Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P<0.01), and KLF4 protein was lower in the cancer tissues than in the adjacent tissue (P<0.01). The bioinformatic analysis showed there is a theoretical binding site (5'-UGCCUUAA-3') of hsa-miRNA124-3p in 3'-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3'-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA12472 h after transfection (P<0.05 ). Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced (P<0.05) There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group (P>0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.

Objective To develop the quality of life questionnaire applicable to the Chinese patients with dysphagia by the translation and modification,as well as psychometric evaluation of the original swallowing quality of life questionnaire (SWAL-QOL).Methods The English version of the SWAL-QOL was translated into Chinese according to the well-accepted scale translation procedure.Then 103 patients with dysphagia were tested using the translated Chinese SWAL-QOL.The inter-rater reliability,test-retest reliability,internal consistency reliability,the content validity and construct validity were tested.Results The inter-rater reliability correlation coefficient of the SWAL-QOL was between 0.945-0.990 (P＜0.01).Its test-retest coefficients ranged from 0.965 to 0.992 (P＜0.01).The Cronbach coefficients ranged from 0.708 to 0.933 (P＜0.01).There revealed significant correlation between each item of SWAL-QOL and its domain,with correlation coefficients between 0.723 and 0.982 (P＜0.01).Factor analysis of each item of the 10 domains of SWAL-QOL extracted 10 common factors,which were with a cumulative contribution of 79.029％.Factor analysis of the total score of the 10 dimensions extracted 2 common factors including dysphagia-related quality of life and general quality of life,with a cumulative contribution of 54.718％.Conclusions The Chinese version of SWAL-QOL is reliable and valid.It can be used as an effective measuring tool to evaluate the quality of life of dysphagia patients.

Aim To observe the effects of icariin on depressive-like behavior in prenatally stressed offsprings and explore its mechanism.Methods The model of depression was established by prenatal restraint stress on late pregnant mother, then male offspring rats were randomly divided into five groups of eight: Control, PS model, Saline group(NS), ICA(80 mg·kg-1), ICA(40 mg·kg-1).The effects of icariin on PS-induced depressive-like behaviors in male offsprings were examined by forced swimming test(FST) and open-field test(OFT).Furthermore, the protein expressions of group I metabotropic glutamate receptors(I mGluRs) and excitatory amino acid transporter 2(EAAT2) in prefrontal cortex were detected by Western blot.Results PS group exhibited depressive-like behaviors with decreased struggling time(P<0.01) and center crossing numbers(P<0.05) compared to CON, with increased protein expressions of group I mGluRs(P<0.01,P<0.05) and decreased EAAT2(P<0.05) in prefrontal cortex compared to CON.Treatment with icariin(80 mg·kg-1, 40 mg·kg-1)significantly increased struggling time(P<0.05,P<0.01) and center crossing numbers(P<0.05) in PS-exposed male offspring rats.Treatment with icariin markedly modulated the protein levels of mGluR1, mGluR5 and EAAT2 in prefrontal cortex.Conclusion Icariin has significant antidepressant effects in prenatally stressed rats, and the mechanism might be associated with the modulation of Ⅰ mGluRs.

BACKGROUND:Neuroblastoma is a common solid tumor in children. Pediatric tumors are little affected by environmental factors, but closely related to child development. The suspension method is an effective and reliable method to harvest neoplastic stem cel s.
OBJECTIVE:To culture the cel clones of human neuroblastoma cel line SK-N-SH and to assess the biological properties of the cel clones.
METHODS:Using the suspension method with no serum media, tumor cel clones were obtained. Immunofluorescence method was used to identify whether tumor cel clones exhibit stem cel properties. SK-N-SH neuroblastoma was labeled by luciferase, and tumor cel clones and tumor cel s were seeded onto the back of nude mice to monitor cel proliferative properties in nude mice using in vivo imaging.
RESULTS AND CONCLUSION:Using the suspension culture method, SK-N-SH neuroblastoma cel s could successful y develop into cloning bal s. Under serum-free culture, cloning bal s were immunofluorescently used to detect molecular markers that showed strong positive expression. Cloning bal s subcutaneously implanted into
nude mice showed the strong ability of self-renewal and differentiation as stem cel s. The cel clones cultured by the suspension method strongly expressed Nestin, but weakly expressed glial fibril ary acidic protein, neuron-specific tubulin, possessing stem cel characteristics and strong proliferation and metastasis in nude mice.

Anelectrochemicallyreducedgrapheneoxide/goldnanoparticle-chitosan(ERGO/AuNP-CS) composite film modified glassy carbon electrode ( GCE) was constructed by directly electrochemical reduction of GO, and then assembly of AuNP-CS polycation on the surface. The surface morphologies of different modified electrodes including bare GCE, GCE/GO, GCE/ERGO and GCE/ERGO/AuNP-CS were characterized by scanning electron microscopy ( SEM ) . The differential pulse voltammetric behaviors of the electrodes were investigated, and the results indicated that the composite of ERGO/AuNP-CS exhibited excellent electrocatalytic oxidation activity to uric acid ( UA) molecule. In 0. 10 mol/L of phosphate buffer solution (pH=6. 5) with a scanning rate of 100 mV/s, the proposed composite film modified electrode showed a linear electrochemical response to UA in the range of 0 . 05-110 μmol/L with a detection limit of 12. 4 nmol/L ( S/N = 3 ). The electrode displayed good selectivity, reproducibility and stability in the determination of UA in human serum and urine samples with a recovery of 93 . 8%-104 . 1%. The detection results were agreed with those of conventional spectrophotometry and uricase Kit methods.

Objective To evaluate the relationship between genetic polymorphism of transforming growth factor (TGF)-β1 and susceptibility of liver cirrhosis after hepatitis B virus infection in Chinese population. Methods CBM, VIP, CNKI, Wanfang technological periodical full-text databases and Pubmed from set up to July, 2013 were electronically searched to identify case-control studies on the relationship between genetic polymorphism of TGF-β1 promoter 509 site, co-don 869 site and liver cirrhosis after hepatitis B virus infection. The data were quantitatively analyzed by RevMan 5.1 soft-ware after assessing the quality of included studies. Results Six case-control studies were selected for Meta-analysis based on our inclusion and exclusion standards. The results of Meta-analysis showed that the pooled OR value for liver cir-rhosis among Chinese patients after hepatitis B virus infection with T allele of TGF-β1 gene at promoter 509 was 1.02 (95%CI:0.67-1.54), the pooled OR values for patients with TT and CT genotypes were 0.80 (95%CI:0.36-1.78). OR values for pa-tients with C allele of TGF-β1 gene at codon 869 was 1.05 (95%CI:0.69-1.62), the pooled OR values for patients with CC and CT genotypes were 0.98 (95%CI:0.48-2.00). No significant publication bias was found. Conclusion The genetic poly-morphism of TGF-β1 at promoter 509 and codon 869 showed no association with susceptibility of liver cirrhosis after hepati-tis B virus infection in Chinese population.

ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.

Objective To generate and identify the Itgbl1(integrin beta-like)promoter-driven Cre knock-in mouse line.Methods Itgbll-Cre knock-in mice were generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing.The Itgbl1-Cre mice were crossed with the Cre reporter ROSALSL-tdTomato)mice to detect the expression profile of Cre activity.The tdTomato expression pattern across tissues and cell-specific markers were used to identify the cell types of Itgbl1-expressing cells and their progeny.Results and Conclusion tdTomato was specifically expressed in mucous acinar cells of the sublingual gland,pancreatic islet cells,and gastric endocrine cells.In addition,tdTomato expression was also found in some of the neurons of the retina and brain,as well as in a few cells in the serosal layer of the intestine,articular cartilage,periosteum,and bone marrow.The first Itgbl1-Cre recombinase transgenic mouse line was established,which can specifically label the mucous acinar cells of the sublingual gland.

Objectives:This study aimed to assess the efficacy and safety of bendamustine in combination with rituximab (BR regimen) for the treatment of newly diagnosed indolent B-cell non-Hodgkin's lymphoma (B-iNHL) and elderly mantle cell lymphoma (eMCL) .Methods:From December 1, 2020 to September 10, 2022, a multi-center prospective study was conducted across ten Grade A tertiary hospitals in Shandong Province, China. The BR regimen was administered to evaluate its efficacy and safety in newly diagnosed B-iNHL and eMCL patients, and all completed at least four cycles of induction therapy.Results:The 72 enrolled patients with B-iNHL or MCL were aged 24-74 years, with a median age of 55 years. Eastern Cooperative Oncology Group (ECOG) performance status scores of 0-1 were observed in 76.4% of patients, while 23.6% had scores of 2. Disease distribution included follicular lymphoma (FL) (51.4% ), marginal zone lymphoma (MZL) (33.3% ), eMCL (11.1% ), and the unknown subtype (4.2% ). According to the Ann Arbor staging system, 16.7% and 65.3% of patients were diagnosed with stage Ⅲ and stage Ⅳ lymphomas, respectively. Following four cycles of BR induction therapy, the overall response rate was 98.6%, with a complete response (CR) rate of 83.3% and a partial response (PR) rate of 15.3%. Only one eMCL patient experienced disease progression during treatment, and only one FL patient experienced a relapse. Even when evaluated using CT alone, the CR rate was 63.9%, considering the differences between PET/CT and CT assessments. The median follow-up duration was 11 months (range: 4-22), with a PFS rate of 96.8% and an OS rate of 100.0%. The main hematologic adverse reactions included grade 3-4 leukopenia (27.8%, with febrile neutropenia observed in 8.3% of patients), grade 3-4 lymphopenia (23.6% ), grade 3-4 anemia (5.6% ), and grade 3-4 thrombocytopenia (4.2% ). The main non-hematologic adverse reactions such as fatigue, nausea/vomiting, rash, and infections occurred in less than 20.0% of patients.Conclusion:Within the scope of this clinical trial conducted in China, the BR regimen demonstrated efficacy and safety in treating newly diagnosed B-iNHL and eMCL patients.

BACKGROUND: The blocking of the programmed cell death protein (PD-1)/programmed death-ligand 1 (PD-L1) axis has been found to have an anticancer activity against various types of cancer by enhancing T cell immunity, while there are no studies linking the PD-1/PD-L1 axis to chemotherapy drugs in osteosarcoma (OS). The present study aimed to investigate the effects of blocking PD-1/PD-L1 axis on the cisplatin chemotherapy in OS in vitro and in vivo. METHODS: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to detect PD-L1 mRNA in OS tissues. Cell proliferation and apoptosis were measured by Cell Counting Kit-8 (CCK-8) and flow cytometry assays, respectively. In vivo, the syngeneic mice were treated with cisplatin and anti-PD-1 antibody alone or jointly. RESULTS: In this study, it revealed that PD-L1 mRNA was highly expressed in OS tissues. Further inhibitory evaluation showed that the K7M2-LV cells (PD-L1 overexpression) co-cultured with PD-1 lymphocytes could promote K7M2 cell proliferation. Meanwhile, the combination of anti-PD-1 antibody and cisplatin significantly decreased the proliferation and increased the apoptosis of K7M2 cells in a co-culture system. In vivo, the combination of anti-PD-1 antibody and cisplatin significantly inhibited tumor growth, while the mechanisms did not involve regulatory T cells. CONCLUSION The present data suggested that the blocking of PD-1/PD-L1 axis had a positive prognostic value, which can enhance the chemotherapeutic effect of cisplatin in OS. These findings provide a rationale for utilizing PD1/PD-L1 blocking antibodies as a single agent to cure refractory OS in patients receiving cisplatin treatment.