BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

Objective To investigate the safety and clinical effect of prophylactic ileostomy using right lower abdominal specimen extraction incision in laparoscopic rectal low anterior resection and its impact on ileos-tomy closure.Methods A retrospective analysis was conducted on 127 patients who underwent laparoscopic low anterior resection of rectal cancer combined with prophylactic ileostomy in Department of General Surgery in our hospital from January 2020 to December 2023.Patients were divided into observation group(n=60)and control group(n=67)based on whether with prophylactic ileostomy using the right lower abdominal specimen extraction incision or not.Relevant data and clinical data of ileostomy closure were compared between the two groups.Results(1)There was no significant difference in baseline clinical data between the two groups(P>0.05).(2)In terms of operation time,pain score within three days after surgery and postoperative first exhaust time,the observation group was better than the control group,showing statistical difference(P<0.05).In the observation group,the average length of the incision was longer than that in the control group,and it was significantly different between the two groups(P<0.05).There was no significant difference between the two groups in terms of blood loss,post-operative feeding time and postoperative hospital stay(P>0.05).(3)There was no significant difference between the two groups in postoperative and ostomy-related complication S(P>0.05).(4)The observation group was better than the control group in terms of postoperative pain score and postoperative first exhaust time,indicating statistical significance(P<0.05).There were no significant differences in surgical time,abdominal adhesion,blood loss,postoperative hospital stay and postoperative complications between the two groups(P>0.05).Conclusion The patients undergoing laparoscopic rectal low anterior resection who needs prophylactic ileostomy using right lower abdominal specimen extraction incision can improve the appearance of the incision,shorten operation time,reduce postoperative pain of the patients but does not increase postoperative complications and the difficulty of ileostomy closure.It is safe and feasible,which worthy for clinical promotion.

OBJECTIVE: To evaluate the application of surgical strategies for the treatment of cervical ossification of the posterior longitudinal ligament (OPLL) involving the C ₂ segment. METHODS: The literature about the surgery for cervical OPLL involving C ₂ segment was reviewed, and the indications, advantages, and disadvantages of surgery were summarized. RESULTS: For cervical OPLL involving the C ₂ segments, laminectomy is suitable for patients with OPLL involving multiple segments, often combined with screw fixation, and has the advantages of adequate decompression and restoration of cervical curvature, with the disadvantages of loss of cervical fixed segmental mobility. Canal-expansive laminoplasty is suitable for patients with positive K-line and has the advantages of simple operation and preservation of cervical segmental mobility, and the disadvantages include progression of ossification, axial symptoms, and fracture of the portal axis. Dome-like laminoplasty is suitable for patients without kyphosis/cervical instability and with negative R-line, and can reduce the occurrence of axial symptoms, with the disadvantage of limited decompression. The Shelter technique is suitable for patients with single/double segments and canal encroachment >50% and allows for direct decompression, but is technically demanding and involves risk of dural tear and nerve injury. Double-dome laminoplasty is suitable for patients without kyphosis/cervical instability. Its advantages are the reduction of damage to the cervical semispinal muscles and attachment points and maintenance of cervical curvature, but there is progress in postoperative ossification. CONCLUSION OPLL involving the C ₂ segment is a complex subtype of cervical OPLL, which is mainly treated through posterior surgery. However, the degree of spinal cord floatation is limited, and with the progress of ossification, the long-term effectiveness is poor. More research is needed to address the etiology of OPLL and to establish a systematic treatment strategy for cervical OPLL involving the C ₂ segment.
Humans ; Longitudinal Ligaments/surgery* ; Ossification of Posterior Longitudinal Ligament/surgery* ; Treatment Outcome ; Osteogenesis ; Decompression, Surgical/methods* ; Cervical Vertebrae/surgery* ; Laminoplasty/methods* ; Kyphosis/surgery* ; Retrospective Studies

Objective:To fundamentally solve the problems of valve dysfunction caused by limited size of artificial valve, high requirements for implantation technology, destruction of subvalvular structure and proliferation of fibrous endothelial tissue. We have developed a prosthetic valve with zero left ventricular occupancy. The valve frame is appropriately higher than the existing products, slightly chimney shaped, fully accommodates the movement of the valve leaves in the valve frame, and completely supraannular surgical implantation. This study uses this valve for preclinical animal test.Methods:Seven sheep underwent the replacement of the artificial valve under the direct vision of cardiopulmonary bypass. Warfarin was used for anticoagulation for 6 months 24 hours after operation. The blood biochemistry, anticoagulation and cardiac color Doppler ultrasound were detected after operation. The sheep were killed 180 days later for autopsy to observe whether the artificial valve caused thromboembolism. The efficacy and safety of valve were observed.Results:Two of the seven sheep died within 24 hours after operation, and five survived for a long time. Later, they were killed at 180 days. The mechanical properties of the artificial valve were good, and there was no thromboembolism around the artificial valve and important organs. The tissue layer of suture ring has been completely organized and endothelial cells, and the thickness of tissue layer is about 0.6-1.0 mm. The mitral valve of experimental animals can be opened and closed freely, with smooth surface and no abnormal echo. The follow-up of color Doppler ultrasound showed that the left ventricular ejection fraction was normal (0.60-0.70) before and 6 months after operation. The results of cross valve pressure difference and blood flow velocity of artificial valve were good (vmean was far lower than 5 mmHg(1 mmHg=0.133 kPa) and pmean was lower than 1 m/s) , suggesting that the hemodynamic performance of artificial valve was good. Left ventricular retrograde angiography showed that the artificial valve was completely located in the left atrium with good position and normal opening and closing. There was no obvious perivalvular leakage and other abnormalities. Within 7 days of blood test, the indexes of surgical stress were more abnormal than those before operation. At 12 and 24 weeks, there were no obvious abnormalities in blood routine test, liver and kidney function and other blood test indexes of animals in each group.Conclusion:The new chimney shaped mechanical valve implanted completely above the mitral annulus has good wear resistance, histocompatibility, antithrombotic and hemodynamic performance.

Objective:To investigate the expressions and clinical significance of tetraspanin CO-029 and integrin αv in intrahepatic cholangiocarcinoma (ICC ).Methods:Tissue microarray (TMA) was used to detect the expression of CO-029 and αv in 254 cases of intrahepatic cholangiocarcinoma. The relationship between the two factors and clinicopathological features, recurrence, metastasis and prognosis was analyzed.Spearman method was used to analyze their correlation.Relationship between αv and CO-029 was studied by mass spectrometry and database search,immunoprecipitation and Western blot were used to detect the coexistence.Results:Tissue microarray analysis showed that the positive expression rate of CO-029 was 51.6% (131/254), and the positive expression rate of αv was 61.4% (156/254). The expression of CO-029 and αv were closely correlated with tumor envelope, size, number and TNM stage ( P<0.05). According to the time of recurrence (TTR), the expressions of CO-029 and αv in early postoperative recurrence group (TTR <1 year) were significantly higher than those in non recurrence group (TTR ≥ 1 year). The patients with high CO-029 expression were more likely to relapse ( HR=2.01, 95% CI=1.45-2.79; P<0.001) and had shorter survival time ( HR=2.03, 95% CI=1.46-2.81; P<0.001). The patients with high expression of αv had shorter recurrence time ( HR=1.85, 95% CI=1.38-2.47; P<0.001) and shorter survival time ( HR=1.95, 95% CI=1.40-2.71; P<0.001). Co immunoprecipitation and Western blot confirmed that αv and CO-029 formed a complex. There was a positive correlation between CO-029 and αv in intrahepatic cholangiocarcinoma ( r=0.401, P<0.01). Conclusions:The differential expression of CO-029 and αv were closely related to the recurrence, metastasis and prognosis of intrahepatic cholangiocarcinoma, and CO-029 may couple with αv to form a complex to promote the invasion and metastasis of intrahepatic cholangiocarcinoma.

Objective:To investigate the correlations between CO-029 expression and cholangiocarcinoma invasion and metastasis, and the further explore the potential mechanism involved.Methods:The constructed lentiviral vector of vshRNA-CO-029 (LV/GFP/CO-029) was used to transfect and screen the stable transfected cholangiocarcinoma cell line HCCC-9810-vshRNA-CO-029 as the silence group, HCCC-9810 cells transfected with the mock plasmid were used as the mock group, and the untransfected cells were used as the control group. Cell scratch assay, Transwell assay and in vivo implantation assay were used to detect the migration, invasion and metastasis of the three groups of cells. Immunoprecipitation and tumor necrosis factor (TNF)-α stimulation assay were used to detect the effect of CO-029 on the expression of EMT-related genes.Results:The scratch healing rate of the silence group was (27.11±4.58)%, which was lower than that in mock group (92.84±6.24)%, the number of cells passing through Matrigel in silence group was (57.15±6.10), which was significantly lower than that in mock group (108.20±9.21) and control group (112.00±10.45), the differences were statistically significant ( all P<0.01). The volume of liver tumors in the silence group of orthotopic xenograft mouse model was (2.17±0.54) cm 3, while the volume of liver tumors in the transplanted simulation group was (0.74±0.15) cm 3, the differences were statistically significant ( P<0.05). The incidence of lung metastasis and the number of lung metastases in the simulated group was 100%(6/6) and (214.17±35.64), respectively, while that in the silence group was 16.7% (1/6) and (41.56±14.15), respectively, the differences were statistically significant (all P<0.05). Co-immunoprecipitation showed that CO-029 can form a complex with TNF-αR1. TNF-α induced the down-regulation of E-cadherin and up-regulation of vimentin and N-cadherin in the mock group, but no significant changes were observed in the silence group. Conclusion:CO-029 expression is positively correlated with tumor invasion and metastasis of cholangiocarcinoma, and could couple with TNF-α to induce EMT, which is a novel well-established potential prognostic and therapeutic target for cholangiocarcinoma metastasis and prognosis intervention.

Objective:To investigate the value and molecular mechanism of gastric cancer stem cells in invasion and metastasis of gastric cancer.Methods:Gastric cancer stem cells were isolated by spheroid culture method, and the biological characteristics were identified. The role of gastric cancer stem cells in multidrug resistance, invasion and metastasis was analyzed. The important molecules involved in the biological behavior of gastric cancer stem cells were identified by gene chip. Screening of the signaling pathway revealed that anthrax toxin receptor 2 (ANTXR2)plays an important role in invasion, metastasis, glomerization and tumor formation.Results:Sox2 and Bmi1 expression in SGC7901 and MGC803 cells were significantly higher than monolayer culture cells (15.39±3.23). Oct4 express increased to (4.19±0.62). The expression levels of the dry-related genes Bmi1, Sox2 and Oct4 in SGC7901-SC cells were (3.29±0.52), (3.12±0.49), (2.58±0.35), respectively, which were higher than those of SGC7901-MN cells, Bmi1, Sox2 and Oct4, respectively. The differences were statistically significant ( t=5.392, 7.316, 6.449, all P<0.05) for (1.41±0.39), (2.04±0.33), (1.39±0.32); in primary cells XN0422-SC and in SGC7901-SC cells, miR-638 was in high expression (0.69±0.11), and miR-181b and miR-181a in low expression (0.12±0.05) and (0.15±0.07). ANTXR2 expression in SGC7901-SC cells was higher than that in SGC7901-MN cells ( t=6.216, P<0.05). The ANTXR2 positive cells in SGC7901-SC was 85.48%. The proportion of ANTXR2 positive cells in SGC7901 was 4.98%. Gastric cancer cells XN0422 and SGC7901 were affected by PLVT713, and ANTXR2 expression protein decreased. Conclusion:ANTXR effects a regulatory role by activating the Src/ERk signaling pathway, which can be used to predict the biological beharious of gastric cancer.

Objective To investigate the clinical value of plasmatic heparin-binding protein in early diagnosis and severity gradation of neonatal sepsis.Methods Thirty-nine patients with general sepsis,37 patients with severe sepsis and 16 patients with septic shock were recruited as corresponding study groups respectively,who all had been admitted to the Neonatal Intensive Care Units(NICU)of Hunan Children′s Hospital from December 2016 to August 2017,meanwhile,34 patients with local infection and 35 patients with non infection were enrolled as relevant control group respectively who all had been admitted to each neonatal ward in the retrospective study.The level of the heparin-binding protein(HBP), procalcitonin (PCT)and high sensitive C-reactive protein(hs-CRP)of all patients were detected respectively at the beginning of hospitalization.The difference of each group was compared by use of nonparametric statistics and the efficacy of every index on diagnosis of infection and sepsis was assessed with the receiver operating characteristic curve(ROC).Results The level of HBP in sepsis group,severe sepsis group and septic shock group HBP(H=91.764,P<0.01), PCT(H=51.757,P<0.01)and hs-CRP(H=28.418,P<0.01)are significantly higher than those in local infection group and non infection group;Plasmic HBP levels of severe sepsis group[52.35(33.65,88.15)(ng/ml)]and septic shock group[73.55(60.61,145.51)(ng/ml)]are statistically higher than general sepsis group[34.12(23.04,41.79)(ng/ml)](H=24.092, P<0.01).There are no statistically differences of serum PCT and hs-CRP among these three groups[(HPCT=1.909,Hhs-CRP=0.292),P>0.05].The area under the curve(AUC)of HBP in diagnosis of neonatal sepsis and infection are 0.885 and 0.904 respectively,more higher than PCT and hs-CRP;With the cut off value of 19.8 ng/ml,the sensitivity and specificity of HBP on diagnosis of infection are 85.7%and 82.9%respectively;the sensitivity and specificity 80.4% and 88.4% for neonatal sepsis with the cut-off value of 28.0 ng/ml respectively.Conclusion HBP probably has the better clinical value than PCT and hs-CRP in the early diagnosis and severity gradation of neonatal sepsis.

Objective To investigate the level of 25-hydroxyvitamin D[25(OH)D]and relationship to C-reactive protein(CRP) and procalcitonin(PCT)in children with severe pneumonia.Methods Serum 25 (OH)D levels of 86 cases of children with severe pneumonia,81 cases of children with mild pneumonia and 85 cases of children in healthy control group on admission were deter-mined by ELISA respectively and compared.At the same time,CRP and PCT were tested and the correlation to 25(OH)D in chil-dren with severe pneumonia was analyzed.Results There are statistical differences among three groups(P <0.05,α=0.05 ).The mean values of serum 25 (OH)D were (62.03 ±17.94),(68.90 ±15.22)and (82.21 ±17.89)μmol/L in severe pneumonia pa-tients,mild pneumonia patients and healthy children respectively,furthermore,prevalence of vitamin D deficiency of the sever pneu-monia group was significantly higher than that of the case-control group and normal group (P < 0.05,α= 0.05 ).The level of 25(OH)D was not significantly related to CRP and PCT level in children with severe pneumonia(P <0.05,α=0.05).Conclusion The mean level of vitamin D in children with severe pneumonia was significantly lower,which may be associated with the occurrence and develop-ment of severe pneumonia in children.