Background::Intrauterine growth restriction (IUGR) is associated with adverse metabolic outcomes during adulthood. Histone modifications and changes in DNA methylation-affected genes are important for fetal development. This study aimed to investigate the epigenetic mechanisms in IUGR.Methods::IUGR models were established in Sprague–Dawley rats using a maternal nutritional restriction approach during pregnancy. The abundance of insulin-like growth factor 2 (IGF2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 2 (AKT2), and peroxisome proliferators-activated receptor gamma coactivator 1 alpha (PGC-1α) was examined by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Chromatin immunoprecipitation RT-PCR was employed to analyze histone modification in CCCTC-binding factor (CTCF) 1–4 binding sites of the Igf2/H19 imprinting control region (ICR). The methylation states of CTCF1–4 binding sites were studied by pyrosequencing. Results::The IUGR models were constructed successfully. Igf2 mRNA abundance in the placenta, fetal liver, and newborn liver was decreased in the IUGR group ( P <0.01). Meanwhile, as compared with the control group, the expression levels of AKT2, PI3K, and PGC-1α were lower in newborn and 8-week-old livers in the IUGR group ( P <0.05). In addition, knocking down Igf2 reduced the protein expression levels of AKT2-phosphorylation and PGC-1α ( P <0.05). In CTCF binding sites 1-4 of the Igf2/ H19 ICR, acetylated histones H3 (AcH3) enrichment was significantly lower in CTCF1-3 in newborn and 8-week-old IUGR rats. Histone H3 tri-methylated lysine 4 (H3K4me3) enrichment was significantly lower in the CTCF1–4 of newborn and 8-week-old IUGR groups ( P <0.01). H3K9me2 enrichment was significantly higher in the IUGR group ( P <0.01). The CpG dinucleotide methylation levels of CTCF1 and CTCF3, but not those of CTCF2 and CTCF4 binding sites in IUGR rat fetal, 4-week old, and 8-week-old livers decreased significantly ( P <0.05). Conclusion::The methylation status and histone modification in the Igf2/H19 ICR are related to growth and lipid metabolism via the PGC-1α/PI3K/AKT2 pathway in IUGR rats.

Objective To investigate the expression and prognostic significance of serum protease C1 inhib-itor(SERPING1)and plasminogen activator inhibitor type 1(SERPINE1)in patients with sepsis.Methods A total of 132 patients with sepsis treated in the hospital from March 2018 to March 2020 were se-lected as the sepsis group.According to whether they died within 28 days of admission,they were divided into a death group(n=34)and a survival group(n=98).Enzyme linked immunosorbent assay was used to detect the expression of serum SERPING1 and SERPINE1.Multivariate Logistic regression model and receiver oper-ating characteristic curve were used to study the value of serum SERPING1 and serpine1 in evaluating the prognosis of patients'death.Results[Compared with the control group,serum SERPING1(331.12±51.80 ng/L vs.639.04±91.12 ng/L)was lower and serum serpine1(412.67±64.84 ng/L vs.42.33±10.32 ng/L)was higher in the sepsis group,and the differences were statistically significant(P＜0.05).[Compared to the survival group,the levels of serum SERPINE1,procalcitonin,C-reactive protein,Acute Physiology and Chronic Health Evaluation Ⅱ(APACHE Ⅱ)score and Sequential Organ Failure Assessment(SOFA)score in the death group were higher,while serum SERPING1 was lower,and the differences were statistically significant(all P＜0.05).Serum SERPING1 showed negative correlation with APACHE Ⅱ and SOFA scores(r=-0.779,-0.653,P＜0.05),while serum SERPINE1 showed positive correlation with APACHE Ⅱ and SO-FA scores(r=0.740,0.685,P＜0.05).APACHE Ⅱ score,SOFA score,and serum SERPINE1 were risk fac-tors affecting the prognosis of sepsis patients,while serum SERPING1 was a protective factor.The area under the curve of serum SERPING1 and SERPINE1 combined for the evaluation of the death in sepsis patients was 0.938(95％CI:0.893-0.968),which was significantly higher than 0.860(95％CI:0.812-0.899)and 0.838(95％CI:0.781-0.868)of the single detection,and the differences were statistically significant(Z=3.861,4.015,P＜0.001).Conclusion The elevated levels of serum SERPING1 and SERPINE1 in patients with sepsis are related to the severity of the patient's condition.The combination of the two has high prognos-tic value for sepsis patients.

Objective To investigate the efficacy and safety of camrelizumab monoclonal antibody combined with molecular-targeted therapy in elderly patients with unresectable or advanced hepatocellular carcinoma(HCC).Methods A retrospective analysis was performed for the patients with unresectable/advanced HCC who attended six hospitals from January 1,2019 to March 31,2021,and all patients received camrelizumab monoclonal antibody treatment,among whom 84.8%also received targeted therapy.According to the age of the patients,they were divided into elderly group(≥65 years)and non-elderly group(＜65 years).The two groups were assessed in terms of overall survival(OS),progression-free survival(PFS),objective response rate(ORR),disease control rate(DCR),and immune-related adverse events(irAE).The chi-square test or the Fisher's exact test was used for comparison of categorical data between groups;the independent samples t-test was used for comparison of normally distributed continuous data,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups.The Kaplan-Meier method was used for survival analysis,and the log-rank test was used for comparison of survival curves.Univariate and multivariate Cox proportional hazards regression analyses were used to determine the independent influencing factors for PFS and DCR at 6 months.Results A total of 99 HCC patients were enrolled,with 27 in the elderly group and 72 in the non-elderly group.The elderly group had an OS rate of 67.8%,an ORR of 44.4%,and a DCR of 74.1%at 12 months and a median PFS of 6.4(95%confidence interval[CI]:3.0-12.4)months,with no significant differences compared with the non-elderly group(all P＞0.05).The median OS was unavailable for the elderly group,while the non-elderly group had an OS of 18.9(95%CI:13.0-24.8)months;there was no significant difference between the two groups(P=0.485).The univariate and multivariate Cox regression analyses showed that major vascular invasion(MVI)was an independent risk factor for PFS(hazard ratio[HR]=2.603,95%CI:1.136-5.964,P=0.024)and DCR(HR=3.963,95%CI:1.671-9.397,P=0.002)at 6 months,while age,sex,etiology of HBV infection,presence of extrahepatic metastasis,Child-Pugh class B,and alpha-fetoprotein＞400 ng/mL were not associated with PFS or DCR at 6 months.For the elderly group,the incidence rates of any irAE and grade 3/4 irAE were 51.9%and 25.9%,respectively,with no significant differences compared with the non-elderly group(P＞0.05),and skin disease was the most common irAE in both groups(39.4%).Conclusion Camrelizumab monoclonal antibody combined with molecular-targeted therapy has similar efficacy and safety in patients with unresectable/advanced HCC aged≥65 years and those aged＜65 years.MVI is associated with suboptimal response to immunotherapy and poor prognosis.

Background/Aims: Total proctocolectomy with ileal pouch-anal anastomosis (IPAA) is widely accepted as a radical surgery for refractory ulcerative colitis (UC). Definite results on the appropriate pouch length for an evaluation of the risk-to-benefit ratio regarding technical complications and long-term quality of life (QOL) are still scarce. Methods: Data on UC patients who underwent IPAA from 2008 to 2022 in four well-established pouch centers affiliated to China UC Pouch Center Union were collected. Results: A total of 208 patients with a median follow-up time of 6.0 years (interquartile range, 2.3 to 9.0 years) were enrolled. The median lengths of the patients’ short and long pouches were 14.0 cm (interquartile range, 14.0 to 15.0 cm) and 22.0 cm (interquartile range, 20.0 to 24.0 cm), respectively. Patients with a short J pouch configuration were less likely to achieve significantly improved long-term QOL (p=0.015) and were prone to develop late postoperative complications (p=0.042), such as increased defecation frequency (p=0.003) and pouchitis (p=0.035). A short ileal pouch was an independent risk factor for the development of late postoperative complications (odds ratio, 3.100; 95% confidence interval, 1.519 to 6.329; p=0.002) and impaired longterm QOL improvement (odds ratio, 2.221; 95% confidence interval, 1.218 to 4.050, p=0.009). Conclusions The length of the J pouch was associated with the improvement in long-term QOL and the development of late post-IPAA complications. A long J pouch configuration could be a considerable surgical option for pouch construction.

Skin avulsion injuries of the lower leg are common in clinical practice, which can easily lead to skin necrosis and infection of the lower leg, and have a significant impact on the appearance and function of the lower limb. Among them, the open avulsion injury has the highest incidence and is often accompanied by multiple tissue injuries. Therefore, improper diagnosis and treatment may cause skin and soft tissue defects of the lower leg combined with bone exposure or even bone defects, seriously impairing patients′ life and health. In order to have a better understanding of the open skin avulsion injury of the lower leg, achieve precise diagnosis and treatment and improve prognosis, the Chinese Society of Traumatology of Chinese Medical Association and the Chinese Association of Microsurgeons of Chinese Medical Doctor Association organized experts in the related fields to formulate Clinical guideline for the diagnosis and treatment of open skin avulsion injuries of the lower leg ( version 2024) based on evidence-based medicine principles. A total of 16 recommendations were proposed on the diagnosis, treatment, postoperative rehabilitation of open lower leg skin avulsion injury, so as to provide a reference for its diagnosis and treatment.

BACKGROUND: Intrauterine growth restriction (IUGR) is associated with adverse metabolic outcomes during adulthood. Histone modifications and changes in DNA methylation-affected genes are important for fetal development. This study aimed to confirm the epigenetic mechanisms in IUGR. METHODS: IUGR models were established in Sprague-Dawley rats using a maternal nutritional restriction approach during pregnancy. The abundance of insulin-like growth factor 2 (IGF2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 2 (AKT2), and PPAR gamma coactivator 1 alpha (PGC-1α) was examined by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Chromatin immunoprecipitation RT-PCR was employed to analyze histone modification in CCCTC-binding factor (CTCF)1-4 binding sites of the IGF2/H19 imprinting control region (ICR). The methylation states of CTCF1-4 binding sites were studied by pyrosequencing. RESULTS: The IUGR models were constructed successfully. IGF2 mRNA abundance in the placenta, fetal liver, and newborn liver was decreased in the IUGR group (P <0.01). Meanwhile, as compared with the control group, the expression levels of AKT2, PI3K, and PGC-1α were lower in newborn and 8-week-old livers in the IUGR group (P <0.05). In addition, knocking down IGF2 reduced the protein expression levels of AKT2-P and PGC-1α (P <0.05). In CTCF binding sites 1-4 of the IGF2/H19 ICR, AcH3 enrichment was significantly lower in CTCF1-3 in newborn and 8-week-old IUGR rats. H3K4me3 enrichment was significantly lower in the CTCF1-4 of newborn and 8-week-old IUGR groups (P <0.01). H3K9me2 enrichment was significantly higher in the IUGR group (P <0.01). The CpG dinucleotide methylation levels of CTCF1 and CTCF3, but not those of CTCF2 and CTCF4 binding sites in IUGR rat fetal, 4-week old, and 8-week-old livers decreased significantly (P <0.05). CONCLUSION The methylation status and histone modification in the IGF2/H19 ICR are related to growth and lipid metabolism via the PGC-1α/PI3K/AKT2 pathway in IUGR rats.

Skin avulsion injuries of the lower leg are common in clinical practice, which can easily lead to skin necrosis and infection of the lower leg, and have a significant impact on the appearance and function of the lower limb. Among them, the open avulsion injury has the highest incidence and is often accompanied by multiple tissue injuries. Therefore, improper diagnosis and treatment may cause skin and soft tissue defects of the lower leg combined with bone exposure or even bone defects, seriously impairing patients′ life and health. In order to have a better understanding of the open skin avulsion injury of the lower leg, achieve precise diagnosis and treatment and improve prognosis, the Chinese Society of Traumatology of Chinese Medical Association and the Chinese Association of Microsurgeons of Chinese Medical Doctor Association organized experts in the related fields to formulate Clinical guideline for the diagnosis and treatment of open skin avulsion injuries of the lower leg ( version 2024) based on evidence-based medicine principles. A total of 16 recommendations were proposed on the diagnosis, treatment, postoperative rehabilitation of open lower leg skin avulsion injury, so as to provide a reference for its diagnosis and treatment.

Background and Objectives: Although human-induced pluripotent stem cells (hiPSC) can be efficiently differentiated into cardiomyocytes (CMs), the heterogeneity of the hiPSC-CMs hampers their applications in research and regenerative medicine. Retinoic acid (RA)-mediated signaling pathway has been proved indispensable in cardiac development and differentiation of hiPSC toward atrial CMs. This study was aimed to test whether RA signaling pathway can be manipulated to direct the differentiation into sinoatrial node (SAN) CMs. Methods: and Results: Using the well-characterized GiWi protocol that cardiomyocytes are generated from hiPSC via temporal modulation of Wnt signaling pathway by small molecules, RA signaling pathway was manipulated during the differentiation of hiPSC-CMs on day 5 post-differentiation, a crucial time point equivalent to the transition from cardiac mesoderm to cardiac progenitor cells in cardiac development. The resultant CMs were characterized at mRNA, protein and electrophysiology levels by a combination of qPCR, immunofluorescence, flow cytometry, and whole-cell patch clamp. The results showed that activation of the RA signaling pathway biased the differentiation of atrial CMs, whereas inhibition of the signaling pathway biased the differentiation of sinoatrial node-like cells (SANLCs). Conclusions Our study not only provides a novel and simple strategy to enrich SANLCs but also improves our under-standing of the importance of RA signaling in the differentiation of hiPSC-CMs.

Objective:To assess the association of hemoglobin (Hb) levels with the prevalence of diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM).Methods:A cross-sectional study. From January 2017 to December 2018, 707 patients with T2DM who were hospitalized in the Department of Internal Medicine of Chu Hsien-I Memorial Hospital & Tianjin Medical University, were included in the study. All patients underwent color photography of the fundus of both eyes with dilated pupils. According to DR diagnostic criteria, patients were divided into DR group and non-DR (NDR) group, with 210 and 497 cases, respectively; DR group was further divided into non-proliferative DR group (NPDR) group and proliferative DR (PDR) group, about 186, 24 cases, respectively. Hb level was detected, single factor analysis of its correlation with DR; logistic regression analysis was used to evaluate the relationship between Hb level and DR risk.Results:The Hb levels of the patients in the NDR group and the DR group were 140.58±17.26 and 132.35±23.48 g/dl; compared with the NDR group, the Hb level of the DR group was significantly lower, and the difference was statistically significant ( t=5.107, P=0.000). In the NDR group, NPDR group, and PDR group, Hb levels of male patients were 149.3±1.01, 142.6±2.35, 132.9±8.44 g/dl, respectively; Hb levels of female patients were 131.7±0.90, 124.0±2.09, 116.8±5.23 g/dl. With the progress of DR, Hb levels of different sexes decreased significantly, and the difference was statistically significant ( P<0.000 1). The results of correlation analysis showed that Hb reduction was an independent risk factor for DR (odds ratio=4.437, 95% confidence interval 2.590-7.603, P<0.000 1). Conclusion:The reduction of Hb in T2DM patients is positively correlated with the severity of DR.

Background and Objectives: Manipulating different signaling pathways via small molecules could efficiently inducecardiomyocytes from human induced pluripotent stem cells (hiPSC). However, the effect of transcription factors on the hiPSC-directed cardiomyocytes differentiation remains unclear. Transcription factor, p53 has been demonstrated indispensable for the early embryonic development and mesendodermal differentiation of embryonic stem cells (ESC).We tested the hypothesis that p53 promotes cardiomyocytes differentiation from human hiPSC. Methods: and Results: Using the well-characterized GiWi protocol that cardiomyocytes are generated from hiPSC via temporal modulation of Wnt signaling pathway by small molecules, we demonstrated that forced expression of p53 in hiPSC remarkably improved the differentiation efficiency of cardiomyocytes from hiPSC, whereas knockdown endogenous p53 decreased the yield of cardiomyocytes. This p53-mediated increased cardiomyocyte differentiation was mediated through WNT3, as evidenced by that overexpression of p53 upregulated the expression of WNT3, and knockdown of p53 decreased the WNT3 expression. Mechanistic analysis showed that the increased cardiomyocyte differentiation partially depended on the amplified mesendodermal specification resulted from p53-mediated activation of WNT3-mediated Wnt signaling. Consistently, endogenous WNT3 knockdown significantly ameliorated mesendodermal specification and subsequent cardiomyocyte differentiation. Conclusions These results provide a novel insight into the potential effect of p53 on the development and differentiation of cardiomyocyte during embryogenesis.