BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

AIM: To evaluate the application effectiveness of the 3D surgical video system and intraoperative optical coherence tomography(OCT)in idiopathic epiretinal membrane vitrectomy.METHODS: A retrospective analysis was conducted on 61 patients(61 eyes)with idiopathic epiretinal membrane at our hospital from January 2023 to October 2023. The patients were divided into two groups based on the surgical methods: with 31 patients(31 eyes)who underwent surgery using the 3D surgical video system and intraoperative OCT technology in 3D group, and 30 patients(30 eyes)underwent surgery using the Resight non-contact wide-angle lens system in traditional surgery group. Surgical time, membrane peeling time were recorded, followed-up for 6 mo, post-operative best-corrected visual acuity(BCVA), intraocular pressure(IOP), central macular thickness(CMT), and complications were analyzed for the two groups.RESULTS:All patients successfully underwent surgery without experiencing serious intraoperative complications such as major retinal hemorrhage, retinal tears, or retinal detachment. Additionally, no postoperative complications such as endophthalmitis or secondary glaucoma occurred. The surgical duration and membrane peeling time in the 3D group were significantly shorter than those in the traditional surgery group(20.13±1.59 vs 25.97±2.09 min; 3.74±0.89 vs 8.13±1.72 min, respectively; both P<0.001). There were no significant differences in BCVA and CMT between preoperative and 1-month postoperative values for both groups of patients(both P>0.008). However, both BCVA and CMT improved significantly at 3 and 6 mo postoperatively compared to preoperative levels(both P<0.008). At 6 mo postoperatively, the BCVA in the 3D group was significantly better than that in the traditional surgery group(P=0.007). There were no significant differences in CMT or IOP between the two groups at any postoperative time point(all P>0.05).CONCLUSION: Both the 3D surgical video system and the traditional surgical group can treat idiopathic epiretinal membrane, but the 3D surgical video system has advantages in shortening the surgical time, improving surgical efficiency, and enhancing the precision of surgical steps during surgery with the assistance of intraoperative OCT.

ObjectiveTo observe the clinical effectiveness and safety of Dingchan Granule （定颤颗粒） for paroxysmal atrial fibrillation with syndrome of qi stagnation and blood stasis. MethodsUsing a randomised， double-blind， placebo controlled study method， 90 patients with paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome were divided into 45 cases each in the treatment group and the control group. Both groups were given conventional western medicine treatment， and the treatment group was additionally treated with Dingchan Granule， while the control group was treated with Dingchan Granule placebo， both of which were taken orally for 8 g each time twice a day. Both groups were treated for 8 weeks. We compared the clinical effectiveness， the improvement of traditional Chinese medicine （TCM） symptoms and the recovery rate of atrial fibrillation between the two groups. We compared the number and duration of atrial fibrillation episodes， TCM symptoms score， atrial fibrillation symptom classification， 24-hour average ventricular rate， Pittsburgh Sleep Quality Index （PSQI）， anxiety index， depression index before and after treatment， and evaluated the safety of the two groups. ResultsThe total clinical effectiveness rate in the treatment group was 82.22% （37/45）， which was better than 60.00% （27/45） in the control group （P＜0.05）. The total effective rate of TCM syndrome effectiveness in the treatment group was 88.89% （40/45）， which was better than 66.67% （30/45） in the control group （P＜0.05）； and the rate of atrial fibrillation regression in the treatment group was 26.67% （12/45）， better than 6.67% （3/45） in the control group （P＜0.05）. The number and duration of atrial fibrillation episodes in both groups were significantly decreased （P＜0.01）， and the number and duration of atrial fibrillation episodes in the treatment group were lower than those in the control group （P＜0.01）. The TCM syndrome scores of both groups after treatment were significantly lower than before treatment （P＜0.01）， and the scores of the treatment group was lower than those of the control group （P＜0.05）. The severity of atrial fibrillation symptoms and the grading of atrial fibrillation symptoms in both groups after treatment were improved （P＜0.01）， and the degree of symptom improvement in the treatment group was better than that in the control group （P＜0.01）. The 24-hour average ventricular rate of both groups after treatment was significantly lower （P＜0.01）. The PSQI， anxiety index and depression index of the treatment group were all lower than before treatment （P＜0.01）， while the PSQI and anxiety index of the control group were both lower than before treatment （P＜0.01 or P＜0.05）， the PSQI， anxiety index and depression index of the treatment group being lower than those of the control group （P＜0.05 or P＜0.01）. No adverse events occurred in both groups， and no abnormalities were observed in blood， urine， stool routine， liver and kidney function， and coagulation function indexes. ConclusionDingchan Granule for paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome can alleviate clinical symptom， improve TCM symptom scores， increase atrial fibrillation recovery rate， stabilise the average ventricular rate， and significantly improve the quality of sleep， alleviate the anxiety and depression， with a good safety profile.

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.

Obiective:To compare characteristics of alveolar macrophages(AMs)or AM-like cells in vitro cultured from different progenitors in mice of different ages in presence of GM-CSF,TGF-β and PPAR-γ agonist rosiglitazone(in brief,GTR),according to a previously reported optimized culture procedure.Methods:After bronchoalveolar lavage fluid(BALF)and bone marrow were isolated from mice of different ages and fetal livers were isolated from embryonic-day-15～17 mice,mononuclear cells were seeded into 12-well plates and cultured in presence of GTR.Generated cells were subjected to flow cytometry analysis to analyze expressions of AM markers F4/80,CD11b,CD170 and CD11c.Results:After culture,almost all BALF-derived cells were round-shaped under light microscope,about half bone marrow-derived cells appeared round-shaped,and about 85% fetal liver-derived cells showed round-shaped morphology.Frequency of BALF-AMs from 4-week-old mice was nearly 90%.Frequency and number of BALF-AMs increased with age of mice.Number of BALF-AMs from 12-week-old mice was nearly 1×106.Frequency of bone marrow-derived AM-like cells from 4-week-old mice was lower than 20% and increased to about 40%～45% for 8-week-old and 12-week-old mice.Number of bone marrow-derived AM-like cells increased with age of mice,and about 0.8×106 cells/well bone marrow-derived AM-like cells were produced from 12-week-old mice.Frequency of fetal liver-derived AM-like cells was around 84% and their number could reach 4×106 cells/well.Conclusion:Purity and number of AMs or AM-like cells in vitro cultured from different progenitors in mice of different ages vary dramatically.Detailed condition should be chosen according to specific study requirements.

Objective:To observe the clinical efficacy and safety of hypomethylating agent therapy in chronic myelomonocytic leukemia(CMML).Methods:From February 2014 to June 2021,the clinical data,efficacy,survival time and safety of CMML patients diagnosed in the Second Hospital of Hebei Medical University and treated with hypomethylating agent therapy were retrospectively analyzed.Results:A total of 25 CMML patients received hypomethylating agent therapy,including 18 cases treated with decitabine(DEC)and 7 cases treated with azacytidine(AZA)as the basic treatment.Among them,20 patients responded,and 7 patients got complete remission(CR).All patients with CR were treated with DEC as the basic treatment.Five cases of CR occurred in the first 4 courses of treatment.After a median follow-up of 16.4(9.4-20.5)months,4 patients with CR progressed to acute myeloid leukemia(AML).The median overall survival(OS)time of 25 CMML patients was 17.4 months(95％CI:12.437-22.363).According to MD Anderson prognostic scoring system(MDAPS),CMML-specific prognostic scoring system(CPSS),CPSS molecular(CPSS-mol),Mayo molecular model(MMM),risk stratification of patients was performed,and the difference only between different risk stratification of MDAPS and survival time was statistically significant.Common adverse reactions of hypomethylating agent therapy in CMML patients included infection,gastrointestinal reaction,hematological toxicity,skin allergy and liver function damage.All patients'symptoms were improved after corresponding treatment.Conclusion:Hypomethylating agent therapy is effective and safe for CMML patients.CR mostly occurs in the first 4 courses of treatment,and hypomethylating agent therapy combined with low-dose chemotherapy can be used for patients who do not respond.Hypomethylating agent therapy can delay the disease,but can't prevent progression.

Objective:To explore the diagnosis and treatment of acquired hemophilia A (AHA ) based on the analysis of clinical data.Methods:A retrospective analysis was conducted on the clinical manifestations,laboratory characteristics,treatment,and outcomes of 25 patients diagnosed with AHA who were admitted to the Second Hospital of Hebei Medical University. Results:Among all patients,11 cases had secondary factors,including 5 cases of autoimmune diseases,3 cases of pregnancy-related disease,1 case of pemphigoid,1 case of Graves'disease,and 1 case of monoclonal gammaglobulinemia of unknown significance (MGUS ).The bleeding sites include the skin,mucous membrane,muscle,joint cavity and brain tissue.Twenty-three patients were treated with prednisone combined with cyclophosphamide (CP regimen),one patient with rituximab combined with cyclophosphamide because of femoral head necrosis,and one case with rituximab monotherapy because of gastrointestinal bleeding after prednisone treatment. Among them,23 cases achieved complete remission (CR),2 cases were partial remission (PR),and 8 cases relapsed after CR.All of 10 patients including 2 cases with PR and 8 relapsed cases after CR were treated with rituximab.At last,8 patients achieved CR,and 2 patients (both were patients with recurrence after first CR)achieved PR.These two patients achieving PR were treated with low-dose rituximab combined with bortezomib (RB regimen ).One patient reached CR after 4 cycles and the other reached CR after 6 cycles of RB regimen.After CR,4 of the 10 patients treated with rituximab received maintenance therapy with rituximab monotherapy for 1.5 to 2 years,in which,none of them relapsed.Among the 6 patients who did not receive maintenance therapy,4 patients relapsed after CR,and the median time to relapse was 15 months.Eight patients treated with CP regimen developed common infections,and two patients treated with rituximab developed severe pneumonia.All 25 patients survived until the end of follow-up.Conclusion:Skin ecchymosis,mucous hemorrhage and muscle hematoma are the most common hemorrhagic manifestations in AHA,and joint hemorrhage and cerebral hemorrhage can also occur.CP regimen is the preferred option of first-line therapy for AHA.Rituximab can be used for patients with steroid contraindication or who failed to respond to the above therapy or relapsed after effective treatment,and maintenance therapy is recommended to reduce the risk of recurrence.Meanwhile,close attention should be paid to the occurrence of infection events during rituximab treatment.Rituximab in combination with bortezomib can also be used in patients with refractory or relapsing AHA.

Objective To observe the effect of electroacupuncture(EA)on the expression of NKCC1,KCC2 and activation of microglia in spinal dorsal horn of paclitaxel(PTX)-induced neuropathic pain rats and its possible mechanism.Methods Male SD rats were randomly divided into the vehicle group(vehicle),the PTX group,the PTX+EA group and the PTX+sham EA group,with 12 rats in each group.The rat model of PTX-induced neuropathic pain was established by intraperitoneal injection of PTX.After modeling,EA was applied to"Zusanli"and"Yanglingquan"for 7 days in the PTX+EA group.Paw withdrawal threshold and paw withdrawal latency were tested at 2 days before and 1,3,5,7,14 and 21 days after PTX injection.Immunofluorescence and Western blot assay were used to detect expression levels of sodium-potassium-chloride cotransporter 1(NKCC1),potassium-chloride cotransporters 2(KCC2)and microglia markers-ionized calcium binding adapter molecule 1(Iba1)in spinal dorsal horn.Results Compared with the vehicle group,mechanical and thermal hyperalgesia of both hind feet were found in the PTX group,and the expression of NKCC1 and the number of activated microglia in dorsal horn tissue of spinal cord were increased.Compared with the PTX group,mechanical and thermal hyperalgesia were significantly improved in the PTX+EA group at day 14 and 21,and the expression levels of NKCC1 and Iba1 in dorsal horn tissue of spinal cord were decreased.There was no significant difference in KCC2 expression between the four groups.Conclusion Electroacupuncture can effectively relieve paclitaxol-induced neuropathic pain,which may be related to the inhibition of NKCC1 expression and microglia activation in spinal dorsal horn of rats.

Field effect transistor(FET)biochemical sensors show great potential in the fields of environmental monitoring,food safety,disease diagnosis and clinical treatment due to their low noise,low power consumption,label-free,easy integration and miniaturization characteristics.Two-dimensional(2D)materials,as a new generation of channel materials for FET biochemical sensors,have atomic-level thickness,high carrier mobility,high specific surface area and tunable bandgap,which can further improve the performance of FET biochemical sensors,extend their application areas,and promote the rapid development of FET biochemical sensors.This review focused on the development and latest progress of 2D material-based FET biochemical sensors,along with the challenges and prospects of 2D material-based FET biochemical sensors,which aimed to provide new device design conceptions and promote the further development of biochemical sensing technology.

Objective To explore whether ferulic acid can inhibit the progression of T-cell acute lymphoblastic leukemia in vivo and in vitro by regulating PTEN/PI3K/AKT signaling pathway.Methods The T-cell acute lymphoblastic leukemia Jurkat cells were divided into the control group,the ferulic acid treatment group and the LY294002 treatment group for in vitro experiment.The cells in the control group were given normal culture;cells in the ferulic acid treatment group were given different concentrations(1.25,2.5,5,10,20,40,80,160 μmol/L)of ferulic acid,respectively,and the cell proliferation was detected by CCK-8 method,to screen the experimental concentration;cells in the LY294002 treatment group were given 50 μmol/L PI3K/AKT inhibitor LY294002.The cells proliferation,apoptosis and invasion were detected by clone formation assay,flow cytometry and Transwell assay.The relative expression levels of nuclear protein Ki67,proliferating cell nuclear antigen(PCNA),cleaved caspase-3,cleaved caspase-9,E-cadherin,N-cadherin,Vimentin,PTEN,p-PI3K,PI3K,p-AKT and AKT proteins were detected by Western blot.The nude mice models of transplanted tumors were constructed by 30 male BALB/c nude mice,and they were averagely divided into the normal group and the ferulic acid treatment group for in vivo experiment.The normal group was given normal saline by gavage,while the ferulic acid treatment group was given 75 mg/kg ferulic acid by gavage after inoculating Jurkat cells.The weight and volume changes of transplanted tumors were compared,and the levels of Ki67,cleaved caspase-3/caspase-3,E-cadherin,N-cadherin,PTEN,p-PI3K,PI3K,p-AKT and AKT in tumor tissues were detected.Results In vitro experiment,compared with the control group,the clone formation rate of cells,number of invasion cells,Ki67,PCNA,N-cadherin,Vimentin,p-PI3K/PI3K and p-AKT/AKT in the 5,10,20 μmol/L ferulic acid treatment group and the LY294002 treatment group were significantly decreased(P<0.05),while the apoptosis rate,cleaved caspase-3/caspase-3,cleaved caspase-9/caspase-9,E-cadherin and PTEN were significantly increased(P<0.05).In vivo experiment,compared with the normal group,the weight and volume of tumors were reduced in the ferulic acid treatment group,Ki67,N-cadherin,p-PI3K/PI3K and p-AKT/AKT in tumor tissues were significantly decreased,cleaved caspase-3/caspase-3,E-cadherin and PTEN were significantly increased,with statistically significant differences(P<0.05).Conclusion Ferulic acid can inhibit the proliferation and invasion of T-cell acute lymphoblastic leukemia Jurkat cells in vivo and in vitro,and induce apoptosis,its mechanism may be related to the regulation of PTEN/PI3K/AKT signaling pathway.