Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.

AIM:To investigate the mammalian target of rapamycin ( mTOR) signaling pathway as the center playing a role in the crizotinib-induced apoptosis of non-small cell lung cancer (NSCLC) cell line H2228, which represents positive echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene. METHODS:H2228 cells were processed according to different purposes .Fluorescence quantitative PCR is used to ob-serve the gene states .MTT assay is used to detect the cell inhibition rates .The cell apoptosis and cell cycle were analyzed by flow cytometry .The expression and activation levels of the key proteins in the mTOR signaling pathway were determined by Western blotting .RESULTS:Crizotinib promoted the apoptosis of H 2228 cells in a time-and dose-dependent manner . Crizotinib blocked the H2228 cells staying at the G1 phase.In apoptotic H2228 cells processed with crizotinib, the activa-tion level of mTOR was decreased , and the activation levels of the key proteins in upstream and downstream of mTOR path -way were both declined .The expression level of the fusion protein EML 4-ALK variant 3 was not affected , but its active form of p-ALK was significantly suppressed .CONCLUSION:mTOR signaling pathway has a certain relationship with the crizotinib-induced apoptosis of lung cancer cell H 2228, which represents positive EML4-ALK fusion gene.

OBJECTIVE: The aim of this study was to build a model to predict the risk of lymphovascular space invasion (LVSI) in women with endometrial cancer (EC). METHODS: From December 2010 to June 2013, 211 patients with EC undergoing surgery at Shanghai First Maternity and Infant Hospital were enrolled in this retrospective study. Those patients were divided into a positive LVSI group and a negative LVSI group. The clinical and pathological characteristics were compared between the two groups; logistic regression was used to explore risk factors associated with LVSI occurrence. The threshold values of significant factors were calculated to build a risk model and predict LVSI. RESULTS: There were 190 patients who were negative for LVSI and 21 patients were positive for LVSI out of 211 patients with EC. It was found that tumor grade, depth of myometrial invasion, number of pelvic lymph nodes, and International Federation of Gynecology and Obstetrics (FIGO) stage (p<0.05) were associated with LVSI occurrence. However, cervical involvement and age (p>0.05) were not associated with LVSI. Receiver operating characteristic (ROC) curves revealed that the threshold values of the following factors were correlated with positive LVSI: 28.1 U/mL of CA19-9, 21.2 U/mL of CA125, 2.58 mg/dL of fibrinogen (Fn), 1.84 U/mL of carcinoembryonic antigen (CEA) and (6.35×10⁹)/L of white blood cell (WBC). Logistic regression analysis indicated that CA125 ≥21.2 (p=0.032) and Fn ≥2.58 mg/dL (p=0.014) were significantly associated with LVSI. CONCLUSION: Positive LVSI could be predicted by CA125 ≥21.2 U/mL and Fn ≥2.58 mg/dL in women with EC. It could help gynecologists better adapt surgical staging and adjuvant therapies.
CA-125 Antigen ; Carcinoembryonic Antigen ; Endometrial Neoplasms* ; Female ; Fibrinogen* ; Gynecology ; Humans ; Infant ; Leukocytes ; Logistic Models ; Lymph Nodes ; Obstetrics ; Retrospective Studies ; Risk Factors ; ROC Curve

Objective To investigate the effects of siRNA-mediated knockdown of S100A4 expression on the inva?sion and migration of SNB19 glioma cells. Methods The S100A4 expression was knockdowned using S100A4 siRNA in SNB19 glioma cells. Glioma cells were assigned into control group,siRNA-negative control treated group (siRNA-NC) and siRNA-S100A4 group. RT-PCR and western blot were used to detect the mRNA and protein expression of S100A4, respectively. The wound-healing assay and transwell invasion assay were used to determine the ability of migration and invasion of SNB19 glioma cells, respectively. The expression of matrix metalloproteinase 9 (MMP-9), matrix metallopro?teinase 2 (MMP-2) and E-cadherin proteins were evaluated by using western blot. Moreover, the morphology of lamellipo?dia of glioma cells were examined by using inverted phase-contrast microscopy. Results The mRNA and protein expres?sion levels of S100A4 was obviously down-regulated after transfection of S100A4 siRNA. Compared with control group, the mRNA expression levels of S100A4 in siRNA-NC group and siRNA-S100A4 group were 0.97±0.07 and 0.21±0.04,respectively(P<0.01). The protein expression levels of S100A4 in control, siRNA-NC and siRNA-S100A4 groups were 78.12%±2.63%, 77.16%±3.00%and 37.95%±2.71%, respectively(P<0.01). The migration and invasiveness capability were decreased up to 46% and 55% in the siRNA-S100A4 group compared with the control group(P<0.01). The pro?tein expression levels of MMP-9 and MMP-2 were inhibited up to 62% and 68%(P<0.01)whereas the expression of E-cadherin was increased up to 154%(P<0.01)in the siRNA-S100A4 group. The lamellipodia became smaller or unex?tended in siRNA-S100A4-treated SNB19 glioma cells. Conclusion S100A4 plays an important role in the invasion and migration of glioma cells, suggesting that S100A4 might be a potential candidate for anti-glioma strategy to prevent the invasion and migration of glioma cells.

Objective To investigate the inhibitory effect of a disintegrin and metalloprotease 12 silenced by shR?NA on self-renewal capacity of CD133 positive giloma cells. Methods The shRNA recombinant lentivirus aimed at si?lencing ADAM12 was prepared. Human glioma cells U87 were employed in this study and assigned into three groups:shRNA-ADAM12, shRNA-NCandshRNA-C. ADAM12 expression was detected at mRNA and protein level using Re?al-time quantitative-PCR and western bloting, respectively. U87 cells were cultured with stem cell culture medium, to obtain cell sphere formation in which CD133 positive glioma cells were enriched. Immunofluorescence was employed to detect the expression of ADAM12 and CD133 in cell spheres and U87 cells; Self-renewal was tested by using tumor sphere formation assay. Molecular markers for differentiated or undifferentiated cells (CD133,GFAP and Tuj1) were de?tected at protein using western blotting. Western blotting was employed to test protein expression of HES1. Results AD?AM12 shRNA significantly down-regulated the mRNA and protein expression levels of ADAM12. Compared with shRNA–C group, the relative expression levels of mRNA in shRNA-ADAM12 group and shRNA-NC group were 0.22 ± 0.03 and 0.98 ± 0.06 (F=425.37,P<0.01). The relative expression levels of protein in shRNA-ADAM12 group, shRNA-NC group and shRNA-C group were 28.72%±2.36%, 69.21%±3.92%and 69.04%±3.57%, respectively (F=145.42,P<0.01). Immunofluorescence staining showed that expression levels of ADAM12 and CD133 in cell spheres were significantly higher than those in normal cells. The number of spheres in three groups were 45.5±2.3、104.2±5.8 and 109.6±6.2, tumor sphere formation ability of shRNA-ADAM12 group was lower than that of shRNA-NC group and shRNA-C group (F=147.03,P<0.01). Compared with the shRNA-NC group and shRNA-C group, the protain expression of GFAP and Tuj1 were increased up to 166% and 146% (P<0.01) whereas the protein expression levels of CD133 and HES1 were down-regulated by 54% and 50% (P<0.01). Conclusion Knockdown of ADAM12 may suppress self-renewal ability of CD133 positive glioma cells by inhibiting the Notch pathway activity.

Abscopal effect (AE) is defined as the phenomenon that the non-irradiated lesions shrink in addition to the directly irradiated lesions when the tumors are treated with radiotherapy. With widespread application of programmed cell death receptor-1 (PD-1), programmed cell death ligand-1 (PD-L1), cytotoxic T-lymphocyte-associated protein 4 (CTL-4) antibody, the treatment of malignant tumors has entered the era of immunotherapy. With the increasing application of radiotherapy combined with immunotherapy, AE has received unprecedented attention from scholars. The combination of radiotherapy and immunotherapy can promote the occurrence of systemic immune response and enhance the anti-tumor response of radiotherapy, which has deepened the understanding of AE. In this article, the history of AE in different stages of radiotherapy was reviewed, and the factors, possible mechanisms and clinical research status affecting the incidence of AE were emphasized, aiming to increase the incidence of AE in the era of immunotherapy, promote the objective effectiveness of radiotherapy combined with immunotherapy in clinical application, and further improve clinical prognosis of patients with malignant tumors.

Objective:To explore the relative concentration changes of oxygenated hemoglobin (Oxy Hb) in the prefrontal cortex (PFC) and brain region activation during emotional face recognition tasks in women with perimenopausal depression.Methods:From February to April 2023, forty perimenopausal women were recruited, including 20 women with perimenopausal depression (experimental group) and 20 women with non-perimenopausal depression (control group). All participants were evaluated by the modified Kupperman score, 24-item Hamilton depression scale (HAMD-24), and patient health questionnaire (PHQ-9). Functional near-infrared spectroscopy (fNIRS) equipment was used to measure the relative concentration of Oxy-Hb in the PFC in two groups under the emotional face recognition task. Statistical analysis was performed by SPSS 26.0 software. Data were analyzed by a t-test, rank sum test, and Pearson correlation. Results:There were statistically significant differences in the results of the modified Kupperman score((23.20±3.66), (18.10±1.28)), HAMD-24((15.95±5.47), (3.35±1.84)), and PHQ-9(7.00(5.00, 10.75), 1.50(1.00, 3.00)) scales between the the experimental group and control group ( P<0.05). There was a positive correlation between the modified Kupperman score and the HAMD-24 score in the experimental group ( r=0.685, P=0.01). The reaction time of the experimental group in identifying negative and neutral emotional faces was statistically significant compared to the control group( t=4.01, 4.80, both P<0.05). Compared with identifying neutral emotions, PFC activation was stronger in the experimental group and control group when identifying negative emotions ( P<0.05). The PFC activation in the experimental group was stronger than that in the control group when identifying negative emotions ( P<0.05). There was no statistically significant difference in the activation level between the two groups when identifying neutral emotions ( P>0.05). Conclusion:Women with perimenopausal depression exhibit specificity in emotional processing, with increased PFC activation when identifying negative emotions, impaired emotional processing function of PFC, and dysfunction of aerobic metabolism.

Objective To investigate the regulating mechanism of Notch1 signaling pathway on the invasion and migration ofglioma initiating cells (GICs).Methods (1) Box-plotting was conducted to analyze the mRNA expression of Notch1 in normal brain tissue and glioblastoma tissue using Bredel Brain,Sun Brain and TCGA databases;Kaplan-Meier survival analysis was conducted to analyze the association between the prognosis of glioma patients with the expression of Hes1 in TCGA database;Heatmap was conducted to analyze the expression of Notch1 and CXCR4 in GICs and common cell line in GEO database.(2) Magnetic activated cell sorting was adopted to establish cell lines of U87 GICs and U251 GICs;immunofluorescence staining was used to detect expression of CXCR4 and Notch1.After the cell lines of U87 GICs and U251 GICs were divided into DMSO,shNC,MK0752 and shNotchl groups,the shNotch1 and shNC groups were transfected respectively with recombinant lentivirus of Notch1-shRNA and its control sequence while the MK0752 and DMSO groups were added respectively with MK-0752 of 80 nmol/mL and the same amount of DMSO.The protein expression of Notch1,CXCR4 and p-mTOR was detected by Westem blotting in the 4 groups.The capabilities of invasion and migration of the GICs were detected by Transwell assay in the shNotch1 and shNC groups.Results (1) The box-plotting showed the mRNA expression of Notch 1 in the glioblastoma tissue was significantly higher than in the normal brain tissue (P＜0.05).The Kaplan-Meier survival analysis showed that the life span ofglioma patients with high expression of Hes1 was significantly shorter than that of those with low expression of Hes1 (P＜0.05).Heatmaps showed that the expression levels of Notch1 and CXCR4 in GICs were higher than in the common cell line.(2) The immunofluorescence staining showed that Notch1 and CXCR4 were highly expressed and colocalized in cell lines of U87 GICs and U251 GICs.The Western blotting showed that the protein expression of Notch1,CXCR4 and p-mTOR in the cell lines of U87GICs and U251 GICs in the MK0752 and shNotch1 groups was lower than that in the DMSO and shNC groups.Transwell assay showed that the penetrating-membrane cells per visual field in the shNotch1group were significantly fewer than those in the shNC group (P＜0.05).Conclusion Notch1 signaling pathway can promote invasion and migration of GICs through regulating CXCR4 expression.

[Objective]Based on the propensity score matching method to explore the therapeutic effect of lever positioning manipulation in the treatment of L4/5 different types of lumbar disc herniation.[Methods]A prospective cohort design was used to select 122 patients with L4/5 lumbar disc herniation and divide them into unilateral group,bilateral group and central group.Samples with balanced covariates were obtained after matching,including 36 patients in unilateral group,35 patients in bilateral group and 35 patients in central group.All patients in the three groups were treated with leverage positioning technique,and all patients in the three groups were treated 3 times a week,once every other day,for a total of 2 weeks.Pain Visual Analogue Scale(VAS)score and Japanese Orthopaedic Association(JOA)score were observed before and after treatment to evaluate the differences in the postoperative effects of the three groups.[Results]The covariates of age,sex,course of disease,VAS score and JOA score were all balanced after matching(P>0.05).The results of following-up after 2 weeks and 6 months of treatment showed that the VAS score and JOA score were statistically significant(P<0.05),the distant and near term curative effect of lever positioning manipulation in the treatment of unilateral lumbar disc herniation was better than bilateral and central disc herniation(P<0.05).[Conclusion]The clinical efficacy of unilateral lumbar disc herniation is superior to bilateral and central disc herniation in the treatment of the same segment of lumbar disc herniation with leverage positioning manipulation.

Objective    To systematically evaluate the risk factors for postoperative pulmonary infection in patients with esophageal cancer. Methods    CNKI, Wangfang Data, VIP, CBM, PubMed, EMbase, The Cochrane Library were searched from inception to January 2021 to collect case-control studies, cohort studies and cross-sectional studies about risk factors for postoperative pulmonary infection in patients with esophageal cancer. Two researchers independently conducted literature screening, data extraction and quality assessment. RevMan 5.3 software and Stata 15.0 software were used for meta-analysis. Results    A total of 20 articles were included, covering 5 409 patients of esophageal cancer. The quality score of included studies was 6-8 points. Meta-analysis results showed that age (MD=1.99, 95%CI 0.10 to 3.88, P=0.04), age≥60 years (OR=2.68, 95%CI 1.46 to 4.91, P=0.001), smoking history (OR=2.41, 95%CI 1.77 to 3.28, P<0.001), diabetes (OR=2.30, 95%CI 1.90 to 2.77, P<0.001), chronic obstructive pulmonary disease (OR=3.69, 95%CI 2.09 to 6.52, P<0.001), pulmonary disease (OR=2.22, 95%CI 1.16 to 4.26, P=0.02), thoracotomy (OR=1.77, 95%CI 1.32 to 2.37, P<0.001), operation time (MD=14.08, 95%CI 9.64 to 18.52, P<0.001), operation time>4 h (OR=3.09, 95%CI 1.46 to 6.55, P=0.003), single lung ventilation (OR=3.46, 95%CI 1.61 to 7.44, P=0.001), recurrent laryngeal nerve injury (OR=5.66, 95%CI 1.63 to 19.71, P=0.006), and no use of patient-controlled epidural analgesia (PCEA) (OR=2.81, 95%CI 1.71 to 4.61, P<0.001) were risk factors for postoperative pulmonary infection in patients with esophageal cancer. Conclusion    The existing evidence shows that age, age≥60 years, smoking history, diabetes, chronic obstructive pulmonary disease, pulmonary disease, thoracotomy, operation time, operation time>4 h, single lung ventilation, recurrent laryngeal nerve injury, and no use of PCEA are risk factors for postoperative pulmonary infection in patients with esophageal cancer. Due to the limitation of the quantity and quality of included literature, the conclusion of this study still needs to be confirmed by more high-quality studies.