Hepatocellular carcinoma (HCC) expresses abundant glycolytic enzymes and displays comprehensive glucose metabolism reprogramming. Aldolase A (ALDOA) plays a prominent role in glycolysis; however, little is known about its role in HCC development. In the present study, we aim to explore how ALDOA is involved in HCC proliferation. HCC proliferation was markedly suppressed both in vitro and in vivo following ALDOA knockout, which is consistent with ALDOA overexpression encouraging HCC proliferation. Mechanistically, ALDOA knockout partially limits the glycolytic flux in HCC cells. Meanwhile, ALDOA translocated to nuclei and directly interacted with c-Jun to facilitate its Thr93 phosphorylation by P21-activated protein kinase; ALDOA knockout markedly diminished c-Jun Thr93 phosphorylation and then dampened c-Jun transcription function. A crucial site Y364 mutation in ALDOA disrupted its interaction with c-Jun, and Y364S ALDOA expression failed to rescue cell proliferation in ALDOA deletion cells. In HCC patients, the expression level of ALDOA was correlated with the phosphorylation level of c-Jun (Thr93) and poor prognosis. Remarkably, hepatic ALDOA was significantly upregulated in the promotion and progression stages of diethylnitrosamine-induced HCC models, and the knockdown of A ldoa strikingly decreased HCC development in vivo. Our study demonstrated that ALDOA is a vital driver for HCC development by activating c-Jun-mediated oncogene transcription, opening additional avenues for anti-cancer therapies.

Objective:To evaluate clinical prognosis and prognostic factors of patients with early stage (Ⅰ stage) and locally advanced (Ⅱ/Ⅲ stage) lung cancer treated with carbon ion radiotherapy (CIRT).Methods:Clinical data, treatment, adverse reactions, survival and so on of 54 lung cancer patients who received CIRT and follow-up in the Heavy Ion Center of Wuwei Cancer Hospital of Gansu Province from March 2020 to September 2022 were retrospectively analyzed. The survival curve was plotted using Kaplan-Meier method. Difference tests were performed using log-rank test. Logistic regression analysis was used to identify prognostic factors.Results:According to inclusion and exclusion criteria, 54 patients were enrolled in the study, including 10 patients with early stage lung cancer and 44 patients with locally advanced lung cancer. The median follow-up time for 10 patients with early stage lung cancer was 11.0 (6.75, 17.25) months, and the median dose of irradiation was 60 Gy [relative biological effect (RBE)]. Upon the last follow-up, 3 patients had complete response (CR) and 3 patients had partial response (PR). Four patients had stable disease (SD) and no progressive disease (PD). The 1-year and 2-year local control rates (LCR), progression-free survival (PFS) rates and overall survival (OS) rates were 100%. During treatment and follow-up, 2 patients developed grade 1 radiation pneumonia, 1 case of grade 2 radiation pneumonia, 1 case of chest wall injury (chest wall pain), and there were no adverse reactions greater than grade 2. The median follow-up time of 44 patients with locally advanced stage was 12.5 (4.25, 21.75) months, and the median irradiation dose was 72 Gy (RBE). Thirty-two (73%) patients received concurrent chemotherapy during treatment, 20 (45%) patients received sequential chemotherapy after treatment, 14 (32%) patients received immune maintenance therapy and 3 (7%) patients obtained PD and received targeted drugs. Upon the last follow-up, 3 (7%) patients had CR, 17 (39%) patients had PR, 19 (43%) patients obtained SD, and 5 (11%) patients had PD. The 1-year and 2-year LCR were 96.0% and 87.3%, 90.9% and 84.1% for the 1-year and 2-year PFS rates, and 93.2% and 86.4% for the 1-year and 2-year OS rates, respectively. The median OS and PFS of patients were not reached. Multivariate logistic regression analysis showed that maintenance therapy after radiotherapy ( P=0.027) and clinical target volume (CTV) irradiation volume ( P=0.028) were the factors affecting PFS. Simultaneous chemoradiotherapy ( P=0.042) and maintenance therapy after radiotherapy ( P=0.020) were the factors affecting OS. And gross tumor volume (GTV) ≥215 ml ( P=0.068) might be an independent risk factor for grade 2 and above radiation pneumonia. Conclusions:The domestic carbon ion system has definite clinical effect and controllable toxic and side effects in the treatment of early stage and locally advanced lung cancer. The combination of synchronous chemotherapy and further maintenance treatment can significantly improve clinical prognosis of patients without significantly increasing the risk of toxic and side effects.

Objective To establish an immortalized tree shrew corneal stromal cells(CSCs)line and to study its response to virus infection.Methods Primary tree shrew CSCs were isolated and cultured by the tissue block adhesion method.CSCs were then transfected with a lentivirus carrying the SV40T gene and monoclonal cells were selected for passage culture.The characteristics of the CSCs were investigated by morphological observation and compared with 40 generations until the 50 generations or more,immunofluorescence identification of vimentin and SV40T genes,karyotype examination,and cell proliferation curve.The CSCs were infected with herpes simplex virus-1(HSV-1)(McKrae strain),Zika virus(ZIKV,GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8).Results The immortalized tree shrew CSCs after＞50 passages appeared spindle-shaped with good cell morphology and structure compared with 40 generations.Positive immunofluorescence expression of vimentin and SV40T genes.The cell growth curve showed that the cells were in logarithmic-phase growth on days 4～5 and grew vigorously.The number of chromosomes in the primary cells was stable at 62,while immortalized CSCs had 64 chromosomes at P21 and P56.The virus titer results showed that the immortalized tree shrew CSCs were sensitive to HSV-1(McKrae strain),ZIKV(GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8),with virus titers of 1.32×105,5.62×106,2.69×107,and 7.76×104 CCID50/mL,respectively.Conclusions The immortalized tree shrew CSCs were established successfully,suggesting that this cell line is suitable for studies of the mechanisms of HSV,ZIKV,Dengue virus,and influenza A virus infection in relation to corneal diseases and antiviral drugs.

In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.

Objective:To compare the changes in pelvic floor electrophysiology and imaging in female patients with myofascial pelvic pain (MFPP), and to explore the characteristics and significance of these changes.Methods:A total of 49 MFPP patients who were admitted to the of Dalian Women′s and Children′s Medical Center (Group) from January 2019 to October 2021 were randomly selected as the research group, and 41 healthy women during the same period were selected as the control group. Both groups filled in the center′s medical history and general condition survey form. French PHENIX series pelvic floor muscle potential detection instrument was used to detect the resting vaginal muscle potential and maximum muscle potential of the two groups. The static tension, dynamic tension and pelvic floor muscle contraction force of the two groups were measured by French PHENIX series electronic tensioning apparatus with 5° and 10° opening respectively. Two dimensional transperineal ultrasound and three dimensional transvaginal ultrasound produced by B-K Company in Denmark were used to measure the length between the lower margin of the bladder neck from the symphysis pubis and the bladder neck and the bladder bottom (BND, BSD), the diameter of the genital tract hiatus and the angle of the anus and rectum. The area, anterior-posterior diameter, transverse diameter and different damage degrees of levator ani levator were measured.Results:The resting muscle potential of the study group was higher than that of the control group: 2 μV vs. 1 μV ( P<0.05); the maximum vaginal myopotential was higher than that of the control group: 7 μV vs. 6 μV ( P<0.05). The static tension, dynamic tension and contractile force: 204 g/m 2 vs. 175 g/m 2, 450 g/m 2 vs. 410 g/m 2 and 237 g/m 2 vs. 51 g/m 2 of pelvic floor muscle in the study group were higher than those in the control group when the tensioner was opened for 5° ( P<0.05). In resting state, BND, BSD and reproductive tract hiatus diameter in the study group were smaller than those in the control group: 14.0 mm vs. 16.7 mm, 15.3 mm vs. 19.7 mm, 46.7 mm vs. 49.5 mm ( P<0.05). The anal angle was greater than that of the control group: 129° vs. 112° ( P<0.05). The anal right angle in the study group was greater than that in the control group: 113° vs. 109° ( P<0.05). In the resting state, the area of levator ANI hiatus: 10.1 cm 2 vs. 11.6 cm 2, anterior and posterior diameters: 44.2 mm vs. 47.4 mm, transverse diameters and the defect scores of levator ani in the study group were all smaller than those in the control group ( P<0.05). Conclusions:MFPP presents with persistent pelvic floor muscle spasm and loss of coordination. MFPP can be treated by spasmolysis of pelvic floor muscle and fascia, which provides reference value for clinical treatment.

Objective To explore the relationship between the use of proton pump inhibitors(PPIs)and the short-term and long-term prognosis of patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD).Methods Clinical data of AECOPD patients admitted to the intensive care unit(ICU)from January 2008 to December 2019 were extracted from the MIMIC-Ⅳ database.Patients were divided into PPIs group and non PPIs group based on whether PPIs were used during ICU treatment.Compare the general conditions of two groups of patients and plot survival curves using Kaplan-Meier method to compare the differences in survival rates between the two groups at 28 d and 90 d,respectively.Cox proportional hazards regression was used to analyze the association between PPIs usage and 28 d and 90 d mortality risk in two groups of patients.Results A total of 447 patients were included,including 358 in the PPIs group and 89 in the non PPIs group.The 28 d mortality rate and 90 d mortality rate of the PPIs group were 15.64％and 23.46％,respectively,which were lower than those of the non PPIs group(31.46％and 40.45％,respectively)(P＜0.05).The Kaplan-Meier curve analysis showed that the 28 d and 90 d survival rates of the PPIs group were higher than those in the non PPIs group(P＜0.001).The Cox proportional hazards regression analysis showed that after adjusting for all included variables,the hazard ratio(HR)for 28 d and 90 d mortality in the PPIs group were 0.58(95％CI 0.35 to 0.94,P=0.030),0.63(95％CI 0.41 to 0.96,P=0.022),respectively,compared to the non PPIs group.Conclusion In AECOPD patients,the use of PPIs may be reduce the 28 d and 90 d mortality risks.

Objective:To compare the adverse reactions, efficacy and survival rate of carbon ion beam irradiation in the elective lymph node (ENI) drainage area of locally advanced non-small cell lung cancer (LA-NSCLC) with relative biological effect (RBE) dose of 48 Gy using 16 and 12 fractions.Methods:A total of 72 patients with pathologically confirmed LA-NSCLC admitted to Wuwei Heavy Ion Center of Gansu Wuwei Tumor Hospital from June 2020 to December 2021 were enrolled and simple randomly divided into groups A and B, with 36 patients in each group. Patients in groups A and B were treated with carbon ion beam irradiation to the lymph node drainage area with 48 Gy (RBE) using 16 and 12 fractions. The acute and chronic adverse reactions, efficacy and survival rate were observed. The survival curve was drawn by Kaplan-Meier method. Difference test was conducted by log-rank test.Results:The median follow-up time was 13.9 (8.8-15.7) months in group A and 14.6 (6.3-15.9) months in group B. Sixteen (44.4%) patients were effectively treated in group A and 9 (25%) patients in group B. Thirty-four (94.4%) cases achieved disease control in group A and 30 (83.3%) cases in group B. Statistical analysis showed that the overall survival rate in group B was similar to that in group A ( χ2=1.192, P=0.275). Comparison of planning parameters between two groups showed CTV volume, D mean, V 5 Gy(RBE), V 20 Gy(RBE) and V 30 Gy(RBE) of the affected lung, cardiac V 20 Gy(RBE), V 30 Gy(RBE) and D mean, esophageal V 30 Gy(RBE), V 50 Gy(RBE), D max and D mean, D max of the trachea and spinal cord had no significant difference (all P>0.05). No grade 3 or 4 adverse reactions occurred in the enrolled patients during treatment and follow-up. No statistical differences were observed in the acute radiation skin reaction ( χ2=5.134, P=0.077), radiation esophagitis ( χ2=1.984, P=0.371), and advanced radiation pneumonia ( χ2=6.185, P=0.103) between two groups. Conclusions:The two dose fractionation modes of carbon ion therapy system are equally safe in the mediastinal lymphatic drainage area of LA-NSCLC, and the adverse reactions are controllable. The long-term efficacy still needs further observation.

Objective:To analyze the mediating effects of emotional changes in self-management interventions on the relationship between behavioral management and quality of life in elderly patients with type 2 diabetes and therefore to provide a reference for improving the emotional status of elderly diabetes patients and future community-based self-management interventions.Methods:From 2016 to 2020, a total of 69 self-management groups were formed in Hangzhou, each consisting of 10-15 patients with type 2 diabetes.Based on the construction of a medical consortium and family doctors signing up to provide services, intervention teams were established to conduct a series of group self-management activities for each group.Surveys via questionnaires were conducted before and after the intervention, as well as 6 months after the intervention, to collect patient data on demographics, disease status, emotions, quality of life, and self-management behaviors.The intervention effects were evaluated, and the correlations between emotional changes, self-management behaviors, and changes in quality of life were analyzed.Bootstrap analysis was used to test the mediating effects.Results:A total of 707 elderly diabetes patients were included.There were statistically significant differences in the scores of the Self-Rating Anxiety Scale(SAS), the Self-Rating Depression Scale(SDS), physical functioning, role-physical, bodily pain, general health, vitality, social functioning, role-emotional, mental health, physical component summary, mental component summary, the dietary control dimension of the type 2 diabetes self-care scale(2-DSCS), regular exercise, medication adherence, blood glucose monitoring, foot care, the dimension of prevention and management of hyperglycemia and hypoglycemia, and the total score of 2-DSCS( P<0.05 for all).Compared with pre-intervention, changes in SAS, SDS, physical component summary and mental component summary scores were all correlated with each dimension of 2-DSCS right after intervention and 6 months after intervention( r=-0.336-0.333, P<0.05), with the exception of changes in the blood glucose monitoring dimension score and changes in the emotional status score.The direct effect of self-management behavior on the quality of life in elderly patients with type 2 diabetes was 0.159, and the indirect effect through emotions was 0.229, with the mediating effect accounting for 59.02% of the total effect. Conclusions:Community-based group self-management activities can effectively improve the emotions and management behaviors of elderly diabetes patients and enhance their quality of life, Emotional changes play a certain mediating role between self-management behaviors and improvement in quality of life.

Objective To explore the effects of low centrifugal force on sperm morphology analysis of normal and non-liquefied semen samples.Methods A total of 169 patients who underwent semen routine examination and sperm morphology analysis at the Reproduc-tive Center of the 904th Hospital of the Joint Logistics Support Force of PLA from January 2021 to December 2021 were collected as the study subjects.Among them,126 were the patients with normal semen routine parameters,and 43 with non-liquefied semen.The mor-phological analysis results of 126 normal semen samples treated by the direct smear method and centrifugation methods under two differ-ent of low centrifugal force were compared.The results of morphological analysis for the 43 non-liquefied semen samples treated by the direct smear method after promoting liquefaction and centrifugation were compared.Results The percentage of normal morphological sperm[(9.39±0.50)％]obtained by centrifugation of 340xg centrifugal force for 10 min was significantly lower than that[(11.08± 0.41)％]obtained by the direct smear method,and the abnormal rate of sperm in the middle segment[(62.05±1.44)％]obtained by centrifugation of 340xg centrifugal force was significantly higher than that[(57.17±0.82)％]obtained by the direct smear method.The percentage of normal morphological sperm[(9.41±0.57)％]obtained by centrifugation at 151×g centrifugal force was significantly lower than that obtained by the direct smear method,and the abnormal rate of sperm in the middle segment[(60.95±1.17)％]ob-tained by centrifugation at 151×g centrifugal force was significantly higher than that obtained by the direct smear method.For non-lique-fied semen samples,the percentage of normal morphological sperm obtained by centrifugation at 340xg centrifugal force for 10 min was[(9.61±0.60)％],while that obtained by the direct smear method after promoting liquefaction was[(11.10±0.73)％].The difference was not statistically significant between the two methods.The abnormal rate of sperm in the middle segment[(60.21±1.51)％]ob-tained by the centrifugation method was also not significantly different from that[(57.36±1.34)％]obtained by the direct smear meth-od after promoting liquefaction.Conclusion Both kinds of low centrifugal force affected the sperm morphology.However,the low cen-trifugal force has no significant effect on the non-liquefied semen.In the clinical application of sperm morphology analysis,the nature of male semen should be referred to determine whether the centrifugation is need or not and the magnitude of centrifugal force.

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) positively affect the initial control of non-small cell lung cancer (NSCLC). Rapidly acquired resistance to EGFR-TKIs is a major hurdle in successful treatment. However, the mechanisms that control the resistance of EGFR-TKIs remain largely unknown. RNA structures have widespread and crucial functions in many biological regulations; however, the functions of RNA structures in regulating cancer drug resistance remain unclear. Here, the psoralen analysis of RNA interactions and structures (PARIS) method is used to establish the higher-order RNA structure maps of EGFR-TKIs-resistant and -sensitive cells of NSCLC. Our results show that RNA structural regions are enriched in untranslated regions (UTRs) and correlate with translation efficiency (TE). Moreover, yrdC N⁶-threonylcarbamoyltransferase domain containing (YRDC) promotes resistance to EGFR-TKIs. RNA structure formation in YRDC 3' UTR suppresses embryonic lethal abnormal vision-like 1 (ELAVL1) binding, leading to EGFR-TKI sensitivity by impairing YRDC translation. A potential therapeutic strategy for cancer treatment is provided using antisense oligonucleotide (ASO) to perturb the interaction between RNA and protein. Our study reveals an unprecedented mechanism through which the RNA structure switch modulates EGFR-TKI resistance by controlling YRDC mRNA translation in an ELAVL1-dependent manner.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; ErbB Receptors/metabolism* ; GTP-Binding Proteins/therapeutic use* ; Lung Neoplasms/genetics* ; Mutation ; Protein Kinase Inhibitors/therapeutic use* ; RNA ; RNA-Binding Proteins/genetics* ; Tyrosine Kinase Inhibitors/therapeutic use*