BACKGROUND:Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital,Beijing University of Chinese Medicine,with clear clinical efficacy,but the specific mechanism needs to be elucidated. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group(n=12)and a model group(n=52).An animal model was established by the Rice-Vannucci method.After successful modeling,52 model rats were randomly divided into control model group(n=12),minocycline group,and the low-,medium-,and high-dose groups of the Shujin Jiannao Prescription(n=10 per group).Rats in the minocycline group were given 40 mg/kg·d minocycline by gavage;rats in the low-,medium,and high-dose groups were given 4,8,and 16 g/kg·d Shujin Jiannao Prescription granules by gavage,respectively;and rats in the normal group and control model group were given an equal dose of normal saline by gavage.Medication in each group was given once a day for 1 week.The rats in each group were evaluated behaviorally using suspension test,abnormal involuntary movement score,and Bederson score.The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining.The levels of tumor necrosis factor α,interleukin 1β,and interleukin 10 in the cerebral cortex were determined using ELISA.The positive expressions of Janus kinase 2(JAK2),phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)in the cerebral cortex were detected using immunohistochemistry.The protein expression levels of JAK2,p-JAK2,and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION:Compared with the normal group,the suspension test score and involuntary movement score were decreased in the control model group(P＜0.01 or P＜0.05).The pathological results showed structural disruption of nerve cells,formation of large numbers of vacuoles,cell swelling,and increased intercellular space in the control model group.In addition,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased(P＜0.01),the expression of interleukin 10 was decreased(P＜0.05),and the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly increased(P＜0.01)in the control model group compared with the normal group.Compared with the model group,minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats(P＜0.01 or P＜0.05)and ischemic-hypoxic pathological changes were attenuated,with only a small amount of necrotic nerve cells and a few vacuoles,and reduced intercellular space.Moreover,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group(P＜0.05),while the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly decreased(P＜0.01).The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group.To conclude,Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury.The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway.

BACKGROUND:At present,there are some limitations in evaluating the severity of acute spinal cord injury,and a rapid and accurate evaluation method is urgently needed. OBJECTIVE:To analyze the correlation between the expression levels of serum oxidative stress and nerve injury indexes and the severity of the disease in patients with acute spinal cord injury. METHODS:A total of 65 patients were included in the study from August 2020 to May 2022,including 32 patients in the experimental group(acute spinal cord injury)and 33 patients in the control group(simple spinal fracture).American Spinal Injury Association(ASIA)Impairment Scale and neurological function score were evaluated within 8 hours of admission.Meanwhile,serum levels of superoxide dismutase,malondialdehyde,glutathione,nitric oxide,glial fibrillary acidic protein and neuron-specific enolase were detected and compared between the two groups.The correlation between the expression levels of the above serological indicators in serum and ASIA impairment grade and AISA neural function score was analyzed. RESULTS AND CONCLUSION:The average serum levels of superoxide dismutase and glutathione in the experimental group were significantly lower than those of the control group(P<0.001),while the average serum levels of malondialdehyde,nitric oxide,glial fibrillary acidic protein and neuron-specific enolase in the experimental group were higher than those of the control group(P<0.01).The serum levels of superoxide dismutase and glutathione in the experimental group were positively correlated with the damage grade of AISA(r=0.862 4,0.849 3,P<0.01),while the serum levels of malondialdehyde,nitric oxide,glial fibrillary acidic protein and neuron-specific enolase were negatively correlated with the damage grade of AISA(r=-0.866 1,-0.638 1,-0.746 6,P<0.001),and the serum level of nitric oxide was not significantly correlated with the damage grade of AISA(r=-0.177 5,P>0.05).The serum level of glutathione in the experimental group was positively correlated with AISA sensory function scores(r=0.569 9,P<0.001),while the serum levels of malondialdehyde,glial fibrillary acidic protein and neuron-specific enolase were negatively correlated with AISA sensory function scores(r=-0.574 1,-0.099 2,-0.708 6,P<0.05),and the serum levels of superoxide dismutase and nitric oxide were not significantly correlated with AISA sensory function scores(r=0.230 8,-0.376 2,P>0.05).The serum levels of superoxide dismutase and glutathione in the experimental group were positively correlated with ASIA motor function scores(r=0.380,0.524 7,P<0.05);the serum levels of malondialdehyde,glial fibrillary acidic protein and neuron-specific enolase were negatively correlated with AISA motor function scores(r=-0.577 9,-0.452 2,-0.662 8,P<0.05);and the level of nitric oxide had no significant correlation with AISA motor function scores(r=-0.049 7,P>0.05).To conclude,the serum levels of superoxide dismutase,malondialdehyde,glutathione,nitric oxide,glial fibrillary acidic protein and neuron-specific enolase in serum of patients with acute spinal cord injury are significantly correlated with ASIA impairment grade and ASIA neural function score,which could be used as biomarkers for early clinical assessment of disease severity.

ObjectiveTo explore the mechanism of Tangbikang granules （TBK） against diabetic peripheral neuropathy （DPN） based on network pharmacology and in-vivo experiment. MethodThe active components in medicinals of TBK and their target genes were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. The active components of the medicinals which are not included in TCMSP were searched from previous research. After the analysis of drug-likeness by SwissADME， the target genes of them were predicted with SwissTargetPrediction. DPN-related target genes were retrieved from GeneCards. The common targets of the disease and the prescription were the hub genes of TBK against DPN， which were uploaded to Metascape for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. High-sugar and high-fat diet and low-dose streptozotocin （STZ， ip） were employed to induce diabetes in rats， and then the model rats were respectively treated with low-dose （0.625 g·kg^-1）， medium-dose （1.25 g·kg^-1）， and high-dose （2.5 g·kg^-1） TBK for 12 weeks. Sensory nerve conduction velocity （SNCV） was evaluated. After hematoxylin and eosin （HE） staining， the sciatic nerve was observed under light microscope to examine the nerve damage. Real-time PCR was performed to detect the gene expression of adenosine monophosphate-activated protein kinase （AMPK） pathway-related targets in rat sciatic nerve， and Western blot to measure the protein expression of AMPK and phosphorylated （p）-AMPK in rat sciatic nerve. ResultThe main active components of TBK， such as quercetin， kaempferol， β-sitosterol， leech pteridine A， stigmasterol， and baicalein were screened out， mainly acting on interleukin-6 （IL-6）， tumor necrosis factor （TNF）， protein kinase B （Akt）， JUN， and HSP90AA1 and signaling pathways such as AMPK， nuclear factor-κB （NF-κB）， and Janus kinase/signal transducer and activator of transcription （JAK/STAT）. Molecular docking results showed that β-sitosterol and stigmasterol had high binding affinity with IL-6， TNF， JUN， and HSP90AA1. As for the animal experiment， compared with the normal group， model group had low SNCV of sciatic nerve （P<0.01）， disordered and loose myelinated nerve fibers with axonotmesis and demyelinization， low mRNA expression of AMPKα， AMPKβ， peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α）， Sirtuin 3 （SirT3）， mitochondrial transcription factor A （TFAM）， and low p-AMPK/AMPK ratio in sciatic nerve （P<0.05， P<0.01）. Compared with the model group， TBK of the three doses raised the SNCV （P<0.01）， restored nerve morphology and nerve compactness， and increased the mRNA expression of AMPKα， AMPKβ， PGC-1α， SirT3， and TFAM （P<0.05， P<0.01）. The ratio of p-AMPK/AMPK in the high-dose and medium-dose TBK groups was higher than that in the model group （P<0.01）， while the protein expression in the low-dose TBK group was insignificantly different from that in the model group. ConclusionTBK exerts therapeutic effect on DPN through multiple pathways and targets. The mechanism is that it activates and regulates AMPK/PGC-1α/SirT3 signaling， which lays a basis for further study of TBK in the treatment of DPN.

With the development of researches on the relationship between periodontal health and general health, more and more evidences showed that periodontitis was closely related to oculopathy, while the mechanisms were not very clear at present. This article will focus on the influences of periodontitis on the occurrence and development of various oculopathy such as diabetic retinopathy and senile macular degeneration, and discuss the possible mechanisms of the influence by periodontitis. This will provide a theoretical basis for the new ideas on prevention and treatment of oculopathy.

BACKGROUND: The periosteum is involved in fracture healing and bone tissue repair because it contains cells with osteogenic potential, and its role in repairing bone defects has become a focus of current research. OBJECTIVE: To review the role of periosteum in fracture healing and bone tissue repair. METHODS: A computer-based online search of China National Knowledge Infrastructure database and PubMed was performed for articles regarding the role of periosteum in fracture healing and bone tissue repair published from January 1964 to December 2019. Key words included “periosteum, bone healing, periosteum cells” in English and Chinese. Finally, a total of 48 articles were included to review. RESULTS AND CONCLUSION: (1) The integrity of periosteum plays an important role in fracture healing and bone tissue repair, and has been widely used in bone tissue engineering due to its good osteogenic properties, excellent material properties and barrier effect. (2) The mechanism of periosteum in the process of distraction ostegogenesis is still controversial. It has been confirmed that absence of periosteum does not affect the osteogenic effect of mandibular defects, but the role of periosteum in osteogenic effect of limb defects remains to be further studied.

Objective: To explore the feasibility of establishment of a artificial joint aseptic loosening mouse model by cobalt-chromium particles stimulation. Methods: Twenty-four 8-week-old male severe combined immunodeficient (SCID) mice were divided into experimental group ( n=12) and control group ( n=12). The titanium nail was inserted into the tibial medullary cavity of mouse in the two groups to simulate artificial joint prosthesis replacement. And the cobalt-chromium particles were injected into the tibial medullary cavity of mouse in experimental group. The survival of the mouse was observed after operation; the position of the titanium nail and the bone mineral density of proximal femur were observed by X-ray film, CT, and Micro-CT bone scanning; and the degree of dissolution of the bone tissue around the tibia was detected by biomechanical test and histological staining. Results: Two mice in experimental group died, and the rest of the mice survived until the experiment was completed. Postoperative imaging examination showed that there was no obvious displacement of titanium nails in control group, and there were new callus around the titanium nails. In experimental group, there was obvious osteolysis around the titanium nails. The bone mineral density of the proximal tibia was 91.25%±0.67%, and the maximum shear force at the tibial nail-bone interface was (5.93±0.85) N in experimental group, which were significantly lower than those in control group [102.07%±1.87% and (16.76±3.09) N] ( t=5.462, P=0.041; t=3.760, P=0.046). Histological observation showed that a large number of inflammatory cells could be seen around the titanium nails in experimental group, while there was no inflammatory cells, and obvious bone tissue formation was observed in control group. Conclusion: The artificial joint aseptic loosening mouse model can be successfully established by cobalt-chromium particles stimulation.

Objective To investigate the effect of periosteum deficiency on the treatment of segmental bone defect of lower extremity by Ilizarov bone transport Technique.Methods From April 2009 to April 2016,data of 54 cases with lower limb segmental bone defect who were treated by Ilizarov bone transport technique were retrospectively analyzed.According to intraoperative observation of the distribution of periosteum at the osteotomy site,all cases were divided into two groups.There were 10 cases in absent periosteum group,including 6 males and 4 females,aged from 23 to 57 years old (mean,38.20±9.90 years old).There were 3 cases locating at femur,and 7 cases at tibia.4 cases had non-infective bone defect,while the other 6 had infective bone defect.Segmental bone defect ranged from 4.0 to 14.0 cm (mean,6.85±2.87 cm).There were 44 cases in normal periosteum group,including 32 males and 12 females,aged from 22 to 65 years old (mean,38.90±10.10 years old).There were 9 cases locating at femur and 35 cases at tibia.12 cases had non-infective bone defect,while 32 cases had infective bone defect.Segmental bone defect ranged from 4.0 to 9.0 cm (mean,6.09±1.54 cm).Visual analogue scale (VAS) was used to estimate the pain during bone transport.The distraction index,bone healing index,external fixator index was collected to evaluate the distraction osteogenesis and mineralization.The Paley method was utilized to appraisal the function of proximal joints in bone transport.Results All the 54 cases were followed up for 24-48 months,with an average of 28.50±5.70 months.All skin and soft tissue defects were satisfied healed.All limbs were restored to their original lengths,and bone healing was eventually achieved.There were no statistically significant differences in the distraction index,bone healing index,external fixator index,VAS between absent periosteum group (12.29± 1.04 d/cm,39.00±3.96 d/cm,51.25±3.69 d/cm,3.30± 1.77 score) and normal periosteum group (11.67±0.96 d/cm,38.07±4.22 d/cm,49.74±4.25 d/cm,3.36± 1.66 score)(P ＞ 0.05).Paley adjacent joint function evaluation showed no statistically difference between absent periosteum group [good rate 100％(10/10)] and normal periosteum group [good rate 97.7％ (43/44)] (P=0.901).The overall incidence of complications was 20.0％ (2/10) in the absence of periosteum group,with 1 case of mild infection and 1 cases of tissue contracture incarcerated at the docking site.The overall incidence of complications was 27.3％ (12/44) in the normal of periosteum group,with 2 cases of mild infection,3 cases of poor limb strength alignment,6 cases of tissue contracture incarcerated at the docking site and 1 case of severe foot drop deformity.There was no statistically significant difference in the overall incidence of complications between the two groups (P=0.636).Conclusion The absence of periosteum at the epiphyseal osteotomy site hasno significant effect on Ilizarov bone transport technique in the treatment of segmental bone defects of lower limbs in bone regeneration and healing,mineralization and bone remodeling,and does not increase the incidence of complications.

Objective: To investigate the clinical efficacy of minimally invasive plate oseoynthesis (MIPO) assisted by a self-designed lower limb axial tractor in the treatment of complex tibial plateau fractures. Methods: The data were retrospectively analyzed of the 31 patients who had undergone surgical treatment for complex tibial plateau fractures from June 2015 to April 2018 at Department of Orthopeadics, Hospital Affiliated to Binzhou Medical College. MIPO was conducted with the assistance of the axial tractor in 17 of them. They were 12 men and 5 women with an age of 50.8±14.7 years, with 8 cases of Schatzker type Ⅴ and 9 ones of Schatzker type Ⅵ. MIPO was conducted without the assistance of the axial tractor but with the assistance of manipulation reduction in the other 14 patients. They were 9 men and 5 women with an age of 48.9±9.5 years, with 6 cases of Schatzker type Ⅴ and 8 ones of Schatzker type Ⅵ. The 2 groups were compared in terms of operation time, intraoperative bleeding, and Rasmussen scores for knee function and medial angle of the proximal tibia at the last follow-up. Results: The 2 groups were comparable due to insignificant differences in the preoperative general data between them (P>0.05). The tractor and manipulation groups were followed up for a mean of 9.8 months and 10.9 months respectively. Bone healing was achieved in all patients. For the tractor and manipulation groups, respectively, the operation time was127.8±40.8 min and 174.1±66.8 min, and the intraoperative bleeding 115.6±41.7 mL and 184.3±91.4 mL, showing significant differences between them (P<0.05). In the tractor group at the last follow-up, the knee flexion was 126.1°±9.3°, the medial angle of the proximal tibial 87.8°±1.4°, the posterior inclination angle 8.7°±3.8° and the Rasmussen score 26.5±2.9 points; in the manipulation group at the final follow-up, the knee flexion was 124.7°±8.9°, the medial angle of the proximal tibial 86.9°±2.1°, the posterior inclination angle 9.8°±4.1° and the Rasmussen score 25.6±3.2 points. There were no significant differences between the 2 groups in the above items (P>0.05). Loss of force line was observed in none of the patients at the last follow-up. Conclusion Compared with conventional manipulation reduction, use of our self-designed lower limb axial tractor can shorten operation time, reduce bleeding and facilitate reduction of fracture fragments in the treatment of tibial plateau fractures of Schatzker types Ⅴ and Ⅵ.

Objective To analyze the sensitivity of auxiliary examinations in different periods of sporadic Creutzfeldt-Jakob disease (sCJD).Methods The clinical data of 53 sCJD patients were retrospectively analyzed including the different stages of skull diffusion-weighted magnetic resonance imaging (DWI),24-hour ambulatory electroencephalogram (EEG),18F-FDG PET/CT (PET-CT)and cerebrospinal fluid 14-3-3 protein.When calculating the sensitivity of an auxiliary examination,the diagnostic criteria were defined by combining the specific clinical manifestations with two or more positive results of other auxiliary examinations.Results There were 24,53 and 22 sCJD patients,respectively,met the criterion of early (E),middle (M) and later (L) stage of disease (some patients fit 2 or 3 stages).The sensitivity ofDWl (E:58.3％ M:85.4％,L:94.7％),EEG (E:45.8％,M:62.7％,L:77.8％),14-3-3 protein in cerebrospinal fluid (E:11.1％,M:52.9％) and PET-CT (E:80％,M:100％) increased gradually with disease progression,The sensitivity of PET-CT was higher than the other auxiliary examinations for E and M stages;no PET-CT was conducted in L stage.High signal regions mainly distributed in the cortex in E and M stages,but in L stage,no significant difference was found on the distribution of high signal regions between cortex and basal ganglia.Conclusions The sensitivities of the auxiliary examinations were different for sCJD patients in different stages.Reexaminations in different periods may improve the sensitivity for sCJD diagnosis.The sensitivity of PET-CT was high,and the combination of PET-CT and other auxiliary examinations may play a key role in the diagnosis of sCJD.

BACKGROUNDThe molecular and cellular origins of migraine headache are among the most complex problems in contemporary neurology. Up to now the pathogenesis of migraine still remains unclearly defined. The objective of this study was to explore new factors that may be related to the mechanism of migraine.

METHODSThe present study performed a comprehensive analysis of gene expression in the trigeminal nucleus caudalis induced by electrical stimulation of dura mater surrounding the superior sagittal sinus in conscious rats using microarray analysis followed by quantitative real-time reverse-transcribed polymerase chain reaction (qRT-PCR) verification. Student's two sample t-test was employed when two groups were compared. A P value <0.05 was considered to be statistically significant.

RESULTSComparing the placebo and the electrical stimulation groups, 40 genes were determined to be significantly differentially expressed. These significantly differentially expressed genes were involved in many pathways, including transporter activity, tryptophan metabolism, G protein signaling, kinase activity, actin binding, signal transducer activity, anion transport, protein folding, enzyme inhibitor activity, coenzyme metabolism, binding, ion transport, cell adhesion, metal ion transport, oxidoreductase activity, mitochondrion function, and others. Most of the genes were involved in more than 2 pathways. Of particular interest is the up-regulation of Phactr3 and Akap5 and the down-regulation of Kdr.

CONCLUSIONThese findings may provide important clues for a better understanding of the molecular mechanism of migraine.

Animals ; Dura Mater ; physiology ; Electric Stimulation ; Gene Expression ; physiology ; Male ; Microarray Analysis ; Migraine Disorders ; genetics ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Superior Sagittal Sinus ; physiology