ObjectiveTo establish a quality evaluation method for Zhuye Shigao granules（Zhuye Shigaotang） based on fingerprint and determination of index components， and to investigate the therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome. MethodsThe fingerprint of Zhuye Shigao granules was established by high performance liquid chromatography（HPLC）， and the methods for determination of total calcium， orientin， isoorientin， ginsenosides Rg₁， Re and Rb₁ and other 2 index components were established. Fifty ICR mice were randomly divided into the blank group， model group， Zhuye Shigao granules low， medium and high dose groups（9.3， 18.6， 37.2 g·kg^-1·d^-1）， with 10 mice in each group. In addition to the blank group， the model mice with cold-dampness pestilence attacking lung syndrome was prepared by nasal drip of lipopolysaccharide combined with cold-dampness environment. Each administration group was given the corresponding liquid by gavage according to the dose， while the blank group and model group were given the same volume of normal saline by gavage. Then， the body temperature and organ index of mice in each group were measured， hematoxylin-eosin（HE） staining was used to investigate the lung tissue injury of mice in each group， and enzyme-linked immunosorbent assay（ELISA） was used to detect the changes of tumor necrosis factor-α（TNF-α）， interleukin（IL）-lβ， IL-6， IL-10 levels in serum and lung tissue， as well as immunoglobulin（Ig）A and IgM levels in serum. ResultsThe fingerprint similarity of 10 batches of Zhuye Shigao granules was>0.950， and 20 common peaks were calibrated. Seven of them were identified， including peak 11（isoorientin）， peak 12（orientin）， peak 14（apioside liquiritin）， peak 15（liquiritin）， peak 17（apioside isoliquiritin）， peak 19（isoliquiritin） and peak 20（liquiritigenin）. The results of quantitative analysis showed that the content range of each index component in 10 batches of Zhuye Shigao granules was as follows：Total calcium of 9.978-11.294 mg·g^-1， isoorientin of 0.033-0.041 mg·g^-1， orientin of 0.046-0.055 mg·g^-1， ginsenoside Rg₁+ginsenoside Re of 0.748-0.762 mg·g^-1， ginsenoside Rb₁ of 0.151-0.197 mg·g^-1， liquiritin of 1.106-1.366 mg·g^-1， glycyrrhizic acid of 0.904-1.182 mg·g^-1_. Compared with the blank group， the body temperature of mice in the model group was significantly increased， the organ indexes of liver， lung and spleen were significantly decreased， the organ index of thymus was significantly increased， HE staining of lung tissue showed infiltration of inflammatory cells， a small amount of serous exudation was observed in the alveoli， and lung tissue was damaged. After the intervention of Zhuye Shigao granules， the pathological changes were improved compared with the model group. The expression levels of IL-1β， IL-6 and TNF-α were significantly increased， the expression level of IL-10 was significantly decreased in serum and lung tissue. The levels of IgA and IgM in serum were significantly decreased（P<0.01）. Compared with the model group， the body temperature， the organ indexes and immune factor levels in serum and lung tissue of mice in the Zhuye Shigao granules medium and high dose groups were significantly reduced（P<0.05， P<0.01）. ConclusionIn this study， the quality evaluation of Zhuye Shigao granules was carried out based on fingerprint combined with determination of index components， and the fingerprint of four herbs（Lophatheri Herba， Ophiopogonis Radix， Pinelliae Rhizoma and Glycyrrhizae Radix et Rhizoma） in this formula and the determination of 8 index components were established. The therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome may be related to inhibiting inflammatory response and mediating immune regulation.

ObjectiveTo clarify the neuroprotective effect of black Panacis Quinquefolii Radix（PQR） and explore its active ingredients， with the aim of establishing an activity-oriented quality evaluation method. MethodsTransgenic Tg（HuC∶EGFP） zebrafish was used to establish a neuronal injury model by aluminum chloride immersion. Different doses（10， 20 mg·L^-1） of PQR and black PQR ethanol extracts were administered. The neuroprotective effects of PQR and black PQR were compared by analyzing the fluorescent area and intensity of zebrafish neurons. Based on ultra-performance liquid chromatography（UPLC）， a fingerprint profile of black PQR was established， followed by principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Differential components were screened using the criteria of variable importance in the projection（VIP） value>1 and P<0.05. The neuroprotective activity of 14 batches of black PQR was assessed， and Spearman correlation analysis was used to identify saponins related to neuroprotective activity， which were then validated. Based on the above results， active marker components were determined， and an UPLC method was established for their quantitation with clear content limits. ResultsPharmacological efficacy results showed that both PQR and black PQR at different doses could significantly improved neuronal damage in zebrafish. At a dose of 20 mg·L^-1， black PQR demonstrated superior efficacy（P<0.05）. The fingerprint similarities of 14 batches of black PQR were>0.94， with 26 common peaks identified. Through comparison with the reference standards， 8 components were confirmed， including peak 1（ginsenoside Rg₁）， peak 2（ginsenoside Re）， peak 5（ginsenoside Rb₁）， peak 9（ginsenoside Rd）， peak 16［ginsenoside 20（S）-Rg₃］， peak 17［ginsenoside 20（R）-Rg₃］， peak 18（ginsenoside Rk₁）， and peak 19（ginsenoside Rg₅）. The results of PCA and OPLS-DA indicated that there were differences in saponins among black PQR samples from different origins， and 12 differential components were screened. All 14 batches of black PQR exhibited good protective effects on zebrafish neurons， with Shaanxi-produced black PQR showing superior protective effects compared to the other three production regions. Spearman correlation analysis revealed that a total of 11 components， including ginsenosides 20（S）-Rg₃， 20（R）-Rg₃， Rk₁ and Rg₅， showed a significant positive correlation with the neuroprotective effect in zebrafish（P<0.05）. The activity validation results indicated that ginsenosides 20（S）-Rg₃， 20（R）-Rg₃， Rk₁ and Rg₅ were the primary components responsible for the neuroprotective effects of black PQR. Quantitative analysis showed that the content of ginsenoside 20（S）-Rg₃ in 14 batches of black PQR ranged from 0.17% to 0.52%， and the repair rate of neuronal damage ranged from 42.77% to 97.83%. ConclusionBased on the fingerprint and neuronal protective activity， the spectrum-effect related quality control model of black PQR was established， with ginsenoside 20（S）-Rg₃ as the quality control index， and the neuronal damage repair rate≥60% as the evaluation standard， the minimum limit of ginsenoside 20（S）-Rg₃ in black PQR should be≥0.20%.

Objective:To explore the effect of melatonin on mitophagy in the brain tissue of rats with early subarach-noid hemorrhage(SAH).Methods:The rat model of SAH was established by internal carotid artery puncture and trea-ted with melatonin by intraperitoneal injection.At 24 hours after the operation,the neurological function of rats was de-tected by mNSS score and balance beam test.The contents of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)and in brain tissue were detected by commercial reagent kits.The expression changes of PINK1/Parkin,microtubule-associated protein1A/1B-light chain3(LC3)and p62 in brain tissue were detected by Western blot.Results:Melatonin can significantly improve the neurological function of SAH rats,reduce the con-tents of MDA(P<0.05)and ROS(P<0.05),and increase the content of GSH(P<0.05).Meanwhile,Western blot detection indicated that melatonin could increase the expression of PINK1 and Parkin(P<0.05),increase the ratio of LC3-II/LC3-I(P<0.05),and reduce the expression of p62(P<0.05).Conclusion:Melatonin alleviates early brain injury in rats with subarachnoid hemorrhage through PINK1/Parkin-mediated mitophagy.

Objective:To evaluate the effect of deep brain stimulation(DBS)of the lateral cerebellar nucleus(LCN)on synaptic plasticity in a traumatic brain injury(TBI)rat model of the medial prefrontal cortex(mPFC).Methods:A controllable mPFC injury rat model was prepared using a hydraulic impact method.Two weeks after inju-ry,stimulation electrodes were implanted into the left LCN.Electrical stimulation was initiated at the 4th week after injury and continued for 30 days.The learning and memory function of the rats was evaluated using the Morris water maze.RT-qPCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor(BDNF),postsynaptic density protein 95(PSD95),and growth-associated protein 43(GAP43)in the mPFC region.Results:The learning and memory function of the mPFC-injured rats significantly decreased.The expressions of BDNF,PSD95,and GAP43 in mPFC decreased.DBS treatment significantly enhanced the learning and memory ability of mPFC-injured rats,and simultaneously upregulated the expressions of BDNF,PSD95,and GAP43 in mPFC.Conclusion:LCN DBS is an effective method for treating cognitive impairment caused by TBI,and it may achieve this through activating the neural plasticity mechanism of the mPFC.

Objective:To evaluate the effect of deep brain stimulation(DBS)of the lateral cerebellar nucleus(LCN)on synaptic plasticity in a traumatic brain injury(TBI)rat model of the medial prefrontal cortex(mPFC).Methods:A controllable mPFC injury rat model was prepared using a hydraulic impact method.Two weeks after inju-ry,stimulation electrodes were implanted into the left LCN.Electrical stimulation was initiated at the 4th week after injury and continued for 30 days.The learning and memory function of the rats was evaluated using the Morris water maze.RT-qPCR and Western blot were used to detect the mRNA and protein expressions of brain-derived neurotrophic factor(BDNF),postsynaptic density protein 95(PSD95),and growth-associated protein 43(GAP43)in the mPFC region.Results:The learning and memory function of the mPFC-injured rats significantly decreased.The expressions of BDNF,PSD95,and GAP43 in mPFC decreased.DBS treatment significantly enhanced the learning and memory ability of mPFC-injured rats,and simultaneously upregulated the expressions of BDNF,PSD95,and GAP43 in mPFC.Conclusion:LCN DBS is an effective method for treating cognitive impairment caused by TBI,and it may achieve this through activating the neural plasticity mechanism of the mPFC.

Objective:To explore the effect of melatonin on mitophagy in the brain tissue of rats with early subarach-noid hemorrhage(SAH).Methods:The rat model of SAH was established by internal carotid artery puncture and trea-ted with melatonin by intraperitoneal injection.At 24 hours after the operation,the neurological function of rats was de-tected by mNSS score and balance beam test.The contents of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)and in brain tissue were detected by commercial reagent kits.The expression changes of PINK1/Parkin,microtubule-associated protein1A/1B-light chain3(LC3)and p62 in brain tissue were detected by Western blot.Results:Melatonin can significantly improve the neurological function of SAH rats,reduce the con-tents of MDA(P<0.05)and ROS(P<0.05),and increase the content of GSH(P<0.05).Meanwhile,Western blot detection indicated that melatonin could increase the expression of PINK1 and Parkin(P<0.05),increase the ratio of LC3-II/LC3-I(P<0.05),and reduce the expression of p62(P<0.05).Conclusion:Melatonin alleviates early brain injury in rats with subarachnoid hemorrhage through PINK1/Parkin-mediated mitophagy.

Objective To compare the effects of using three different filling appliances,CompothixoTM resin filler,conventional resin filler,and SonicFill sonic handpiece,on marginal microleakage after auxiliary resin filling of class Ⅱ cavities.Methods A total of 40 permanent molars extracted from the Department of Stomatology,Affiliated Hospital of Shandong Second Medical University(Weifang Medical University Affiliated Hospital)from October 2020 to February 2021 were collected and randomly divided into four groups:A,B,C,and D.Standard class Ⅱ cavities were prepared in the proximal and distal middle of the isolated teeth,filled according to the standard filling requirements of the instruments in each grouping,dissected after temperature cycling and methylene blue staining,and the dye penetration depths of the dental tissues and restorative axial and gingival walls were observed and graded under the body-view microscope to calculate the microleakage degree scores.Results Microleakage occurred in four groups,and tested by the Mann-Whitney U test,the microleakage scores of group A was higher than those of the other three groups(P<0.05);There was no statistically significant difference between groups B and C(P>0.05);The scores of groups B and C were higher than those of group D(P<0.05).Group A had higher microleakage scores than group D(P<0.05),and there was no statistically significant difference between the remaining groups(P>0.05).Conclusion The use of the CompothixoTM resin filler is beneficial in reducing microleakage between the resin and the tooth tissue when compared to conventional filling appliances;Compare with the SonicFill handpiece,the use of the CompothixoTM resin filler is not conducive to the reduction of microleakage between the SonicFill ultrasonic resin and the tooth tissue.

Objective The purpose of this study was to investigate the differences in the anti cyclic fatigue perfor-mance of Woride KS(WKS),Proteper Gold(PTG),and Hyflex CM(HCM)nickel titanium instruments with different tip diameters in curved root canal models,and to provide reference for the targeted selection of suitable nickel titanium in-struments in clinical preparation of curved root canals.Methods Three kinds of new nickel titanium files with 20#and 25#(0.20 mm and 0.25 mm)tip diameters were selected,including WKS(20/0.06),WKS(25/0.06),PTG(20/0.07),PTG(25/0.08),HCM(20/0.06),and HCM(25/0.06),each with 20 files.According to the recommended speed and torque of the motor,the anti cycle fatigue performance of the nickel titanium file was tested in 30° and 60° stainless steel root canal models.The time from rotation to fatigue fracture(TTF)of the nickel titanium file was recorded with a camera and timer,and the fragment length(FL)was measured and recorded with a Vernier scale.Results Com-parison of TTF of the same type of file.① WKS:the TTF of files with the same tip diameter in a 30° curved root canal was longer than 60°(P＜0.05);The TTF of 25# nickel titanium files in the same angle curved root canal was longer than that of 20# nickel titanium files(P＜0.05).②PTG:the 20# nickel titanium files had a TTF longer than 60° in a 30° curved root canal(P＜0.05),while the 25# nickel titanium files had no statistically significant difference in TTF be-tween the 30° curved root canal and 60° curved root canal(P＞0.05);In a 30° curved root canal,the TTF of 20# nickel titanium files was longer than that of 25# nickel titanium files(P＜0.05).In a 60° curved root canal,there were no sta-tistically significant difference between the TTF of 20# nickel titanium files and 25# nickel titanium files(P＞0.05).③HCM:the TTF of files with the same tip diameter in a 30° curved root canal was longer than 60°(P＜0.05);The TTF of 20# nickel titanium files in the same angle curved root canal is longer than that of 25# nickel titanium filea.Compari-son of TTF of different files:in a 30° curved root canal,there was no statistically significant difference in TTF among the three types of 20# nickel titanium files(P＞0.05).The TTF of 25# WKS was longer than that of other files(P＜0.05);In a 60 ° curved root canal,the TTF of 20# HCM was longer than other files with the same tip diameter(P＜0.05),and the TTF of 25# WKS was longer than HCM with the same tip diameter(P＜0.05).The FL of 20# PTG,25#PTG,and 20# HCM in the 30° curved root canal model is significantly longer than that of 60°(P＜0.05),while there is no statistical difference in FL among 20# WKS,25# WKS,and 25# HCM in the 30° and 60° curved root canal models.Conclusion The anti cyclic fatigue performance of WKS is significantly superior to PTG and HCM in a 30° curved root canal.In a 60° curved root canal,20# HCM had a significant advantage in terms of anti cyclic fatigue performance compared to other files,while only 25# WKS had a significant advantage in anti cyclic fatigue performance compared to HCM.

Objective:To study the role of DDX3X/NF-κB pathway in early neuronal apoptosis in subarachnoid hemorrhage(SAH)mice.Methods:The mouse model of SAH was established by internal carotid artery puncture,and the neurological function score of the mice was evaluated.The DDX3X expression was knocked down using recombinant lentivirus expressing DDX3X targeted shRNA(Lv-shDDX3X),or the NF-κB pathway was inhibited by NF-κB-IN-1(IN-1).Western Blot was used to detect the expression of DDX3X and NF-κB(p65)in mouse cortex.TUNEL/NeuN staining was used to detect the apoptosis of cerebral cortex neurons.Results:Twenty-four hours after SAH operation,the neurological function of mice was significantly impaired(P＜0.05).While the expression of DDX3X was signifi-cantly increased and the expression of NF-κB(p65)was significantly decreased in the cortex(P＜0.05).When the DDX3X expression is knocked down firstly,then SAH surgery is performed.The neurological function of mice was sig-nificantly recovered,and the expression of NF-κB(p65)protein was significantly higher than that in SAH group(P＜0.05);If the NF-κB activity was inhibited by IN-1 while DDX3X knockdown,there is no significant recovery of neuro-logical function in SAH mice.TUNEL/NeuN staining showed that the number of TUNEL-positive neurons in the brain tissue after DDX3X knockdown was less than that in the SAH group(P＜0.05),while the number of TUNEL-positive neurons was not significantly reduced when IN-1 was used to inhibit NF-κB activity at the same time of DDX3X knock-down.Conclusion:DDX3X/NF-κB mediated cell death in mice with early brain injury after SAH.

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.