BACKGROUND:Primary cortical neurons and microglial cells play a crucial role in exploring cell therapies for neurological disorders,and most of the current methods for obtaining the two types of cells are cumbersome and require separate extraction.It is therefore crucial to find a convenient and rapid method to extract both types of cells simultaneously. OBJECTIVE:To explore a novel method for simultaneous extraction of primary cortical neurons and microglial cells. METHODS:Newborn suckling SD rats were taken within 24 hours.The brain was removed and placed in a dish with DMEM,and the pia mater was removed for later use.Primary neurons were extracted from the same brain tissue,and then the remaining brain tissue was used to extract microglial cells.The whole process was performed on ice.Extraction and culture steps of primary cortical neurons:The cerebral cortex was taken 2.0-3.0 mm with forceps,and the tissue was digested with papain for 20 minutes.After aborting digestion,the blown tissue presented an adherent tissue suspension.The supernatant cell suspension was obtained,filtered,and dispensed into 15 mL centrifuge tubes.After centrifugation and re-suspension,the cells were inoculated onto 6-well plate crawls coated with L-polylysine.Neuronal morphology was observed at 1-day intervals,and staining could be performed for identification using immunofluorescence staining of MAP2 and β-Tubulin by day 7.Microglia extraction and culture steps:The remaining brain tissue at 8-10 mm thick was subjected to microglial cell extraction,digested by trypsin for 20 minutes.After digestion was stopped,the tissue was blown to a homogenate,and then the homogenate was transferred to the culture bottle for culture.On day 14,the culture flasks were sealed and subjected to constant temperature horizontal shaking for 2 hours.Microglial cells were shed in the supernatant.Purified microglial cells were taken and continued to be cultured for 3 days for identification by Iba1 immunofluorescence staining. RESULTS AND CONCLUSION:(1)After 24 hours of culture,the neurons were adherent to the wall,the cytosol was enlarged,and some neurons developed synapses.After 3 and 5 days of culture,the cytosol was further enlarged,and most of the neurons were in the form of synapses,and some neurons were growing in clusters.On day 7,neuronal synapses were prolonged and thickened,and they were connected with each other to form a network.The neurons were identified by β-Tubulin and MAP2 immunofluorescence staining.(2)The cells grew close to the wall on day 1 of culture.On days 3,5,and 7,the density of microglial cells was small,and the cell morphology was bright oval or round,but the cells basically grew in clumps on the upper layer of other cells.On day 10,the density of microglial cells increased significantly.On day 14,microglial cells grew in dense clumps on the upper layer of other cells,and then they could be isolated and purified.The isolated and purified cells were taken and re-cultured to day 3 and identified as microglial cells by Iba1 immunofluorescence;their purity was greater than 95%.(3)The results show that primary cortical neurons and microglial cells obtained by this method after extraction and culture are of high purity,good morphology,and high viability.

ObjectiveTo study the characteristics of the five tones of Parkinson's disease （PD） patients based on the theory of "five-viscera phonology"， and provide references for the syndrome differentiation and treatment of PD. MethodsA total of 272 cases of PD patients were collected as the PD group， and 240 individuals， including patient family members and hospital staff， were recruited as the control group. The 25-tone analyzer was used to collect the five-tone characteristics of both PD patients and control group participants. The participants were then stratified into three age groups， 41~55， 56~70， and 71~85 years old， and categorized by gender （male and female） for analysis. The frequency and composition ratio of the five tones were analyzed for both groups across the different age ranges and genders. Additionally， the average voice frequency of each participant was calculated to compare differences between groups， stratified by age range and gender. ResultsIn the 41~55 and 56~70 age groups， male participants in the PD group and the control group exhibited the highest frequency of Yu （羽） tone， with the PD group showing a significantly higher composition ratio of Yu tone compared to the control group （P＜0.05）； for males in the 56~70 age group， the composition ratios of Shang （商） and Zhi （徵） tone in the PD group were lower than those in the control group （P＜0.05）. For males in the 71~85 age group， both the PD group and the control group had the highest frequency of Yu tone， but there was no statistically significant difference in the composition ratios of the five tones between groups （P＞0.05）. For female participants in the PD group across all age groups， Yu tone was the most frequent， whereas for the control group， Jue （角） tone was the most frequent in all age groups， and the composition ratio of Yu tone in the PD group was significantly higher than that in the control group across all age groups （P＜0.05）； in the 56~70 age group， the composition ratio of Jue tone was lower in the PD group compared to that in the control group （P＜0.05）. Regarding voice frequency， males in the PD group aged 41~55 and 56~70 had higher voice frequency than those in the control group of the same age range， and similarly， females in the PD group aged 56~70 and 71~85 had higher voice frequency than their counterparts in the control group （P＜0.05）. ConclusionPD patients have a voice with a higher frequency and an increased proportion of Yu tone in their five-tone distribution. According to the theory of five-viscera phonology， PD patients may have disease mechanism of kidney essence deficiency.

Objective:To analyze the general chapter for lotions 0127 of the Chinese Pharmacopoeia 2020 Vol-ume Ⅳ,and discuss how to improve the general technical requirements of lotions 0127 in the Chinese Pharma-copoeia.Methods:By comparing the general chapter for lotions in domestic and international pharmacopoe-ias,the definition,classification,process,storage and corresponding inspection requirements were analyzed.Results and Conclusions:The general chapter for lotions 0127 of the Chinese Pharmacopoeia should be revised,including improvement of the definition,increasement of forms of preparations,and expansion of included varieties,so as to promote scientific regulation for drugs and exhibit a guiding role of the Chinese pharmacopoeia in drug control.

Objective To describe the spatiotemporal patterns of genital chlamydia trachomatis(GCT)infections in Guangdong,and to explore social-economic factors associated with the risk of GCT infections.Methods Data of GCT infections among women of childbearing age in Guangdong province from 2014 to 2019 and social-economic factors were collected.Bayesian spatiotemporal hierarchy model with Poisson distribution was established including temporal and spatial terms.Integrate nested Laplace approximates(INLA)was used to estimate the parameters.Results GCT infections among women of childbearing age in Guangdong had no obviously temporal effect,but had significantly spatial heterogeneity.After adjusting for fixed covariate effects and temporal random effect,high-risk regions concentrated in Pearl Delta region.Comprehensive economic development(RR=1.185,95%CI:1.117～1.257)and population and transportation level(RR=1.058,95%CI:1.012～1.106)were positively associated with the prevalence of GCT infections,while the health service level had negatively association(RR=0.942,95%CI:0.924～0.960).Conclusion The prevalence of GCT infections among women of childbearing age shows spatial clustering characteristics.Social-economic factors are significantly related to the prevalence of GCT infections.

Patients with metabolic syndrome(MS)are at potential risk for cardiovascular disease and have received increasing public and medical attention.Studies have shown that regular physical exercise can effectively regulate meta-bolic indicators such as blood pressure,blood sugar and blood lipids,and play a positive role in reducing the risk of cardio-vascular disease and improving the prognosis of patients.Exercise intensity has been identified as the most important as-pect in reducing the risk of cardiovascular death and all-cause mortality in exercise intervention.Therefore,the design of exercise prescription which is both scientific and satisfying individual differences has become the focus of research.Most of the current clinical studies are based on the percentage of exercise intensity as the basis for the formulation of standard-ized exercise prescription for MS patients,while the studies on the individualized threshold of exercise intensity based on cardiopulmonary exercise test(CPET)are still few.CPET has shown that individualized exercise prescription can effec-tively reduce body composition index,blood pressure and blood glucose,improve cardiorespiratory function,exercise en-durance and quality of life in MS patients.This paper reviewed the development of individualized exercise programs with different intensification according to threshold indexes in CPET,analyzed the intervention effects and possible mechanisms for MS patients and subgroups,and provided certain reference for the formulation and implementation of personalized exer-cise prescriptions for MS patients,and also provided references for in-depth research on individualized exercise intervention for MS.

Objective To observe the value of dual domain(projection domain and image domain)joint learning reconstruction network(DDRNet)for reconstructing chest limited angle CT images.Methods Totally 4 300 chest enhanced CT images of 65 patients with chest tumors were retrospectively enrolled and reconstructed with DDRNet,and 3D and 2D projection information fusion were performed.The reconstruction effect of DDRNet was evaluated and compared with that of single domain reconstruction and filtered back projection(FBP),residual encoder-decoder convolutional neural network(RED-CNN),Resnet and deconvolution network(RDN),as well as of generative adversarial network(GAN).Results The peak signal to noise ratio(PSNR)of DDRNet reconstructed images tended to stabilize after approximately 60 iterations,while the projection domain and image domain learning networks tended to stabilize after approximately 90 and 80 iterations.After stable training,compared to the projection domain learning network,the fluctuation of output results of DDRNet and image domain learning networks were less.After 200 rounds of training,PSNR of DDRNet reconstructed images was significantly higher than that of projection domain and image domain learning networks.The quality of DDRNet reconstructed image was significantly better than that of FBP,RED-CNN,RDN and GAN.Conclusion DDRNet could be used to effectively reconstruct high-quality chest limited angle CT images.

Objective:To investigate the dosimetric impact of tumor treating fields (TTF) transducer arrays on concurrent radiotherapy for patients with glioblastoma (GBM).Methods:A strategy was developed to accurately simulate the dosimetric impact of TTF arrays on radiotherapy, including the establishment of accurate auto-segmentation technique for TTF arrays, determination of the relative electron density (RED) of the transducer arrays and validation of the dose calculation accuracy in the treatment planning system (TPS) for TTF arrays. Based on this strategy, the dosimetric impact of TTF arrays on clinical treatment plans of 10 patients with GBM was evaluated. Furthermore, the dosimetric comparison between the clinical plans with different beam energies were investigated when TTF arrays were used. The methods of analysis of variance were paired t-test or Wilcoxon signed-rank test based on whether the differences followed a normal distribution. Results:The auto-segmentation technique for TTF arrays was established by designing a workflow in Mim software and achieved a Dice coefficient of 0.93 and a Jaccard index of 0.87 compared to the standard contours. The RED of TTF arrays was 3.3 which was derived from the comparison between the measured and simulated percentage depth dose (PDD) with and without TTF arrays on phantom. Measured and calculated dose distributions were compared using the 2D gamma analysis. The gamma passing rates on the coronal plane of 4 mm and 5.1 cm depth were 96.64% and 94.55% at the criteria of 3% /3 mm, indicating that the calculation accuracy of algorithm in TPS for TTF arrays could meet clinical requirements. In the clinical treatment plans of patients with GBM, the presence of TTF arrays caused a mean reduction of planning target volume (PTV) dose of approximately 1%, and an increase in scalp dose of approximately 5%, with minimal impact on other organs at risk (OAR). The 10 MV plans resulted in a higher dose of PTV by 0.3% and lower dose of scalp by approximately 3% compared to the 6 MV plans, when considering TTF arrays.Conclusions:The accurate simulation strategy for the dosimetric impact of TTF arrays on radiotherapy established in this study ensures the accuracy and precision of the calculations. In TTF therapy combined with concurrent radiotherapy for GBM, TTF arrays have slight effect on PTV dose, but significantly increase scalp dose. High-energy beam can reduce the impact of TTF arrays.

Objective:To compare the plan quality between coplanar and non-coplanar volumetric modulated arc therapy (co-VMAT and nco-VMAT) techniques for the whole brain radiotherapy with hippocampus and hypothalamic-pituitary (HT-P) axis sparing.Methods:A total of 15 patients who underwent prophylactic cranial irradiation in Tianjin Medical University General Hospital from November 2021 to August 2023 were retrospectively selected. The hippocampus and HT-P axis were delineated according to Radiation Therapy Oncology Group (RTOG) 0933 and contouring guidelines for hypothalamus. Co-VMAT and nco-VMAT plans were generated for each patient with a prescription dose of 30 Gy in 10 fractions. Then, dosimetric parameters, plan robustness, plan complexity, and delivery efficiency for both plans were compared using paired t-test. Results:Both co-VMAT and nco-VMAT plans could achieve dosimetric objectives. There were no significant differences in D 2%, D 95% and conformity index (CI) of planning target volume (PTV) between the two plans. The D 98% and homogeneity index (HI) of PTV in co-VMAT showed a slight inferiority compared to that in nco-VMAT (D 98%: 26.37 Gy vs. 26.96 Gy, P=0.001; HI: 0.25 vs. 0.24, P=0.002). The D min of bilateral hippocampus in co-VMAT were 8.55 Gy (left) and 8.32 Gy (right), which were lower than 9.31 Gy (left) and 9.26 Gy (right) in nco-VMAT. In addition, the D mean of the hypothalamus and pituitary in the co-VMAT plan were lower than those in the nco-VMAT plan (hypothalamus: 11.54 Gy vs. 12.27 Gy; pituitary: 11.72 Gy vs.12.1 Gy, both P<0.001). The doses to the hippocampus and HT-P axis were highly sensitive to errors in both co-VMAT and nco-VMAT plans, while the sensitivity of dose to errors in the PTV and other organs at risk was low. The co-VMAT plan had lower complexity compared to the nco-VMAT plan, with γ passing rate at 3%/3 mm criteria of 99.06%±0.60% and 98.05%±2.89%, respectively. The average beam-on time of the co-VMAT plan was 4.8 min, approximately 2/3 of the time for nco-VMAT, while the average treatment time was 6.3 min, approximately half of the treatment time for nco-VMAT. Conclusions:Both co-VMAT and nco-VMAT can achieve hippocampus and HT-P axis sparing in the whole brain radiotherapy. In the co-VMAT plan, the D 98% of the PTV is slightly smaller, but it provides better protection for the hippocampus and HT-P axis. The doses to the hippocampus and HT-P axis are sensitive to errors in both plans. However, the co-VMAT plan has lower complexity, higher delivery efficiency, and is more suitable for clinical treatment.

BACKGROUND:Studies have found that activation of nuclear factor-erythroid 2-related factor 2/heme oxidase-1(Nrf2/HO-1)pathway can alleviate oxidative stress caused by cerebral ischemia-reperfusion injury,but whether human bone marrow mesenchymal stem cells(hBMSC)can activate Nrf2/HO-1 pathway to alleviate cerebral ischemia-reperfusion injury is still lacking relevant studies. OBJECTIVE:To investigate whether intracranial transplantation of hBMSC alleviates oxidative stress injury in cerebral ischemia-reperfusion animal models by activating Nrf2/HO-1 pathway. METHODS:Totally 40 male SPF SD rats were randomly divided into sham operation group,model group,hBMSC transplantation group,hBMSC+solvent group and hBMSC+Nrf2 inhibitor group.Each group consisted of eight animals.In the model group and the hBMSC transplantation group,middle cerebral artery occlusion model was prepared by thread embolization method.The thread embolization was removed 1 hour later,and 30 μL PBS or hBMSC cultured to at least passage 5 was injected into the right cortex and striatum of rats.In the hBMSC+Nrf2 inhibitor group and hBMSC+solvent group,the left ventricle was injected with Nrf2 inhibitor Brusatol and its solvent dimethyl sulfoxide respectively 24 hours before model establishment,then the middle cerebral artery occlusion model was prepared,and hBMSC was injected.Relevant indexes were detected 3 days after transplantation. RESULTS AND CONCLUSION:(1)CT and TTC staining showed the same area and volume of cerebral infarction:model group>hBMSC+Nrf2 inhibitor group>hBMSC+solvent group>hBMSC transplantation group>sham operation group.(2)Hematoxylin-eosin staining and Nissl's staining showed that the ischemic brain tissue was intact and the neurons were normal in the sham operation group.Compared with the model group,the pathological morphology and neuronal injury of the hBMSC transplantation group and the hBMSC+solvent group were significantly improved.Compared with the hBMSC+solvent group,the hBMSC+Nrf2 inhibitor group had more serious pathological morphology and neuronal damage.(3)Western blot assay and oxidative stress index detection results showed that compared with the sham operation group,Nrf2 and HO-1 proteins were decreased(all P<0.05),malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the model group.Compared with the model group,the expression levels of Nrf2 and HO-1 proteins were increased(all P<0.05),malondialdehyde was decreased and superoxide dismutase was increased(all P<0.05)in the hBMSC transplantation group and the hBMSC+solvent group.Compared with the hBMSC+solvent group,the expression levels of Nrf2 and HO-1 proteins were simultaneously decreased(all P<0.05),and malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the hBMSC+Nrf2 inhibitor group.(4)These results indicate that hBMSC can alleviate cerebral ischemia-reperfusion injury possibly by activating Nrf2/HO-1 pathway.

Objective: To examine the mediating effects of mindfulness level on resilience and symptoms of anxiety and depression among healthcare workers, so as to provide the reference for developing effective psychological intervention. Methods: The clinical doctors, nurses and public health professionals were selected using the stratified random cluster sampling method from hospitals, disease prevention and control centers, and health departments in five cities in Shandong Province, including Qingdao, Jinan, Rizhao, Jining and Liaocheng in January 2023. Basic information, mindfulness level, resilience and symptoms of anxiety and depression among healthcare workers were collected using general demographic questionnaires, the 5-item Mindful Attention Awareness Scale, 10-item Connor-Davidson Resilience Scale, Generalized Anxiety Disorder Scale and Patient Health Questionnaire, respectively. The Process program was used to analyze the mediating effects of mindfulness level on resilience and symptoms of anxiety and depression. Results: A total of 1 836 healthcare workers were investigated, including 472 males (25.71%) and 1 364 females (74.29%), and the median age was 39 (interquartile range, 12) years. There were 629 clinical doctors (34.26%), 963 nurses (52.45%) and 244 public health professionals (13.29%). The median scores of mindfulness level and resilience were 22 (interquartile range, 7) and 20 (interquartile range, 4) points, respectively. The detection rates of anxiety and depression symptoms were 49.78% and 72.28%, respectively. The mediation analysis showed that mindfulness level exerted a partial mediating effect between resilience and anxiety symptoms (β=-0.510, P<0.001), with a direct effect value of -0.130 and a mediating effect value of -0.046, and the mediating effect accounted for 26.14% of the total effect; mindfulness level also exerted a partial mediating effect between resilience and depression symptoms (β=-0.575, P<0.001), with a direct effect value of -0.120 and a mediating effect value of -0.052, and the mediating effect accounted for 30.23% of the total effect. Conclusion Mindfulness level plays a mediating effect between resilience and symptoms of anxiety and depression among healthcare workers.