Microglia, the resident immune cells in the central nervous system (CNS), rapidly transition from a resting to an active state in the acute phase of ischemic brain injury. This active state mediates a pro-inflammatory response that can exacerbate the injury. Targeting the pro-inflammatory response of microglia in the semi-dark band during this acute phase may effectively reduce brain injury. Shionone (SH), an active ingredient extracted from the dried roots and rhizomes of the genus Aster (Asteraceae), has been reported to regulate the inflammatory response of macrophages in sepsis-induced acute lung injury. However, its function in post-stroke neuroinflammation, particularly microglia-mediated neuroinflammation, remains uninvestigated. This study found that SH significantly inhibited lipopolysaccharide (LPS)-induced elevation of inflammatory cytokines, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS), in microglia in vitro. Furthermore, the results demonstrated that SH alleviated infarct volume and improved behavioral performance in middle cerebral artery occlusion (MCAO) mice, which may be attributed to the inhibition of the microglial inflammatory response induced by SH treatment. Mechanistically, SH potently inhibited the phosphorylation of serine-threonine protein kinase B (AKT), mammalian target of rapamycin (mTOR), and signal transducer and activator of transcription 3 (STAT3). These findings suggest that SH may be a potential therapeutic agent for relieving ischemic stroke (IS) by alleviating microglia-associated neuroinflammation.
Animals ; Microglia/immunology* ; Mice ; Male ; Mice, Inbred C57BL ; Brain Ischemia/immunology* ; Neuroinflammatory Diseases/drug therapy* ; Neuroprotective Agents/administration & dosage* ; Interleukin-1beta/genetics* ; STAT3 Transcription Factor/genetics* ; TOR Serine-Threonine Kinases/genetics* ; Tumor Necrosis Factor-alpha/genetics* ; Proto-Oncogene Proteins c-akt/immunology* ; Nitric Oxide Synthase Type II/genetics* ; Lipopolysaccharides

Objective:To investigate the clinical efficacy of 3D printed metal augment or tibial prosthesis for reconstruction of large bone defects in total knee arthroplasty (TKA) and knee revision surgery.Methods:A total of 7 patients (7 knees) with TKA or knee revision who were admitted to the Department of Orthopaedics of the Second Affiliated Hospital of Zhejiang University School of Medicine with large bone defects from July 2018 to December 2023 were retrospectively analyzed, including 4 patients with TKA and 3 patients with knee revision. There were 3 males and 4 females, aged 58.7±7.6 years (range, 54-68 years), 3 patients with left knee and 4 patients with right knee. All the patients had bone defects in the knee joint (AORI type III), 2 cases had bone defects only in the femur, 4 cases had bone defects only in the tibia, and 1 case had bone defects in both the tibia and femur, which were treated with personalized reconstruction using 3D printing. Hip-knee-ankle angles, American Knee Society score (KSS) before and after surgery were compared, and postoperative complications were observed.Results:All patients successfully completed the operation, and the operation time was 189.3±35.5 min (range, 125-240 min). Complex TKA was performed in 4 cases with surgical times of 175, 195, 210, and 240 min, and revision surgery was performed in 3 cases with surgical times of 125, 180, and 200 min, respectively. Intraoperative blood loss was 114±24.4 ml (range, 100-150 ml). Five cases used 3D printed metal augment, and two used 3D printed one-piece tibial components. All patients were followed up for 2, 2, 5, 6, 7, 20, 57 months, respectively. The KSS of the five patients at 3 months postoperatively were 56, 61, 66, 56, and 56 points, respectively, greater than the preoperative scores of 35, 44, 36, 27, and 41 points. The KSS functional scores of the five patients at 3 months postoperatively were 45, 45, 45, 30, and 45 points, respectively, which were greater than the preoperative scores of 30, 30, 15, 20, and 20 points. The hip-knee-ankle angle was 181.8°±3.4° (range, 177.9° to 188.0°) at the final follow-up and 175.8°±12.4° (range, 153.3° to 192.1°) before surgery, with no significant difference ( t=-1.230, P=0.242). At the final follow-up, the 3D printed component was well integrated with the bone surface, the prosthesis was securely positioned, and the force lines of the lower limbs were normal. There were no postoperative complications such as poor wound healing, infection, fat liquefaction, nerve injury, deep vein thrombosis of lower limbs, knee joint stiffness, periprosthesis infection and loosening. Conclusion:Using 3D printed metal augment or tibial prosthesis to reconstruct the huge bone defect in TKA and revision has a satisfactory early clinical effect, satisfactory joint function and good surgical safety.

Objective To investigate the influence of Nei endonuclease Ⅷ-like protein 1(NEIL1)on H2O2-induced apoptosis and autophagy in lens epithelial cells(LECs).Methods Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of NEIL1 messenger ribonucleic acid(mRNA)in the anterior lens capsule and LEC line SRA01/04 cells from age-related cataract(ARC)patients and macular epiretinal membrane patients who underwent the clear lens extraction.SRA01/04 cells were then divided into the control group,OE-Vector group and OE-NEIL1 group.RT-PCR and Western blot(WB)were used to detect the overexpression efficiency.Subsequently,the cells were divided into the control group,H2O2 group,OE-Vector H2O2 group and OE-NEIL1 H2O2 group;cell viability was detected by Cell Count-ing Kit-8,WB was used to evaluate the expression of autophagy-related proteins(ATG7,P62,Beclinl,LC3-Ⅰ and LC3-Ⅱ)and apoptosis-related proteins(Bax and Bcl-2),and fluorescent staining was adopted to measure cell apoptosis and mito-chondrial membrane potential.Results The expression of NEIL1 mRNA in the anterior lens capsule samples from ARC patients was significantly lower than that of macular epiretinal membrane patients,and the expression of NEIL1 mRNA in the SRA01/04 cells in the H2O2 group was also lower than that in the control group(both P＜0.05).The results of RT-PCR and WB revealed that the expressions of NEIL1 mRNA and protein in the SRA01/04 cells of the OE-NEIL1 group were signif-icantly higher than those in the OE-Vector group(both P=0.000).Compared with the control group,the viability of SRA01/04 cells in the H2O2 group decreased,the expression of the Bax protein was up-regulated,the Bcl-2 protein was down-regulated,viable cells labeled with Mito-Tracker decreased,and apoptotic cells labeled with Annexin V-FITC in-creased,with statistical significances(all P＜0.05).Compared with the OE-Vector H2O2 group,in the OE-NEIL1 H2O2 group,the viability of SRA01/04 cells significantly improved,the autophagy-related proteins(ATG7,P62,Beclin1,LC3-Ⅰand LC3-Ⅱ)and Bcl-2 protein were significantly up-regulated,the Bax protein was down-regulated,viable cells marked with Mito-Tracker increased,and apoptotic cells labeled with Annexin V-FITC decreased(all P＜0.05).Conclusion Under the oxidative stress induced by H2O2,NEIL1 can promote autophagy of LECs,maintain homeostasis of LECs,and then increase cell viability and reduce apoptosis of LECs,participating in the occurrence and development of ARC.

Chronic cerebral hypoperfusion leads to white matter injury (WMI), which subsequently causes neurodegeneration and even cognitive impairment. However, due to the lack of treatment specifically for WMI, novel recognized and effective therapeutic strategies are urgently needed. In this study, we found that honokiol and magnolol, two compounds derived from Magnolia officinalis, significantly facilitated the differentiation of primary oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes, with a more prominent effect of the former compound. Moreover, our results demonstrated that honokiol treatment improved myelin injury, induced mature oligodendrocyte protein expression, attenuated cognitive decline, promoted oligodendrocyte regeneration, and inhibited astrocytic activation in the bilateral carotid artery stenosis model. Mechanistically, honokiol increased the phosphorylation of serine/threonine kinase (Akt) and mammalian target of rapamycin (mTOR) by activating cannabinoid receptor 1 during OPC differentiation. Collectively, our study indicates that honokiol might serve as a potential treatment for WMI in chronic cerebral ischemia.
Magnolia ; White Matter ; Brain Ischemia/metabolism* ; Oligodendroglia/metabolism*

Objective:To analyze the effect of spine-pelvis sagittal parameters and sagittal orientation of facet joint on degeneration of cranial L 3,4 facet joint (facet joint degeneration, FJD) after L 4-S 1 posterior lumbar interbody fusion (PLIF). Methods:Patients with lumbar degenerative diseases who underwent L 4-S 1 PLIF from January 2012 to December 2016 were retrospectively investigated, there were 54 cases, including 28 males and 26 females. Age: 54.59±5.48 years (range, 45-60 years). X-ray, CT, MRI and Weishuapt grade was used to evaluate the degeneration of L 3,4 facet joint at the cranial adjacent segment. The general information and the sagittal parameters of spine pelvis at the last follow-up were compared between the two groups. The former included age, gender, body mass index (BMI), bone mineral density (BMD), follow-up time and preoperative diagnosis. The latter included lower lumbar lordosis angle (LLL), lumbar lordosis angle (LL), pelvis incidence (PI), pelvis tilt (PT), sacrum slope (SS), the height of the intervertebral space (HD), the angle of cranial facet joint, Oswestry disability index (ODI), Japanese Orthopedic Association (JOA) lumbar function score and improvement rate were compared at the same time. Independent sample t-test was used to compare continuous variables between groups; comparison of categorical variable components χ 2 test or Fisher's exact test. Multivariate logistic regression analysis was used to predict the risk factors of adjacent FJD. Results:Postoperative follow-up was 33.44±6.85 months (range, 24-36 months), there were 17 patients in the degenerative group and 37 patients in the non degenerative group. There were no significant differences in age, gender, BMI, BMD, follow-up time or preoperative diagnosis between the two groups. LLL, LL and SS also showed no significant difference. At the last follow-up, PI (56.28°±6.03° vs. 47.87°±8.30°, t=3.74, P=0.001), PT (17.90°±7.06° vs. 14.41°±5.51°, t=1.97, P=0.042) and the joint angle of the cephalic facet (58.48°±2.00° vs. 54.69°±3.01°, t=4.72, P=0.072) in the degenerative group were greater than those in the non-degenerative group. In the subgroup analysis of lumbar lordosis distribution, the difference between the two groups was statistically significant (χ 2=9.90, P=0.006). The HD in the degenerative group 7.50±3.60 mm was significantly lower than that in the non degenerative group 9.30±2.79 mm ( t=2.00, P=0.031). Multivariate logistic regression analysis showed that increase of PI ( OR=1.22, P=0.005) and magnified cephalic facet joint angle ( OR=2.04, P=0.008) were risk factors for adjacent segment facet degeneration. At the last follow-up, the ODI improvement rate in the degenerative group (58.14%±13.41% vs. 70.18%±8.03%, t=4.11, P<0.001) and the JOA score improvement rate (44.72%±9.53% vs. 68.86%±8.55%, t=0.43, P=0.001) were lower than those in the non degenerative group. Conclusion:The increase of PI and sagittal facet (increased joint angle of proximal facet) are risk factors of adjacent segment FJD after lumbar fusion; The abnormal distribution of lower lumbar lordosis and poor PT recovery in adjacent segment FJD patients after lumbar fusion are more obvious, which may be related to the increase of PI; After lumbar fusion, the orientation of adjacent facet joint tended to be sagittal.

Objective:To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.

To explore the effect of connexin 43 (Cx43) silence on the apoptosis in mouse chondrocyte under mechanical stress.
 Methods: Mouse chondrocyte ATDC5 cells were divided into a control group, a mechanical stress group, a Cx43 siRNA transfection group, a scramble siRNA transfection group, a mechanical stress+scramble group, and a mechanical stress+siCx43 group. Flexcell FX-5000 system was used to produce mechanical stress on ATDC5 cells cultured in vitro. The mRNA and protein level of Cx43 was detected by quantitative RT-PCR (RT-qPCR) and Western blot. The cell activity and cell apoptosis was detected by cell counting kit-8 (CCK-8) method and flow cytometry, respectively. Caspase-3 activity was detected by colorimetric assay. The protein expression of Bcl-2, Bax, p-JNK and JNK was detected by Western blot.
 Results: Mechanical stress upregulated the mRNA and protein expression of Cx43 (both P<0.05). Transfection of Cx43 siRNA significantly decreased Cx43 mRNA and protein level (both P<0.05). After stimulation with mechanical stress, chondrocyte viability was significantly decreased, whereas cell apoptosis and caspase-3 activity were increased (both P<0.05). Mechanical stress obviously upregulated Bax protein level, and downregulated Bcl-2 protein expression and Bcl-2/Bax (both P<0.05). Cx43 siRNA transfection significantly increased cell viability, inhibited cell apoptosis and caspase-3 activity (both P<0.05). Cx43 siRNA also inhibited Bax expression, and increased the Bcl-2 protein expression and Bcl-2/Bax (both P<0.05). Furthermore, Cx43 siRNA significantly suppressed the p-JNK expression induced by mechanical stress (P<0.05).
 Conclusion: Cx43 silence inhibits mechanical stress-induced apoptosis in chondrocyte, which might be mediated by JNK signaling pathway.
Animals ; Apoptosis ; Chondrocytes ; Connexin 43 ; Mice ; Proto-Oncogene Proteins c-bcl-2 ; Stress, Mechanical ; bcl-2-Associated X Protein

Objective To evaluate the biomechanical effects of lateral meniscus posterior root (LMPR) tears fixed at different suture positions, so as to investigate the optimal suture method for repairing LMPR tears. Methods Eight fresh cadaveric knees were used. Each knee was tested under 6 conditions: intact knee, ruptur of LMPR, suture of LMPR to the center point of root insertion, suture of LMPR posterior, interior and later 5 mm to the center point of root insertion, respectively. The peak contact pressure, the average contact pressure and contact area were evaluated using a Tek-scan sensor positioned between the meniscus and tibial plateau, under 1 kN compressive loading, at 0 degree knee extension. Results In the lateral compartment, the average contact pressure and peak pressure significantly increased under rupture of LMPR compared with the intact state (P<0.01), and the contact area decreased significantly (P<0.05). For LMPR tears fixed by four different suture methods, both the average pressure and peak contact pressure reduced, and the contact area increased. The average contact pressure, peak pressure and contact area were closer to the knee joint in the intact state when the suture positions of LMPR tears was posterior 5 mm to the center point of root insertion (P<0.05). In the medial compartment, there were no significant differences in contact pressure, peak contact pressure and contact area with the knee joint at 0 degree (P>0.05). Conclusions The average contact pressure, the peak contact pressure and the contact area between the lateral meniscus and the tibial plateau changed obviously due to the LMPR tears. When the suture position was 5 mm lateral to the center point of root insertion, similar biomechanical function with the intact knee could be obtained.

Objective To discuss the risk factors for coagulopathy in acute isolated traumatic brain injury (TBI).Methods A retrospective study was performed on 191 patients with acute isolated TBI hospitalized from July 2012 to June 2015.There were 70 patients with coagulopathy (coagulopathy group) and 121 patients without coagulopathy (control group).Age,gender,injury type,midline shift on CT and injury severity (Glasgow Coma Scale,GCS) were analyzed to identify the independent risk factors for coagulopathy using the logistic regression analysis.Correlation between the independent risk factors and coagulation indices was analyzed.Results Injury severity,acute subdural hematoma,intraventricular bleeding and midline shift on CT were identified as the independent risk factors for coagulopathy(P ＜ 0.05,OR ＞ 1).Furthermore,injury severity and acute subdural hematoma were respectively associated with abnormalities of international normalized ratio (INR) and fibrinogen (Fg) (P ＜0.05 or P ＜ 0.01),intraventricular bleeding with abnormalities of prothrombin time (PT) and platelet count (PC) (P ＜ 0.01),and midline shift on CT with abnormalities of Fg and PC (P ＜ 0.05).Conclusions Injury severity,acute subdural hematoma,intraventricular bleeding and midline shift on CT are independent risk factors for coagulopathy in patients with acute isolated TBI,and correlate with abnormalities of several coagulation indices.Changes in coagulation indices should be monitored accurately after TBI,and timely treatment of coagulopathy can improve the prognosis.

Objective To explore the correlation between the glucose variability and the severity of acute isolated traumatic brain injury(TBI). Method According to the inclusion/exclusion criteria,a total of 125 cases of acute isolated TBI admitted in Department of Neurosurgery of China Medical University from July 2012 to June 2015 were included. According to Glasgow coma scale(GSC),the patients were divided into five groups including control(GCS 15),mild(GCS 13?14),moderate(GCS 9?12),severe(GCS 6?8),and extra?severe(GCS 3?5)groups. Blood glucose control(including relief of the stress and the application of insulin)were carried out immediately. The average,standard deviation,and variation co?efficient of blood glucose of all groups were recorded at admission,48 hours and 3?7 days after hospitalization. The clinical records and glycemic in?dex were compared among different groups and during different periods,so as to analyze the relationship of the variability of glucose and the duration of hyperglycemia with the severity of TBI and the effects of glycemic intensive care management. Results The results of Kruskal Wallis test and Mann?Whitney Utest showed that the average,standard deviation,and variation coefficient of glucose in the extra?severe group and the severe group were statistically higher than those in the control group(P<0.05)during the same period. Meanwhile,the average,standard deviation,and variation coefficient of glucose at admission,48 hours and 3?7 days after hospitalization were also different among each group(P<0.05). The duration of hy?perglycemia and conscious disturbance in both the extra?severe group and the severe group were longer than those in the control(P<0.05). The analyses using rank correlation indicated that glucose variability,the level and duration of hyperglycemia were positively correlated with the severity of TBI(r>1). Conclusion The glucose variability in acute isolated TBI patients could be considered as the index of the severity of TBI.