Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

The incidence of osteoporotic thoracolumbar vertebral fracture (OTLVF) in the elderly is gradually increasing. The kyphotic deformity caused by various factors has become an important characteristic of OTLVF and has received increasing attention. Its clinical manifestations include pain, delayed nerve damage, sagittal imbalance, etc. Currently, the definition and diagnosis of OTLVF with kyphotic deformity in the elderly are still unclear. Although there are many treatment options, they are controversial. Existing guidelines or consensuses pay little attention to this type of fracture with kyphotic deformity. To this end, the Lumbar Education Working Group of the Spine Branch of the Chinese Medicine Education Association and Editorial Committee of Chinese Journal of Trauma organized the experts in the relevant fields to jointly develop Clinical guidelines for the diagnosis and treatment of osteoporotic thoracolumbar vertebral fractures with kyphotic deformity in the elderly ( version 2024), based on evidence-based medical advancements and the principles of scientificity, practicality, and advanced nature, which provided 18 recommendations to standardize the clinical diagnosis and treatment.

OBJECTIVE: To explore the regulatory effects of GABAergic neurons in the zona incerta (ZI) on sevoflurane and propofol anesthesia. METHODS: Forty-eight male C57BL/6J mice divided into 8 groups (n=6) were used in this study. In the study of sevoflurane anesthesia, chemogenetic experiment was performed in 2 groups of mice with injection of either adeno-associated virus carrying hM3Dq (hM3Dq group) or a virus carrying only mCherry (mCherry group). The optogenetic experiment was performed in another two groups of mice injected with an adeno-associated virus carrying ChR2 (ChR2 group) or GFP only (GFP group). The same experiments were also performed in mice for studying propofol anesthesia. Chemogenetics or optogenetics were used to induce the activation of GABAergic neurons in the ZI, and their regulatory effects on anesthesia induction and arousal with sevoflurane and propofol were observed; EEG monitoring was used to observe the changes in sevoflurane anesthesia maintenance after activation of the GABAergic neurons. RESULTS: In sevoflurane anesthesia, the induction time of anesthesia was significantly shorter in hM3Dq group than in mCherry group (P < 0.05), and also shorter in ChR2 group than in GFP group (P < 0.01), but no significant difference was found in the awakening time between the two groups in either chemogenetic or optogenetic tests. Similar results were observed in chemogenetic and optogenetic experiments with propofol (P < 0.05 or 0.01). Photogenetic activation of the GABAergic neurons in the ZI did not cause significant changes in EEG spectrum during sevoflurane anesthesia maintenance. CONCLUSION Activation of the GABAergic neurons in the ZI promotes anesthesia induction of sevoflurane and propofol but does not affect anesthesia maintenance or awakening.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Propofol/pharmacology* ; Sevoflurane/pharmacology* ; Zona Incerta ; Anesthesia, General ; GABAergic Neurons

Objective: To investigate the effect of co-culture of adipose-derived stem cells(ADSC) and endothelial progenitor cells(EPC) on the activity of EPC and its related mechanism． Methods: Rat ADSC and EPC were isolated，cultured，expanded and identified in vitro ．The experiment was divided into three groups : EPC group，EPC + ADSC co-culture group，and EPC + ADSC + PI3K-inhibitor group．After 48 hours of co-culture，the cells of the three groups were treated with Transwell．The effects of ADSC and EPC co-culture and PI3K / AKT pathway on EPC activity were evaluated by CCK-8 assay，scratch assay and angiogenesis assay，respectively．Western blot was used to detect vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) ，vascular endothelial-cadherin ( VE-cadherin) ，CD133 ，phospho-phosphatidylinositol 3-kinase ( phospho-phosphatidylinositol 3-kinase(p-PI3K) and phospho-protein Kinase B (p-AKT) expression levels in EPC to detect the effects of ADSC and EPC co-culture and PI3K / AKT pathway on the differentiation ability of EPC into mature endothelial cells. Results: CCK-8 results showed that the absorbance at 450 nm of EPC in EPC + ADSC co-culture group was higher than that in EPC group and EPC + ADSC + PI3K-inhibitor group at different time points，and the difference was statistically significant (P＜0. 01) ．The scratch test showed that the relative scratch distance of EPC + ADSC co-culture group was smaller than that of EPC group and EPC + ADSC + PI3K-inhibitor group after 24 hours ，and the difference was statistically significant (P＜0. 01) ．Tube formation assay showed that the average number of tubelike structures formed in EPC + ADSC co-culture group was higher than that in EPC group and EPC + ADSC + PI3K-inhibitor group，and the difference was statistically significant (P＜0. 01) ．Western blot showed that the expression levels of VEGFA，eNOS，VE-cadherin，p-PI3K and p-AKT of EPC in EPC + ADSC co-culture group were higher than those in EPC group and EPC + ADSC + PI3K-inhibitor group．The expression level of CD133 in EPC group was lower than that in EPC + ADSC + PI3K-inhibitor group，and the difference was statistically significant (P ＜0. 01) ． Conclusion Co-culture of ADSC and EPC can improve the proliferation，migration，differentiation and vasogenic activity of EPC through the regulation of PI3K / AKT pathway.

COVID-19 has been prevalent for three years. The virulence of SARS-CoV-2 is weaken as it mutates continuously. However, elderly patients, especially those with underlying diseases, are still at high risk of developing severe infections. With the continuous study of the molecular structure and pathogenic mechanism of SARS-CoV-2, antiviral drugs for COVID-19 have been successively marketed, and these anti-SARS-CoV-2 drugs can effectively reduce the severe rate and mortality of elderly patients. This article reviews the mechanism, clinical medication regimens, drug interactions and adverse reactions of five small molecule antiviral drugs currently approved for marketing in China, so as to provide advice for the clinical rational use of anti-SARS-CoV-2 in the elderly.

Objective:To investigate the factors for and prevention of muscular calf vein thrombosis (MCVT) after unilateral total knee arthroplasty (TKA).Methods:Between January 2018 and October 2020, 551 patients were admitted to Department of Orthopedics, The First Affiliated Hospital to Zhengzhou University for unilateral TKA. They were 187 males and 364 females, aged from 32 to 90 years (average, 64.6 years) and with 234 left and 317 right knees affected. They were assigned into a MCVT group ( n=77) and a non-MCVT group ( n=474) according to whether or not MCVT had happened at 7 days after operation. Recorded were the patients’ baseline information, tourniquet time, intraoperative blood loss, postoperative prothrombin time (PT), postoperative thrombin time (TT), postoperative fibrinogen (FIB), D-dimer, platelet count (PLT), postoperative bed time, knee society score (KSS), erythrocyte sedimentation rate (ESR) fall time, and C-reactive protein (CRP) fall time so as to analyze the risk factors for MCVT. Results:There were significant differences between the 2 groups in age [(66.8±7.0) versus (64.2±9.6) years], body mass index (BMI) [(28.7±2.2) versus (25.0±2.4) kg/m 2], smoking (20/57 versus 41/433), diabetes (56/21 versus 172/302), primary hypertension (45/32 versus 174/300), coronary heart disease (50/27 versus204/270), hyperlipidemia (33/44 versus 149/325), varicosity (50/27 versus 166/308), tourniquet time [(97.9±22.6) versus (83.1±10.6) min], intraoperative blood loss [(73.2±40.6) versus (62.4±11.5) mL], postoperative PT [(10.7±0.8) versus (11.9±1.0) s], TT [(15.2±1.3) versus (17.2±2.4) s], FIB [(3.7±0.8) versus (3.2±0.5) g/L], D-dimer [(1.1±1.0) versus (0.8±0.3) μg/L], PLT [(233.4±68.5) versus (178.5±27.8) 10 9/L], postoperative bed time [(17.3±2.6) versus (14.6±3.8) h], KSS [(3.32±0.7) versus (3.61±0.56) points], ESR fall time [(2.90±0.74) versus (1.55±0.64) d] and CRP fall time [(2.90±0.74) versus (1.55±0.64) d] (all P<0.05). Multivariate logistic regression analysis showed that old age (95% CI: 0.890 to 1.112, P=0.034), high BMI (95% CI: 1.012 to 1.214, P=0.046), diabetes (95% CI: 1.002 to 2.590, P=0.020), D-dimer (95% CI: 1.239 to 10.292, P=0.001) and postoperative PLT (95% CI: 1.012 to 1.112, P=0.014) were independent risk factors for MCVT. Reduced postoperative bed time (95% CI: 1.009 to 1.469, P=0.040) was a protective factor. Conclusions:As old age, high BMI, diabetes, and high postoperative levels of D-dimer and PLT may be independent risk factors for MCVT, patients with such characteristics should be alert to MCVT. Early ambulation should be encouraged in patients after unilateral TKA to reduce postoperative bed time for prevention of the disease.

Dopaminergic neurons in the ventral tegmental area (VTA) play an important role in cognition, emergence from anesthesia, reward, and aversion, and their projection to the cortex is a crucial part of the "bottom-up" ascending activating system. The prelimbic cortex (PrL) is one of the important projection regions of the VTA. However, the roles of dopaminergic neurons in the VTA and the VTA^DA-PrL pathway under sevoflurane anesthesia in rats remain unclear. In this study, we found that intraperitoneal injection and local microinjection of a dopamine D1 receptor agonist (Chloro-APB) into the PrL had an emergence-promoting effect on sevoflurane anesthesia in rats, while injection of a dopamine D1 receptor antagonist (SCH23390) deepened anesthesia. The results of chemogenetics combined with microinjection and optogenetics showed that activating the VTA^DA-PrL pathway prolonged the induction time and shortened the emergence time of anesthesia. These results demonstrate that the dopaminergic system in the VTA has an emergence-promoting effect and that the bottom-up VTA^DA-PrL pathway facilitates emergence from sevoflurane anesthesia.
Anesthesia ; Animals ; Dopaminergic Neurons/metabolism* ; Rats ; Receptors, Dopamine D1/metabolism* ; Sevoflurane/pharmacology* ; Ventral Tegmental Area/metabolism*

Objective:To investigate the feasibility of replacing the femoral prosthesis and implanting antibiotic calcium sulfate carriers in a two-stage revision for periprosthetic infection following total knee arthroplasty (TKA).Methods:Between May 2017 and January 2020, 35 patients were admitted to Department of Orthopaedic Surgery, The First Affiliated Hospital to Zhengzhou University for periprosthetic infection after TKA. They were 12 males and 23 females, aged from 49 to 84 years (average, 67.9 years). The two-stage revision for periprosthetic infection was performed for all of them and replacement of femoral prosthesis and implantation of antibiotic calcium sulfate carriers were carried out in stage-one revision. Recorded were postoperative culture of micro-organisms, white blood cell count (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) before stage-one and stage-two revisions; the Hospital for Special Surgery (HSS) knee score, range of motion (ROM) and American Knee Society Score (KSS) were compared between preoperation and the last follow-up.Results:Postoperative negative culture was found in 22 cases (62.9%), and positive one in 13 cases (37.1%) of which 4 were caused by Staphylococcus aureus, 2 by Staphylococcus epidermidis, 2 by Candida glabrata, 2 by Candida parapsilosis, one by Candida albicans, one by Mycobacterium tuberculosis and one by Escherichia coli. WBC, ESR and CRP decreased on average from 13.67×10 9/L, 49.71 mm/h and 45.13 mg/L before stage-one revision to 6.44×10 9/L, 18.79 mm/h and 7.82 mg/L before stage-two revision. All patients were followed up for an average of 22.4 months (from 8 to 41 months). At the last follow-up, ROM, HSS and KSS were significantly increased from preoperative 73.2°±15.9°, 59.5±14.6 and 36.1±6.0 to 105.6°±13.2°, 84.3±10.0 and 86.1±5.6, respectively ( P<0.05). None of the patients showed any sign of re-infection at the last follow-up. Conclusion:For patients with periprosthetic infection following total knee arthroplasty, replacing femoral prosthesis and implantation of antibiotic calcium sulfate carriers can well control infection, facilitating recovery of range of motion and function after surgery.

Due to good mechanical properties and biocompatibility, tissue engineering scaffolds have become the vital method for repairing and regenerating articular cartilage defects. With the continuous development of tissue engineering technology, many scaffolds preparation and formation methods have been developed and tested in the past decade, however, the preparation of ideal regenerative scaffolds remain controversial. As load-bearing tissue inside the body joints, the matrix structure and cell composition of articular cartilage are hierarchical, and there are several smooth natural gradients from the cartilage surface to the subchondral bone layer, including cell phenotype and number, specific growth factors, matrix composition, fiber arrangement, mechanical properties, nutrient and oxygen consumption. Therefore, in the design of regenerative scaffolds, it is necessary to achieve these gradients to regenerate articular cartilage in situ. In recent studies, many new biomimetic gradient scaffolds have been used to simulate the natural gradient of articular cartilage. These scaffolds show different mechanical, physicochemical or biological gradients in the structure, and have achieved good repair effects. The related articles on tissue engineering for the treatment of articular cartilage defects were retrieved by searching databases with key wordsarticular cartilage injury, cartilage repair and gradient scaffolds. In this work,the structural, biochemical, biomechanical and nutrient metabolism gradients of natural articular cartilage were studied and summarized firstly. Then, the latest design and construction of articular cartilage gradient scaffolds were classified. Besides that, the material composition (such as hydrogels, nanomaterials, etc.) and the preparation process (such as electrospinning, 3D printing, etc.) of grandient scaffolds were further enhanced. Finally, the prospect and challenge of biomimetic gradient scaffolds in cartilage engineering are discussed, which provides a theoretical basis for the successful application of gradient scaffolds in clinical transformation.