0.05) . No significant differences were observed between the expansion folds of hNSC derived from various fetal brain samples when cultured under the same conditions. LIF played great roles on cell proliferation,in LIF + groups, hNSC cell number increased ranging from 4 000-8 400 folds, no cell differentiation occurred; and in LIF" groups,only 43 to 96 folds.The differentiation phenomenons were watched when cultured more than two months. In the course of cell culturing, observed that the effects of LIF on hNSC expansion were obviously demonstrated 50-60 days after inoculation.The number of neurons and astrocytes differentiated from the cultured hNSC were respectively identified by means of Immuno-cytochemical fluorescent assay, and the percentages of neurons(as a proportion of neuron and astrocyte number) were calculated,which were ranging from 12% to 83% in LIF+ cultures, significantly higher than 8% to 23% in LIF- ones(P

Objective To detect the expression levels of vascular endothelial growth factor D ( VEGF-D) , D2-40 and MMP-7 in papillary thyroid carcinoma ( PTC) and to explore their relationship with micro lymphat-ic density, distribution,and cervical lymph node metastasis .Methods SP immunohistochemistry was used to study the expression of VEGF-D, D2-40 and MMP-7 in 38 cases of PTC with cervical lymph node metastasis , 45 cases of PTC without cervical lymph node metastasis , and 22 cases of normal thyroid tissues .Micro lymphatic density ( MLD) was counted .Results The expression level of VEGF-D in PTC with cervical lymph node metasta-sis and normal thyroid tissues had significant difference (χ2 =13.074, P=0.000, P<0.0167).The expression level of D2-40 in PTC with and without cervical lymph node metastasis had significant difference ( P<0.05 ) . MMP-7 expression in PTC with and without cervical lymph node metastasis had statistical significance ( 91.67%vs 73.33%, P<0.05).MLD in PTC with and without cervical lymph node metastasis and in normal thyroid tis-sues had statistical difference (11.7 ±3.5 vs 8.9 ±3.1 vs 3.9 ±2.7, P<0.05).Conclusion VEGF-D, D2-40 and MMP-7 are highly expressed in PTC and they are possibly related with cervical lymph node metastasis of PTC.

Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.

Objective To explore the predictive value of the expression of CD44v6 and EGFR on the efficacy of neoadjuvant chemotherapy (NACT) in stageⅡ-Ⅲ cervical cancer. Methods A total of 53 patients with stageⅡ-Ⅲ cervical cancer diagnosed by pathology were selected. All patients received two cycles of paclitaxel+platinum NACT. The pathological tissue samples of cervical tumors before NACT treatment were collected. The expression of CD44v6 and EGFR were detected by the immunohistochemical SP method, and we analyzed their predictive value of NACT in stageⅡ-Ⅲ cervical cancer. Results Among the 53 patients, 38 were in the NACT effective group (CR+PR), and 15 were in the NACT ineffective group (SD+PD). The expression of CD44v6 in the ineffective group was significantly higher than that in the effective group (P < 0.05). The expression of CD44v6 was significantly different in patients with CR, PR, and SD (P < 0.05). The AUC of CD44v6 to NACT effect on stage Ⅱ-Ⅲ cervical cancer was 0.74 (P < 0.05). The patients in the high expression group of CD44v6 had worse efficacy in NACT than those in the low expression group of CD44v6 (P < 0.05). Pearson test showed that CD44v6 and EGFR expression were correlated (R=0.34, P < 0.05). Conclusion High expression of CD44v6 may reduce the efficacy of NACT in stageⅡ-Ⅲ cervical cancer, suggesting that the expression of CD44v6 has a certain predictive value and clinical significance in the efficacy of paclitaxel+platinum NACT on cervical cancer. Moreover, CD44v6 is positively correlated with EGFR expression.

Objective:To study the biological traits and mutations of the influenza A (H3N2) virus in order to produce a vaccine and offer references for controlling and preventing influenza epidemics.Methods:Four strains of the influenza A(H3N2) virus were chosen from the Huai′an surveillance network laboratory. Nucleic acid extraction, library building, and sequencing (CridION x5 MKI Nanopore) were used to produce the whole-genome sequences. Using homologous alignments of whole-genome sequences, phylogenetic tree construction, and amino acid variant screening, bioinformatics analysis was carried out.Results:The nucleotide identity between 8 gene segments ranged from 97.1% to 100.0%. The gene that differed the most from the reference sequences was HA (97.1%-99.9%), and the gene that differed the least was MP (98.6%-99.9%). The HA gene (3.06%) and MP gene (1.43%) were the regions with the greatest and lowest frequencies of nucleotide site change, respectively. The rates of nucleotide change varied significantly between the genes ( χ2=14.293, P=0.046). Four influenza A(H3N2) virus strains′ whole-genome phylogenies from each of the eight gene segments maintained a roughly consistent topological structure. One strain was linked to the 3C.2a1b.1b clade, which was lost at the 142NWT, 149NGT(HA1), and 436NLS(NA). Three strains were linked to the 3C.2a1b.2a.1a clade lineage. Amantadine and NA inhibitors were effective against all Huai′an strains. Conclusions:The antigenicity of one strain of Huai'an strain changed and its matching with the vaccine strain of that year was low. It is suggested that the genetic surveillance of H3N2 influenza virus should be continuously strengthened to provide scientific basis for influenza prevention and control and influenza vaccine screening.

BACKGROUND:Cannabidiol is effective in ameliorating the body's inflammatory response,but no clear mechanistic studies have been conducted to ameliorate skeletal muscle inflammation induced by exhaustive exercise. OBJECTIVE:To explore the mechanism by which cannabidiol improves skeletal muscle inflammation during exhaustive exercise by using transcriptome sequencing technology. METHODS:Thirty-six Sprague-Dawley rats were randomly divided into six groups:blank control group,exercise coconut oil group,exercise control group,50 mg/kg cannabidiol group,60 mg/kg cannabidiol group,and 70 mg/kg cannabidiol group,with six rats in each group.Except for rats in the blank control group,rats in each group were subjected to swimming exercise for 9 days to produce the exhaustive exercise model.At the end of each swimming exercise,rats in the cannabidiol groups were given 2 mL of fat-soluble cannabidiol at different concentrations(50,60,and 70 mg/kg)by gavage;rats in the exercise coconut oil group were given the same volume of coconut oil by gavage until the end of the exercise on the 9th day;and rats in the blank control group and the exercise control group were not given any special treatment.The levels of inflammatory factors and differentially expressed genes in the skeletal muscle of rats in each group were determined using ELISA and transcriptome sequencing techniques.Differentially expressed genes obtained were subjected to KEGG analysis,and the accuracy of the sequencing data was verified by fluorescence quantitative PCR. RESULTS AND CONCLUSION:The results of ELISA showed that the contents of interleukin-6(P<0.05),tumor necrosis factor-α(P<0.01),interleukin-10 and other inflammatory factors in the exercise group increased significantly compared with the blank control group and the coconut oil group.After cannabidiol intervention,the mass concentrations of interleukin-6 and tumor necrosis factor-α showed a sequential decrease with increasing cannabidiol concentration.By comparing GO and KEGG databases,the functional properties of differentially expressed genes were analyzed,and the results showed that the differentially expressed genes were mainly involved in the tumor necrosis factor signaling pathway and the Toll-like receptor signaling pathway.RT-qPCR results showed that the trends of five randomly selected differentially expressed genes were in agreement with the transcriptome sequencing results.To conclude,cannabidiol can improve skeletal muscle inflammation caused by exhaustive exercise.

Objective:To identify the factors influencing acute kidney injury (AKI) following abdominal surgery in elderly patients and develop a predictive model based on the LASSO regression.Methods:The medical records of American Society of Anesthesiologists (ASA) Physical Status classificationⅠ-Ⅳ patients, aged ≥60 yr, with operation time ≥ 2 h, undergoing elective abdominal surgery under anesthesia in the General Hospital of Ningxia Medical University from May 2021 to May 2023, were retrospectively collected. AKI was diagnosed based on the Kidney Disease: Improving Global Outcomes organization guidelines. The patients were divided into 2 groups based on whether AKI occurred within 7 days after surgery: AKI group and non-AKI group. The least absolute shrinkage and selection operator algorithm was performed to reduce the dimension of unbalanced factors between AKI group and non-AKI group and the known risk factors for AKI. A nomogram prediction model was developed by integrating the optimized features derived from the LASSO regression model into multivariate logistic regression analysis. Internal validation was performed using the Bootstrap method, and the predictive ability and accuracy of the prediction model were assessed through the calibration curve, area under the receiver operating characteristic curve, Brier index and decision curve analysis.Results:Five hundred and ninety patients were finally included in this study, with 62 cases (10.5%) suffered postoperative AKI. The results of multivariate logistic regression analysis showed that increased age ( OR=1.06, 95% confidence interval [ CI] 1.01-1.11, P=0.048), higher ASA classification ( OR=2.32, 95% CI 1.21-4.45, P=0.011), preoperative coronary heart disease ( OR=1.89, 95% CI 1.01-3.61, P=0.049), and longer surgical duration ( OR=1.01, 95% CI 1.01-1.02, P=0.004) were risk factors for AKI after abdominal surgery, and the intraoperative use of dexmedetomidine ( OR=0.22, 95% CI 0.08-0.59, P=0.003) and increased postoperative albumin concentrations ( OR=0.91, 95% CI 0.85-0.98, P=0.017) were protective factors for postoperative AKI in elderly patients ( P<0.05). A risk prediction model was constructed based on the 9 identified factors of age, ASA classification, Charlson Comorbidity Index, preoperative coronary heart disease, preoperative hemoglobin concentration, preoperative estimated glomerular filtration rate, surgical duration, intraoperative use of dexmedetomidine and postoperative albumin concentration. A nomogram was plotted to visualize the model and verify it, showing that the Brier score of the model was 0.079, with a discrimination of 0.844, sensitivity of 84.4%, and specificity of 70.2%. Two hundred bootstrap resamples were used for internal validation, yielding a receiver operating characteristic curve of 0.821 with a 95% confidence interval of 0.79 to 0.90. The clinical decision curve indicated significant net benefits when the threshold probability of the model was between 0.03 and 0.45. Conclusions:Increased age, higher ASA classification, preoperative coronary heart disease, and longer surgical duration are risk factors, and the intraoperative use of dexmedetomidine and increased postoperative albumin concentrations are protective factors for postoperative AKI in elderly patients. The AKI prediction model following abdominal surgery developed based on age, ASA classification, Charlson Comorbidity Index, preoperative coronary heart disease, preoperative hemoglobin concentration, preoperative estimated glomerular filtration rate, surgical duration, intraoperative use of dexmedetomidine and postoperative albumin concentration has good predictive value in elderly patients.

OBJECTIVE: To explore the therapeutic mechanism of Liushen Wan (LSW) against colitis-associated colorectal cancer (CAC) by network pharmacology. METHODS: TCMSP, BATMAN-TCM, CNKI, PubMed, Genecards, OMIM, and TTD databases were used to obtain the related targets of LSW and CAC. The common targets of LSW and CAC were obtained using Venny online website. The PPI network was constructed using Cytoscape 3.8.2 to screen the core targets of LSW in the treatment of CAC. GO and KEGG enrichment analysis were conducted using DAVID database. The therapeutic effect of LSW on CAC was evaluated in a C57BL/6J mouse model of AOM/DSS-induced CAC by observing the changes in body weight, disease activity index, colon length, and size and number of the tumor. HE staining and RT-qPCR were used to analyze the effect of LSW on inflammatory mediators. Immunohistochemistry and TUNEL staining were used to evaluate the effect of LSW on the proliferation and apoptosis of AOM/DSS-treated colon tumor cells. Immunohistochemistry and Western blotting were used to detect the effects of LSW on the expression of TLR4 proteins in CAC mice. RESULTS: Network pharmacology analysis identified 69 common targets of LSW and CAC, and 33 hub targets were screened in the PPI network. KEGG pathway enrichment analysis suggested that the effect of LSW on CAC was mediated by the Toll-like receptor signaling pathway. In the mouse model of AOM/DSS-induced CAC, LSW significantly inhibited colitis-associated tumorigenesis, reduced tumor number and tumor load (P < 0.05), obviously improved histopathological changes in the colon, downregulated the mRNA levels of proinflammatory cytokines, and inhibited the proliferation (P < 0.01) and promoted apoptosis of colon tumor cells (P < 0.001). LSW also significantly decreased TLR4 protein expression in the colon tissue (P < 0.05). CONCLUSION LSW can inhibit CAC in mice possibly by regulating the expression of TLR4 to reduce intestinal inflammation, inhibit colon tumor cell proliferation and promote their apoptosis.
Mice ; Animals ; Toll-Like Receptor 4 ; Colitis-Associated Neoplasms ; Network Pharmacology ; Mice, Inbred C57BL ; Colonic Neoplasms/pathology*