BACKGROUND:Needle-knife release of the ligamentum flavum can effectively improve symptoms in patients with lumbar degeneration,and ultrasound guidance can increase the precision of needle-knife release;however,the specific effects of needle-knife release of the ligamentum flavum on the degenerated intervertebral discs and the possible mechanisms remain to be clarified. OBJECTIVE:To investigate the effect of ultrasound-guided needle-knife release of the ligamentum flavum. METHODS:Twenty-four New Zealand rabbits were randomized into control(n=6)and model(n=18)groups.A rabbit model of lumbar disc degeneration model was established in the model group by cutting the supraspinous and interspinous ligaments of the L5/6 and L6/7 segments to maintain a standing posture and apply axial load to the lumbar spine.After successful modeling,the model rabbits were subdivided into a control group,a model group,an ultrasonic needle-knife group,and a sham needle-knife group according to a random number table method,with six animals in each group.The ultrasonic needle-knife group underwent ultrasound-guided needle-knife release of the right yellow ligament of L7/S1,once every week,for a total of four times.The needle-knife approach in the sham needle-knife group was the same as that in the ultrasound needle-knife group,but the ligamentum flavum was not released.At 30 days after the intervention,MRI was used to observe the changes in the signal intensity of the nucleus pulposus within the L7/S1 segment.Hematoxylin-eosin staining was used to observe the morphological changes of the L7/S1 segment.Immunohistochemical staining was used to detect the expression of type I and II collagen in the nucleus pulposus of the L7/S1 segment.RT-PCR and western blot were used to detect the expression of integrin α5 and β1,p38,and nuclear factor κB in the L7/S1 segment. RESULTS AND CONCLUSION:MRI findings indicated that the nucleus pulposus of the intervertebral disc of rabbits in the model group was gray-black in color,and the gray value of the nucleus pulposus was significantly lower than that of the control group(P<0.01).The brightness of the nucleus pulposus of the intervertebral disc of the rabbits in the ultrasonic needle-knife group was elevated compared with that of the model group,and the gray value of the nucleus pulposus was higher than that of the model group(P<0.01).Results from hematoxylin-eosin staining showed that in the model group,the shape of the nucleus pulposus was irregular,the number of nucleus pulposus cells was reduced,the extracellular matrix was compressed,the fibrous ring was ruptured,the structure and boundary of the end plate were unclear,and the chondrocytes were arranged disorderly.Compared with the model group,the ultrasonic needle-knife group showed an increase in the number of the nucleus pulposus,an improvement in the rupture of the fibrous ring,and more regular arrangement of cartilage endplate cells.Results from immunohistochemical staining showed an increase in positive expression of type I collagen(P<0.01)and a decrease in positive expression of type II collagen in the nucleus pulposus of the model group compared with the control group as well as a decrease in positive expression of type I collagen and an increase in positive expression of type II collagen in the nucleus pulposus of the ultrasonic needle-knife group compared with the model group(P<0.01).RT-PCR and western blot assays showed that the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the model group were increased compared with that in the control group(P<0.01);the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the ultrasonic needle-knife group was decreased compared with that in the model group(P<0.01).To conclude,ultrasound-guided needle-knife release of the ligamentum flavum can improve the degree of lumbar disc degeneration in rabbits,which may be related to the inhibition of p38 and nuclear factor-κB expression by modulating integrin α5 and β1 expression.

Lipid nanovehicles are currently the most advanced vehicles used for RNA delivery, as demonstrated by the approval of patisiran for amyloidosis therapy in 2018. To illuminate the unique superiority of lipid nanovehicles in RNA delivery, in this review, we first introduce various RNA therapeutics, describe systemic delivery barriers, and explain the lipid components and methods used for lipid nanovehicle preparation. Then, we emphasize crucial advances in lipid nanovehicle design for overcoming barriers to systemic RNA delivery. Finally, the current status and challenges of lipid nanovehicle-based RNA therapeutics in clinical applications are also discussed. Our objective is to provide a comprehensive overview showing how to utilize lipid nanovehicles to overcome multiple barriers to systemic RNA delivery, inspiring the development of more high-performance RNA lipid nanovesicles in the future.

ObjectiveHemoglobin is the iron-containing protein in the red blood cells of many animals. The primary function of hemoglobin is to transport oxygen from lung to tissues. It is composed of two identicalα‑globin subunits and two identical β‑globin subunits. Hemoglobin has unique magnetic properties. The paramagnetism of deoxyhemoglobin, and the diamagnetism of oxyhemoglobin and carboxyhaemoglobin have been reported previously. Studies have also shown that external magnetic field affected blood flow rate, but whether magnetic field may affect the oxygenation rate of hemoglobin remains unknown. Here in this study, we are aiming to address this question with recombinant hemoglobin. Human hemoglobin and yak hemoglobin were selected as the research objects, and a recombinant protein expression and purification system was established to explore the magnetic field effects on the oxygenation rate of hemoglobin, as well as the differences in the oxygenation rate between human hemoglobin and yak hemoglobin under external magnetic field. MethodsThe recombinant expression and purification system of human and yak hemoglobin was established. The recombinant hemoglobin expression was further optimized and appropriated inducing temperature and IPTG concentration were screened. Recombinant human hemoglobin and yak hemoglobin were purified to homogeneity by affinity chromatography and further by size-exclusion chromatography. SDS-PAGE was used to validate the purification, and UV-Vis spectrum and EPR were used to characterize the biochemical properties of recombinant hemoglobin. Deoxyhemoglobin of human and yak were placed under 0.3 T external magnetic field to test the magnetic field effects on oxygenation rate, and geomagnetic field condition was used as a sham control. The UV-Vis spectrum data were measured every 10 min, and the concentration and proportion of oxygenated hemoglobin, deoxyhemoglobin and methemoglobin were calculated to analyze the effects of magnetic field on the oxygenation rate of hemoglobin. The magnetic properties of human oxygenated hemoglobin and human deoxygenated hemoglobin have been measured by SQUID, a superconducting quantum interference magnetic measurement system. Three biological replications were performed for each experiment. The possible mechanism of the effect of magnetic field on the oxygenation rate of hemoglobin has been investigated and discussed. ResultsHuman and yak hemoglobin were successfully expressed and purified by E.coli prokaryotic expression system. The optimal expression temperature was 30℃, and the most suitable IPTG concentration was 1 mmol/L. EPR results suggested that trace amount of methemoglobin existed both in the purified human hemoglobin and yak hemoglobin proteins. The oxygenation rate of yak hemoglobin appeared to be faster than that of human hemoglobin, and the additional magnetic field treatment significantly increased the oxygenation rate of both human and yak hemoglobin, and yak hemoglobin was more sensitive to magnetic field than human hemoglobin. The paramagnetism of deoxyhemoglobin was verified by SQUID measurement. However, the diamagnetism of oxygenated hemoglobin remains uncertain, probably due to the presence of trace amount of methemoglobin in the sample of oxygenated hemoglobin, which was consistent with EPR results. ConclusionIn this study, human and yak hemoglobin were successfully expressed and purified. The purified hemoglobin proteins have similar function and conformational states as native protein. External static magnetic field facilitates hemoglobin oxygenation, and yak hemoglobin seems more sensitive to magnetic field compared with human hemoglobin. These findings provide theoretical basis for the potential applications of applying magnetic field to improve hypoxia symptoms in clinical practice in the future.

OBJECTIVE To study the influencing factors for proteinuria in patients with malignant tumors treated with apatinib， then establish and evaluate a risk prediction model based on it. METHODS A total of 120 patients with malignant tumors treated with apatinib in our hospital from January 2020 to December 2022 were selected as the training set， and the clinical data was collected. Univariate analysis and multivariate Logistic regression analysis were used to identify independent risk factors for proteinuria associated with apatinib and then construct a risk prediction model. The predictive value of the model was evaluated by using the receiver operator characteristic （ROC） curve. A total of 34 patients with malignant tumors treated with apatinib from January to December 2023 in our hospital were selected as the validation set， and their clinical data were obtained to cross-validate the accuracy of the prediction model. RESULTS The incidence of proteinuria in the training set of 120 patients was 26.67%. The proportions of patients with smoking history， combined hypertension， apatinib daily dose of ≥500 mg， and alanine aminotransferase level were significantly higher in proteinuria group than those in non-proteinuria group. At the same time，the neutrophilic granulocyte count was significantly lower than that in non-proteinuria group （P＜0.05）. Patients with smoking history and combined hypertension were the independent risk factors for apatinib-induced proteinuria （odds ratios were 5.005 and 5.342， respectively； with 95% confidence intervals of 1.806- 13.872 and 1.227-9.602， respectively； P＜0.05）. The binary Logistic regression model equation for the probability （P） of apatinib- induced proteinuria is expressed as LogitP＝1.610XMH+1.233XSH－1.483 （MH for combined hypertension， SH for the smoking history）， with a model accuracy of 80.0%. ROC curve analysis demonstrated the area under the ROC curve of 0.771， the maximum Youden’s index of 0.474， and the optimal cut-off value for LogitP was 0.159 9， with a sensitivity of 90.6% and specificity of 56.8%. Cross-validation results indicated an overall prediction accuracy of 88.24% for the 34 patients. CONCLUSIONS Combined hypertension and smoking history are independent risk factors for apatinib-induced proteinuria. The constructed risk prediction model has moderate predictive value and can be used to predict the risk of proteinuria in patients with malignant tumors induced by apatinib.

The gastrointestinal motility disorder of the patients with Parkinson's disease(PD)occurs in the early stages of this disease,even before the onset of motor symptoms.The gastrointestinal motility disorder is one type of gastrointestinal dysfunction,not only affect the absorption of medication,exacerbating the progression of PD,but also severely impact the quality of life of the patients.Therefore,it is essential to find new therapeutic targets to alleviate PD-induced gastrointestinal dysmotility in order to improve the progression of the disease and the quality of life of the patients.The gastrointestinal motility function is highly dependent on the health of the gut and central nervous regulating the gastrointestinal movements.A healthy gut is closely related to the integrity of the intestinal barrier,gut microbiota,neuroinflammation,and the normal function of enteric neurons responsible for the contraction and relaxation of the gastrointestinal tract.The gut function of the PD patients is compromised to some extent.This review summarizes the effects of the enteric nervous system,central nervous system,and gut microbiota on the development of gastrointestinal motility disorder of the PD patients;it also outlines the current therapeutic methods available and their limitations,with the aim of providing the new insights into the treatment of gastrointestinal motility disorder of the PD patients.

Acute cerebral infarction(ACI)has the characteristics of onset nasty and high mortality,and thus the rapid determination of the occurrence and development of ACI plays a key role in the diagnosis,treatment and prognosis of ACI patients.It has shown that the serum level of human haptoglobin(Hp)is related to ACI.In this study,surface enhanced Raman scattering(SERS)combined with immune recognition was applied to establish a quantitative analysis method for serum Hp.Firstly,the SERS substrate of silver nanoparticles was prepared on silicon wafer,and 4-mercaptobenzoic Acid(MBA)was used as a Raman probe by forming Ag—S bond and connecting it on the surface of nanoparticles.The carboxyl group of MBA was linked to amino group of self-made high-affinity antibody through forming CO—NH structure thus forming a SERS self-assembled chip of Hp(Ag/MBA/anti-Hp).Hp in serum could be specifically captured by antibodies on SERS substrate,which caused the shift of SERS characteristic peak of MBA.The results showed that there was a good linear relationship between the logarithm of Hp concentration and the SERS characteristic peak shift of MBA.The detection range was 1-1000 ng/mL(R2=0.988).The Hp concentrations in serum of 90 ACI patients were determined by this method,and the results were consistent with those of ELISA method,which proved the practicability and accuracy of this method.This method was highly specific,simple and convenient,which could realize the specific recognition and quantitative analysis of serum Hp,so as to be an effective means for clinical detection of serum Hp,thus providing a reference for the treatment and prognosis of ACI.

Objective To explore the efficacy and safety of ticagrelor de-escalation and nicorandil therapy in elderly patients with acute coronary syndrome(ACS)after percutaneous coronary intervention(PCI).Methods A total of 300 elderly patients with ACS were selected from the Sixth and Seventh Medical Center of Chinese PLA General Hospital and Beijing Chaoyang Integrative Medicine Emergency Rescue and First Aid Hospital from November 2016 to June 2019,including 153 males and 147 females,aged>65 years old.All the patients received PCI,and all had double antiplatelet therapy(DAPT)scores≥2 and a new DAPT(PRECISE-DAPT)score of≥25.All patients were divided into two groups by random number table method before operation:ticagrelor group(n=146,ticagrelor 180 mg load dose followed by PCI,and ticagrelor 90 mg bid after surgery)and ticagrelor de-escalation + nicorandil group(n=154,ticagrelor 180 mg load dose followed by PCI,ticagrelor 90 mg bid+nicorandil 5 mg tid after surgery,changed to ticagrelor 60 mg bid+ nicorandil 5 mg tid 6 months later).Follow-up was 12 months.The composite end points of cardiovascular death,myocardial infarction and stroke,the composite end points of mild hemorrhage,minor hemorrhage,other major hemorrhage and major fatal/life-threatening hemorrhage as defined by the PLATO study,and the composite end points of cardiovascular death,myocardial infarction,stroke and bleeding within 12 months in the two groups were observed.Results The comparison of general baseline data between the two groups showed no statistically significant difference(P>0.05).There was also no significant difference in the composite end points of cardiovascular death,myocardial infarction and stroke between the two groups(P>0.05).The cumulative incidence of bleeding events in ticagrelor de-escalation + nicorandil group was significantly lower than that in ticagrelor group(P<0.05),while the composite end points of cardiovascular death,myocardial infarction,stroke and bleeding were also significantly lower than those in tecagrelor group(P<0.05).Conclusion In elderly patients with ACS,the treatment of ticagrelor de-escalation + nicorandil after PCI may not increase the incidence of ischemic events such as cardiovascular death,myocardial infarction or stroke,and it may reduce the incidence of hemorrhagic events.

ObjectiveThis study aims to investigate the effectiveness and potential mechanism of Erxian Decoction （二仙汤） for late-onset depression （LOD）. MethodsForty Wistar male rats of 7-8 weeks were divided into 20 each of normal group and youth depression group. Sixty Wistar male rats of 20 months were divided into 20 each of elder group， LOD group and Erxian Decoction group. The rats in youth depression group were modelled with chronic unpredictable mild stress （CUMS） for 6 weeks to build a depression model， and the elder rats in LOD group and Erxian Decoction group were modelled with CUMS for 6 weeks to build a LOD model， with no interventions in the normal group and the elder group. Gastric administration was carried out at the same time of modelling， rats in Erxian Decoction group were given 8 g/（kg·d） of Erxian Decoction by gavage， and rats in the normal group， youth depression group， elder group， and LOD group were given 4 ml/（kg·d） of pure water by gavage， for 6 weeks in each group. Sugar water preference experiment and forced swimming experiment were used to evaluate the depression-like behaviour of rats， absent field experiment was used to evaluate the spontaneous activity ability of rats， and T-maze experiment was used to evaluate the cognitive function of rats； Western blot assay was used to detect neuronal nuclei （NeuN）， nestin， doublecortin （DCX） in hippocampal tissue， and zonula occludens-l （ZO-1）， folate receptor α （FRα）， reduced folate carrier （RFC）， and protoncoupledfolate transporter （PCFT） protein expression in choroid plexus； immunofluorescence was used to detect the number of double-positive cells for Ki-67/nestin， the number of double-positive cells for 5-bromodeoxyuracil nucleoside （BrdU）/DCX in hippocampal dentate gyrus， and the protein expression of ZO-1， FRα， RFC， and PCFT in choroid plexus tissues； ELISA technique was used to detect plasma， cerebrospinal fluid， 5-methyltetrahydrofolate （5-MTHF） in hippocampal tissues； Pearson correlation analysis was used to correlate the 5-MTHF content in cerebrospinal fluid and hippocampal tissues and the expression of nestin， DCX， and NeuN proteins in hippocampal tissues of rats in youth depression group and LOD group. ResultsCompared with normal group， rats in youth depression group and LOD group showed reduced sugar water preference， reduced total distance travelled in the absent field， reduced number of times through the grid， prolonged forced swimming immobilisation， reduced hippocampal NeuN， nestin and DCX protein expression， reduced number of Ki-67/nestin double-positive cells in hippocampal dentate gyrus， reduced number of BrdU/DCX double-positive cells in hippocampal dentate gyrus， and reduced expression of ZO-1 and FRα proteins and fluorescence intensity （P＜0.05 or P＜0.01）； the correct rate of T-maze on days 3 and 4 in the rats of youth depression group and on days 2 to 4 in the rats of LOD group significantly decreased， and the content of 5-MTHF in the cerebrospinal fluid and hippocampal tissues of the rats of LOD group significantly decreased as well （P＜0.05 or P＜0.01）. Compared with youth depression group， rats in LOD group showed a decrease in total distance travelled in absenteeism， number of times through the grid， a significant decrease in the correct rate of the T-maze on day 1-4， a decrease in NeuN， nestin， DCX protein expression of hippocampal tissue and the number of Ki-67/nestin double-positive cells， BrdU/DCX double-positive cells of hippocampal dentate gyrus （P＜0.05 or P＜0.01）. Compared with LOD group， rats in Erxian Decoction group had elevated sugar-water preference and total distance travelled in absenteeism and number of times through the grid， shorter forced swimming immobility time， significantly higher correct rate of the T-maze on day 3-4， elevated expression of NeuN， nestin， and DCX proteins in hippocampal tissues， increased number of Ki-67/nestin double-positive cells and BrdU/DCX double-positive cells in hippocampal dentate gyrus， and increased 5-MTHF content in cerebrospinal fluid and hippocampal， and ZO-1， FRα， RFC， PCFT protein expression and fluorescence intensity increased in choroid plexus （P＜0.05 or P＜0.01）. Correlation analysis showed that there was a positive correlation between 5-MTHF content in cerebrospinal fluid （r = 0.466）， hippocampal tissue （r = 0.522） and nestin expression in hippocampal tissue （P＜0.05）. ConclusionErxian Decoction could improve depressive-like behaviour and cognitive impairment in LOD model rats， and its mechanism may promote hippocampal neurogenesis by alleviating structural damage to the choroid plexus of the brain tissue， and improving impaired choroid plexus folate transport.

BACKGROUND:Host immune response triggered by plaque biofilm is an initiator of periodontitis progression and destruction.Macrophages are an important component of the innate immune response,which play an important role in inflammation occurrence and development. OBJECTIVE:To review the relationship between macrophage polarization and periodontitis and the related progress in the treatment of periodontitis by regulating macrophage polarization. METHODS:PubMed and CNKI databases were searched for relevant literature published from 1990 to 2023,with"macrophage polarization,M1/M2 macrophage,periodontitis,periodontitis treatment,macrophage polarization and periodontitis,osteoimmunology,ferroptosis,macrophage polarization and ferroptosis,periodontitis and ferroptosis"as the English and Chinese search terms.After the initial screening,96 articles were selected for review. RESULTS AND CONCLUSION:The switch between different phenotypes of macrophages is closely related to the tissue destruction caused by periodontitis,and various cytokines and inflammatory mediators secreted from macrophages are involved in the destruction and repair of periodontal tissue.Therefore,regulation of macrophage polarization and cytokine secretion in inflammatory state helps to alleviate periodontitis inflammation and improve the periodontal microenvironment,thereby reducing tissue destruction or promoting periodontal tissue regeneration.Many studies have been conducted to develop drugs or biomaterials to modulate macrophage function for the purpose of immunomodulatory treatment of periodontitis.However,macrophages act throughout the development of periodontitis and play an important role in the process of anti-infection,bone destruction and bone repair,and polarization is a complex and dynamic process influenced by many factors.Therefore,further exploration on possible mechanisms is still needed to clarify the interaction between materials or drugs and macrophages.

Objective To investigate the effect of Jiaotaiwan on brain insulin-PI3K/AKT pathway in a mouse model of Alzheimer's disease(AD).Methods Fifty 3-month-old male APP/PS1 double transgenic mice were randomized into AD model group,low-,medium-and high-dose Jiaotaiwan treatment groups,and donepezil treatment group.Cognitive functions of the mice were assessed using water maze and open field tests,and neuronal pathologies were observed with HE staining and Nissl staining;immunohistochemistry was used to detect amyloid Aβ deposition in the brain.Fasting serum insulin levels of the mice were measured,and the expressions of Aβ42,insulin-PI3K/AKT pathway components and downstream glucose transporters in the brain tissue were detected with RT-qPCR and Western blotting.Results The AD mouse models exhibited obvious impairment of learning and memory abilities,significantly reduced hippocampal neurons,and obvious Aβ amyloid plaques in the brain tissue with increased Aβ42 protein expression(P<0.05)and insulin resistance index,decreased hippocampal PI3K expressions,lowered expressions of AKT and InR,reduced expressions of GLUT1,GLUT3,and GLUT4,and increased expression of GSK3β in both the hippocampus and cortex.Treatment with Jiaotaiwan and donepezil both effectively improved memory ability of the mouse models,increased the number of hippocampal neurons,reduced Aβ amyloid plaques and increased the expressions of PI3K,AKT,InR,GLUT1,GLUT3 and GLUT4 in the hippocampus and cortex.Conclusion Jiaotaiwan improves learning and memory abilities of APP/PS1 double transgenic mice and delay the development of AD by activating the PI3K/AKT pathway and regulating the expression levels of its downstream GLUTs in the brain.