Objective:To establish the reference intervals (RIs) of systemic immune inflammatory index (SII), platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), lymphocyte to monocyte ratio (LMR) and monocyte to lymphocyte ratio (MLR) in healthy pregnant women in Henan province, China.Methods:A retrospective analysis was conducted on the data of the healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from August 2016 to February 2019. A total of 4 016 healthy pregnant women were selected for establishing RIs. Data from healthy adult control group were derived from the healthy adult cohort in Henan established earlier by our team, and the Propensity Score Matching analysis was used and 3 595 healthy adult women and 3 595 healthy pregnant women to compare the indicators between the two groups. The RIs of the above indicators were established using the indirect method with a 95% confidence interval. The Tukey Rule was used to identify and remove outliers. The RIs were stratified and grouped based on the differences in each indicator during the pregnancy: SII: 3 929 cases, including 712 in the first trimester, 1 947 in the second trimester, and 1, 270 in the third trimester; PLR: 3 927 cases, no grouping; NLR: 3 925 cases, including 712 in the first trimester and 3 213 in the second and third trimesters; LMR: 3 925 cases, including 723 in the first trimester, 1 942 in the second trimester, and 1 260 in the third trimester; MLR: 3 904 cases, including 721 in the first trimester, 1 928 in the second trimester, and 1 255 in the third trimester. After the RIs were established, another 396 healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from February to April 2019 were selected for the validation of the RIs.Results:SII, NLR, LMR, MLR, and PLR differ significantly between healthy adult women and healthy pregnant women. There were significant differences in SII, LMR, and MLR among the three trimesters ( P<0.05). NLR in the first trimester was significantly lower than that in the second and third trimesters ( P<0.05), while there was no significant difference between the second and third trimester ( P=0.124). PLR only showed significant differences between the second and third trimester ( P<0.05), while no significant differences were found among the other groups. Based on the above results, the stratified RIs of each index in healthy pregnant population were established and verified. SII: first trimester (341-1 426)×10 9/L, second trimester (437-1 680)×10 9/L, third trimester (379-1 580)×10 9/L; PLR: 73-215; NLR: first trimester 1.78-5.60, second and third trimester 2.21-6.74; LMR: first trimester 2.20-6.61, second trimester 1.85-5.42, third trimester 1.63-4.82; MLR: first trimester 0.14-0.42, second trimester 0.17-0.49, third trimester 0.18-0.55. The rejection rate of 396 cases was less than 10%. Conclusions:The RIs of SII, NLR, LMR, MLR and PLR for healthy pregnant women in Hernan province of China were established and validated, and4 could be used in clinical practice.

Objective:To establish the reference intervals (RIs) of systemic immune inflammatory index (SII), platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), lymphocyte to monocyte ratio (LMR) and monocyte to lymphocyte ratio (MLR) in healthy pregnant women in Henan province, China.Methods:A retrospective analysis was conducted on the data of the healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from August 2016 to February 2019. A total of 4 016 healthy pregnant women were selected for establishing RIs. Data from healthy adult control group were derived from the healthy adult cohort in Henan established earlier by our team, and the Propensity Score Matching analysis was used and 3 595 healthy adult women and 3 595 healthy pregnant women to compare the indicators between the two groups. The RIs of the above indicators were established using the indirect method with a 95% confidence interval. The Tukey Rule was used to identify and remove outliers. The RIs were stratified and grouped based on the differences in each indicator during the pregnancy: SII: 3 929 cases, including 712 in the first trimester, 1 947 in the second trimester, and 1, 270 in the third trimester; PLR: 3 927 cases, no grouping; NLR: 3 925 cases, including 712 in the first trimester and 3 213 in the second and third trimesters; LMR: 3 925 cases, including 723 in the first trimester, 1 942 in the second trimester, and 1 260 in the third trimester; MLR: 3 904 cases, including 721 in the first trimester, 1 928 in the second trimester, and 1 255 in the third trimester. After the RIs were established, another 396 healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from February to April 2019 were selected for the validation of the RIs.Results:SII, NLR, LMR, MLR, and PLR differ significantly between healthy adult women and healthy pregnant women. There were significant differences in SII, LMR, and MLR among the three trimesters ( P<0.05). NLR in the first trimester was significantly lower than that in the second and third trimesters ( P<0.05), while there was no significant difference between the second and third trimester ( P=0.124). PLR only showed significant differences between the second and third trimester ( P<0.05), while no significant differences were found among the other groups. Based on the above results, the stratified RIs of each index in healthy pregnant population were established and verified. SII: first trimester (341-1 426)×10 9/L, second trimester (437-1 680)×10 9/L, third trimester (379-1 580)×10 9/L; PLR: 73-215; NLR: first trimester 1.78-5.60, second and third trimester 2.21-6.74; LMR: first trimester 2.20-6.61, second trimester 1.85-5.42, third trimester 1.63-4.82; MLR: first trimester 0.14-0.42, second trimester 0.17-0.49, third trimester 0.18-0.55. The rejection rate of 396 cases was less than 10%. Conclusions:The RIs of SII, NLR, LMR, MLR and PLR for healthy pregnant women in Hernan province of China were established and validated, and4 could be used in clinical practice.

BACKGROUND:Anterior cruciate ligament injury tends to lead to secondary meniscus injury and osteoarthritis.At present,there are few studies on the mechanics of meniscus and articular cartilage injury caused by anterior cruciate ligament injury. OBJECTIVE:To study the effect of partial rupture of the anterior cruciate ligament on the stress of medial and lateral meniscus and articular cartilage of knee joint by finite element analysis. METHODS:The CT and MRI images of the knee joint of a healthy volunteer were selected,and the scan data were imported into Mimics,Geomagic and SolidWorks software.After registration and fusion,four kinds of three-dimensional knee joint models were established:models of intact anterior cruciate ligament,rupture of the posterior external tract of anterior cruciate ligament,rupture of the anterior internal tract of anterior cruciate ligament,and absence of anterior cruciate ligament.Finally,data were imported into Ansys software to apply four different modes of loads to the knee joint:Longitudinal loads of 750 N were applied to the top of the femur;longitudinal load of 750 N to the top of the femur and forward thrust of 134 N behind tibia;a longitudinal load of 750 N and a varus moment of 10 Nm were applied to the top of the femur to simulate genu varus;750 N longitudinal load and 4 Nm internal rotation moment were applied to the proximal end of the femur to simulate knee internal rotation.The finite element analysis of biomechanical stress changes of the meniscus and articular cartilage of the knee joint was carried out. RESULTS AND CONCLUSION:(1)In the straight position of the knee joint,when the anterior medial tract of the anterior cruciate ligament was broken and the anterior cruciate ligament was missing under longitudinal loads of 750 N at the top of the femur,the total stress and peak value of meniscus increased significantly,but the stress distribution of the meniscus and the stress of articular cartilage did not change significantly.In longitudinal load of 750 N to the top of the femur and forward thrust of 134 N behind tibia,the fracture of the anterior internal tract of the anterior cruciate ligament increased the tibia forward,the compressive stress of posterior angle of the meniscus increased,and the stress of the articular cartilage did not change significantly.During simulating genu varus,the posterior angular stress of the lateral meniscus decreased,the stress of the medial meniscus increased,and the stress of articular cartilage slightly decreased when anterior cruciate ligament injuries were complete.When the anterior internal tract of the anterior cruciate ligament was broken or absent under knee internal rotation,the equivalent stress peak value of femoral cartilage and tibia cartilage shifted from medial cartilage to lateral cartilage,and the stress peak value of meniscus increased significantly.At this time,the anterior internal tract of the anterior cruciate ligament played a leading role in the rotational stability of the knee joint.(2)These results indicate that the risk of secondary meniscus injury in patients with anterior and medial anterior cruciate ligament band rupture was much higher than that in patients with posterior and external anterior cruciate ligament band rupture when the knee was in the upright standing position,varus and pronation,and there was no significant difference in the impact on articular cartilage.

Pyran- and furanocoumarins are key representatives of tetrahydropyrans and tetrahydrofurans, respectively, exhibiting diverse physiological and medical bioactivities. However, the biosynthetic mechanisms for their core structures remain poorly understood. Here we combined multiomics analyses of biosynthetic enzymes in Peucedanum praeruptorum and in vitro functional verification and identified two types of key enzymes critical for pyran and furan ring biosynthesis in plants. These included three distinct P. praeruptorum prenyltransferases (PpPT1-3) responsible for the prenylation of the simple coumarin skeleton 7 into linear or angular precursors, and two novel CYP450 cyclases (PpDC and PpOC) crucial for the cyclization of the linear/angular precursors into either tetrahydropyran or tetrahydrofuran scaffolds. Biochemical analyses of cyclases indicated that acid/base-assisted epoxide ring opening contributed to the enzyme-catalyzed tetrahydropyran and tetrahydrofuran ring refactoring. The possible acid/base-assisted catalytic mechanisms of the identified cyclases were theoretically investigated and assessed using site-specific mutagenesis. We identified two possible acidic amino acids Glu303 in PpDC and Asp301 in PpOC as vital in the catalytic process. This study provides new enzymatic tools in the epoxide formation/epoxide-opening mediated cascade reaction and exemplifies how plants become chemically diverse in terms of enzyme function and catalytic process.

Objective To investigate the levels of soluble tyrosine kinase-1(sFlt-1)and chemokine C-C ligand 3(CCL3)in serum of patients with coronary heart disease(CHD)and their correlation with the severity of the disease.Methods A total of 230 elderly CHD patients admitted to the De-partment of Cardiovascular Medicine of Xinxiang Central Hospital from November 2020 to No-vember 2022 were collected as the study subjects(CHD group),and according to their Gensini score,they were divided into mild(n=89),moderate(n=95),and severe(n=46)CHD sub-groups.Another 230 healthy individuals who taking physical examination during the same period served as the control group.ELISA was applied to measure serum levels of sFlt-1 and CCL3.ROC curve was plotted to analyze the diagnostic values of serum sFlt-1 and CCL3 levels for CHD.Pear-son correlation analysis was employed to analyze the relationship between serum sFlt-1 and CCL3 levels and the CHD severity.Results The serum levels of sFlt-1 and CCL3 were obviously higher in the CHD group than the control group(121.71±29.80 ng/L vs 98.70±17.57 ng/L,18.22± 5.41 ng/L vs 13.68±3.89 ng/L,P＜0.01).ROC curve analysis showed that the AUC value of the two indicators combined together was significantly greater than that of them alone in diagnosis of CHD(0.886 vs 0.791,0.775,P＜0.01).The serum levels of sFlt-1 and CCL3 were increased along with the severity of the disease and Gensini score when the levels and the score were compared among the mild,moderate and severe subgroups(P＜0.05).Pearson correlation analysis indicated that the serum levels of sFlt-1 and CCL3 were positively correlated with the Gensini score(r=0.420,r=0.479,P＜0.01).Conclusion The levels of serum sFlt-1 and CCL3 are obviously ele-vated in CHD patients,and closely associated with the severity of coronary lesions.

Objective:To investigate the expression profile of IL-36 family members in patients with coronary atherosclerotic heart disease (CAHD) and to assess the regulatory effects of exogenous IL-36 on CD8 + T cell function in CAHD patients. Methods:Twenty controls and 82 CAHD patients including 31 with stable angina pectoris (SAP), 27 with unstable angina pectoris (UAP) and 24 with acute myocardial infarction (AMI) were enrolled in this study. Anti-coagulant peripheral blood samples were collected. Plasma and peripheral blood mononuclear cells were isolated. The levels of IL-36α, IL-36β, IL-36γ and IL-36 receptor antagonist (IL-36RA) in plasma were measured by ELISA. CD8 + T cells were enriched. The expression of IL-36 receptor subunits at mRNA level was semi-quantified by real time PCR. Flow cytometry was used to detect the expression of programmed death-1 (PD-1), cytotoxic T lymphocytes associated protein-4(CTLA-4) and lymphocyte-activation gene-3 (LAG-3) in CD8 + T cells. Levels of periforin, granzyme B, granulysin, IFN-γ and TNF-α in the culture supernatants of CD8 + T cells were measured by ELISA. Purified CD8 + T cells from controls and AMI patients were stimulated with recombinant human IL-36RA. Changes in the expression of immune checkpoint molecules and the secretion of cytotoxic molecules and cytokines after IL-36RA stimulation were analyzed. One-way analysis of variance or paired t-test was used for statistical analysis. Results:There were no significant differences in plasma IL-36α, IL-36β or IL-36γ level between the control, SAP, UAP and AMI groups ( P>0.05). Plasma IL-36RA level was significantly down-regulated in the AMI group as compared with that in the control, SAP and UAP groups[(1 159.57±297.83) pg/ml vs (1 773.47±754.29) pg/ml, (1 600.12±740.48) pg/ml and (1 578.72±720.42) pg/ml; P<0.05]. The expression of IL-1 receptor 6 (IL-1R6) and IL-1 receptor accessory protein (IL-1RAcP) at mRNA level, the expression of PD-1 and CTLA-4, and the secretion of IFN-γ and TNF-α by CD8 + T cells showed no significant differences between the four groups ( P>0.05). Periforin, granzyme B and granulysin levels secreted by CD8 + T cells of the AMI group were significantly higherthan those of the control, SAP and UAP groups ( P<0.05). In the control group, recombinant human IL-36RA stimulation did not affect the expression of immune checkpoint molecule or the secretion of cytotoxic molecules and cytokines by CD8 + T cells ( P>0.05). In the AMI group, the percentage of PD-1 + CD8 + T cells increased after recombinant human IL-36RA stimulation ( P=0.033), but no significant change in the percentage of CTLA-4 + CD8 + T cells was observed ( P=0.288). Moreover, recombinant human IL-36RA stimulation suppressed the CD8 + T cells of AMI patients to secrete periforin, granzyme B and granulysin ( P<0.05), but not affect the secretion of IFN-γ and TNF-α ( P>0.05). Conclusions:The reduced IL-36RA level in AMI patients might induce the enhancement of CD8 + T cell activity by promoting CD8 + T cells to secrete cytotoxic molecules, which was involved in the immunopathogenesis of AMI.

There are a variety of post-transcriptional modifications in mRNA, which regulate the stability, splicing, translation, transport and other processes of mRNA, followed by affecting cell development, body immunity, learning and cognition and other important physiological functions. m⁶A modification is one of the most abundant post-transcriptional modifications widely existing in mRNA, regulating the metabolic activities of RNA and affecting gene expression. m⁶A modified homeostasis is critical for the development and maintenance of the nervous system. In recent years, m⁶A modification has been found in neurodegenerative diseases, mental diseases and brain tumors. This review summarizes the role of m⁶A methylation modification in the development, function and related diseases of the central nervous system in recent years, providing potential clinical therapeutic targets for neurological diseases.
Methylation ; Central Nervous System/metabolism* ; RNA, Messenger/metabolism* ; RNA

Objective:To investigate the clinical effect of medial plantar flap combined with free groin flap in the reconstruction of heel defect.Methods:The patients with heel skin and soft tissue defects admitted to the Department of Burns & Plastic and Hand & Foot Surgery of Yulin No.2 Hospital from October 2015 to December 2020 were retrospectively analyzed. After emergency debridement, a plantar medial island flap was used to repair the foot heel defect, a free groin flap was used to repair the medial plantar donor site, and the groin donor site was closed primarily. Postoperatively routine anti-infection, spasmolysis, anticoagulation, expanding treatment were performed after the procedure. The blood supply, survival of the flap, and the healing of the donor area of the flap were observed. The shape and function of the heel were observed in follow-up.Results:Eight patients were enrolled, including 7 males and 1 female, aged from 20 to 71 years, with an average of 32.2 years. There were 5 cases of heel trauma, 1 case of heel squamous cell carcinoma, 1 case of heel frostbite, and 1 case of heel ulcer. The wound area of the heel was 4 cm×3 cm-7 cm×6 cm. The surgical procedure was smooth, and the incision range of the heel island flap and groin flap was 0.5-1.0 cm larger than that of the heel wound. All 8 patients had primary healing after the operation. Follow-up for 3-12 months showed that all patients were satisfied with heel shape, sensory function and walking function. There was no depression, scar hyperplasia, and contracture in the medial plantar donor area, and no local skin ulcer. There is only a linear scar in the groin donor area.Conclusions:Medial plantar island flap combined with a free groin flap can repair the defect of the heel, and the affected foot has good healing, certain sensory function, and satisfactory curative effect.

Background The mechanisms of silicon dioxide (SiO₂)-induced inflammation and cell injury in pulmonary macrophages are not fully characterized. Objective To investigate the potential roles of inhibition of toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling in inflammation and macrophage polarization in mouse Raw264.7 cells in response to SiO₂ stimulation. Methods Sixteen 6- to 8-week-old C57BL/6 mice, half male and half female, were intratracheally instilled with 50 µL of SiO₂ (50 mg·mL⁻¹ in saline) or normal saline via oropharyngeal route, and the lungs of mice were harvested at 14 d and 28 d post the first challenge of SiO₂. HE staining of mouse lung was used for histopathological analysis. The expressions of TLR4 signaling-related proteins were detected by Western blotting (WB) and immunofluorescent (IF) assay, including TLR4, myeloid differentiation factor 88 (Myd88), and TNF receptor associated factor 6 (TRAF6). Raw264.7 cells were stimulated with SiO₂ (100 μg·cm²) for 12 h in absence or presence of TLR4 inhibitor M62812 for 13 h before the culture supernatants and cell lysates were harvested for analysis. The expressions of key components of TLR4 signaling cascade including TLR4, Myd88, and phosphorylated nuclear factor-kappa B P65 (P-NF-κB P65), P-1NF-kappa-B inhibitor α (P-1κbα), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6), M1 phenotype markers inducible nitric oxide synthase (iNOS) and cluster of differentiation 86 (CD86), as well as M2 phenotype arginase-1 (Arg-1) were accessed by WB and IF. The expressions of inflammation factors IL-6 and TNF-α in supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Results After SiO₂ intratracheal instillation for 14 d, the HE staining results showed obvious fibrotic nodules in the lung tissues of mice. The results of WB analysis revealed more abundant TLR4, Myd88, and TRAF6 in the silicosis mouse lung samples than in the controls. The results of IF assay showed an increased abundance of TLR4 and Myd88 proteins in the lung samples of silicosis mice at 14 d post the silica challenge, compared to the controls, indicating TLR4 signaling activation. As seen in the in vitro experiment, significant upregulations after the exposure to 100 μg·cm² SiO₂ were observed in TLR4 and P-1κbα at 6, 12, and 24 h (P<0.05); Myd88 at 12 and 24 h (P <0.05); and P-NF-κB P65 at 12 h (P<0.05). The inhibitor significantly suppressed the expressions of TLR4, Myd88, TRAF6, P-NF-κB P65, TNF-α, and IL-6 in Raw264.7 cells. In addition, the SiO₂-induced M1 phenotype marker iNOS was significantly suppressed, but the M2 phenotype marker Arg-1 was increased in the Raw264.7 cells. Conclusion The inhibition of TLR4/NF-κB signaling could result in a reduction of the inflammation response and the transition of M1 toward M2 phenotypes of macrophages in response to SiO₂ challenge.

Objective:To investigate the clinical effect of medial plantar flap combined with free groin flap in the reconstruction of heel defect.Methods:The patients with heel skin and soft tissue defects admitted to the Department of Burns & Plastic and Hand & Foot Surgery of Yulin No.2 Hospital from October 2015 to December 2020 were retrospectively analyzed. After emergency debridement, a plantar medial island flap was used to repair the foot heel defect, a free groin flap was used to repair the medial plantar donor site, and the groin donor site was closed primarily. Postoperatively routine anti-infection, spasmolysis, anticoagulation, expanding treatment were performed after the procedure. The blood supply, survival of the flap, and the healing of the donor area of the flap were observed. The shape and function of the heel were observed in follow-up.Results:Eight patients were enrolled, including 7 males and 1 female, aged from 20 to 71 years, with an average of 32.2 years. There were 5 cases of heel trauma, 1 case of heel squamous cell carcinoma, 1 case of heel frostbite, and 1 case of heel ulcer. The wound area of the heel was 4 cm×3 cm-7 cm×6 cm. The surgical procedure was smooth, and the incision range of the heel island flap and groin flap was 0.5-1.0 cm larger than that of the heel wound. All 8 patients had primary healing after the operation. Follow-up for 3-12 months showed that all patients were satisfied with heel shape, sensory function and walking function. There was no depression, scar hyperplasia, and contracture in the medial plantar donor area, and no local skin ulcer. There is only a linear scar in the groin donor area.Conclusions:Medial plantar island flap combined with a free groin flap can repair the defect of the heel, and the affected foot has good healing, certain sensory function, and satisfactory curative effect.