Objective Studies on the expression and location of zinc finger protein A20 (A20) and connective tissue growth factor (CTGF) in liver tissues of patients with chronic hepatitis B were conducted, and the relationship between them and liver fibrosis was determined by FibroScan. Methods Studies on A20 and CTGF in liver tissues of 160 patients with chronic hepatitis B were conducted in accordance with the stage of pathological fibrosis and inflammation of the liver, and quantitative immunohistochemistry test was conducted, and statistical analysis was conducted by FibroScan. Results The expressions of A20 and CTGF in liver tissues increased with the aggravation of liver pathological fibrosis and inflammation, and there were significant differences between each stage and the control group (P<0.05), and there were significant differences between adjacent groups (P<0.05). Studies have shown that FibroScan increases along with pathological fibrosis and inflammation in the liver. There are significant differences between the stage and the control group (P<0.05), and no significant differences between the adjacent groups (P>0.05). There was positive correlation between liver A20 and CTGF, r=0.796 (P<0.05). Conclusions In patients with chronic hepatitis B, A20, CTGF and FibroScan are positively correlated with the degree of liver fibrosis, and A20 and CTGF are also positively correlated with the degree of liver inflammation, which can be used as indicators to evaluate the degree of liver inflammation and fibrosis, and further guide the anti-inflammatory and anti-fibrosis treatment of patients.

Objective:To explore the value of radiomics and deep learning in predicting the efficacy of initial transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC).Methods:This was a cohort study. The imaging and clinical information of HCC patients treated with TACE in the Second Affiliated Hospital of Harbin Medical University from January 2015 to January 2021 were collected retrospectively. A total of 265 patients were divided into response group (175 cases) and non-response group (90 cases) according to the modified solid tumor efficacy evaluation criteria (mRECIST) 1 to 2 months after initial TACE. According to the proportion of 8∶2, the patients were randomly divided into training group (212 cases, 140 responders and 72 non-responders) and test set (53 cases, 35 responders and 18 non-responders). Univariate and multivariate logistic regression was used to screen clinical variables and construct a clinical model. The radiomics features were extracted from the preoperative CT images, and radiomics model was constructed after feature dimensionality reduction. Using the deep learning method, three residual network (ResNet) models (ResNet18, ResNet50 and ResNet101) were established, and their effectiveness was compared and integrated to build a deep learning model with best performance. Univariate and multivariate logistic regression was used to combine pairwise three models to establish the combined model. The receiver operating characteristic curve was used to evaluate the performance of the model to distinguish between TACE response and non-response groups.Results:In the test set, the area under the curve (AUC) of the clinical model and the radiomics model in the differentiation between response and non-response after TACE were 0.730 (95% CI 0.569-0.891) and 0.775 (95% CI 0.642-0.907). The AUC of ResNet18, ResNet50 and ResNet101 were 0.719, 0.748 and 0.533, respectively. The AUC for deep learning model obtained by integrating ResNet18 and ResNet50 was 0.806 (95% CI 0.665-0.946). After pairwise fusion, the combined deep learning-radiomics model showed the highest performance, with an AUC of 0.843 (95% CI 0.730-0.956), which was better than those of the deep learning-clinical model (AUC of 0.838, 95% CI 0.719-0.957) and the radiomics-clinical model (AUC of 0.786, 95% CI 0.648-0.898). Conclusions:The combined model of radiomics and deep learning has high performance in predicting the curative effect of TACE in patients with HCC before operation.

Objective:To explore any effect of repeated transcranial magnetic stimulation (rTMS) on the upper limb motor function and cerebral cortex activation of stroke survivors.Methods:Sixty stroke survivors were randomly divided into an intervention group and a control group, each of 30. In addition to routine rehabilitation training (including drug therapy, comprehensive hemiplegic limb training and physical factor therapy), the intervention group received 15 minutes of rTMS daily, five days a week for 4 weeks while the control group was given false rTMS. Upper limb motor function was evaluated before and after the treatment using the Fugl Meyer upper limb motor function rating scale (FMA-UE). Functional near-infrared spectroscopy was used to detect and compare the activation (β values) of the prefrontal cortex, the motor cortex and the primary somatosensory cortex in the 2 groups. The correlation between the FMA-UE scores and the β values was quantified.Results:①There was no significant difference in the average FMA-UE scores between the two groups before the treatment. Afterward, though both groups′ average scores had increased significantly, there was significantly greater improvement in the treatment group. ②There was also no significant difference in average β value between the two groups before the experiment, but afterward the average βs of channels 27 and 13 in the intervention group were significantly higher than in the control group. Moreover, in patients with lesion in the left brain, the β-values of CH27 and CH13 were also significantly higher than the control group ( P<0.05). ③The FMA-UE scores of the intervention group were moderately correlated with the CH27 and CH13 β values, but those of the control group were only weakly correlated with the β values of CH27. Conclusion:Transcranial magnetic stimulation activates a lesioned left brain region, improving upper limb motor function. The improvement is correlated with the activation of the left prefrontal cortex and the left primary somatosensory cortex.

Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.

OBJECTIVE To observe the clinical efficacy of Erdong Xiaoke Formula in the treatment of type 2 diabetes dry eyes with yin deficiency and heat excess syndrome and its effect on serum interleukin 17(IL-17)and interleukin 1β(IL-1β),and to ex-plore the therapeutic mechanism of Erdong Xiaoke Formula.METHODS 110 cases of type 2 diabetes patients with dry eyes of yin deficiency and heat excess syndrome from Jiangsu Province Hospital of Chinese Medicine were enrolled and randomly divided into a treatment group and a control group,55 cases each.5 cases dropped out of the treatment group and 4 cases dropped out of the control group.The control group was given a basic hypoglycemic regimen combined with topical sodium hyaluronate eye drops.The treatment group was given Erdong Xiaoke Formula in addition to the treatment in the control group.The treatment course for both groups was 4 weeks.Changes in TCM syndrome scores of the two groups of patients were observed before and after treatment,and the clinical effica-cy of TCM was evaluated.Blood glucose and pancreatic islet function-related indicators[fasting blood glucose(FBG),fasting insulin(FINS),fasting C-peptide(FCP),postprandial 2 h blood glucose(PBG),insulin secretion function index(HOMA-β),insulin re-sistance index(HOMA)-IR)],ocular surface indicators[tear break up time(BUT),corneal sodium fluorescein staining(FL),Schirmer Ⅰ test(SⅠT)],and inflammation-related indicators(IL-17,IL-1β)were detected.The occurrence of adverse reactions in the two groups of patients was observed during the treatment period.RESULTS After treatment,the total scores of TCM syn-dromes in both groups were significantly reduced(P＜0.01),the treatment group was better than the control group(P＜0.01),and the total effective rate of TCM syndromes in the treatment group was higher than that of the control group(P＜0.01);SⅠT and BUT of pa-tients in both groups increased significantly(P＜0.01),and the treatment group was better than the control group(P＜0.05,P＜0.01);the FL of patients in both groups significantly reduced(P＜0.01),and there was no significant difference between the groups;the ser-um IL-17 and IL-1β of the patients in the treatment group decreased significantly(P＜0.01),which was significantly better than that of the control group(P＜0.05).There were no significant changes in blood glucose and pancreatic islet function-related indicators in the two groups before and after treatment(P＞0.05).During the treatment,no obvious adverse reactions were observed in the two groups.CONCLUSION Erdong Xiaoke Formula can improve SⅠT,BUT,FL and eye symptoms in patients with diabetic dry eye,effectively treat diabetic dry eye,and reduce ocular surface inflammation.Its mechanism may be related to reducing serum inflammato-ry factors IL-17 and IL-1β.

Objective To investigate the mechanism of RNF7 for regulating myeloid-derived suppressor cells(MDSCs)in non-small cell lung cancer(NSCLC).Methods TIMER2.0 database and immunohistochemistry were used to analyze RNF7 expression level and its correlation with immune cell infiltration in non-small cell lung cancer.The impact of RNF7 expression levels on prognosis of lung cancer patients was analyzed using Kaplan-Meier survival analysis.CMT-167 cells with RNF7 overexpression or knockdown were inoculated subcutaneously in C57BL/6 mice,and the mice in RNF7 knockdown group were treated with anti-PD-1 or IgG isotype control 7 days after the inoculation.The tumor tissues were harvested after 30 days for tumor volume measurement,detection of S100A8+A9 and Gr-1 expressions with immunohistochemistry,and analysis of MDSC infiltration.Gene set enrichment analysis(GSEA)was performed to identify the potential pathways regulated by RNF7 in NSCLC.Western blotting and luciferase assays were used to assess the impact of RNF7 on the NF-κB signaling pathway.ELISA and RT-qPCR were used to measure chemokine(C-X-C motif)ligand 1(CXCL1)expression.Results RNF7 expression was significantly upregulated in NSCLC,and high RNF7 expression levels were associated with poor prognosis of the patients(P<0.001).TIMER2.0 analysis revealed a positive correlation between RNF7 expression and MDSC infiltration(P<0.001).GSEA suggested that RNF7 was enriched in the NF-κB signaling pathway.In NSCLC cells,RNF7 knockdown significantly inhibited NF-κB activation and reduced CXCL1 expression.In the tumor-bearing mice,RNF7 overexpression significantly increased MDSC infiltration in the tumor tissue,and RNF7 knockdown obviously reduced MDSC infiltration and enhanced the efficacy of anti-PD-1 therapy.Conclusion High expression of RNF7 in NSCLC cells promotes CXCL1 expression by activating the NF-κB signaling pathway,thus leading to the chemotactic recruitment of MDSCs,which contributes to tumor resistance to anti-PD-1 therapy.

Objective:To explore the genetic etiology and clinical outcome of a child with co-morbid progressive IgA nephropathy and COQ8B-associated glomerulopathy. Methods:A child who was admitted to Peking University First Hospital on March 2, 2021 was selected as the study subject. Genomic DNA was extracted from peripheral blood samples from the child and his parents and sister. Whole exome sequencing was carried out, and candidate variant was verified by Sanger sequencing. This study was approved by Medical Ethics Committee of the Peking University First Hospital (Ethics No. 2016[1029]).Results:The child, a 7-year-old boy who had developed proteinuria 8 months before, was diagnosed with IgA nephropathy (M1E1S1T1C1). With steroid, cyclophosphamide, cyclosporine and angiotensin-converting enzyme inhibitor therapy, partial remission of proteinuria was achieved. However, his serum creatinine level had increased from 53.8 mol/L at the onset of disease to 86.7 mol/L after 3.9 years, along with massive proteinuria. Kidney biopsy still indicated IgA nephropathy (M0E0S1T0C0). The child was found to harbor a homozygous c. 737G>A (p.Ser246Asn) missense variant of the COQ8B gene, for which his parents and sister were heterozygous carriers. The variant was predicted to be pathogenic (PS1+ PM2_Supporting+ PM3+ PP3+ PP4) based on the guidelines from the American College of Medical Genetics and Genomics. The child was treated with high-dose coenzyme Q10 in combination with steroid and/or mycophenolate mofetil, though his serum creatinine level still increased to 286 mol/L after 7.3 years, which conformed to a chronic kidney disorder with glomerular filtration rate category of G3b. Conclusion:The homozygous c.737G>A missense variants of the COQ8B gene probably underlay the progressive kidney dysfunction in this child. For children with IgA nephropathy presenting with atypical clinical manifestations, unsatisfactory therapeutic effect, and/or early onset of kidney function decline, coexistence of other diseases should be suspected.

The killing effect of radiation therapy on healthy cells has led to the creation of targeted radionuclide therapy,which effectively reduces the damage to surrounding normal cells.At present,alpha(α)and beta(β)radionuclides are the research hotspots of targeted therapy.Numerous preclinical and clinical studies have shown that radiation therapy not only has local anti-tumor effects,but also exerts systemic anti-tumor effects by triggering the body's immune response.This paper describes in detail the characteristics and clinical applications of commonly used radionuclides,and discusses the mechanism of radiation-triggered body immune response as well as the related research on the combined use of radiation therapy,targeted radionuclide therapy and immunotherapy.

Objective To investigate the association between microRNA(miR)-23a,miR-150,and miR-150-5p levels and diabetic osteoporosis(DOP)through correlation analysis.Methods A total of 200 patients with diabetes who visited the hospital between March 2020 and March 2022 were selected and followed up for a duration of 2 years.Patients who developed DOP and those who did not were respectively assigned to the DOP group and the non-DOP group.The general information and serum levels of miR-23a,miR-150,and miR-150-5p between these two groups were compared.Pearson correlation analysis was conducted to examine the relationship between serum levels of miR-23a,miR-150,and miR-150-5p,as well as bone density.Logistic regression model was employed to identify factors influencing DOP.Results The incidence rate of DOP was 28.89%.The serum levels of miR-23a,miR-150,and miR-150-5p were significantly elevated in the DOP group compared to the non-DOP group(P＜0.05).Pearson correlation analysis revealed a negative association between the serum levels of miR-23a,miR-150,and miR-150-5p in diabetic patients and bone density(P＜0.05).Logistic regression model analysis demonstrated that age,body mass index(BMI),as well as miR-23a,miR-150,and miR-150-5p were influential factors contributing to the development of DOP(P＜0.05).Conclusion The levels of serum miR-23a,miR-150,and miR-150-5p are elevated in patients with DOP,and all three exhibit a negative correlation with bone density.Furthermore,these factors collectively contribute to the development of DOP.

Objective To investigate the mechanism of RNF7 for regulating myeloid-derived suppressor cells(MDSCs)in non-small cell lung cancer(NSCLC).Methods TIMER2.0 database and immunohistochemistry were used to analyze RNF7 expression level and its correlation with immune cell infiltration in non-small cell lung cancer.The impact of RNF7 expression levels on prognosis of lung cancer patients was analyzed using Kaplan-Meier survival analysis.CMT-167 cells with RNF7 overexpression or knockdown were inoculated subcutaneously in C57BL/6 mice,and the mice in RNF7 knockdown group were treated with anti-PD-1 or IgG isotype control 7 days after the inoculation.The tumor tissues were harvested after 30 days for tumor volume measurement,detection of S100A8+A9 and Gr-1 expressions with immunohistochemistry,and analysis of MDSC infiltration.Gene set enrichment analysis(GSEA)was performed to identify the potential pathways regulated by RNF7 in NSCLC.Western blotting and luciferase assays were used to assess the impact of RNF7 on the NF-κB signaling pathway.ELISA and RT-qPCR were used to measure chemokine(C-X-C motif)ligand 1(CXCL1)expression.Results RNF7 expression was significantly upregulated in NSCLC,and high RNF7 expression levels were associated with poor prognosis of the patients(P<0.001).TIMER2.0 analysis revealed a positive correlation between RNF7 expression and MDSC infiltration(P<0.001).GSEA suggested that RNF7 was enriched in the NF-κB signaling pathway.In NSCLC cells,RNF7 knockdown significantly inhibited NF-κB activation and reduced CXCL1 expression.In the tumor-bearing mice,RNF7 overexpression significantly increased MDSC infiltration in the tumor tissue,and RNF7 knockdown obviously reduced MDSC infiltration and enhanced the efficacy of anti-PD-1 therapy.Conclusion High expression of RNF7 in NSCLC cells promotes CXCL1 expression by activating the NF-κB signaling pathway,thus leading to the chemotactic recruitment of MDSCs,which contributes to tumor resistance to anti-PD-1 therapy.