ObjectiveTo explore the effect and mechanism of tumor necrosis factor alpha （TNF-α）-regulated circular RNA Williams Beuren syndrome chromosomal region 22 （circWBSCR22） on the malignant behaviors of cervical cancer cells. MethodsHuman cervical cancer HeLa cells were treated with TNF-α. Reverse transcription quantitative polymerase chain reaction （RT-qPCR） was used to detect the expression level of circWBSCR22. Nuclear-cytoplasmic RNA separation experiments were conducted to determine the localization of circWBSCR22 in cells. Cell migration， invasion， viability， and proliferation were assessed through Transwell assay， tetrazolium salt （MTT） assay， and colony formation assay， respectively. Dual-luciferase reporter gene assays were performed to verify the targeting relationship between microRNA-512-5p （miR-512-5p） and circWBSCR22 or family with sequence similarity 60A （FAM60A）. RT-qPCR was used to detect the RNA levels of miR-512-5p and FAM60A. Western blot was used to detect the protein level of FAM60A. ResultsCompared with the Control group， the expression level of circWBSCR22 in HeLa cells treated with TNF-α increased （P<0.05）. About 66% of circWBSCR22 was found in the cytoplasm. Compared with the control group， the migration （P<0.01） and invasion （P<0.01） of HeLa cells transfected with circWBSCR22 were enhanced， but the cell activity and cell proliferation were not significantly affected （P>0.05）. The double luciferase reporter gene assay showed that the relative luciferase activity of the co transfected circWBSCR22-WT and miR-512-5p groups was lower than that of the co transfected circWBSCR22-WT and miR-NC groups （P<0.01）. Compared with the control group， the expression level of miR-512-5p in HeLa cells transfected with circWBSCR22 decreased （P<0.01）. Compared with the control group， the migration （P<0.01） and invasion （P<0.05） ability of HeLa cells transfected with miR-512-5p decreased. Double luciferase reporter gene assay showed that compared with the co transfected FAM60A 3´-UTR-WT and miR-NC groups， the fluorescence activity of the co transfected FAM60A 3´-UTR-WT and miR-512-5p groups decreased （P<0.01）.Compared with the control group， the expression levels of FAM60A mRNA （P<0.05） and protein （P<0.01） in HeLa cells transfected with miR-512-5p decreased. Compared with the control group， the migration （P<0.01） and invasion （P<0.01） of cervical cancer HeLa cells transfected with FAM60A were enhanced. Compared with the control group， the expression levels of FAM60A mRNA （P<0.01） and protein （P<0.05） in the cells transfected with circWBSCR22 increased. The results of rescue experiment showed that compared with the pcDNA+miR-NC group， the expression levels of FAM60A RNA （P<0.01） and protein （P<0.001） in the pcDNA+miR-512-5p group decreased， and the ability of migration （P<0.01） and invasion （P<0.05） decreased； Compared with pcDNA+miR-512-5p group， the expression levels of FAM60A RNA （P<0.01） and protein （P<0.001） in circWBSCR22+miR-512-5p group increased， and the ability of migration （P<0.01） and invasion （P<0.05） was enhanced. ConclusionTNF-α promotes the migration and invasion of cervical cancer cells through the circWBSCR22/miR-512-5p/FAM60A regulatory axis.

OBJECTIVES: To investigate the protective effects and underlying mechanisms of Gynostemma pentaphyllum (GP)ethanol extract on motor dysfunction in a mouse model of Parkinson's disease (PD). METHODS: Eighty C57BL/6 male mice were randomly divided into five groups: control group, model group, levodopa group (positive control group), low-dose GP group, and high-dose GP group, with 16 mice per group. The PD model was induced by injection of 6-hydroxydopamine into the substantia nigra pars reticulata of the mice. Two weeks after 6-hydroxydopamine, positive control group received intraperitoneal injection of levodopa 10 mg·kg^－1·d^－1, while low-dose GP and high-dose GP groups received GP extract 100 or 200 mg·kg^－1·d^－1 orally for three weeks. After a 3-week-treatment, the effects of GP on motor dysfunction in 6-hydroxydopamine-induced PD were assessed using open field and CatWalk gait tests, while the effects on muscle strength were evaluated by forelimb grip strength. Immunofluorescence staining was used to detect the number of tyrosine hydroxylase (TH) positive neurons. The levels of dopamine and serotonin in the midbrain were determined by enzyme-linked immunosorbent assay. In addition, Western blotting was performed to detect the expression of mitogen-activated protein kinase (MAPK) family proteins such as p-extracellular signal-regulated kinase (ERK)1/2, p-p38 and p-c-Jun N-terminal kinase (JNK)1/2, and mitochondrial apoptosis pathway proteins such as B-cell lymphoma (Bcl)-2, Bcl-2 associated X protein (Bax), and cleaved-cysteine aspartic acid specific protease (caspase)-3. RESULTS: Behavioral experiments showed that GP significantly improved the spontaneous activity and motor coordination of PD mice (P<0.05). The forelimb grip strength was also increased by GP treatment (P<0.05), compared to the PD model group. In addition, compared with the model group, the number of TH-positive neurons in substantia nigra pars reticulata region, the levels of dopamine and serotonin in midbrain and the expression of p-ERK1/2 were significantly increased by GP treatment (all P<0.05), whereas the expression of p-p38 and p-JNK1/2, the ratio of Bax/Bcl-2 and cleaved-caspase-3/caspase-3 were significantly decreased (all P<0.05). CONCLUSIONS The results indicate that GP might increase dopamine and serotonin levels in the midbrain and promote the survival of dopaminergic neurons in substantia nigra pars reticulata by regulating the expression of phosphorylation of MAPK family proteins and the expression of mitochondrial apoptosis-related proteins, thereby ameliorating motor deficits in PD mice.
Animals ; Mice ; Male ; Gynostemma/chemistry* ; Mice, Inbred C57BL ; Apoptosis/drug effects* ; Plant Extracts/therapeutic use* ; Parkinson Disease/metabolism* ; Disease Models, Animal ; Neurons/pathology*

Objective To explore the application value of low-temperature plasma bipolar forceps in open thyroid cancer surgery.Methods The clinical data of 30 patients with differentiated thyroid cancer treated with low-temperature plasma bipolar forceps by our team from May 2021 to October 2024 were retrospectively analyzed.And 30 patients with differentiated thyroid cancer treated with monopolar electrocautery+ultrasonic scalpel were selected as the control group.The surgical procedures for both groups were thyroid lobectomy,isthmus resection+ipsilateral level Ⅵ lymph node dissection.The age,gender,body mass index(BMI),tumor TNM stage,operation time,intraoperative blood loss,48 h postoperative drainage volume,and surgical complications of the two groups were compared.Results There were no significant differences in age,gender,BMI,and tumor TNM stage between the two groups.The operation time,intraoperative blood loss,48 h postoperative drainage volume,and the incidence of postoperative transient hypoparathyroidism in the low-temperature plasma bipolar forceps group were significantly less than those in the control group(P＜0.05).There were no occurrences of postoperative subcutaneous hematoma,permanent recurrent laryngeal nerve palsy,and permanent hypoparathyroidism in both groups.Conclusion In thyroid cancer surgery,low-temperature plasma bipolar forceps can not only achieve fine detailed operations,but also effectively protect the blood supply of the recurrent laryngeal nerve,superior laryngeal nerve,and parathyroid glands.It has less trauma and a faster postoperative recovery,and is worthy of clinical promotion and application.

Objective To explore the application value of low-temperature plasma bipolar forceps in open thyroid cancer surgery.Methods The clinical data of 30 patients with differentiated thyroid cancer treated with low-temperature plasma bipolar forceps by our team from May 2021 to October 2024 were retrospectively analyzed.And 30 patients with differentiated thyroid cancer treated with monopolar electrocautery+ultrasonic scalpel were selected as the control group.The surgical procedures for both groups were thyroid lobectomy,isthmus resection+ipsilateral level Ⅵ lymph node dissection.The age,gender,body mass index(BMI),tumor TNM stage,operation time,intraoperative blood loss,48 h postoperative drainage volume,and surgical complications of the two groups were compared.Results There were no significant differences in age,gender,BMI,and tumor TNM stage between the two groups.The operation time,intraoperative blood loss,48 h postoperative drainage volume,and the incidence of postoperative transient hypoparathyroidism in the low-temperature plasma bipolar forceps group were significantly less than those in the control group(P＜0.05).There were no occurrences of postoperative subcutaneous hematoma,permanent recurrent laryngeal nerve palsy,and permanent hypoparathyroidism in both groups.Conclusion In thyroid cancer surgery,low-temperature plasma bipolar forceps can not only achieve fine detailed operations,but also effectively protect the blood supply of the recurrent laryngeal nerve,superior laryngeal nerve,and parathyroid glands.It has less trauma and a faster postoperative recovery,and is worthy of clinical promotion and application.

Objective This study aimed to explore the effect of pituitary tumor-transforming gene 1(PTT-G1)on the invasion and proliferation of oral squamous cell carcinoma(OSCC)cell lines under the action of miR-362-3p.Methods The bioinformatics online database was used to query the expression of PTTG1 in head and neck squamous cell carcinoma(HNSCC).The expression of PTTG1 in the Cal-27,HN-30,and HOK cell lines was detected by Western blot.A wound-healing assay was used to determine the effect of PTTG1 on the migration ability of the OSCC cells.The Transwell assay was used to examine the changes in cell-invasion ability.5-ethynyl-2'-deoxyuridine(EdU)cell-proliferation assay was used to detect changes in cell-proliferation ability.Bioinformatics approach predicted the upstream miRNA of PTTG1.The targeting relationship between miR-362-3p and PTTG1 was examined by the dual luciferase assay,and quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the expression of miRNA in OSCC tissues.Results The ENCORI database showed that PTTG1 expression was up-regulated in OSCC tissues.Western blot confirmed that PTTG1 expression was up-regulated in Cal-27 and HN-30 cells than HOK cells.PTTG1 knockout can inhibit the migration,invasion,and prolif-eration of Cal-27 and HN-30 cells(P<0.05).Bioinformatics prediction websites predicted that the upstream miRNA of PTTG1 was miR-362-3p,and PTTG1 can bind to miR-362-3p.Results of qRT-PCR showed that miR-362-3p expression was downregulated in OSCC tissues compared with normal tissue(P<0.05).Transwell and EdU experiments confirmed that miR-362-3p knockdown can promote the invasion and proliferation of Cal-27 and HN-30 after PTTG1 knockdown.Conclusion miR-362-3p can inhibit the invasion and proliferation of Cal-27 and HN-30 cells by targeting PTTG1.

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 screen plasma exosomal protein molecular markers in patients with spinal cord injury (SCI) by applying Label-Free quantification and bioinformatics analysis.Methods:Fifty plasma specimens from the First Affiliated Hospital of Nanjing Medical University (from January 2021 to June 2022) were collected from SCI patients and healthy people, respectively. Plasma exosomes were isolated using ultracentrifugation and identified by transmission electron microscopy, nanoparticle tracking analysis and western blot. Plasma exosomal differentially expressed proteins (DEPs) were analyzed using Label-Free quantitative proteomics, and DEPs were characterized, annotated, and enriched based on Gene Ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) databases. The screened DEPs were validated by western blot and enzyme linked immunosorbent assay (ELISA) using plasma exosomal specimens.Results:According to the spinal cord injury classification of the American Spinal Injury Association, 14 cases were grade A, 19 cases were grade B, 12 cases were grade C, and 5 cases were grade D. Plasma exosomes of SCI patients and control groups showed typical cup-like morphology, with diameters mainly ranging from 30-200 nm. A total of 493 exosomal proteins were identified by Label-Free quantification, and 126 proteins were screened for differential expression, of which 38 were up-regulated and 88 were down-regulated. GO annotation revealed that DEPs were mainly involved in functions such as protein activation cascade, complement activation and immune response. KEGG pathway analysis revealed that DEPs were involved in biological pathways such as complement and coagulation cascade reactions, proteasome and neurodegenerative disease pathways. Two candidate proteins, APOB and S100A9, were initially screened based on quantitative results from proteomics and bioinformatics analyses. Western blot results showed that the relative expression of S100A9 protein in plasma exosomes of 30 SCI patients (1.62±0.19) was elevated compared with that of 30 control groups (0.86±0.24), and the difference was statistically significant ( t=8.55, P<0.001), while the relative expression of APOB protein (1.06±0.13 and 1.02±0.23) were not statistically significant ( t=0.46, P=0.653). The results of ELISA analysis showed that the expression of S100A9 in plasma exosomes of patients with different degrees of SCI (grade A 197.7±11.7 pg/ml, grade B 151.7±15.2 pg/ml, grade C 136.3±14.7 pg/ml) had statistical significance ( F=69.94, P<0.001), the higher the severity of SCI, the higher the expression of S100A9 in plasma exosomes (A vs. B, q=13.11, P<0.001; A vs. C, q=15.66, P<0.001; B vs. C, q=4.19, P=0.005). Conclusion:S100A9 is a potentially valid plasma exosomal molecular marker for assessing the severity of SCI.

Osteoporotic thoracolumbar fracture in the elderly will seriously reduce their quality of life and life expectancy. For osteoporotic thoracolumbar fracture in the elderly, spinal reconstruction is necessary, which should comprehensively consider factors such as the physical condition, fracture type, clinical characteristics and osteoporosis degree. While there lacks relevant clinical norms or guidelines on selection of spinal reconstruction strategies. In order to standardize the concept of spinal reconstruction for osteoporotic thoracolumbar fracture in the elderly, based on the principles of scientificity, practicality and progressiveness, the authors formulated the Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients ( version 2022), in which suggestions based on evidence of evidence-based medicine were put forward upon 10 important issues related to the fracture classification, non-operative treatment strategies and surgical treatment strategies in spinal reconstruction after osteoporosis thoracolumbar fracture in the elderly, hoping to provide a reference for clinical treatment.

Objective:To explore the effect of improving oral care based on cluster management in ventilator-associated pneumonia (VAP).Methods:Totally 126 VAP patients from February 2019 to February 2020 in our hospital were selected. The patients admitted from February 2019 to August 2019 were the control group, and the patients from September 2019 to February 2020 were the observation group, 63 cases in each group. The control group was treated with cluster nursing, and the observation group was treated with cluster nursing with improved oral care. The EICU hospitalization time, mechanical ventilation time, plaque index, pathogen infection, oral cleaning score, oral bleeding ulcer and aspiration were compared between the two groups.Results:After nursing, the EICU hospitalization time and mechanical ventilation time in the observation group were (18.34±4.15), (8.56±2.14) days, which were shorter than (23.56±4.82), (12.04±3.10) days in the control group ( t value was -6.514, -7.333, P<0.05). During the nursing process, the incidence of bleeding ulcer and aspiration in the observation group were 12.70% (8/63), 6.35% (4/63), which were lower than 30.16% (19/63), 20.63% (13/63) in the control group ( χ 2 values were 5.704, 5.508, P<0.05). After nursing, the plaque index of the two groups was lower than that before nursing ( t values were 12.516, 6.654, P<0.05), and the plaque index of the observation group was lower than that of the control group ( t value was -6.860, P<0.05). During the nursing process, the incidence of pathogenic bacteria infection in the observation group was 28.57% (18/63), which was lower than 49.21% (31/63) in the control group ( χ 2 value was 5.644, P<0.05). After nursing, the oral cleaning score of the two groups was lower than that before nursing ( t values were 11.118, 6.240, P<0.05), and the score of the observation group was (14.38±3.60) points, which was lower than (18.20±4.11) points of the control group ( t value was -5.549, P<0.05). Conclusion:Auricular pressure therapy can effectively improve the constipation symptoms, shorten the time required to take effect for the main symptoms, and improve the quality of life of schizophrenic patients.

Chimeric antigen receptor T (CAR-T) cell therapy is a novel cellular immunotherapy that is widely used to treat hematological malignancies, including acute leukemia, lymphoma, and multiple myeloma. Despite its remarkable clinical effects, this therapy has side effects that cannot be underestimated. Cytokine release syndrome (CRS) is one of the most clinically important and potentially life-threatening toxicities. This syndrome is a systemic immune storm that involves the mass cytokines releasing by activated immune cells. This phenomenon causes multisystem damages and sometimes even death. In this study, we reported the management of a patient with recurrent and refractory multiple myeloma and three patients with acute lymphocytic leukemia who suffered CRS during CAR-T treatment. The early application of tocilizumab, an anti-IL-6 receptor antibody, according to toxicity grading and clinical manifestation is recommended especially for patients who suffer continuous hyperpyrexia, hypotensive shock, acute respiratory failure, and whose CRS toxicities deteriorated rapidly. Moreover, low doses of dexamethasone (5-10 mg/day) were used for refractory CRS not responding to tocilizumab. The effective management of the toxicities associated with CRS will bring additional survival opportunities and improve the quality of life for patients with cancer.