ObjectiveThe nucleolar protein PES1 (Pescadillo homolog 1) plays critical roles in ribosome biogenesis and cell cycle regulation, yet its involvement in cellular senescence remains poorly understood. This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role. MethodsInitially, we assessed PES1 expression patterns in two distinct senescence models: replicative senescent mouse embryonic fibroblasts (MEFs) and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells. Subsequently, PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types. Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays, respectively. The expression of senescence-associated proteins (p53, p21, and Rb) and SASP factors (IL-6, IL-1β, and IL-8) were analyzed by Western blot or qPCR. Furthermore, Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology. ResultsPES1 expression was significantly downregulated in senescent MEFs and HepG2 cells. PES1 knockdown resulted in decreased EdU-positive cells and increased SA‑β‑gal-positive cells, indicating proliferation inhibition and senescence induction. Mechanistically, PES1 suppression activated the p53-p21 pathway without affecting Rb expression, while upregulating IL-6, IL-1β, and IL-8 production. Notably, PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress, as evidenced by aberrant nucleolar morphology. ConclusionOur findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent (but Rb-independent) cellular senescence, highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.

Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Objective To explore the clinical efficacy of low molecular weight heparin combined with insulin in the treatment of hyper-triglyceridemic-acute pancreatitis(HTG-AP).Methods A total of 106 patients diagnosed with HTG-AP who were admitted to the department of gastroenterology of Huaibei People's Hospital from May 2022 to July 2023 were selected as the research objects.According to the random number table method,the low-molecular heparin group(35 cases,received a 5 000 U subcutaneous injection low-molecular heparin once every 12 hours for 6 days),the insulin group(35 cases,received intravenous insulin pumping at a rate of 2 U/h,with careful monitoring of the patient's random blood glucose levels to prevent hypoglycemia),and the combination therapy group(36 cases,received both low-molecular heparin and insulin).Before treatment and at 1,2,and 6 days after treatment,the difference of serum triacylglycerol(TG),total cholesterol(TC),blood amylase,inflammatory factors[C-reactive protein(CRP),interleukin-6(IL-6)],calcium ions,and creatinine levels among the three groups were compared.The modified computed tomography severity index(MCTSI)scores,acute physiology and chronic health evaluationⅡ(APACHEⅡ),hospital length of stay,and hospital costs before and after 6 days of treatment were observed.Results After treatment,the TC of all three groups significantly decreased compared to before treatment(P<0.05),but there was no significant difference among the three groups.The calcium ion levels of the three groups did not show a statistically significant difference before and after treatment.After 6 days of treatment,the creatinine levels of the three groups significantly decreased compared to before treatment,but there was no significant difference among the three groups.After 2 days of treatment,serum TG levels were significantly lower in the combination therapy group and insulin group compared to the low-molecular heparin group(mmol/L:4.6±1.7,4.4±1.8 vs.5.6±2.0,both P<0.05).However,there was no statistically significant difference between the combination therapy group and the insulin group.After 6 days of treatment,the combination therapy group showed significantly lower levels of serum TG,blood amylase,CRP,and IL-6 compared to the insulin group and the low-molecular heparin group[TG(mmol/L):2.8±1.9 vs.4.3±1.9,5.0±2.2,blood amylase(U/L):36.0(32.0,45.0)vs.59.0(43.0,71.0),52.0(45.0,64.0),CRP(mg/L):12.9(8.8,29.7)vs.35.3(21.7,50.3),31.4(23.0,45.1),IL-6(ng/L):15.4(9.8,23.5)vs.25.6(16.4,51.5),32.9(14.7,41.4),all P<0.05].After 6 days of treatment,the APACHEⅡscores of all three groups decreased significantly(all P<0.05).The MCTSI scores of the insulin group and the combined treatment group also decreased significantly compared to before treatment.Furthermore,the MCTSI and APACHEⅡscores of the combination therapy group were significantly lower than those of the low-molecular heparin group and the insulin group(MCTSI score:2.3±0.7 vs.3.3±1.7,2.9±1.3,APACHEⅡscore:1.3±1.2 vs.2.5±2.4,2.6±2.5,all P<0.05).The combination therapy group had significantly lower length of hospital stay and treatment cost compared to the low molecular heparin and insulin groups[length of hospital stay(days):6.9±1.6 vs.8.8±3.4,8.5±2.8,and cost of treatment(yuan):6 040.5(5 239.4,7 105.9)vs.6 696.4(5 791.5,11 026.2),6 918.5(6 087.9,10 080.8),all P<0.05].Conclusions The combination of low-molecular heparin and insulin treatment can significantly reduce serum TG and inflammatory factor levels,as well as the severity and duration of the disease.This approach can also reduce the cost of treatment.Therefore,it is worth promoting and applying in clinical settings.

Objective To investigate the risk factors for bronchopulmonary dysplasia(BPD)in twin preterm infants with a gestational age of<34 weeks,and to provide a basis for early identification of BPD in twin preterm infants in clinical practice.Methods A retrospective analysis was performed for the twin preterm infants with a gestational age of<34 weeks who were admitted to 22 hospitals nationwide from January 2018 to December 2020.According to their conditions,they were divided into group A(both twins had BPD),group B(only one twin had BPD),and group C(neither twin had BPD).The risk factors for BPD in twin preterm infants were analyzed.Further analysis was conducted on group B to investigate the postnatal risk factors for BPD within twins.Results A total of 904 pairs of twins with a gestational age of<34 weeks were included in this study.The multivariate logistic regression analysis showed that compared with group C,birth weight discordance of>25%between the twins was an independent risk factor for BPD in one of the twins(OR=3.370,95%CI:1.500-7.568,P<0.05),and high gestational age at birth was a protective factor against BPD(P<0.05).The conditional logistic regression analysis of group B showed that small-for-gestational-age(SGA)birth was an independent risk factor for BPD in individual twins(OR=5.017,95%CI:1.040-24.190,P<0.05).Conclusions The development of BPD in twin preterm infants is associated with gestational age,birth weight discordance between the twins,and SGA birth.

Objective To observe the clinical efficacy and safety of traditional Chinese medicine(TCM)constitution correction based on the theory of preventive treatment of disease on the patients with early asymptomatic diabetic peripheral neuropathy(DPN).Methods A total of 87 patients with early asymptomatic DPN were randomly divided into the control group with 43 cases and the trial group with 44 cases.The control group was required to have standard diet,do exercises and take medicines for diabetes.On the basis of treatment for the control group,the trial group was given the corresponding TCM constitution correction regimen according to the predominated constitution types of the patients.The course of treatment for the two groups lasted for 48 weeks.Before and after intervention,the two groups were observed in the alteration of TCM constitution types,and the changes of body mass index(BMI),systolic blood pressure(SBP),diastolic blood pressure(DBP),fasting plasma glucose(FPG),glycosylated hemoglobin(HbA1c),total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C)and nerve conduction velocity(NCV).Moreover,the safety of the two groups was evaluated.Results(1)After the intervention,the constitution of 17 patients in the trial group was transformed into the balanced constitution,while only the constitution of 5 patients in the control group was transformed into balanced constitution.There was a statistically significant difference in the distribution of TCM constitution between the two groups(P＜0.05).(2)Before intervention,there were no significant differences in the levels of BMI,SBP,DBP,FPG,HbA1c,TC,TG,HDL-C and LDL-C between the two groups(P＞0.05).After intervention,the levels of BMI,SBP and DBP in the two groups and the levels of HbA1c,TC,TG and LDL-C in the trial group were lower than those before intervention(P＜0.05 or P＜0.01),while the levels of FPG and HDL-C in the control group were higher than those before intervention(P＜0.05 or P＜0.01).The intergroup comparison showed that the levels of FPG,HbA1c,TG and LDL-C in the trial group were significantly lower than those in the control group(P＜0.05 or P＜0.01).(3)After the intervention,10 cases in the control group had typical clinical symptoms of DPN,while only 6 cases in the trial group had the clinical symptoms of DPN.Before intervention,there was no significant difference in the motor nerve conduction velocity(MNCV)of bilateral tibial nerve and common peroneal nerve and in the sensory nerve conduction velocity(SNCV)of bilateral sural nerve and superficial peroneal nerve between the two groups(P＞0.05).After intervention,the MNCV of bilateral tibial nerve and common peroneal nerve and the SNCV of bilateral sural nerve and superficial peroneal nerve in the control group were slower than those before intervention(P＜0.05),while the above indicators of nerve conduction velocity in the trial group were mildly increased compared with those before intervention,but the differences were not significant(P＞0.05).The intergroup comparison showed that the MNCV of bilateral tibial nerve and SNCV of bilateral sural nerve and superficial peroneal nerve in the trial group were higher than those in the control group,and the differences were statistically significant(P＜0.05).(4)During the intervention,no obvious adverse reactions occurred in the two groups,with high safety.Conclusion The TCM constitution correction can adjust the biased constitution of patients with early asymptomatic DPN,and has certain protective effect on the glucose and lipid metabolism,MNCV of tibial nerve,and SNCV of bilateral sural nerve and superficial peroneal nerve.

Objective To observe the effect of neuromuscular electrical stimulation(NMES)on interleukin-6(IL-6)/STAT3 signaling pathway in mice after spinal cord injury,and to explore the mechanism of its effect on motor function recovery.Methods Seventy-two SPF grade mice were randomly divided into sham operation group,spinal cord injury(SCI)group and NMES group.BMS score,inclined plane test and neuromuscular electrophysiology(EMG)were used to evaluate the recovery of spinal cord injury in mice.Western blotting and Real-time PCR were used to detect the expression of inflammatory factors,IL-6/STAT3,glial fibrillary acidic protein(GFAP)and brain-derived neurotrophic factor(BDNF)in spinal cord tissues of three groups of mice.HE staining was used to observe the pathological changes of spinal cord injury.Results BMS scores and the inclined plane test of mice in the NMES group were higher than those in SCI group(P＜0.05).The maximum amplitude of motor evoked potential in NMES group was higher than that in SCI group(P＜0.05).The expressions of TNF-α,IL-12A and GFAP in the spinal cord of NMES group were lower than that of SCI group(P＜0.05),while the expressions of TGF-β,IL-10 and BDNF were higher than that of SCI group(P＜0.05).The protein expressions of IL-6/STAT3 signaling pathway of NMES group were lower than that of SCI group(P＜0.05).Conclusion Neuromuscular electrical stimulation plays an anti-inflammatory role by inhibiting the IL-6/STAT3 signaling pathway,thereby promoting the recovery of hind limb motor function in mice after spinal cord injury.

Objective:To analyze the clinical characteristics and prognosis of patients with CD5+diffuse large B-cell lymphoma(DLBCL).Methods:The clinical data of 161 newly treated DLBCL patients in Gansu Provincial Hospital from January 2013 to January 2020 were retrospectively analyzed.According to CD5 expression,the patients were divided into CD5+group and CD5-group.The clinical characteristics and prognosis of the two groups were statistically analyzed.Results:The median age of patients in CD5+group was 62 years,which was higher than 56 years in CD5-group(P=0.048).The proportion of women in CD5+group was 62.96％,which was significantly higher than 41.79％in CD5-group(P=0.043).The proportion of patients with IPI score＞2 in CD5+group was 62.96％,which was higher than 40.30％in CD5-group(P=0.031).Survival analysis showed that the median overall survival and progression-free survival time of patients in CD5+group were 27(3-77)and 31(3-76)months,respectively,which were both shorter than 30(5-84)and 32.5(4-83)months in CD5-group(P=0.047,P=0.026).Univariate analysis showed that advanced age,positive CD5 expression,triple or double hit at initial diagnosis,high IPI score and no use of rituximab during chemotherapy were risk factors for the prognosis of DLBCL patients.Further Cox multivariate regression analysis showed that these factors were also independent risk factors except for advanced age.Conclusion:CD5+DLBCL patients have a worse prognosis than CD5-DLBCL patients.Such patients are more common in females,with advanced age and high IPI score,which is a special subtype of DLBCL.

OBJECTIVE: To investigate the effects of methionine restriction on proliferation, cell cycle and apoptosis of human acute leukemia cells. METHODS: Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of methionine restriction on HL-60 and Jurkat cells proliferation. The effect of methionine restriction on cell cycle of HL-60 and Jurkat cells was examined by PI staining. Annexin V-FITC / PI double staining was applied to detect apoptosis of HL-60 and Jurkat cells following methionine restriction. The expression of cell cycle-related proteins cyclin B1, CDC2 and apoptosis-related protein Bcl-2 was evaluated by Western blot assay. RESULTS: Methionine restriction significantly inhibited the proliferation of HL-60 and Jurkat cells in a time-dependent manner (HL-60: r =0.7773, Jurkat: r =0.8725), arrested the cells at G2/M phase (P < 0.001), and significantly induced apoptosis of HL-60 and Jurkat cells (HL-60: P < 0.001; Jurkat: P < 0.05). Furthermore, Western blot analysis demonstrated that methionine restriction significantly reduced the proteins expression of Cyclin B1 (P < 0.05), CDC2 (P < 0.01) and Bcl-2 (P < 0.001) in HL-60 and Jurkat cells. CONCLUSION Acute leukemia cells HL-60 and Jurkat exhibit methionine dependence. Methionine restriction can significantly inhibit the proliferation, promote cell cycle arrest and induce apoptosis of HL-60 and Jurkat cells, which suggests that methionine restriction may be a potential therapeutic strategy for acute leukemia.
Humans ; Cyclin B1/pharmacology* ; Cell Proliferation ; Methionine/pharmacology* ; Cell Cycle ; Apoptosis ; Leukemia, Myeloid, Acute ; Cell Division ; Cell Cycle Proteins ; Jurkat Cells ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; HL-60 Cells

Objective:To evaluate the feasibility of using bedside ultrasound and serum biomarkers for the prediction of sepsis-induced myocardial dysfunction(SIMD)and mortality in septic shock patients.Methods:The patients diagnosed as septic shock were enrolled in the study from January 2019 to July 2021 in PICU at Shanghai Children′s Medical Center Affiliated to Shanghai Jiaotong University School of Medicine.Bedside ultrasound results were recorded at day 1, 2, 3, 7 and 10.Blood samples were collected at the same time, markers of myocardial injury were detected, and prognosis was recorded at 28 days.According to the left ventricular ejection fraction (LVEF), children with septic shock were divided into SIMD group and non-SIMD group.Those with LVEF <50% or decreased by ≥10% from baseline level were defined as SIMD.Differences in cardiac ultrasound parameters and biomarkers between two groups were compared.Logistic regression analysis was performed to determine the independent risk factors for SIMD and the independent risk factors for death at 28 days after septic shock.The area under the receiver operating characteristic curve (AUC) was used to evaluate the efficacy of different indicators in predicting SIMD and the death outcome of children with septic shock on 28 days.Results:A total of 57 children were enrolled, including 28 cases in SIMD group and 29 cases in non-SIMD group.Univariate analysis showed that there were statistically significant differences in pediatric critical illness score, N-terminal B-type natriuretic peptide(NT-proBNP), LVEF and left ventricular short axis shortening rate between two groups ( P<0.05). Logistic analysis demonstrated that LVEF( OR=0.890, 95% CI 0.818-0.969, P=0.007)and NT-proBNP ( OR=1.000, 95% CI 1.000-1.000, P=0.015)could independently predict SIMD.There were 42 cases in survival group and 15 in non-survival group according to the prognosis on 28 days.Univariate analysis showed that there were significant differences in pediatric risk mortality score Ⅲ, pediatric sequential organ failure assessment, cardiac troponin I, and mitral annular plane systolic excursion(MAPSE)( P<0.05). Logistic analysis showed that only MAPSE independently predicted mortality( OR=85.670, 95% CI 1.685-4 356.736, P=0.026). Compared with MAPSE(AUC=0.727), MAPSE combined with pediatric risk mortality score Ⅲ, pediatric sequential organ failure assessment, cardiac troponin I(AUC=0.926) could be better to predict the 28 days prognosis of patients with septic shock on 28 days. Conclusion:NT-proBNP increases significantly in the early stage of SIMD.MAPSE shows no difference between SIMD and non-SIMD patients.MAPSE is correlated with the prognosis of patient with septic shock.

Objective: To analyze the short-term effect of individual atmospheric PM_2.5 exposure on the diversity, enterotype, and community structure of gut microbiome in healthy elderly people in Jinan, Shandong province. Methods: The present panel study recruited 76 healthy elderly people aged 60-69 years old in Dianliu Street, Lixia District, Jinan, Shandong Province, and followed them up five times from September 2018 to January 2019. The relevant information was collected by questionnaire, physical examination, precise monitoring of individual PM_2.5 exposure, fecal sample collection and gut microbiome 16S rDNA sequencing. The Dirichlet multinomial mixtures (DMM) model was used to analyze the enterotype. Linear mixed effect model and generalized linear mixed effect model were used to analyze the effect of PM_2.5 exposure on gut microbiome α diversity indices (Shannon, Simpson, Chao1, and ACE indices), enterotype and abundance of core species. Results: Each of the 76 subjects participated in at least two follow-up visits, resulting in a total of 352 person-visits. The age of 76 subjects was (65.0±2.8) years old with BMI (25.0±2.4) kg/m². There were 38 males accounting for 50% of the subjects. People with an educational level of primary school or below accounted for 10.5% of the 76 subjects, and those with secondary school and junior college or above accounting for 71.1% and 18.4%. The individual PM_2.5 exposure concentration of 76 subjects during the study period was (58.7±53.7) μg/m³. DMM model showed that the subjects could be divided into four enterotypes, which were mainly driven by Bacteroides, Faecalibacterium, Lachnospiraceae, Prevotellaceae, and Ruminococcaceae. Linear mixed effects model showed that different lag periods of PM_2.5 exposure were significantly associated with a lower gut α diversity index (FDR<0.05 after correction). Further analysis showed that PM_2.5 exposure was significantly associated with changes in the abundances of Firmicutes (Megamonas, Blautia, Streptococcus, etc.) and Bacteroidetes (Alistipes) (FDR<0.05 after correction). Conclusion: Short-term PM_2.5 exposure is significantly associated with a decrease in gut microbiome diversity and changes in the abundance of several species of Firmicutes and Bacteroidetes in the elderly. It is necessary to further explore the underlying mechanisms between PM_2.5 exposure and the gut microbiome, so as to provide a scientific basis for promoting the intestinal health of the elderly.
Aged ; Humans ; Male ; Middle Aged ; Feces/microbiology* ; Gastrointestinal Microbiome ; Particulate Matter ; RNA, Ribosomal, 16S/genetics* ; Female