Objective To explore the method of primary culture and biological characteristics of aortic vascular smooth mus-cle cells (VSMC)in mice,providing experimental material for cellular and molecular scientific research of vascular disease. Methods Thoracic and abdominal aortas in mice were isolated and VSMC were obtained by using improved method of tissue piece inoculation.Digested with trypsin and passaged,VSMC were purified with differential adherence method.The conditions of cellular morphology and growth were observed under inverted phase contrast microscope,and VSMC were identified with hematoxylin-eosin (HE)staining and immunofluorescence.Results VSMC were isolated successfully and grown vigorously with good bioactivity,the cells had a radial or typicalpeak-valleylike growth,and showed fusiform,abundant cytoplasm with large and round or oval nucle-us by HE staining,the expressions of specific cytoplasmicα-smooth muscle actin were positive by immunofluorescence stain.Conclu-sion It can isolate and cultivate VSMC with high purity and good activity under in vitro conditions with simple,economical,reliable method.

Objective:To investigate the ethical practices in individual supervision of counseling in China.Method:Forty-one graduate students and 38 participants of the China American Psychoanalytic Alliance (CAPA) were included in this study.The Ethical Practices in Supervision Scale (EPSS) was used for the investigation.The scale included two parts.The first part was to investigate supervisors'ethical practices,and the second part focused on supervisees'impact from unethical supervisors.Results:The findings showed that 7 out of 28 items in the scale were significantly different between two groups.CAPA participants' supervisors performed better than graduate students' supervisors (P ＜ 0.05) in supervision scheduling,ethics discussion,and counselor/therapist role.Graduate students' supervisors performed better than CAPA participants' supervisors (P ＜ 0.05) in live supervision,audio/video review,informing clients of supervisee's status as trainee,and supervision contract.If individuals experience their supervisors being unethical or unprofessional,more than 55 percent of the individuals responded to negatively affect their thoughts toward their supervisor and the supervisory alliance,and almost 50 percent of the individuals responded to negatively impact their emotion and quality of their counseling for clients.Conclusion:Both graduate students and CAPA participants respond that their supervisors perform well in both monitoring supervisees' performance and maintaining appropriate boundaries.However,both of them should work more on reviewing video or audiotapes of supervisees' sessions regularly.

Objective To explore the expression of serine/threonine kinase 15 (STK15) gene and its significance for papillary thyroid carcinoma. Methods SP immunohistochemical method was used to detect the expression of STK15 gene in 71 cases of papillary thyroid carcinoma and 45 cases nodular goiter tissue. Results The positive expression rates of STK15 gene in 71 cases of papillary thyroid carcinoma were 100%, and the adjacent of papillary thyroid carcinoma expressions of STK15 gene 8.5%, nodular goiter tissue of STK15 gene expression rates 24.4%. The expression of STK15 gene was positively correlated with that of STK15 gene in papillary thyroid carcinoma (P < 0.01). Conclusion High expression of STK15 gene is confirmed in papillary thyroid carcinoma. The detection of STK15 gene can provide valuable evidence for diagnosis of papillary thyroid carcinoma and evaluation of the malignant potential of nodular goiter.

Objective To observe the effect of budesonide/doxofylline on patients with persistent asthma. Methods Sixty patients with moderate asthma were randomly divided into treatment group and control group. Patients in treatment group accepted the inhalation budesonide 400μg/d and doxofylline tablet 0.2 bid po but those in control group accepted the inhalation budesonide/formoterol 160/4.5μg bid. FeNO, sputum eosinophils, PEF and ACT scores were compared between two groups. Results PEF and ACT scores in treatment group were lower to those in control group (P<0.05), whereas the score of sputum eosinophils in treatment group was higher (P<0.05) one month later. There was no significant difference in FeNO between two groups (P>0.05). After three months, sputum eosinophils in treatment group were better than that in control group (P<0.05), whereas there was no significant difference in PEF, ACT scores and FeNO between two groups(P>0.05). Conclusion There is no significant difference in controlling asthma between budesonide/doxofylline and budesonide/formooterol

Objective To investigate the selection of surgery and clinical outcomes of upper cervical injuries.Methods 25 upper cervical injury patients were involved in this retrospective study from November 2011 to June 2014.Including 20 males and 5 females with mean age of 37.1 years old (range,14-55 years old).Individual operation methods were based on the comprehensive evaluation of specific situations including the clinical manifestation,the type of the injuries and the imaging data.HaloVest distraction was applicated before operation.The surgery by anterior approach were performed for 7 patients and posterior approach were performed for 18 patients.Preoperative and postoperative American Spinal Injury Association (ASIA) grade and Functional Independence Measurement (FIM) score were studied to evaluate the nerve functional restoration.Imaging data before and after the operation were contrasted to evaluate the reduction of the fracture,the bone union,the fusion of the bone graft and the condition of the internal fixation.Wilcoxon Singed Rank Test was applied to compare the FIM score between pre-operation and last follow-up.Results 15 patients presented neurological function deficit because of cervical spinal cord compromise.All cases were followed up for 6-35 months (mean 18.2 months),showing good clinical and radiological effects.Solid fusion was obtained in all patients among 3-12 months.The ASIA grade improved by an average of 1.1 (6 months after operation) and 1.2 (12 months after operation).There was significant difference in FIM score between pre-operation and last follow-up.One patient got cerebrospinal fluid leakage.Conservative treatment was implemented with the Trendelenburg position,rehydration fluids and so on.Removal of drainage tube 8 days later when the drainage was less than 30 ml/24 h.No incision infection,cerebrospinal fluid leakage,migration or breakage of internal fixation was observed at the last follow-up.Conclusion The type of upper cervical injuries are complicated,the characteristics of fracture,dislocation and nerve injury in different patients are different.The specific situation should be evaluated comprehensively to make individual operation methods.The success of the operation requires the proficiency of the anatomic basis,the biomechanical characteristics,precise entrance point and direction in operation,appropriate diameter of the screw and suitable depth of the screw road.

Objective To study the roles of ethyl pyruvate ( EP ) in spinal cord edema after spinal cord injury ( SCI ) and in spinal cord astrocytic swelling after oxygen-glucose deprivation and reoxygenation ( OGD/R) in vitro in rats. Methods After SCI models were established in adult Sprague-Dawley ( SD ) rats, an intraperitoneal injection of EP was conducted to inhibit high mobility group box-1 ( HMGB1 ). Effects of EP on spinal cord edema, HMGB1 expression and astrocyte activation ( glial fibrillary acidic protein ( GFAP ) expression) in SCI rats were analyzed. Spinal cord astrocytes were cultured in post-natal SD rats and incubated under OGD/R procedure. Effects of EP on cell swelling, expression of HMGB1, aquaporin-4 ( AQP4 ) and toll-like receptor 4 ( TLR4 ) , and nuclear expression of nuclear factor-kappa B ( NF-κB ) in spinal cord astrocytes were observed. Results The water content in the spinal cord was increased significantly more at 1 d after SCI than at 12 h and 3 d ( P ＜0.05 ). Intraperitoneal injection of EP at 50 mg/kg reduced spinal cord water content, HMGB1 expression and astrocyte activation ( GFAP expression ) in SCI rats signif-icantly more than that at 25 mg/kg or 100 mg/kg ( P ＜0.05 ). The volume of spinal cord astrocytes cultured in vitro after OGD 6 h/R 24 h was significantly greater than that after OGD 6 h/R 6 h or OGD 6 h/R 12 h ( P ＜0.05 ). EP at 12 μmol/L reduced cell swelling, decreased expression of HMGB1, AQP4 and TLR4, and downgraded nuclear expression of NF-κB in spinal cord astrocytes after OGD/R significantly more than EP at 6 μmol/L( P ＜0.05). Conclusion EP may reduce early spinal cord edema after SCI, attenuate spinal cord astrocyte swelling and decrease AQP4 expression after OGD/R in vitro by inhibiting HMGB1 in rats.

Objective: Based on the information-motivation-behavioral skills (IMB) model theory, the structural equation model(SEM)was used to analyze the effect of using new media to carry out AIDS health education among young students. Methods: Stratified random cluster sampling method was used to select 2 561 students from four colleges in Yantai to conduct a questionnaire survey, and to carry out various forms of new media AIDS health education for the intervention group. Data were analyzed with structural equation model. Results: After intervention，the awareness rate of AIDS were 85.9% and 94.0% respectively(χ2=46.67，P<0.01). The structural equation model fits well, and the fitted indices were: RMSEA=0.04, GFI=0.97, AGFI=0.96, IFI=0.97, CFI=0.98. The direct effects of new media interventions on prevention information and prevention motivation were 0.30 and 0.65 respectively. The influence of information and motivation on preventive behavior was mainly reflected through the indirect effect of behavioral skills. The overall effects of standardization were 0.09 and 0.29 respectively. The overall effect of behavioral skills on preventive behavior was 0.80, which was the main factor affecting HIV prevention behavior. Conclusion The use of new media to carry out AIDS health education among young students had a significant effect. Motivation promotion should be used as a new breakthrough point for future work, and information, motivation, and behavioral skills should be used to promote the prevention of AIDS prevention among young students.

OBJECTIVE: To detect the expression of differential expression genes(DEGs) on microarray chips of macrophages exposed to nano-silicon dioxide(SiO_2) dust, and to screen the leading signaling pathway of nano-SiO_2 dust exposure-related diseases. METHODS: The gene chip GSE13005 of RAW264.7 macrophage intervened by nano-SiO_2 dust was obtained from the public gene chip database developed by the National Center for Biotechnology Information. The macrophages in the control group were cultured in complete medium without adding SiO_2 dust, whereas the macrophages in the exposure group were treated with SiO_2 dust with the final concentrations of 5, 20, and 50 mg/L. The gene expression data of macrophages was analyzed by RMA Express 1.2.0 software and R language 3.5.1. The Kyoto Encyclopedia of Genes and Genomes(KEGG) was used to screen DEGs and perform gene ontology(GO) enrichment analysis on related genes and signaling pathways. RESULTS: A total of 67 DEGs of macrophages were screened after SiO_2 dust treatment, of which 48 genes were up-regulated and 19 genes were down-regulated. GO enrichment analysis results showed that the main functional items of participating DEGs were reaction of amine, regulation of viral genome replication,negative regulation of amino acid transport, ovulation, bronchodilator response, chemokine activity, negative regulation of muscle cell differentiation, response to lack of amino acid, positive regulation of glomerular mesangial cell proliferation, and positive regulation of vascular smooth muscle cell proliferation. KEGG signaling pathway analysis results suggested that DEGs could function through 7 signaling pathways including nuclear transcription factor-κB(NF-κB) signaling pathway, p53 signaling pathway, glioma, melanoma, toll-like receptor signaling pathway, renal cell carcinoma and salmonella infection. Further functional enrichment revealed that NF-κB signaling pathway changed most significantly after macrophages were exposed to nano-SiO_2 dust. CONCLUSION: Exposure to nano-SiO_2 could induce the abnormal expression of 67 genes in macrophages. The genes that participated in macrophage activation process induced by nano-SiO_2 dust exposure are related to NF kappa B signaling pathway.

Background Macrophages are essential components of the natural immune system. They play a significant role in resisting foreign bodies in the respiratory tract and maintaining the homeostasis of the internal environment of lung tissue. Objective To investigate the mechanism of macrophage pyroptosis induced by silica dust with different particle sizes. Methods The modified murine macrophage cell line, RAW-ASC cells, was cultured and divided into a blank control group, a lipopolysaccharide (LPS) group (1 μg·mL⁻¹ LPS), a nano-SiO₂ group (1 μg·mL⁻¹ LPS+100 μg·mL⁻¹ nano-SiO₂), a micro-SiO₂ group (1 μg·mL⁻¹ LPS+750 μg·mL⁻¹ micro-SiO₂), and a positive control group [1 μg·mL⁻¹ LPS+3 mmol·L⁻¹ adenosine triphosphate (ATP)]. Apart from the blank control group, cells in other groups were pretreated with LPS for 6 h, and then exposed to SiO₂ or ATP for 4 h. According to the molecular target NOD-like receptor pyrin domain-containing protein 3 (NLRP3) and reactive oxygen species (ROS), we applied MCC950 (NLRP3 inhibitor) and N-acetyl cysteine (NAC, ROS scavenger) to macrophages. CCK-8 assay was used to detect cell viability; 5-ethynyl-2'-deoxyuridine (EdU) staining was used to detect cell proliferation; lactate dehydrogenase (LDH) assay kit was used to detect LDH in supernatant; calcein AM/PI fluorescent double-staining was applied to evaluate cell rupture; 2',7'-dichlorofluorescin diacetate (DCFH-DA) fluorescent probe was used to measure the content of ROS; Western blotting was used to measure the expressions of NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), Caspase-1, gasdermin D (GSDMD), and interleukin-1β (IL-1β). Results Compared with the blank group, 100 μg·mL^-1nano-SiO₂ and 750 μg·mL^-1micro-SiO₂ dust exposure reduced the cell viability to 40% and 68% (P<0.05), and the cell proliferation rate to 30% and 33% (P<0.01), respectively; they also induced cell lysis and ROS release, upregulated NLRP3, ASC, Caspase-1, GSDMD, and IL-1β at protein level (P<0.05), and induced macrophage pyroptosis. After intervening with MCC950 (10 μmol·L^-1) and NAC (10 mmol·L^-1), the expressions of NLRP3, ASC, Caspase-1, and IL-1β decreased (P<0.05), and, specifically, NAC effectively reduced ROS levels (P<0.05). Conclusion Both nano- and micro-SiO₂ dust have cytotoxicity, can upregulate ROS level, activate NLRP3 inflammasome, and promote the release of cytokines, leading to pyroptosis. These results are helpful to reveal the molecular mechanism of macrophage pyroptosis induced by SiO₂ dust.

Ankylosing spondylitis (AS) combined with spinal fractures with thoracic and lumbar fracture as the most common type shows characteristics of unstable fracture, high incidence of nerve injury, high mortality and high disability rate. The diagnosis may be missed because it is mostly caused by low-energy injury, when spinal rigidity and osteoporosis have a great impact on the accuracy of imaging examination. At the same time, the treatment choices are controversial, with no relevant specifications. Non-operative treatments can easily lead to bone nonunion, pseudoarthrosis and delayed nerve injury, while surgeries may be failed due to internal fixation failure. At present, there are no evidence-based guidelines for the diagnosis and treatment of AS combined with thoracic and lumbar fracture. In this context, the Spinal Trauma Academic Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate the Clinical guideline for the diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture ( version 2023) by following the principles of evidence-based medicine and systematically review related literatures. Ten recommendations on the diagnosis, imaging evaluation, classification and treatment of AS combined with thoracic and lumbar fracture were put forward, aiming to standardize the clinical diagnosis and treatment of such disorder.