BACKGROUND: Fibrosis of the connective tissue in the vaginal wall predominates in pelvic organ prolapse (POP), which is characterized by excessive fibroblast-to-myofibroblast differentiation and abnormal deposition of the extracellular matrix (ECM). Our study aimed to investigate the effect of ECM stiffness on vaginal fibroblasts and to explore the role of methyltransferase 3 (METTL3) in the development of POP. METHODS: Polyacrylamide hydrogels were applied to create an ECM microenvironment with variable stiffness to evaluate the effects of ECM stiffness on the proliferation, differentiation, and expression of ECM components in vaginal fibroblasts. METTL3 small interfering RNA and an overexpression vector were transfected into vaginal fibroblasts to evaluate the effects of METTL3 silencing and overexpression on matrix stiffness-induced vaginal fibroblast-to-myofibroblast differentiation and abnormal modulation of the ECM. Both procedures were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining, Western blotting (WB), quantitative real-time polymerase chain reaction (RT-qPCR), and immunofluorescence (IF). RESULTS: Vaginal fibroblasts from POP patients exhibited increased proliferation ability, increased expression of α-smooth muscle actin (α-SMA), decreased expression of collagen I/III, and significantly decreased expression of tissue inhibitors of matrix metalloproteinases (TIMPs) in the stiff matrix ( P <0.05). Compared with those from non-POP patients, vaginal wall tissues from POP patients demonstrated a significant increase in METTL3 content ( P <0.05). However, silencing METTL3 expression in vaginal fibroblasts with high ECM stiffness resulted in decreased proliferation ability, decreased α-SMA expression, an increased ratio of collagen I/III, and increased TIMP1 and TIMP2 expression. Conversely, METTL3 overexpression significantly promoted the process of increased proliferation ability, increased α-SMA expression, decreased ratio of collagen I/III and decreased TIMP1 and TIMP2 expression in the soft matrix ( P <0.05). CONCLUSIONS Elevated ECM stiffness can promote excessive proliferation, differentiation, and abnormal ECM modulation, and the expression of METTL3 plays an important role in alleviating or aggravating matrix stiffness-induced vaginal fibroblast-to-myofibroblast differentiation and abnormal ECM modulation.
Humans ; Female ; Extracellular Matrix/metabolism* ; Cell Differentiation/genetics* ; Methyltransferases/metabolism* ; Pelvic Organ Prolapse/pathology* ; Fibroblasts/metabolism* ; Myofibroblasts/metabolism* ; Vagina/metabolism* ; Cell Proliferation/physiology* ; Cells, Cultured ; Middle Aged

OBJECTIVE To determ ine the contents of main active constituents in Gynura divaricata leaf with different drying methods and at different harvesting time ，so as to confirm the best drying method and harvesting time. METHODS G. divaricata leaf with different drying methods [drying in the shade ，drying in the sun ，oven drying （60℃，70℃，80℃），microwave drying and freeze drying] and different harvesting time （March to October ）were prepared. The content of water-soluble extract was determined by hot dip method. The contents of total flavonoids and polysaccharides were determined by ultraviolet-visible spectrophotometry. The content of astragalin was determined by HPLC. Analytic hierarchy process was used for comprehensive analysis. RESULTS The time of drying in the shade ，drying in the sun ，drying at 60 ℃，drying at 70 ℃，drying at 80 ℃， microwave drying and freeze drying were 7 d，5 d，8 h，5 h，3.5 h，6 min and 1 d respectively. The average contents of water-soluble extract in G. divaricata leaf were 55.98％，60.78％，52.33％，49.54％，46.87％，59.70％ and 58.24％；those of total flavonoids were 3.27％，3.22％，1.99％，1.70％，1.31％，3.92％ and 2.28％；those of polysaccharides were 4.70％，6.09％， 6.48％，5.45％，5.74％，5.76％ and 7.15％；those of astragalin were 0.48％，0.46％，0.24％，0.23％，0.20％，0.48％，0.29％. The comprehensive score of microwave drying was the highest ，being 0.996 3. The average contents of water-soluble extract from March to October were 41.50％，40.57％，39.16％，40.65％，40.68％，43.30％，45.19％ and 40.12％；those of total flavonoids were 2.24％，2.81％，3.87％，3.92％，3.82％，3.93％，3.66％ and 3.25％；those of polysaccharides were 4.41％，4.61％， 4.98％，5.26％，5.75％，5.94％，5.32％ and 4.47％；those of astragalin were 0.20％，0.21％，0.25％，0.26％，0.25％，0.24％， 0.25％ and 0.21％，respectively. The comprehensive scores of samples collected from May to September exceeded 0.92，and the comprehensive score in August was the highest （0.988 6）. CONCLUSIONS Microwave-dried Gynura divaricata leaf has the best quality ，and the best harvesting time is from May to September.

Ankylosing spondylitis (AS) is a chronic inflammatory disease characterized by inflammation of the sacroiliac joints and the spinal attachment point and is the most common type of spondyloarthritis (SpA). The pathogenesis of AS is related to both the immune system and the skeletal system. The main pathological changes include enthesitis, osteogenesis changes, osteolytic bone destruction, and immune system changes. Bone cells interact with immune cells, secrete a series of inflammatory factors, and jointly regulate the pathogenesis of AS. The imbalance of various immune cells in AS and the changes in inflammatory cytokines lead to a disorder of bone metabolism. At the same time, the osteocytes express various inflammatory cytokines, which leads to an imbalance of the immune system. The interaction between the immune system and the skeletal system has become a hot spot in the pathogenesis of AS. Understanding the bone immunological mechanism of AS will help to understand the exact pathogenesis of the disease and explore new treatment methods for it. In this review, the changes of various immune cells and inflammatory cytokines in AS and their effects on the skeletal system, as well as the changes of various osteocytes in the skeletal system and their effects on the immune system, were reviewed, and the latest progress in the treatment of AS was summarized.

With the appearance of the disadvantages of traditional tumor treatment, immunotherapy has entered people′s horizons as modern emerging treatment strategies, among which plant polysaccharides have received much more attention due to their antitumor activity and significant immunomodulatory effects. Tumor-associated macrophages (TAMs), as a component of tumor microenvironment, are important factors affecting tumors, and the regulation of TAMs by plant polysaccharides is one of the effective immunotherapy to treat tumor. In this review, we mainly described the regulation of TAMs by plant polysaccharides and the underlying mechanisms, and then gave an outlook on the research interests and the development of plant polysaccharides as immune adjuvants, aiming to provide reference for the study of plant polysaccharides in the immunotherapy for tumors.

Objective To evaluate the left atrial(LA) function in patients with severe preeclampsia ( SPE) by full-volume imaging technology. Methods Forty-two patients of SPE were randomly chosen as SPE group,34 healthy pregnant women were selected as the control group. LA related function parameters including LA maximum volume( LAVmax),LA minimum volume( LAVmin),LA pre-atrium contraction volume(LAVpre),LA reservoir volume(LARV),LA passive emptying volume(LAPEV),LA contraction volume(LACV),LA expansion index(LAEI),LA passive emptying fraction(LAPEF),LA passive emptying fraction( LAAEF ) were separately required by biplane area-length method and full-volume imaging technology in antepartum and postpartum. Results Compared with control group before delivery,the parameters of SPE group before delivery such as LAVmax,LAVmin,LAVpre were higher,LAEI,LAPEF, LAAEF indexed to body surface area(BSA) were lower(all P ＜0.01).Compared with control group before delivery,the parameters of control group after delivery such as LAVmax,LAVmin,LAVpre were lower( all P ＜0.05),LAEI, LAAEF calculated as ratio to BSA were somewhat reduced,LAPEF calculated as ratio to BSA was relatively higher(all P ＞0.05).Compared with SPE group before delivery,the parameters of SPE group after delivery such as LAVmax,LAVmin,LAVpre were lower,LAEI,LAPEF,LAAEF indexed to BSA were higher( all P ＜0.01). Compared with control group after delivery,the parameters of SPE group after delivery such as LAVmax,LAVmin,LAVpre were still incresed ( all P ＜ 0.05 ), LAEI,LAPEF, LAAEF indexed to BSA were relatively lower( all P ＞0.05). Conclusions Full-volume imaging technology can quantitatively evaluate the LA function. LA reservoir function,conduit function and booster pump function are reduced in SPE before delivery,LA rereservoir function,conduit function and booster pump function are basically recovery in SPE after delivery.