MRI can be used to quantitatively evaluate redox metabolism of healthy rabbit kidneys in vitro model and normal experimental rabbits.

Objective To investigate the protective effects and underlying mechanisms of Liuwei Dihuang Glucoside(LW-AFC)against fear sensitization induced by traumatic stress.Methods Mice were divided into naive,control,stressed and LW-AFC administration groups.The LW-AFC treated group received LW-AFC(1.6 g/kg daily)via oral gavage for two weeks following exposure to traumatic stress.The non-associative memory dependent fear sensitization responses in mice subjected to trauma were investigated,including behavior in novel environments,social interaction,and observational fear tests.Z-score normalization method was employed to integrate and assess multiple behavioral variables such as travel distance,freezing time,and corner time,and comprehensively examined fear sensitization behaviors across the groups.Additionally,serum concentrations of adrenocorticotropic hormone,corticosterone,aldosterone,renin,angiotensin Ⅱ,and aldosterone were measured using ELISA.Results Compared to the control group,stressed mice exhibited significantly reduced travel distance(P<0.0001)and increased freezing time(P<0.0001)in the new context test.Integrated Z-scores indicated a significant increase in fear behavior among stressed mice during the new context test(P<0.0001).In the social interaction test,stressed mice demonstrated significantly reduced travel distance(P<0.0001),increased freezing time(P<0.0001),increased corner time(P<0.05),and higher integrated Z-scores(P<0.0001).In the observational fear test,stressed mice showed significantly reduced travel distance(P<0.05),increased freezing time(P<0.001),increased corner time(P<0.05),and higher integrated Z-scores(P<0.0001).Compared to the stressed group,mice in the LW-AFC treated group displayed trends toward improvement in travel distance,freezing time,corner time,and integrated Z-scores in the new context tests,albeit without statistical significance.In the social interaction test,the LW-AFC treated group exhibited a significant reduction in freezing time(P<0.05)and integrated Z-scores(P<0.05).In the observational fear test,the LW-AFC treated group showed a significant reduction in freezing time(P<0.05)and integrated Z-scores(P<0.01).Compared to the naive group,control and stressed groups exhibited an increased trend in renin and aldosterone levels after the fear sensitization test.Although there were no significant differences between stressed and control groups,renin and aldosterone levels significantly increased between stressed and naive groups(P<0.05,P<0.05).Following LW-AFC treatment,serum renin levels showed no significant change,while aldosterone levels significantly decreased(P<0.05).Conclusion Stressed mice exhibited significant fear sensitization behavior in new context,social interaction,and observational fear tests,possibly associated with partial activation of the renin-angiotensin-aldosterone system(RAAS)system.LW-AFC treatment significantly mitigated fear sensitization behavior of stressed mice in social interaction and observational fear test,potentially due to its regulatory effects on the RAAS system in mice subjected to traumatic stress.

Endodontic diseases are a kind of chronic infectious oral disease.Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha.However,it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy(RCT).Recent research,encompassing bacterial etiology and advanced imaging techniques,contributes to our understanding of the root canal system's anatomy intricacies and the technique sensitivity of RCT.Success in RCT hinges on factors like patients,infection severity,root canal anatomy,and treatment techniques.Therefore,improving disease management is a key issue to combat endodontic diseases and cure periapical lesions.The clinical difficulty assessment system of RCT is established based on patient conditions,tooth conditions,root canal configuration,and root canal needing retreatment,and emphasizes pre-treatment risk assessment for optimal outcomes.The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT.These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.