OBJECTIVE To investigate the protective effect proanthocyanidin B2(PC-B2) on oxidative damage and apoptosis of mouse astrocytes(AS) induced by hydrogen peroxide(H2O2) and its mechanism. METHODS AS were isolated and cultured from neonatal C57BL/6 mice(1−3 d). The optimal concentration of H2O2 and PC-B2 was divided into four groups: normal group, normal+PC-B2 group(100 μg·mL‒1 PC-B2 treated for 24 h), H2O2 model group(200 μmol·L‒1 H2O2 treated for 24 h), PC-B2 group(200 μmol·L‒1 H2O2 and 100 μg·mL‒1 PC-B2 treated for 24 h). The cell viability of each group was detected by CCK-8 method. Cytotoxicity was detected by LDH method. The antioxidant capacity was detected by ABTS and DPPH. The content of MDA and the activity of SOD, CAT and GSH-Px were detected by ELISA kit. Detection of apoptosis in each group was done by TUNEL staining. The mRNA and protein expression levels of Bax, Bcl-2, Caspase-3, Akt/Stat3, p-Akt, p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting, respectively. RESULTS PC-B2 could significantly enhance cell viability and inhibit AS apoptosis. Compared with the H2O2 model group, PC-B2 intervention could significantly reduce the content of LDH and MDA in AS, and increase the activity of SOD, CAT and GSH-Px. PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3, and up-regulate the mRNA and protein expression of Akt/Stat3, Bcl-2, Nrf2/HO-1. CONCLUSION PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways, therefore reduce H2O2-induced AS oxidative damage and apoptosis.

Alzheimer's disease(AD)is a severe threat to human health and one of the three major causes of human death.Double-stranded RNA-dependent protein kinase(PKR)is an interferon-induced protein kinase involved in innate immunity.In the occurrence and development of AD,PKR is upregulated and continu-ously activated.On the one hand,the activation of PKR triggers an integrated stress response in brain cells.On the other hand,it indirectly upregulates the expression of 3-site amyloid precursor protein cleaving enzyme 1 and facilitates the accumulation of amyloid-β protein(Aβ),which could activate PKR activator to further activate PKR,thus forming a sustained accumulation cycle of Aβ.In addition,PKR can promote Tau phosphorylation,thereby reducing microtubule stability in nerve cells.Inflammation in brain tissue,neurotoxicity resulted from Aβaccumulation,and disruption of microtubule stability led to the progression of AD and the declines of memory and cognitive function.Therefore,PKR is a key molecule in the development and progression of AD.Effective PKR detection can aid in the diagnosis and prediction of AD progression and provide opportunities for clinical treat-ment.The inhibitors targeting PKR are expected to control the activity of PKR,thereby controlling the progression of AD.Therefore,PKR could be a target for the development of therapeutic drugs for AD.

Objective To explore the relationship between the polymorphism of excision repair cross-complementation group 1 (ERCC1) C8092A locus and the efficacy and prognosis of platinum-based chemotherapy for lung cancer (LC), and to provide a theoretical basis for precision treatment of LC. Methods From January 2014 to October 2017, 120 patients from two tertiary hospitals in Haikou City, and with pathologically confirmed lung cancer treated with platinum-based chemotherapy were selected as the research objects. After informed consent was obtained, peripheral blood samples were collected for DNA extraction, and the genotype of ERCC1 C8092A locus was detected by mass spectrometry. WHO's Response Evaluation Criteria in Solid Tumours (RECIST) was used to judge patients' chemotherapy efficacy and patients' survival status was obtained by telephone follow-up and other means. Results Among the 120 LC patients, the genotype frequencies of ERCC1 C8092A locus were 67 cases of CC wildtype (55.8%), 45 cases of CA heterozygous type (37.5%), and 8 cases of AA rare mutation type (6.7%), which conformed to Hardy-Weinberg equilibrium (χ2=0.140, P>0.05). The total effective rate of chemotherapy was 32.5%, with the highest effective rate in patients with the CA genotype (42.2%) at the ERCC1 C8092A locus and the lowest in patients with the CC genotype (25.4%). The overall one-year survival rate was 68.3% and the three-year survival rate was 35.8%. The patients with ERCC1 C8092A AA genotype had the lowest survival rate, with a one-year survival rate of 50.0% and three-year survival rate of only 25.0%. However, there were no statistical differences in the overall survival rate among the three genotypes of carriers of ERCC1 C8092A (χ2=0.328, P=0.849). Conclusions The polymorphism of ERCC1 C8092A locus is associated with the efficacy of platinum-based chemotherapy for LC, and patients with CA genotype have the highest efficacy. The one-year and three-year survival rates of patients with CC genotype are significantly higher than those of CA and AA genotypes.

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*

OBJECTIVE To comprehensively evaluate the quality of Eriobotrya japonica leaves from different producing areas. METHODS The contents of alcohol-soluble extracts were determined by hot-dipping method using 30 batches of E. japonica leaves from different producing areas as samples. The contents of total flavonoids and total triterpene acids were determined by ultraviolet spectrophotometry. The contents of five kinds of triterpenic acids （euscaphic acid，crataegolic acid，corosolic acid，oleanolic acid and ursolic acid） were determined by HPLC. The quality of E. japonica leaves from different producing areas was comprehensively evaluated by using entropy weight technique for order preference by similarity to an ideal solution （TOPSIS）. The bivariate correlation analysis of E. japonica leaves was conducted by SPSS 22.0 software in terms of weight， comprehensive evaluation value， the content of alcohol-soluble extract， the contents of total flavonoids， total triterpene acids and five triterpenic acids. RESULTS The contents of alcohol-soluble extract in 30 batches of E. japonica leaves were （24.56±0.08）%-（34.85±0.13）%； the contents of total flavonoids were （4.69±0.11）-（14.23±0.27） mg/g； the contents of total triterpene acid were （27.58±0.59）- （63.95±1.27） mg/g； the contents of euscaphic acid， crataegolic acid， corosolic acid， oleanolic acid and ursolic acid were （0.728± 0.011）-（6.064±0.063）， （0.526±0.013）-（3.245±0.022）， （1.222±0.025）-（8.807±0.094）， （0.856±0.021）-（2.931±0.075）， （4.704±0.087）-（11.806±0.283） mg/g， respectively. The analysis result of entropy weight TOPSIS method showed that the top three samples with comprehensive evaluation values （No.Kjcx-5） were S14 （Huotian Town， Yunxiao County， Zhangzhou，Fujian）， S19 （Qinnan District， Qinzhou， Guangxi） and S29 （Guoyang County， Bozhou， Anhui）. Comprehensive evaluation 0596-2559522。E-mail：jxrcwxp@163.com of E. japonica leaves was positively correlated with the contents of five kinds of triterpenic acids， such as euscaphic acid， crataegolic acid， corosolic acid， oleanolic acid and ursolic acid （P＜0.01）. The weight of E. japonica leaves was positively correlated with the comprehensive evaluation value （P＜0.01）. CONCLUSIONS The qualities of E. japonica leaves from different producing areas are very different. Among them， the qualities of E. japonica leaves from Huotian Town， Yunxiao County， Zhangzhou of Fujian， Qinzhou Qinnan District of Guangxi， and Bozhou Guoyang County of Anhui are relatively better. The weight of E. japonica leaves is positively correlated with their quality.

Objective:To explore the short-term efficacy of small dose omalizumab in refractory sinusitis with eosinophilia after extended sinus surgery.Methods:A total of 24 patients who met the diagnostic criteria for eosinophilic chronic rhinosinusitis and remained poorly controlled after multiple surgical treatments were included in this study. These patients were admitted to Hunan People′s Hospital between January 2020 and June 2022, and comprised 13 males and 11 females with an average age of (46.43±13.74) years. The patients were randomly divided into experimental group (12 cases) and control group (12 cases), both of which underwent extended sinus opening surgery. The experimental group received a small dose of omalizumab (150 mg/month) for 4 months, while no omalizumab was applied in the control group. All patients were followed up monthly, subjective and objective symptom scores were collected and compared between groups, which included visual analogue scale (VAS) score, sino-nasal outcome test (SNOT)-22 score, Lund-Mackay score, and Lund-Kennedy score. Statistical analysis was performed using SPSS 24.0 software.Results:The baseline was set at 1 month after surgery. There was no significant difference in baseline clinical characteristics between the two groups. After 4 months of treatment with omalizumab, the experimental group showed significant improvements in VAS scores for nasal obstruction, rhinorrhea, hyposmia, SNOT‐22 score, and Lund-Kennedy score (3.11±1.05 vs 6.44±1.13, 2.00±0.87 vs 6.55±1.33, 2.22±0.67 vs 7.00±1.22, 4.44±0.88 vs 15.22±1.20, 1.67±1.00 vs 7.44±0.88, respectively, all P<0.001). Compared to the control group at 4 months after baseline, the experimental group had significantly lower scores for nasal obstruction, rhinorrhea, hyposmia, SNOT-22, and Lund-Kennedy (3.11±1.05 vs 7.11±1.17, 2.00±0.87 vs 7.67±1.41, 2.22±0.67 vs 7.56±0.88, 4.44±0.88 vs 15.33±2.34, 1.67±1.00 vs 9.00±1.41, respectively, all P<0.001). During a 2-month follow-up period after drug withdrawal, the VAS, SNOT-22, and Lund-Kennedy scores of the experimental group were slightly higher than those before drug withdrawal but showed no significant difference (3.44±1.33 vs 3.11±1.05, 2.22±1.09 vs 2.00±0.86, 2.55±0.88 vs 2.22±0.66, 4.77±0.97 vs 4.44±0.88, 2.11±1.05 vs 1.67±1.00, respectively, all P>0.05). Conclusion:For patients of refractory sinusitis with eosinophilia, a combination of extended sinus surgery and postoperative small dosage of omalizumab can effectively control mucous inflammation, promote mucosal epithelization, and play an important role in the critical early stage of disease recovery.

Organoid models are used to study kidney physiology, such as the assessment of nephrotoxicity and underlying disease processes. Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies, but there is a need to accelerate basic and translational research in the field. Here, we developed an automated continuous imaging setup with the "read-on-ski" law of control to maximize temporal resolution with minimum culture plate vibration. High-accuracy performance was achieved: organoid screening and imaging were performed at a spatial resolution of 1.1 μm for the entire multi-well plate under 3 min. We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system. The acquired images were processed via machine learning-based classification and segmentation algorithms, and the toxicity in kidney organoids was determined with 95% accuracy. The results obtained by the automated "read-on-ski" imaging device, combined with label-free and non-invasive algorithms for detection, were verified using conventional biological procedures. Taking advantage of the close-to-in vivo-kidney organoid model, this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.
Humans ; Kidney ; Organoids ; Pluripotent Stem Cells

ObjectiveTo investigate the potential mechanism of serum N-glycan alterations in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) by measuring serum N-glycan profile and comparing glycosyltransferase gene expression between HCC tissue and adjacent tissue. MethodsThe samples of HCC tissue, adjacent tissue, and normal liver tissue were collected from 34 patients with HBV-related HCC who were admitted to Chinese PLA General Hospital, and serum samples were also collected. Among these 34 patients, 8 were randomly selected and their serum samples were established as HCC experimental group, and the serum samples of 20 healthy adults were established as control group. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis was used to analyze serum N-glycan profile in the HCC experimental group and the control group. Quantitative real-time PCR was used to measure the mRNA expression of 8 glycosyltransferase genes (FUT3, FUT4, FUT6, FUT7, FUT8, Gn-TIII, Gn-TIVa, and Gn-TV) in the HCC tissue and adjacent tissue of 34 patients with HBV-related HCC, and Western blot was used to measure the expression of corresponding proteins. The independent samples t-test was used for comparison of continuous data between two groups. ResultsCompared with the control group, the HCC experimental group had a significant increase in the abundance of N-glycan peak9 (NA3Fb) in serum(t=-2.514,P＜0.05). There were significant differences in the mRNA expression of FUT8, Gn-TIVa, and Gn-TV between HCC tissue and adjacent tissue, and the mRNA and protein expression levels of FUT8 and Gn-TV in HCC tissue were significantly higher than those in adjacent tissue (FUT8 mRNA: 1.50±0.34 vs 0.65±0.11, t=-2.354，P=0.022; Gn-TV mRNA: 3.57±0.64 vs 1.33±016, t=-3.384,P=0001; FUT8 protein: 0.70±0.11 vs 0.083±0.017, t=9.555,P=0.001; Gn-TV protein: 1.33±0.19 vs 0.60±0.15, t=5.097,P=0.007). The mRNA expression level of Gn-TIVa in HCC tissue was significantly higher than that in adjacent tissue (2.90±0.47 vs 1.68±0.19, t=-2.403,P=0.019), but there was no significant difference in the protein expression level of Gn-TIVa between HCC tissue and adjacent tissue (052±0.24 vs 0.24±0.11,t=1.833, P=0.141). The changes of glycosyltransferase gene expression in HCC tissue were consistent with the alteration of serum N-glycan profile. ConclusionSerum N-glycan alterations in patients with HBV-related HCC may be closely associated with the upregulated expression of the glycosyltransferase genes FUT8, Gn-TIVa, and Gn-TV in HCC tissue.

Objective:To explore the effects of different approaches for second-trimester multifetal pregnancy reduction on pregnancy outcome in women with dichorionic triamniotic (DCTA) triplet.Methods:A retrospective study was performed on 51 women with DCTA triplet pregnancies who were referred to Guangdong Women and Children Hospital for second-trimester multifetal pregnancy reduction from January 2014 to January 2020. All participants were divided into either preventive group ( n=39) or treatment group ( n=12) according to the indication for multifetal pregnancy reduction, and they were further allocated to three subgroups based on different reduction methods, which were reduction to dichorionic twin by radiofrequency ablation (RFA) (RFA subgroup), reduction to monochorionic singleton (KCl-singleton subgroup) or monochorionic twin (KCl-twin subgroup) by cardiac injection of potassium chloride. Pregnancy loss rate, neonatal birth weight, gestational age at delivery, incidence of intrauterine death, and neonatal death were compared and analyzed between different groups using t-test, analysis of variance, Chi-square test, Fisher's exact test and Bonferroni correction. Results:(1) The mean gestational week at operation in the treatment group was significantly later than that in the preventive group [(18.5±3.1) vs (15.0±2.3) weeks, t=-4.209, P<0.001]. In the preventive group, the mean gestational week at operation in the RFA subgroup was later than the KCl-singleton and KCl-twin subgroup[(17.2±1.6) vs (13.8±1.5) and (12.7±1.0) weeks, t=6.630 and 3.875, respectively, both P<0.05]. (2) The postoperative pregnancy loss rate in the preventive group was decreased compared with the treatment group [10.3%(4/39) vs 5/12, Fisher's exact test, P<0.05], and the live birth ratio was increased [ 85.7%(48/56) vs 10/18, χ2=5.640, P=0.018]. No live birth infants with birth weight <1 500 g was reported in the KCl-singleton subgroup in preventive group, and the statistical significance was observed in the intra-group differences ( P<0.05) rather than the pairwise comparison differences in the preventive group. For the proportion of live births, there was a statistically significant difference in the intra-group comparison in the treatment group, which was higher in the RFA subgroup than that in the KCl-twin subgroup (6/6 vs 1/6, P=0.045). No significant difference was revealed among pregnancy loss rate, gestational weeks at delivery, the mean birth weight, premature delivery <32 gestational weeks, and full-term birth rate among three different approaches within the two groups. (3) No monochorionic twin complications or perinatal death occurred in any RFA or KCl-singleton subgroups in the two groups. In the KCl-twin subgroups including five cases with ten fetuses, including three live birth, four miscarriage, three intrauterine death occured, while no neonatal death was reported. One case with selective fetal uterine growth restriction in the preventive group delivered two live births, and one case with twin-to-twin transfusion syndrome in the treatment group had intrauterine death in one fetus and one survival neonate. Conclusions:The pregnancy outcome of multifetal pregnancy reduction to dichorionic diamniotic twins by RFA or reduction to singleton by cardiac injection of potassium chloride are comparative in women with DCTA triplet, regardless of whether it is a preventive or therapeutic reduction.

Objective:To study the changes of proliferation and oxidation indexes of Cochlear hair cell line (HEI-OC1 cells) exposed to ethylbenzene.Methods:From July to December 2019, 11 groups with ethylbenzene concentrations of 0, 30, 60, 90, 300, 600, 900 μmol/L and 3, 6, 9, 10 mmol/L, were used to determine the proliferation activity of HEI-OC1 cells exposed to ethylbenzene for 24 hours, and the cells were treated with 0, 1, 2, 4, 8, 16, 32, 64 mmol/L ethylbenzene for 24 hours, then the 50% inhibitory concentration of ethylbenzene was calculated. After HEI-OC1 cells were exposed to 0, 6, 9 and 12 mmol/L ethylbenzene for 24 hours, the malondialdehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were measured.Results:Compared with 0 mmol/L concentration group, the survival rate of HEI-OC1 cells at 6, 9, 12 mmol/L concentration was significantly decreased ( P<0.01) . The 50% inhibitory concentration of ethylbenzene on HEI-OC1 cells was 12.86 mmol/L ( R2=99.05) . There were significant differences in SOD and GSH-Px activity in HEI-OC1 cells treated with ethylbenzene at different concentrations (0, 6, 9, 12 mmol/L) for 24 hours ( F=65.11, 6.48, 22.85, P<0.05) . Compared with 0 mmol/L concentration group, the MDA content of HEI-OC1 cells was significantly increased in 9 and 12 mmol/L concentration groups, the SOD activity was significantly decreased in 12 mmol/L concentration group, and the GSH-Px activity was significantly decreased in 6 and 12 mmol/L concentration groups. Conclusion:Ethylbenzene can inhibit the proliferation of HEI-OC1 cells and cause oxidative damage.