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.

BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

OBJECTIVE: To explore the effect of bear bile powder (BBP) on acute lung injury (ALI) and the underlying mechanism. METHODS: The chemical constituents of BBP were analyzed by ultra-high-pressure liquid chromatography-mass spectrometry (UPLC-MS). After 7 days of adaptive feeding, 50 mice were randomly divided into 5 groups by a random number table (n=10): normal control (NC), lipopolysaccharide (LPS), dexamethasone (Dex), low-, and high-dose BBP groups. The dosing cycle was 9 days. On the 12th and 14th days, 20 µL of Staphylococcus aureus solution (bacterial concentration of 1 × 10^-7 CFU/mL) was given by nasal drip after 1 h of intragastric administration, and the mice in the NC group was given the same dose of phosphated buffered saline (PBS) solution. On the 16th day, after 1 h intragastric administration, 100 µL of LPS solution (1 mg/mL) was given by tracheal intubation, and the same dose of PBS solution was given to the NC group. Lung tissue was obtained to measure the myeloperoxidase (MPO) activity, the lung wet/dry weight ratio and expressions of CD14 and other related proteins. The lower lobe of the right lung was obtained for pathological examination. The concentrations of inflammatory cytokines including interleukin (IL)-6, tumour necrosis factor α (TNF-α ) and IL-1β in the bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay, and the number of neutrophils was counted. The colonic contents of the mice were analyzed by 16 sRNA technique and the contents of short-chain fatty acids (SCFAs) were measured by gas chromatograph-mass spectrometer (GC-MS). RESULTS: UPLC-MS revealed that the chemical components of BBP samples were mainly tauroursodeoxycholic acid and taurochenodeoxycholic acid sodium salt. BBP reduced the activity of MPO, concentrations of inflammatory cytokines, and inhibited the expression of CD14 protein, thus suppressing the activation of NF-κB pathway (P<0.05). The lung histopathological results indicated that BBP significantly reduced the degree of neutrophil infiltration, cell shedding, necrosis, and alveolar cavity depression. Moreover, BBP effectively regulated the composition of the intestinal microflora and increased the production of SCFAs, which contributed to its treatment effect (P<0.05). CONCLUSIONS BBP alleviates lung injury in ALI mouse through inhibiting activation of NF-κB pathway and decreasing expression of CD14 protein. BBP may promote recovery of ALI by improving the structure of intestinal flora and enhancing metabolic function of intestinal flora.
Animals ; Acute Lung Injury/pathology* ; Lipopolysaccharides ; Ursidae ; Gastrointestinal Microbiome/drug effects* ; Bile/chemistry* ; Lipopolysaccharide Receptors/metabolism* ; Powders ; Male ; Lung/drug effects* ; Mice ; Peroxidase/metabolism* ; Signal Transduction/drug effects* ; Cytokines/metabolism*

Objective To investigate the biomechanical effects of lingual metal-reinforced denture bases on edentulous implant-supported prostheses by three-dimensional finite element analysis,and provide references for the treatment of edentulous jaws.Methods Two implant-supported Locator-type overdenture models with lingual metal-reinforced and non-reinforced denture bases were constructed.A 150 N vertical load on the fovea of the posterior teeth,a 150 N oblique load on the fovea of the posterior teeth,and a 150 N vertical load on the anterior teeth were applied to simulate the centric occlusion,left and right lateral chewing,and anterior teeth cutting,and stresses on the tissues of two models were analyzed.Results When the posterior and anterior teeth were loaded vertically,the stress distribution on each organization was similar between the two denture base designs.The maximum stress difference was between 0 and 0.16 MPa.Under unilateral masticatory chewing,the range of stress concentration on denture base,implant and mucoperiosteum,and the maximum stress of the implant and peri-implant bone without metal reinforcement were significantly greater than those with metal reinforcement,and the maximum stress difference was between 0.59 MPa and 2.99 MPa.Conclusions Lingual metal-reinforced denture base can play a role in stress dispersion,or reduce the risk of bone resorption and denture base fracture to a certain extent.

Objective To investigate the influence of spectral CT monochromatic imaging technique on image quality and liver vol-ume measurement in computed tomography portal venography(CTPV).Methods A total of 120 patients who underwent contrast-enhanced abdominal CT examination were prospectively selected and randomly divided into group A and group B.The group A(n=60)was scanned with conventional parameters such as 120 kVp,Smart mA mode,and image reconstruction of 60％adaptive statistical iterative reconstruction-Veo(ASIR-V);while the group B(n=60)was with instantaneous switching 80/140 kVp,gemstone spectral imaging(GSI)Assist mode,and image reconstruction of 60％ASIR-V.In group B,six subgroups of images from 75 to 50 keV(with 5 keV interval)were recorded as subgroups B1 to B6.On the axial images,the CT values and standard deviation(SD)values of por-tal vein,liver and erector spinae were measured,and then the contrast-to-noise ratio(CNR)of portal vein and liver were calculated.The Liver Segmentation software was used to segment the liver in groups A and B,the liver volumes by automatic segmentation and manual segmentation(golden standard)were recorded,and then the volume difference rate was calculated.The overall image quality and automatic liver segmentation results were evaluated by two radiologists using a 5-point scale.Results In terms of overall image quality,subgroup B6 achieved the highest score and was superior to group A(P＜0.001).In terms of liver segmentation,subgroup B3 had the highest score and was superior to group A(P＜0.001).With the decrease of keV,the CT values,SD values and CNR of portal vein and liver in group B were gradually increased(P＜0.05),in which subgroup B6 was higher than that in group A(P＜0.001).The volume difference rate initially decreased and then increased with the decrease of keV.Except for subgroups B2 and B3,the differences were statistically significant between other subgroups and group A(P＜O.001),and the subgroup B3 had the lowest volume difference rate.Conclusion Spectral CT monochromatic ima-ging technique has an influence on CTPV image quality and liver volume measurement.The 50 keV images are the best for displaying portal vein,and the automatic liver segmentation volume of 65 keV images is closest to the real liver volume.

Objective To compare the clinical efficacy and imaging results of uniportal interlaminar endoscopy(UIE)and unilateral biportal endoscopy(UBE)for the treatment of lumbar disc herniation.Methods The clinical information for 50 patients diagnosed with lumbar disc herniation was collected,and treated by UIE endoscopic surgery and UBE endoscopic surgery in the The First People's Hospital of Hefei city from March 2021 to October 2022 were retrospectively analyzed.The patients were divided into two groups,UIE group and the UBE group.Perioperative indexes including incision length,operation time,intraoperative blood loss,and surgical complications,clinical efficacy indexes including VAS scores of low back pain and leg pain before surgery,3 days after surgery,3 months after surgery,6 months after surgery,and 12 months after surgery,ODI scores of dysfunc-tion index,and imaging results including spinal canal area,vertebral space height,before surgery and 1 year after surgery were recorded and compared between the two groups.Results Both groups completed the procedure and were followed up for 12～18 months,with an average of 15 months.1 case was dural injury,no nerve root injury,and no nerve root symptoms during the follow-up.The symptoms of lumbar and leg pain were all relieved in both groups after the procedure.The UBE groups hawed larger surgical incisions,more intraoperative blood loss,and shorter operative time compared to the UIE group(P＜0.05,respectively).Both groups had significant develop-ments in the VAS scores,ODI scores of back,and leg pain at 3 days,3 months,6 months,and 12 months after the operation(all P＜0.05).The UIE group showed significant developments in the VAS scores and ODI scores of back and leg pain at 3 days and 3 months after the operation,as compared to the UBE group(P＜0.05).The imaging analysis did not showed significant changes in the height of intervertebral space and the angle of lumbar lordosis,but a significantly larger increase in the dural sac area in both groups one year after the procedure,and the UBE group had even a larger increase than the UIE group(P＜0.05).Conclusion Both UIE and UBE have good clinical efficacy and imaging results in the treatment of lumbar disc herniation via interlaminal approach.However,the UIE group is superior to the UBE group in terms of the operation time,intraoperative blood loss,postoperative VAS score of low back pain as well as the decompression effectiveness.

Objective To investigate the regulatory effect of Mahuang Dingchuan Decoction on airway inflammation,airway remodeling and imbalance of helper T cell 1(Th1)/helper T cell 2(Th2)in children with allergic asthma.Methods A total of 126 children with allergic asthma of phlegm-heat obstructing the lung type at acute attack stage were randomly divided into observation group and control group,with 63 cases in each group.The control group was treated with conventional western medicine,and the observation group was treated with Mahuang Dingchuan Decoction on the basis of treatment for the control group.The total course of treatment for the two groups covered 14 days.Before and after treatment,the two groups were observed in the changes of lung function indicators of forced expiratory volume in one second(FEV1)and peak expiratory flow(PEF),airway inflammation indicators of neutrophil count,lymphocyte count,eosinophil count,monocyte percentage and levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the sputum,airway remodeling indicators of airway wall area(WA),airway lumen area(LA)and airway total area(TA),and levels of Th1/Th2 cytokines of gamma-interferon(IFN-γ)and interleukin 4(IL-4).After treatment,the clinical efficacy and safety of the two groups were evaluated.Results(1)After 14 days of treatment,the total effective rate of the observation group was 93.65％(59/63),and that of the control group was 80.95％(51/63).The intergroup comparison(tested by chi-square test)showed that the efficacy of the observation group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the values of pulmonary function indicators of FEV1 and PEF in the two groups were significantly higher than those before treatment(P＜0.01),and the increase in the observation group was significantly superior to that in the control group(P＜0.05).(3)The analysis of airway inflammation indicators showed that the percentages of neutrophils,eosinophils and monocytes and the levels of IL-6 and TNF-α in the sputum of the two groups after treatment were significantly lower than those before treatment(P＜0.05),and the percentage of lymphocytes was significantly higher than that before treatment(P＜0.05).The decrease of the percentages of neutrophils,eosinophils and monocytes and the levels of IL-6 and TNF-α in the sputum as well as the increase of the percentage of lymphocytes in the observation group was significantly superior to that in the control group(P＜0.05).(4)After treatment,the value of airway remodeling indicator of WA in the two groups was lower(P＜0.05)and the values of LA and TA were higher than those before treatment(P＜0.05).The decrease of WA value and the increase of LA and TA values in the observation group were significantly superior to those in the control group(P＜0.05).(5)The observation of Th1/Th2 cytokines showed that the serum IFN-γ level and IFN-γ/IL-4 ratio in the two groups after treatment were significantly higher than those before treatment(P＜0.05),and the serum IL-4 level was significantly lower than that before treatment(P＜0.05).The increase of serum IFN-γ level and IFN-γ/IL-4 ratio and the decrease of serum IL-4 level in the observation group were significantly superior to those in the control group(P＜0.05).(6)No serious adverse reactions were found in the two groups during the treatment,or no obvious abnormalities were found in the safety indicators such as electrocardiogram,blood routine test,liver function examination,heart rate and blood pressure.Conclusion Mahuang Dingchuan Decoction is beneficial to alleviating airway inflammation and airway remodeling in children with allergic asthma,regulating Th1/Th2 imbalance,and improving lung function in children,with definite effective and high safe.

Objective To identify and conduct antimicrobial susceptibility testing on Vibrio species isolated from blood culture specimens of two patients with bloodstream infection(BSI),analyze the microbiological characteristics of non-O1/non-O139 Vibrio cholerae(NOVC),and provide evidence for the diagnosis,prevention and control of Vibrio cholerae infection.Methods Two Vibrio strains were identified by matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry(MALDI-TOF MS),API bacterial biochemical reaction identification test strip,VITEK 2 Compact identification instrument and 16S rRNA gene sequencing.Serological typing,virulence gene molecular detection,and drug resistance phenotype detection were performed to the Vibrio strains.Results Two strains were identified as Vibrio cholerae,classified as NOVC by serological typing,and were negative for ctxAB virulence gene detection.Antimicrobial susceptibility testing showed that one strain was sensitive to ampici-llin,azi-thromycin,doxycycline and chloramphenicol,while resistant to tetracycline and trimethoprim-sulfametho-xazole.The other strain was sensitive to all tested antimicrobial agents.Conclusion BSI caused by NOVC is rarely reported in China.Accurate identification,typing and drug-resistant phenotype detection of Vibrio cholerae isolated from blood culture specimens are valuable for the diagnosis,treatment,prevention and control of Vibrio cholerae-associated infection.

BACKGROUND:Observational studies have suggested that statin drugs may have a protective effect on bone density,making them a potential treatment option for osteoporosis. OBJECTIVE:To evaluate the causal relationship between drug target-mediated lipid phenotypes and bone mineral density(BMD)using Mendelian randomization methods. METHODS:We obtained single nucleotide polymorphismsrelated to statin drugs and BMD data from the IEU Open GWAS database.The primary analysis method was the inverse variance weighted method,and we also used weighted median,simple median,weighted mode,and MR-Egger regression.We usedβ values and 95%confidence intervals(CI)to assess the causal relationship between statin drugs and BMD.Additionally,we conducted sensitivity analyses to validate the results,assessed heterogeneity using Cochran's Q test,examined for horizontal pleiotropy using the MR-Egger intercept test,and performed leave-one-out analyses to determine if individual or multiplesingle nucleotide polymorphism influenced the results. RESULTS AND CONCLUSION:There was a significant association between the statin target of action,3-hydroxy-3-methyl glutaryl coenzyme A reductase-mediated low-density lipoprotein cholesterol,and heel bone BMD(β=-0.086,95%CI:-0.117 to-0.055,P=5.42×10-8)and whole-body BMD(β=-0.193,95%CI:-0.288 to-0.098,P=7.35×10-5).The findings of this study support the protective effect of statin drugs on BMD.These findings not only deepen our understanding of the relationship between cholesterol-related genes and bone health but also reveal potential therapeutic targets for improving BMD.