Although cancer immunotherapy has made great strides in the clinic, it is still hindered by the tumor immunosuppressive microenvironment (TIME). The stimulator of interferon genes (STING) pathway which can modulate TIME effectively has emerged as a promising therapeutic recently. However, the delivery of most STING agonists, specifically cyclic dinucleotides (CDNs), is performed intratumorally due to their insufficient pharmacological properties, such as weak permeability across cell membranes and vulnerability to nuclease degradation. To expand the clinical applicability of CDNs, a novel pH-sensitive polycationic polymer-modified lipid nanoparticle (LNP-B) system was developed for intravenous delivery of CDNs. LNP-B significantly extended the circulation of CDNs and enhanced the accumulation of CDNs within the tumor, spleen, and tumor-draining lymph nodes compared with free CDNs thereby triggering the STING pathway of dendritic cells and repolarizing pro-tumor macrophages. These events subsequently gave rise to potent anti-tumor immune reactions and substantial inhibition of tumors in CT26 colon cancer-bearing mouse models. In addition, due to the acid-sensitive property of the polycationic polymer, the delivery system of LNP-B was more biocompatible and safer compared with lipid nanoparticles formulated with an indissociable cationic DOTAP (LNP-D). These findings suggest that LNP-B has great potential in the intravenous delivery of CDNs for tumor immunotherapy.

Cold tumors have a poor response to tumor immunotherapy due to low immune cell infiltration and the ability to evade immune attacks. Converting cold tumors into hot tumors can enhance the clinical effectiveness of anti-tumor immunotherapy. High-intensity focused ultrasound (HIFU) as a non-invasive treatment can damage tumors through mechanical effects, but there is a lack of research on its cytotoxic mechanisms at the cellular level and its role in inducing anti-immune responses. In this study, the role of HIFU in triggering tumor ferroptosis by disrupting the GSH/GSSG balance through mechanochemical action and the associated anti-tumor immune priming effect were investigated. The use of a nano-enhancer loaded with PFOB combined with HIFU could enhance ferroptosis in triple-negative breast cancer at a specific stage of tumor growth (UTGR = 0) while promoting the conversion of a cold tumor into a hot tumor, thereby improving the immune response. Overall, this provides valuable guidance for the clinical application of HIFU in tumor immunotherapy.

Objective:The sensory processing measure-preschool scale(SPM-P)was transformed into Chinese,and its re-liability and validity were tested in preschool children. Method:According to Brislin's translation model,the SPM-P source scale was translated into Chinese and the Chinese version of SPM-P scale was formed.From September 2021 to December 2021,395 preschool children were investigated by cluster stratified random sampling method to test the reliability and validity of the scale. Result:The Chinese version of the SPM-P scale includes 8 dimensions of social participation,vision,hear-ing,touch,body awareness,balance and movement,planning and conception,and overall sensory system,with a total of 75 items.Cronbach alpha coefficient for the overall scale was 0.899,the split half reliability coefficient was 0.700;the test-retest reliability coefficient was 0.899.The item content validity(I-CVI)value was 0.920,the average content validity(S-CVI/Ave)value was 0.984,and the content validity was good;the results of confirmatory factor analysis showed that the construct validity was good;The results of confirma-tory factor analysis showed that the construct validity was good(x2/df=2.41,CFI=0.992,TLI=0.960,RMSEA=0.060,SRMR=0.046);The Chinese version of SPM-P scale was negatively correlated with the corresponding evaluation dimensions of the screening questionnaire for children's sensory integration disorder,and the correla-tion coefficient was between-0.585 and-0.399,with good criterion validity. Conclusion:The Chinese version of SPM-P scale has good reliability and validity and can be used for stan-dardized evaluation of sensory integration ability of preschool children in China.

Purpose To investigate the expression of chiti-nase-3-like protein 1(CHI3L1)in esophageal squamous cell carcinoma(ESCC)and its correlations between CHI3L1 and molecules associated with the Wnt/β-catenin signaling pathway.Methods Paraffin-embedded tissue specimens from 119 ESCC patients and 60 normal esophageal tissues were retrospectively selected.Immunohistochemistry was used to detect the protein expression levels of CHI3L1 and Wnt/β-catenin signaling path-way related molecules in the above tissues,and the ectopic pro-tein expression levels of CHI3L1,TCF4,Cyclin D1,APC andβ-catenin in ESCC patients'tissues and normal esophageal tis-sues were compared.The survival of ESCC patients was followed up.Results Compared with normal esophageal tissue,the pos-itive rates of CHI3L1,TCF4,Cyclin D1 and β-catenin in ESCC were higher(63.0％,69.7％,68.1％and 67.2％,respec-tively),and the positive rates of APC were lower(41.2％).The expression levels of CHI3L1,TCF4,Cyclin D1 and β-cate-nin were positively correlated with each other(P＜0.05),but negatively correlated with APC(P＜0.05).CHI3L1 protein was associated with ESCC tumor volume,invasion depth,lymph node metastasis,and pTNM stage(P＜0.05).High expression of CHI3L1,TCF4,Cyclin D1 or abnormal high expression of β-catenin protein was associated with poor prognosis in ESCC pa-tients,suggesting shortened survival time(Log-rank=10.121,Log-rank=34.896,Log-rank=34.923,Log-rank=4.082,all P＜0.05).Conclusion In ESCC,the high expression of CHI3L1 is closely related to the activation of Wnt/β-catenin sig-naling pathway,suggesting that CHI3L1 may participate in the occurrence and development of ESCC through abnormal activa-tion of Wnt/β-catenin signaling pathway.

Purpose To investigate the expression of chiti-nase-3-like protein 1(CHI3L1)in esophageal squamous cell carcinoma(ESCC)and its correlations between CHI3L1 and molecules associated with the Wnt/β-catenin signaling pathway.Methods Paraffin-embedded tissue specimens from 119 ESCC patients and 60 normal esophageal tissues were retrospectively selected.Immunohistochemistry was used to detect the protein expression levels of CHI3L1 and Wnt/β-catenin signaling path-way related molecules in the above tissues,and the ectopic pro-tein expression levels of CHI3L1,TCF4,Cyclin D1,APC andβ-catenin in ESCC patients'tissues and normal esophageal tis-sues were compared.The survival of ESCC patients was followed up.Results Compared with normal esophageal tissue,the pos-itive rates of CHI3L1,TCF4,Cyclin D1 and β-catenin in ESCC were higher(63.0％,69.7％,68.1％and 67.2％,respec-tively),and the positive rates of APC were lower(41.2％).The expression levels of CHI3L1,TCF4,Cyclin D1 and β-cate-nin were positively correlated with each other(P＜0.05),but negatively correlated with APC(P＜0.05).CHI3L1 protein was associated with ESCC tumor volume,invasion depth,lymph node metastasis,and pTNM stage(P＜0.05).High expression of CHI3L1,TCF4,Cyclin D1 or abnormal high expression of β-catenin protein was associated with poor prognosis in ESCC pa-tients,suggesting shortened survival time(Log-rank=10.121,Log-rank=34.896,Log-rank=34.923,Log-rank=4.082,all P＜0.05).Conclusion In ESCC,the high expression of CHI3L1 is closely related to the activation of Wnt/β-catenin sig-naling pathway,suggesting that CHI3L1 may participate in the occurrence and development of ESCC through abnormal activa-tion of Wnt/β-catenin signaling pathway.

OBJECTIVETo investigate the effects of adenoviral vector-mediated over-expression of Wnt3 on the apoptosis of hepatic progenitor cells in vitro.

METHODSHepatic progenitor cells transfected with Ad-GFP-Wnt3 vector or the control vector Ad-GFP were examined for cell apoptosis under fluorescence microscopy with Hoechst 33342 staining, and the proportion of apoptotic cells were determined by flow cytometric analysis with Annexin-PE/7-ADD staining. The mRNA and protein expressions of Bax, Bcl-2 and Bcl-xl in the cells were detected by real-time PCR and Western blotting, respectively.

RESULTSReal-time PCR and Western blotting showed a high expression of Wnt3 in Ad-GFP-Wnt3-transfected hepatic progenitor cells, which exhibited significantly decreased cell apoptosis as compared with the control group. The expressions of Bcl-2 and Bcl-xl mRNA and proteins increased significantly while Bax expression decreased obviously in Ad-GFP-Wnt3-transfected cells (P<0.05).

CONCLUSIONSAdenoviral vector-mediated over-expression of Wnt3 can suppress apoptosis of hepatic progenitor cells possibly through the Bcl-2 pathway.

Apoptosis ; Cells, Cultured ; Genetic Vectors ; Hepatocytes ; cytology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stem Cells ; cytology ; Transfection ; Wnt3 Protein ; metabolism ; bcl-X Protein ; metabolism

Objective To investigate the effects of adenoviral vector-mediated over-expression of Wnt3 on the apoptosis of hepatic progenitor cells in vitro. Methods Hepatic progenitor cells transfected with Ad-GFP-Wnt3 vector or the control vector Ad-GFP were examined for cell apoptosis under fluorescence microscopy with Hoechst 33342 staining, and the proportion of apoptotic cells were determined by flow cytometric analysis with Annexin-PE/7-ADD staining. The mRNA and protein expressions of Bax, Bcl-2 and Bcl-xl in the cells were detected by real-time PCR and Western blotting, respectively. Results Real-time PCR and Western blotting showed a high expression of Wnt3 in Ad-GFP-Wnt3-transfected hepatic progenitor cells, which exhibited significantly decreased cell apoptosis as compared with the control group. The expressions of Bcl-2 and Bcl-xl mRNA and proteins increased significantly while Bax expression decreased obviously in Ad-GFP-Wnt3-transfected cells (P<0.05). Conclusion Adenoviral vector-mediated over-expression of Wnt3 can suppress apoptosis of hepatic progenitor cells possibly through the Bcl-2 pathway.

Objective To investigate the effects of adenoviral vector-mediated over-expression of Wnt3 on the apoptosis of hepatic progenitor cells in vitro. Methods Hepatic progenitor cells transfected with Ad-GFP-Wnt3 vector or the control vector Ad-GFP were examined for cell apoptosis under fluorescence microscopy with Hoechst 33342 staining, and the proportion of apoptotic cells were determined by flow cytometric analysis with Annexin-PE/7-ADD staining. The mRNA and protein expressions of Bax, Bcl-2 and Bcl-xl in the cells were detected by real-time PCR and Western blotting, respectively. Results Real-time PCR and Western blotting showed a high expression of Wnt3 in Ad-GFP-Wnt3-transfected hepatic progenitor cells, which exhibited significantly decreased cell apoptosis as compared with the control group. The expressions of Bcl-2 and Bcl-xl mRNA and proteins increased significantly while Bax expression decreased obviously in Ad-GFP-Wnt3-transfected cells (P<0.05). Conclusion Adenoviral vector-mediated over-expression of Wnt3 can suppress apoptosis of hepatic progenitor cells possibly through the Bcl-2 pathway.

OBJECTIVETo develop the system for the exclusive control substance of plant drug (CSPD) in traditional Chinese herbal medicines (TCHM), this paper investigated the (CSPD) of Radix Stephaniae Tetrandrae as well as its proton nuclear magnetic resonance (1H-NMR) and high performance liquid chromatography (HPLC) analytical methods for the purpose of original identification and quality control of the crude drug.

METHODThe CSPDs and their 1H-NMR and HPLC profiles of Radix Stephaniae Tetrandrae were obtained by standardized procedure. Chemical components were isolated from the CSPD by silica gel column chromatography. The assignments of the characteristic signals in 1H-NMR and HPLC profiles were achieved on the basis of elucidation of the isolates structures.

RESULTFor nine samples from the different sources in this paper, the 1H-NMR and HPLC profiles from eight sources had wonderful reproducibility and characteristics, and the other gave differences compared with the eight samples in the signal strength of the main components. Furthermore, seven compounds were isolated from CSPD and their chemical structures were authenticated by spectral analysis as tetrandrine, fangchinoline, tetrandrine-2'-N-beta-oxide, tetrandrine-2'-N-alpha-oxide, dicentrine, dicentrinone, and adenine, respectively. The 1H-NMR and HPLC profiles of the CSPD of Radix Stephaniae Tetrandrae showed mainly the characteristic signals of the bisbenzylisoquinoline alkaloids isolated in this work.

CONCLUSIONThe 1H-NMR and HPLC profiles of the CSPD of Radix Stephaniae Tetrandrae exhibit the structures and total composition of the main active constituents in it, and can be used for its original identification and quality evaluation.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Quality Control ; Stephania tetrandra ; chemistry