Objective : To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis. Methods : An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression. Results : Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01). Conclusion Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.

Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Objective To investigate the differences of brain imaging changes in patients with mesial temporal lobe epilepsy(mTLE)based on individual structural covariance networks of gray matter volume.Methods A total of 74 mTLE patients,including 39 patients in the left mTLE group and 35 patients in the right mTLE group,along with 46 healthy controls(control group),had completed 3D T1WI structural imaging scans.The network template perturbation approach was used to analyze the individualized structural covariance networks in patients with mTLE.Results Compared with the control group,the left and right mTLE groups showed decreased structural covariance connections for the ipsilateral hippocampus to the contralateral hippocampus and parahippocampal gyrus,orbitofrontal gyrus,middle frontal gyrus,superior parietal gyrus,amygdala and fusiform gyrus.In addition,increased structural covariation connections was mainly distributed in the bilateral frontal lobe,parietal lobe,temporal lobe,occipital lobe and paralimbic system in the right mTLE group,whereas increased structural covariation connections was mainly located within the left frontal lobe,parietal lobe and occipital lobe in the left mTLE group(P＜0.05).Compared with the left mTLE group,the right mTLE group showed decreased structural covariance connections with the ipsilateral hippocampus as a core node(P＜0.05).Conclusion Compared with the left mTLE group,the right mTLE group showed more pronounced decreased structural covariance connections centered around the ipsilateral hippocampus,as well as a more intricate compensatory mechanism.The pattern of the individual structural covariance networks in mTLE patients not only contribute to understanding of its pathogenesis but also served as a potential biomarker for clinical diagnosis.

Objective To investigate the differences of brain imaging changes in patients with mesial temporal lobe epilepsy(mTLE)based on individual structural covariance networks of gray matter volume.Methods A total of 74 mTLE patients,including 39 patients in the left mTLE group and 35 patients in the right mTLE group,along with 46 healthy controls(control group),had completed 3D T1WI structural imaging scans.The network template perturbation approach was used to analyze the individualized structural covariance networks in patients with mTLE.Results Compared with the control group,the left and right mTLE groups showed decreased structural covariance connections for the ipsilateral hippocampus to the contralateral hippocampus and parahippocampal gyrus,orbitofrontal gyrus,middle frontal gyrus,superior parietal gyrus,amygdala and fusiform gyrus.In addition,increased structural covariation connections was mainly distributed in the bilateral frontal lobe,parietal lobe,temporal lobe,occipital lobe and paralimbic system in the right mTLE group,whereas increased structural covariation connections was mainly located within the left frontal lobe,parietal lobe and occipital lobe in the left mTLE group(P＜0.05).Compared with the left mTLE group,the right mTLE group showed decreased structural covariance connections with the ipsilateral hippocampus as a core node(P＜0.05).Conclusion Compared with the left mTLE group,the right mTLE group showed more pronounced decreased structural covariance connections centered around the ipsilateral hippocampus,as well as a more intricate compensatory mechanism.The pattern of the individual structural covariance networks in mTLE patients not only contribute to understanding of its pathogenesis but also served as a potential biomarker for clinical diagnosis.

Objective·To establish a prognostic model for the overall survival(OS)of hepatocellular carcinoma(HCC)based on mitochondrial genes and clinical information.Methods·The gene expression and the clinical data of 369 HCC patients and 50 controls with normal liver were downloaded from The Cancer Genome Atlas(TCGA)database.The nuclear-encoded mitochondrial genes(NEMGs)were obtained from the MitoCarta3.0 database.The"DESeq2"R package and univariate Cox analysis were used to select NEMGs[ubiquinol cytochrome C reductase hinge protein(UQCRH),ATP citrate lyase(ACLY),phosphoenolpyruvate carboxykinase 2(PCK2),Bcl-2 homologous antagonist/killer1(BAK1),Bcl-2-associated X protein(BAX)and Bcl-2/adenovirus E1B interacting protein 3-like(BNIP3L)]in HCC that were associated with OS of HCC and participated in dysregulation of oxidative phosphorylation,tricarboxylic acid cycle and cell apoptosis.Multivariate Cox analysis was applied to select independent risk factors for OS of HCC.A comprehensive prognostic model and a prognostic nomogram with 6-NEMG risk characteristics and TNM staging were established.By using the median of prognostic scores as a cut-off,HCC patients were classified into low-risk and high-risk group.Kaplan-Meier survival curve analysis was conducted and log-rank test was performed to evaluate the survival rates between the low-risk and high-risk group.The area under the curve(AUC)values of receiver operating characteristic(ROC)curve were calculated via using the"timeROC"package.The prognostic model for HCC was validated by using the GEO HCC cohort(GSE14520)for 1,3 and 5 years.Finally,the relative expression level of 6-NEMG was validated in 34 clinical samples of HCC from Xinhua Hospital,Shanghai Jiao Tong University School of Medicine by using real-time quantitative polymerase chain reaction(qPCR)method.Results·Compared to 6-NEMG risk signature only(AUCs for 1,3 and 5 years were 0.77,0.66 and 0.65,respectively)or TNM stage only(AUCs for 1,3 and 5 years were 0.66,0.67 and 0.63,respectively),ROC curve analysis showed that this integrated prognostic model displayed better predictive performance for 1-year(AUC,0.78),3-year(AUC,0.73)and 5-year(AUC,0.69)OS of HCC.The Kaplan-Meier survival curve analysis showed that the OS of HCC patients in the high-risk group was significantly worse than that in the low-risk group(P=0.001).In addition,predictive performance of the prognostic model(AUC for 1,3 and 5 years is 0.67,0.66 and 0.74,respectively)and prognostic differences between the high-risk and low-risk group(P=0.001)were further validated in GEO(GSE14520)external cohort,and these results were consistent with the TCGA data.In addition to BNIP3L,dysregulation of five other NEMGs in the clinical HCC cohort was validated.The correlation analysis in GSE14520 and HCC clinical cohort showed a positive correlation between prognosis score and the size and number of tumors.Conclusion·A new prognostic model that combines 6-NEMG risk characteristics with TNM staging for predicting OS in HCC patients was constructed and validated.This model may help improve the prognosis prediction of HCC patients.

Objective To investigate the effects of pterostilbene on human colon cancer LoVo cells and study the regulatory mechanism of nuclear factor E2-related factor 2(Nrf2)in the process of pterostilbene acting on LoVo cells.Methods LoVo cells were treated with different concentrations(5,10,20,40,60,80,100 panol/L)of pterostilbene.Cell viability,migration,invasion,and apoptosis were examined by CCK-8,scratch,Tran-swell,and TUNEL assays,respectively.The mitochondrial membrane potential was measured by the mitochon-drial membrane potential assay kit with JC-1.The reactive oxygen species level was measured by 2',7'-dichlo-rofluorescein diacetate.The protein levels of Nrf2,phosphorylated Nrf2,heme oxygenase 1,and apoptotic pro-teins(Bcl2 and Bax)were determined by Western blotting.In addition,cell viability,Nrf2 expression,and ap-optosis rate were determined after co-application of the Nrf2-specific agonist sulforaphane.Results Compared with the control group,40,60,80,100 μmol/L pterostilbene reduced the viability of LoVo cells(P=0.014,P＜0.001,P＜0.001,P＜0.001).Pterostilbene at 5,10,20 μmol/L did not show effects on cell viability but inhibited cell migration(P=0.008,P＜0.001,P＜0.001)and invasion(all P＜0.001).Pterostilbene at 40,60,80 μmol/L increased apoptosis(P=0.014,P＜0.001,P＜0.001),promoted mitochondrial membrane potential depolarization(P=0.026,P＜0.001,P＜0.001)and reactive oxygen species accumula-tion(all P＜0.001),and down-regulated the expression of phosphorylated Nrf2(P=0.030,P＜0.001,P＜0.001),heme oxygenase 1(P=0.015,P＜0.001,P＜0.001),and Bc12(P=0.039,P＜0.001,P＜0.001)in LoVo cells.Pterostilbene at 60,80 μmol/L down-regulated Nrf2 expression(P=0.001,P＜0.001)and up-regulated Bax expression(both P＜0.001).The application of sulforaphane reversed the effects of pterostilbene on cell viability(P＜0.001),apoptosis(P＜0.001),and Nrf2 expression(P=0.022).Conclusion Pterostilbene is a compound that can effectively inhibit colon cancer cells by inhibiting the Nrf2 pathway.

Objective:To understand the molecular epidemic characteristics and genes of human astrovirus (HAstV) in infantile diarrhea patients in Jilin province from 2017 to 2020.Methods:Fecal samples of hospitalized infants with diarrhea under 5 years of age from January 2017 to December 2020 were collected. The nucleic acids of fecal samples were amplified by reverse transcription-polymerase chain reaction (RT-PCR), the positive amplification products were sequenced, and the phylogenetic tree was constructed.Results:A total of 2 019 fecal samples were collected, of which 71 were positive for HAstV, with a positive rate of 3.5%. Among the cases with positive samples of HAstV, 42 had mixed infection with other viruses causing diarrhea, accounting for 59.2% of the total number of positive cases, half of which were mixed infection with rotavirus and HAstV. From 2017 to 2020, the positive rates of HAstV were 4.69%, 1.98%, 5.93% and 0.43% per year, respectively. There were two epidemic peaks every year and one epidemic peak year every two years. The positive rate of 36-47 months old was the highest, followed by 0-2 months old. A total of 55 sequences were obtained in this study. Phylogenetic tree analysis showed that they all belonged to classical HAstV, including 42 HAstV-1a subtypes, 8 HAstV-1b subtypes and 5 HAstV-5 subtypes.Conclusions:HAstV is one of the important pathogens of diarrhea in children under 5 years of age in Jilin province. The epidemic law has obvious seasonal and age differences. The dominant strain is HAstV-1a.

Objective:To explore the expression of the nucleotide-binding oligomerization domain-like receptor containing pyrin domain protein 6 (NLR family,pyrin domain containing 6,NLRP6) in the gastric tissue and gastric juice of children with chronic non-atrophic gastritis (CNG), and to analyze the influence of Helicobacter pylori (Hp) infection on the expression of NLRP6.Methods:A case-control study was conducted to select 120 CNG patients in pediatrics of Department of Pediatrics, The Affiliated Hospital of Xuzhou Medical University from October 2020 to July 2021. According to pathological diagnosis, endoscopic gastric mucosal damage and Hp infection, they were divided into 4 groups: mild CNG group Hp negative, Moderate to severe CNG group Hp negative, Mild CNG group Hp positive, Moderate to severe CNG group Hp positive. The enzyme-linked immunosorbent assay (ELISA) was used to detect the expression level of NLRP6 in the four groups of gastric tissue and gastric juice, and Western blot was used to detect the expression of NLRP6 in the gastric tissue of the 4 groups, and the significance of expression in CNG of children is analyzed. Independent sample t-test was used to compare the mean between the two groups. One way ANOVA was used to compare the mean of multiple groups of samples, and LSD t-test was used for pairwise comparison. Comparison between count data groups χ 2 inspection. Results:The positive rate of Hp in the moderate to severe chronic non-atrophic gastritis group was 62.96% (34/54) higher than that in the mild chronic non-atrophic gastritis group 37.04% (20/54), and the difference was statistically significant (χ 2=18.32, P<0.001). Under the same Hp conditions, the expression of NLRP6 in the mild chronic non-atrophic gastritis group (Hp negative mild CNG: gastric tissue (653.73±37.71) ng/L, gastric juice (471.75±38.47) ng/L; Hp positive mild CNG: Gastric tissue (616.69±43.33) ng/L, gastric juice (445.29±36.39) ng/L was higher than the moderate to severe chronic non-atrophic gastritis group (Hp negative moderate to severe CNG: gastric tissue (623.82±52.99) ng/L, gastric juice (446.48±47.49) ng/L; Hp positive Moderate to severe CNG: gastric tissue (580.43±62.75) ng/L, gastric juice (406.88±51.85) ng/L, the difference is statistically significant (under Hp negative, mild compared with moderate to severe CNG: gastric tissue P=0.035; gastric juice P=0.046; Under Hp positive, mild compared with moderate to severe CNG: gastric tissue P=0.010;gastric juice P=0.002); in the same degree of gastric mucosal injury, NLRP6 expression in Hp-negative group (Hp-negative mild CNG: gastric tissue (653.73±37.71) ng/L, gastric juice (471.75±38.47) ng/L; Hp negative moderate to severe CNG: gastric tissue (623.82±52.99) ng/L, gastric juice (446.48±47.49) ng/L higher than the positive group (Hp positive mild CNG: gastric tissue (616.69±43.33) ng/L, gastric juice (445.29±36.39) ng/L; Hp positive moderate to severe CNG: gastric tissue (580.43±62.75) ng/L, gastric juice (406.88±51.85) ng/L, the difference is statistically significant (under mild CNG, Hp negative is compared with positive: Gastric tissue P=0.005; gastric juice P=0.023; under moderate to severe CNG, negative versus positive: gastric tissue P=0.004; gastric juice P=0.003). Conclusion:Under the same Hp conditions, the more severe the gastric mucosal damage, the lower the NLRP6; under the same degree of mucosal damage, the expression level of NLRP6 in the Hp-negative group was significantly higher than that of the Hp-positive group. It is suggested that NLRP6 plays a role in inhibiting inflammation in chronic gastritis, maintaining the integrity of epithelial cells, and Hp can inhibit the expression of NLRP6.

Objective: To analyze the genetic characteristics of norovirus(NoV) in Jilin province from 2014 to 2016. Methods: Stool samples from children under 5 years of age with diarrhea were collected from Changchun Children′s Hospital. Samples were amplified by RT-PCR. The polymerase and capsid gene of the positive specimens were amplified.Sequences were analyzed by homologous comparison and phylogenetic analysis. Results: A total of 1 335 samples were detected, 177 were positive for NoV, and positive rate was 13.26%. The positive rate was 11.83% in 2014, 12.53% in 2015 and 15.75% in 2016. Genotyping of polymerase region revealed GII.P12 types dominated in 2014 and GII.Pe types dominated in 2015 and 2016. Genotyping of polymerase region revealed that GII.3 types dominated in 2014 and GII.4_Sydney2012 types dominated in 2015 and 2016. The types of both regions included GII.P17/GII.17, GII.Pe/GII.4_Sydney2012, GII.P12/GII.3, GII.Pe/GII.3、GII.P16/GII.2 and GII.P7/GII.6. Conclusions Genotypes of NoV infection are diverse in Jilin province. Most of them are recombinant strains. The main epidemic strains changed from GII.P12/GII.3 in 2014 to GII.Pe/GII.4_Sydney2012 in 2015 and 2016.

Objective: To compare the clinical application of three-dimensional laparoscopic pancreatoduodenectomy (3D-LPD) with that of two-dimensional laparoscopic pancreatoduodenectomy (2D-LPD), and to explore the safety and feasibility of 3D-LPD. Methods: A retrospective analysis was made from the data of 45 patients with 3D-LPD and 45 patients with 2D-LPD who underwent total laparoscopic pancreatoduodenectomy from March 2017 to August 2018 at Department of Hepato-Pancreato-Biliary Surgery, the First People′s Hospital of Changzhou.The differences of intraoperative conditions, postoperative complications and postoperative pathological findings between the two methods were compared.Measurement data were compared with independent sample t-test, enumeration data were statistically analyzed with Chi-square test or Fisher exact probability. Results: The operation time of 3D-LPD group was shorter than that of 2D-LPD group ((335±95) min vs. (419±113) min, t=-3.817, P=0.000), which mainly showed that the time of digestive tract reconstruction was reduced ((92±26) min vs. (131±46) min, t=-4.951, P=0.000). The intraoperative blood loss in the 3D-LPD group was significantly less than that in the 2D-LPD group ((242±124) ml vs. (350±176) ml, t=-3.365, P=0.001), and the perioperative blood transfusion in the 3D-LPD group was significantly less than that in the 2D-LPD group (χ²=4.444, P=0.035). Postoperative hospitalization days and ICU stay time were not significantly different between the two groups(both P>0.05). Postoperative complications such as pancreatic fistula, biliary fistula, postoperative bleeding, gastric emptying disorders, abdominal infection, were not significantly different between the two groups(all P>0.05). Conclusions The operation time of 3D-LPD is shorter than that of 2D-LPD, and the amount of bleeding is less. Short-term clinical data showed that, 3D-LPD is effective, safe and worth popularizing.