Background and aims Hepatocellular carcinoma (HCC) cells are metabolically reprogrammed for excessive uptake and metabolism of many nutrients. The tumor suppressive microRNA-148a-3p (miR-148a-3p) is downregulated in HCC, whereas its function in regulating HCC cell metabolism remains obscure. Herein we aimed to delineate the role of miR-148a-3p in HCC cell metabolism by using novel bioengineered miR-148a-3p (BioRNALeu/miR-148a-3p) agent produced in vivo. Methods BioRNALeu/miR-148a-3p was designed by using human leucyl transfer RNA fused hsa-pre-miR-34a carrier, overexpressed in Escherichia coli (E. coli), and purified to high homogeneity. After transfection into HCC cells, the released miR-148a-3p levels were assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell proliferation was determined by CellTiter-Glo assays. Targets were validated by dual-luciferase reporter assays, immunoblotting, and immunofluorescence confocal imaging. Glycolysis capacity was evaluated by Seahorse XF assays, and glucose, lactate, and amino acid levels were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. Results BioRNALeu/miR-148a-3p was efficiently processed into target miR-148a-3p in HCC cells to effectively inhibit cell proliferation in a dose- and time-dependent manner. Mechanistically, miR-148a-3p suppressed the protein levels of glucose transporter GLUT1/SLC2A1 and L-type amino acid transporter LAT1/SLC7A5 via acting on their 3′-untranslated regions, as well as amino acid transporter ASCT2/SLC1A5. These, in turn, led to a reduction of glucose uptake, lactate production, and glycolytic flux in HCC cells, and alteration of intracellular amino acid metabolome including glutamine, leucine, phenylalanine, tyrosine, and methionine. Conclusions Reintroduction of miR-148a-3p into HCC cells modulates glucose and amino acid metabolism via regulating multiple SLC transporters, thereby suppressing HCC cell viability. These findings highlight the role of miR-148a-3p in HCC cell metabolism and potential of bioengineered miRNA molecules for functional studies and therapeutic development.

Objective:To investigate whether hysteroscopic transcervical resection of septum (TCRS) prior to in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) can improve cumulative live birth rates in patients with uterine septum. Methods:A retrospective cohort study was conducted to analyze data from 244 patients with partial uterine septum who underwent IVF/ICSI at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University between January 2016 and August 2022. The patients were divided into a surgical group ( n=171) and a non-surgical group ( n=73) based on whether TCRS was performed prior to IVF/ICSI. The clinical outcomes of IVF/ICSI in the two groups were analyzed, with the primary observation indicator being the cumulative live birth rate. Cox regression analysis was employed to identify determinants. Results:The age of patients in the operated group [(31.20±3.80) years] was younger than that in the non-operated group [(32.92±5.34) years, P=0.005], and the basal antral follicle count [17.0 (11.0, 24.0)] was higher than that in the non-operated group [14.0 (8.0, 21.5), P=0.039]. There were no significant differences in other baseline data (all P>0.05). The cumulative pregnancy rate [79.53% (136/171)] and the cumulative live birth rate [60.23% (103/171)] in the operated group during the 24-month follow-up period were significantly higher than those in the non-operated group [65.75% (48/73), P=0.022; 45.21% (33/73), P=0.030]. Compared with the operated group [296.0 (260.0, 430.0) d], the duration from the start of ovarian stimulation to the first live birth was significantly prolonged in the non-operated group [379.0 (329.5, 471.5) d, P<0.001]. Adjusted Cox-regression analysis showed that whether or not surgery was performed ( HR=1.683, 95% CI: 1.116-2.539, P=0.013) and the basal antral follicle count ( HR=1.032, 95% CI: 1.000-1.065, P=0.048) were independent factors affecting cumulative live birth rate. Conclusion:Performing TCRS before IVF/ICSI can improve cumulative live birth rates of patients with uterine septum.

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

Objective To explore the mediating role of activities of daily living (ADL) in pain and depressive symptoms in the elderly in China. Methods Utilizing the data from 2020 China Health and Retirement Longitudinal Study, 4403 Chinese elderly individuals aged ≥ 60 years old were selected as the research subjects. Depression Scale (CES-D 10) of the Center for Epidemiological Survey and ADL scale were used in the study. The PROCESS4.1 macro was used to test the mediating effect of daily living activities between pain and depressive symptoms, and the Bootstrap method was applied for verification of the mediating variables. Results A total of 2368 cases of depressive symptoms were detected in the elderly in China, with a detection rate of 53.78%. Pain was positively correlated with depressive symptoms (r=0.27, P<0.01), and activities of daily living were negatively correlated with pain and depressive symptoms (r=-0.27, -0.337, P<0.01). The results showed that the total effect value of pain on depressive symptoms was 0.33, the direct effect value was 0.24, and the mediating effect value of daily living activities was 0.09, accounting for 27.27%. Conclusion Pain and activities of daily living are important factors influencing depressive symptoms in the elderly, and activities of daily living play a partial mediating role in the relationship between pain and depressive symptoms in the elderly.

Objective:To investigate whether hysteroscopic transcervical resection of septum (TCRS) prior to in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) can improve cumulative live birth rates in patients with uterine septum. Methods:A retrospective cohort study was conducted to analyze data from 244 patients with partial uterine septum who underwent IVF/ICSI at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University between January 2016 and August 2022. The patients were divided into a surgical group ( n=171) and a non-surgical group ( n=73) based on whether TCRS was performed prior to IVF/ICSI. The clinical outcomes of IVF/ICSI in the two groups were analyzed, with the primary observation indicator being the cumulative live birth rate. Cox regression analysis was employed to identify determinants. Results:The age of patients in the operated group [(31.20±3.80) years] was younger than that in the non-operated group [(32.92±5.34) years, P=0.005], and the basal antral follicle count [17.0 (11.0, 24.0)] was higher than that in the non-operated group [14.0 (8.0, 21.5), P=0.039]. There were no significant differences in other baseline data (all P>0.05). The cumulative pregnancy rate [79.53% (136/171)] and the cumulative live birth rate [60.23% (103/171)] in the operated group during the 24-month follow-up period were significantly higher than those in the non-operated group [65.75% (48/73), P=0.022; 45.21% (33/73), P=0.030]. Compared with the operated group [296.0 (260.0, 430.0) d], the duration from the start of ovarian stimulation to the first live birth was significantly prolonged in the non-operated group [379.0 (329.5, 471.5) d, P<0.001]. Adjusted Cox-regression analysis showed that whether or not surgery was performed ( HR=1.683, 95% CI: 1.116-2.539, P=0.013) and the basal antral follicle count ( HR=1.032, 95% CI: 1.000-1.065, P=0.048) were independent factors affecting cumulative live birth rate. Conclusion:Performing TCRS before IVF/ICSI can improve cumulative live birth rates of patients with uterine septum.

Several types of arthritis share the common feature that the generation of inflammatory mediators leads to joint cartilage degradation. However, the shared mechanism is largely unknown. H2BK120ub1 was reportedly involved in various inflammatory diseases but its role in the shared mechanism in inflammatory joint conditions remains elusive. The present study demonstrated that levels of cartilage degradation, H2BK120ub1, and its regulator WW domain-containing adapter protein with coiled-coil (WAC) were increased in cartilage in human rheumatoid arthritis (RA) and osteoarthritis (OA) patients as well as in experimental RA and OA mice. By regulating H2BK120ub1 and H3K27me3, WAC regulated the secretion of inflammatory and cartilage-degrading factors. WAC influenced the level of H3K27me3 by regulating nuclear entry of the H3K27 demethylase KDM6B, and acted as a key factor of the crosstalk between H2BK120ub1 and H3K27me3. The cartilage-specific knockout of WAC demonstrated the ability to alleviate cartilage degradation in collagen-induced arthritis (CIA) and collagenase-induced osteoarthritis (CIOA) mice. Through molecular docking and dynamic simulation, doxercalciferol was found to inhibit WAC and the development of cartilage degradation in the CIA and CIOA models. Our study demonstrated that WAC is a key factor of cartilage degradation in arthritis, and targeting WAC by doxercalciferol could be a viable therapeutic strategy for treating cartilage destruction in several types of arthritis.

Objective To investigate renal expression level of STING in mice with renal ischemia-reperfusion injury(IRI)and its regulatory role in IRI.Methods C57BL/6 mice were divided into sham operation group,IRI(induced by clamping the renal artery)model group,IRI+DMSO treatment group,and IRI+SN-011 treatment group.Serum creatinine and blood urea nitrogen of the mice were analyzed,and pathological changes in the renal tissue were assessed with PAS staining.RT-qPCR,ELISA,Western blotting,and immunohistochemistry were used to detect the expression levels of STING,KIM-1,Bcl-2,Bax,caspase-3,TLR4,P65,NLRP3,caspase-1,CD68,MPO,IL-1β,IL-6,and TNF-α in the renal tissues.In the cell experiment,HK-2 cells exposed to hypoxia-reoxygenation(H/R)were treated with DMSO or SN-011,and cellular STING expression levels and cell apoptosis were analyzed using RT-qPCR,Western blotting or flow cytometry.Results In C57BL/6 mice,renal IRI induced obvious renal tissue damage,elevation of serum creatinine and blood urea nitrogen levels and renal expression levels of KIM-1,STING,TLR4,P65,NLRP3,caspase-1,caspase-3,Bax,CD68,MPO,IL-1β,IL-6,and TNF-α,and reduction of Bcl-2 expression level.Treatment of the mouse models with SN-011 for inhibiting STING expression significantly alleviated these changes.In HK-2 cells,H/R exposure caused significant elevation of cellular STING expression and obviously increased cell apoptosis rate,which was significantly lowered by treatment with SN-011.Conclusion Renal STING expression is elevated in mice with renal IRI to exacerbate renal injury by regulating the TLR4/NF-κB/NLRP3 pathway and promoting inflammation and apoptosis in the renal tissues.

Objective To investigate renal expression level of STING in mice with renal ischemia-reperfusion injury(IRI)and its regulatory role in IRI.Methods C57BL/6 mice were divided into sham operation group,IRI(induced by clamping the renal artery)model group,IRI+DMSO treatment group,and IRI+SN-011 treatment group.Serum creatinine and blood urea nitrogen of the mice were analyzed,and pathological changes in the renal tissue were assessed with PAS staining.RT-qPCR,ELISA,Western blotting,and immunohistochemistry were used to detect the expression levels of STING,KIM-1,Bcl-2,Bax,caspase-3,TLR4,P65,NLRP3,caspase-1,CD68,MPO,IL-1β,IL-6,and TNF-α in the renal tissues.In the cell experiment,HK-2 cells exposed to hypoxia-reoxygenation(H/R)were treated with DMSO or SN-011,and cellular STING expression levels and cell apoptosis were analyzed using RT-qPCR,Western blotting or flow cytometry.Results In C57BL/6 mice,renal IRI induced obvious renal tissue damage,elevation of serum creatinine and blood urea nitrogen levels and renal expression levels of KIM-1,STING,TLR4,P65,NLRP3,caspase-1,caspase-3,Bax,CD68,MPO,IL-1β,IL-6,and TNF-α,and reduction of Bcl-2 expression level.Treatment of the mouse models with SN-011 for inhibiting STING expression significantly alleviated these changes.In HK-2 cells,H/R exposure caused significant elevation of cellular STING expression and obviously increased cell apoptosis rate,which was significantly lowered by treatment with SN-011.Conclusion Renal STING expression is elevated in mice with renal IRI to exacerbate renal injury by regulating the TLR4/NF-κB/NLRP3 pathway and promoting inflammation and apoptosis in the renal tissues.

Objective:To explore the effect of body fat percent (BFP) on assisted reproductive outcomes in infertile women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) and fresh embryo transfer. Methods:We analyzed clinical data on infertile women in a retrospective cohort study, who underwent IVF/ICSI and embryo transfer at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from March 2022 to October 2023. The study subjects were divided into non-obese group (BFP<35%, 800 cases) and obese group (BFP≥35%, 742 cases) according to BFP. The baseline data, ovulation induction outcomes and clinical pregnancy outcomes were compared between the two groups.Results:1) The body mass index [BMI, 25.85 (24.22, 28.04) kg/m 2], basal testosterone [0.80 (0.45, 1.12) nmol/L], triglyceride [1.29 (1.03, 1.59) mmol/L], serum total cholesterol [4.55 (4.29, 4.81) mmol/L], low-density lipoprotein cholesterol [3.17 (2.90, 3.40) mmol/L], fasting glucose [5.40 (5.10, 5.75) mmol/L], fasting insulin [12.99 (9.01, 18.31) mU/L], homeostasis model assessment-insulin resistance [3.09 (2.14, 4.50)], antral follicle count [16.00 (11.00, 22.00)], the patients combined with polycystic ovary syndrome [9.4% (70/742)] in obese group were significantly higher than those in non-obese group [21.94 (20.32, 23.51) kg/m 2, P<0.001; 0.69 (0.43, 0.98) nmol/L, P<0.001; 1.00 (0.79, 1.21) mmol/L, P<0.001; 4.42 (4.19, 4.66) mmol/L, P<0.001; 2.91 (2.67, 3.15) mmol/L, P<0.001; 5.22 (5.00, 5.45) mmol/L, P<0.001; 11.30 (8.33, 14.82) mU/L, P<0.001; 2.61 (1.86, 3.48), P<0.001; 14.00 (10.00, 20.00) mmol/L, P<0.001; 4.8% (38/800), P<0.001]. Basal follicle-stimulating hormone [6.58 (5.64, 7.73) U/L], basal estradiol [133.01 (102.35, 171.56) pmol/L], basal luteinizing hormone [4.80 (3.62, 6.53) U/L] and high-density lipoprotein cholesterol [1.29 (1.17, 1.39) mmol/L] in obese group were significantly lower than those in non-obese group [6.91 (5.86, 8.33) U/L, P<0.001; 145.52 (105.23, 187.95) pmol/L, P=0.001; 5.16 (3.82, 6.94) U/L, P=0.022; 1.45 (1.36, 1.55) mmol/L, P<0.001]. 2) The initiated dosage of gonadotropin (Gn) used [187.50 (150.00, 225.00) U] and the total dosage of Gn used [2 481.25 (1 856.25, 3 225.00) U] in obese group were significantly higher than those in non-obese group [225.00 (175.00, 250.00) U, P<0.001; 2 925.00 (2 250.00, 3 675.00) U, P<0.001]. Serum estradiol level on the day of human chorionic gonadotropin (hCG) injection [8 984.00 (6 087.75, 11 978.25) pmol/L], luteinizing hormone level on the day of hCG injection [1.23 (0.87, 1.79) U/L], the rate of blastocyst formation [55.56% (33.33%, 75.00%)] in obese group were significantly lower than those in non-obese group [9 378.50 (6 528.50, 12 624.50) pmol/L, P=0.016; 1.37 (0.94, 2.01) U/L, P=0.001; 60.00% (37.86%, 80.00%), P=0.014]. 3) Sustained pregnancy rate in obese group [41.5% (308/742)] was lower than that in non-obese group [47.6% (381/800), P=0.016]. The number of embryo transfer, embryo transfer type, implantation rate, clinical pregnancy rate, abortion rate and live birth rate were not statistically significant between the two groups (all P>0.05). 4) BFP was not an independent factor of clinical pregnancy rate and live birth rate (all P>0.05). 5) The area under the curve (AUC) of BMI and BFP to predict clinical pregnancy was 0.509 and 0.518, and there was no significant difference between them (AUC difference=0.009, 95% CI: -0.010-0.028, P=0.376). The AUC of BMI and BFP for predicting live birth was 0.501 and 0.513, with no statistically significant difference (AUC difference=0.012, 95% CI: -0.007-0.030, P=0.221). Conclusion:High BFP can increase total dosage of Gn used, decrease blastocyst formation rate and continuous pregnancy rate.

Objective:To investigate the factors influencing patients' willingness to have a second child after obtaining their first singleton live birth by in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). Methods:The data of patients who achieved their first single live birth via IVF/ICSI at the Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University were collected between July 2016 and July 2021 in a retrospective cohort study. Until July 2023, the patients were divided into the returning group ( n=1 809) and the non-returning group ( n=5 824) according to whether they returned again to receive assisted reproduction treatment for the birth of a second child. The clinical data of the two groups were compared, and the factors affecting patients' willingness to have a second child were analyzed. Results:The characteristics of the two groups of patients were compared. Variables that were statistically significant after being included in the univariate analysis were employed to construct a multivariate logistic regression equation. The results indicated that the age of the female ( OR=0.93, 95% CI: 0.91-0.94, P<0.001), the duration of infertility ( OR=0.96, 95% CI: 0.93-0.98, P=0.002), male+female factors of infertility factor ( OR=0.82, 95% CI: 0.70-0.97, P=0.023) the previous reproductive history ( OR=0.33, 95% CI: 0.27-0.42, P<0.001), the method of assisted pregnancy ( OR=1.31, 95% CI: 1.11-1.55, P=0.001), the transplantation plan ( OR=0.83, 95% CI: 0.74-0.93, P=0.002), the mode of delivery ( OR=0.63, 95% CI: 0.55-0.71, P<0.001), pregnancy complications ( OR=0.70, 95% CI: 0.60-0.82, P<0.001), the presence or absence of remaining embryos ( OR=2.67, 95% CI: 2.24-3.19, P<0.001), the gender of the first live birth ( OR=0.40, 95% CI: 0.36-0.45, P<0.001), the degree of education ( OR=0.74, 95% CI: 0.64-0.85, P<0.001), and the type of household registration ( OR=0.74, 95% CI: 0.65-0.84, P<0.001) were the influencing factors of the second-child fertility intention of patients after obtaining the first singleton live birth through IVF/ICSI. Conclusion:The age of the female, the duration of infertility, the factors of infertility, the previous reproductive history, frozen-thawed embryo transfer, cesarean section, having pregnancy complications, the gender of the first live birth being a boy, having a high school education or above, and urban household registration are negatively correlated with the patients' return visits. ICSI-assisted pregnancy and having remaining embryos are positively correlated with the patients' return visits.