Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.

Objective:To investigate the role of NIMA-related kinase-7 (NEK7) in breast cancer (BC) and its potential molecular mechanism.Methods:Quantitative real-time reverse-transcription (RT-qPCR) was used to detect the expression of NEK7 in BC tissue and cell lines. The effect of NEK7 on BC cell proliferation was examined by CCK-8. Proteins interacted with NEK7 were screened in Biological database. The effect of overexpression of NOD-like receptor protein 3 (NLRP3) on BC cell proliferation was evaluated. Western blot was used to detect NLRP3 protein expression, and ELISA was employed to evaluate IL-1β and IL-18 expression level.Result:NEK7 was upregulated in BC tissues and cells, and enforced-expression of NEK7 promoted BC cell proliferation[NEK7 over-expression group: 24 h: (0.33±0.02) , 48 h: (0.59±0.02) , 72 h: (0.76±0.02) ; Blank group: 24 h: (0.30±0.02) , 48 h: (0.45±0.02) , 72 h: (0.62±0.03) ; NEK7 empty vector group: 24 h: (0.32±0.02) , 48 h: (0.46±0.02) , 72 h: (0.63±0.03) ]. There was a positive correlation between NEK7 and NLRP3 ( R=0.13) . Overexpression of NLRP3 increased the proliferation ability of BC cell[NLRP3 over-expression group: 24 h: (0.35±0.02) , 48 h: (0.65±0.02) , 72 h: (0.80±0.03) ; Blank group: 24 h: (0.33±0.02) , 48 h: (0.51±0.02) , 72 h: (0.66±0.03) ; NLRP3 empty vector group: 24 h: (0.34±0.02) , 48 h: (0.52±0.03) , 72 h: (0.66±0.03) ]. NEK7 could positively regulate NLRP3 protein and up-regulate IL-1β (NEK7 over-expression group: 129.96±7.62 pg/ml, Blank group: 19.80±2.42pg/ml, NEK7 empty vector group: 21.30±1.77 pg/ml) and IL-18 (NEK7 over-expression group: 144.08±17.20 pg/ml, Blank group: 16.84±2.34pg/ml, NEK7 empty vector group: 17.64±1.94 pg/ml) expression. Conclusion:The upregulation of NEK7 was involved in the process of BC progression by inducing NLRP3 inflammasome activation, suggesting that NEK7 might be a promising therapeutic target for BC.

After years of exploration, Central South University has comprehensively reformed and upgraded the curriculum in both horizontal and vertical integration, including optimizing and constructing a basic clinical core curriculum system with organ system integration as the main line, utilizing the advantages of comprehensive universities to further advance the cross-teaching reform of science, engineering, arts and medicine, strengthening pre-medical education with the goal of early exposure to medicine, promoting the reform of early contact clinical integrated teaching according to the concept of "early, multiple and repeated clinical practice", and accelerating the integration of clinical skills training courses with the support of clinical skills simulation teaching. After the integration, the faculty team has gradually matured and the teaching quality has been significantly improved, which has strengthened the students' medical thinking and overall literacy.

Objective To explore the correlation between serum lipoprotein-related phospholipase A2 (Lp-PLA2), cystatin C (Cys-C) levels and disease severity in patients with hepatitis B cirrhosis (HBC). Methods Clinical data of 110 HBC patients in the hospital from October 2017 to May 2019 were retrospectively analyzed. According to Child-Pugh classification criteria of liver cirrhosis, they were divided into grade A (n=42), grade B (n=37) and grade C groups (n=31). Another 30 healthy controls during the same period were enrolled as control group. The levels of serum Lp-PLA2 and Cys-C were detected. And their correlation with disease severity was analyzed. Results Levels of serum Lp-PLA2 and Cys-C in HBC group were higher than those in control group (P<0.05). The levels of serum Lp-PLA2 and Cys-C were the highest in grade C group, followed by grade B group and grade A group (P<0.05). The areas under the ROC curve (AUC) of serum Lp-PLA2 combined with Cys-C for evaluating grade A and B, grade B and C HBC were 0.875 and 0.837, which were higher than those of Lp-PLA2 (0.772, 0.750) and Cys-C (0.750, 0.691) alone (P<0.05). Spearmann rank correlation analysis showed that levels of serum Lp-PLA2 and Cys-C were positively correlated with disease severity (r=0.659, 0.561, P<0.05). Conclusion The levels of serum Lp-PLA2 and Cys-C are significantly increased in HBC patients, which are gradually increased with the aggravation of HBC. The two indexes are positively correlated with disease severity, which are of diagnostic efficiency for the classification of liver cirrhosis.

Objective:To propose and verify a surgical classification system for the axial primary malignant and aggressive benign tumor.Methods:The CZH surgical classification system was originally developed for the axial primary malignant and aggressive benign tumor. The CZH surgical classification system includes seven types, according to the anatomic features and the extension of tumor violation. A total of 136 patients (79 males and 57 females) with axial primary malignant and aggressive benign tumor from multiple tertiary centers who received surgery from July 2006 to July 2019 were included. The average age was 44.40±17.55 years (8-83 years) old. There were 99 malignant tumors and 37 aggressive benign tumors included. The number of patients with each classification was presented as followed, Type I 13, Type II 15, Type IIIa 3, Type IIIb 20, Type IVa 43, Type IVb 12, Type Va 21, Type Vb 3, Type VI 2, Type VIIa 3 and Type VIIb 1. Surgical procedures were selected according to different types in classification. The inter- and intra-observer consistencies were evaluated by the Kendall's W test. The VAS, Frankel score, overall survival and recurrence free survival were recorded during the follow-up. Results:The inter- and intra-observer consistent coefficient was 0.973 and 0.996, respectively ( P<0.05). The single posterior approach was adopted for the Type II tumors. Other patients underwent surgery by the combined antero-posterior approach. The majority in anterior approach (113 cases) was the modified submandibular approach. The reconstruction modes included anterior "T" shape titanium mesh (112 cases) or the 3D printed prothesis (7 cases) combined with the posterior occipto-cervical fusion (92 cases) or the pedicle screw system (44 cases). The average surgical duration and the volume of intraoperative bleeding was 348.40±136.14 min (60-760 min) and 1 225.69±859.40 ml (80-4 000 ml), respectively. The operation duration and volume of intraoperative bleeding among each type were with statistical difference. The patients with Type IV, V tumors had longer operation duration than those with Type II tumors. Those with Type V and VII tumors had longer operation duration than those with Type I tumors. The patients with Type V tumors had more intraoperative bleeding than those with Type I-IV tumors. The average preoperative VAS score was 4.15±2.25 and then was reduced significantly to 0.62±0.71 and 0.38±0.59 at one and three months after operation, respectively. The Frankel score was also significantly ameliorated at one and three months postoperatively. There were 22 postoperative complications (16.2%). The complications included cerebral spinal fluid leak (12.5%), dysphagia and/or dysphonia (7.4%), dyspnea (5.1%), wound infection (3.7%), wound hemorrhage (2.2%) and pharyngeal dehiscence (1.5%). The incidence of postoperative complication was 25.9% in Type IV-VII tumors, while 11.8% in Type I-III tumors. Conclusion:CZH surgical classification system was verified with high observer consistency. This classification system could assist surgeons to select proper surgical approaches, resection modes and reconstruction modes, and thus ensure the safety of surgery and reduce the recurrence. The tumors in Type IV, V and VII may be with more challenging for surgeons. The incidence of postoperative complication in Type IV-VII tumors may be higher than that in Type I-III tumors.

Objective To explore the effect of PDCA [plan (P), design (D), check (C), act (A)] cycle on primary cardiopulmonary resuscitation (CPR) training for medical assistants in hospitals. Methods PDCA cycle was used to enhance continuous quality improvement (CQI) of team members to carry out brain storming to find out the root causes of the training difficulty, and directing to the 3 main root causes: lack of emergency rescue consciousness, without systematic training system and improper education procedure, it was proposed to arrange 3 great strategies: emergency rescue knowledge training, design of systematic training system and proper arrangement of training process. The changes of medical assistants' subjective willingness to perform the first aid, the accurate rates of answering questions on CPR location, frequency, depth, ratio of compression to breathing and awareness degree of CPR before and after training were observed. Results After training, the medical assistants' subjective willingness to perform the first aid was higher than that before training [91.7% (121/132) vs. 2.3% (3/132), P＜0.05]. The accurate answer rates on questions concerning CPR basic knowledge, such as location, frequency, depth and compression-breathing ratio had been greatly improved after training compared with those before training [location: 65.2% (86/132) vs. 4.5% (6/132), frequency: 40.2% (53/132) vs. 0 (0/132), depth: 90.2% (119/132) vs. 0 (0/132), compression-breathing ratio: 84.8% (112/132) vs. 1.5% (2/132 ), all P＜0.05]. After training, the percentage of medical assistants having very familiar awareness degree of CPR was significantly higher than that before training [65.2% (86/132) vs. 3.0% (4/132), P＜0.05]. Conclusion Via CPR training PDCA cycle, not only the efficiency of CPR training management is greatly improved, but also the training effect of participants is significantly elevated.

Retinoic acid, an active metabolite of vitamin A, exerts multiple effects on regulating embryonic development and inducing differentiation, proliferation, apoptosis as well as resistance in various cancer cells. Apart from the classic genomic action (binding to the nuclear receptors to regulate the expression of its downstream target genes), retinoic acids also play important roles in anti-cancer effect through non-genomic pathways (via extranuclear and non-transcriptional effects).
Apoptosis ; Cell Differentiation ; Genomics ; Humans ; Neoplasms ; drug therapy ; Receptors, Retinoic Acid ; Tretinoin

BACKGROUND: Bone marrow-derived mononuclear cells (BM-MNCs) hold the potential of differentiating into osteoclasts. Polygonatum sibiricum polysaccharide (PSP) may inhibit the differentiation of BM-MNCs into osteoclasts and it is expected to become a new drug for the treatment of osteoporosis. OBJECTIVE: To investigate the effect of PSP on the differentiation of mouse BM-MNCs into osteoclasts induced by receptor activator of nuclear factor kappa-B ligand (RANKL) and bone resorption in vivo. METHODS: Mouse bone marrow-derived macrophages cultured in vitro, the effect of macrophage colony stimulating factor and PSP (5, 10, 20, 40, 80,160, 320, 640, 1280, 2560 mg/L) on the proliferation of mouse BM-MNCs was detected by cell counting kit-8 assay to determine the PSP concentration range; the mouse BMMs were cultured and induced in DMEM medium containing macrophage colony stimulating factor, RANKL and 5, 10, 20, 40, 80,160, 320, 640 mg/L PSP, respectively; those cultured without PSP served as control group. The morphological changes of cells were observed under an inverted microscope.; the number of osteoclasts was detected by tartrate-resistant acid phosphatase staining; the mRNA expression levels of osteoclast-related genes including tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 were evaluated by quantitative real-time PCR. A mouse model of calvarial osteolysis induced by lipopolysaccharide was established to receive PSP intervention, and then micro CT scanning, three-dimensional reconstruction and relevants software were used for quantitative analysis of bone volume/volume percentage, trabecular number, trabecular bone spacing and thickness. The number of osteoclasts was identified by tartrate-resistant acid phosphatase staining and quantitative analysis of bone resorption area was conducted. RESULTS AND CONCLUSION: Compared with the control group, the concentration of PSP below 640 mg/L showed no significant effect on the proliferation of BMMs (P > 0.05). Different concentrations of PSP (40-640 mg/L) significantly reduced the number of osteoclasts, osteoclast differentiation and maturation, and the mRNA expression levels of tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 TRAP, MMP-9, CtsK and NFATc1 (P < 0.05). Compared with lipopolysaccharide, PSP could effectively alleviate the lipopolysaccharide-induced calvarial osteolysis, and the bone volume/volume percentage, trabecular number, and trabecular bone spacing were significantly decreased (P < 0.05); additionally, the number of osteoclasts and the area of bone resorption were decreased significantly (P < 0.01). To conclude, PSP can inhibit the differentiation and maturation of mouse BMMs to osteoclasts and alleviate lipopolysaccharide-induced calvarial osteolysis.

Objective To investigate PSP on bone microstructures,Ca,P,OPG and RANKL of osteoporotic rat model.Methods Thirty female rats randomly divided into five groups:Sham,OVX,H-,M-,L-PSP.Sham and OVX were irrigated stomachsaline;PSP solution was gavaged to other groups.After 8-week,bone microstructures of tibial metaphyseal,Ca,P,OPG and RANKL were measured.Results Body weight,Ca,P,RANKL,Tb.Sp of OVX were significantly increased compared to Sham,OPG,BV/TV,Tb.Th,Tb.N decreased.Body weight of H-,M-PSP,Ca and Tb.Sp of PSP,P and RANKL in H-PSP were decreased compared to OVX,OPG in H-,M-PSP,BV/TV,Tb.Th,Tb.N of PSP group increased.The differences were statistically significant (P ＜ 0.05).Conclusion PSP prevents osteoporosis by improving the microstructure of trabecular bone,reducing bone turnover,increasing OPG and reducing RANKL expression.

BACKGROUND: miRNA plays a critical regulatory role in the development and plasticity of spinal cord, and pathological changes after spinal cord injury. OBJECTIVE: To study the effect of miR-136-5p on the A20 expression in mouse astrocytes stimulated by interleukin-17 (IL-17). METHODS: C57BL/6 mouse astrocytes were cultured in vitro, identified by immunofluorescence staining, and then stimulated by 100 μg/L IL-17 for 0, 3, 6, 12 and 24 hours, respectively. The relative mRNA expression levels of IL-6 and tumor necrosis factor-α were detected by RT-PCR to determine the optimal stimulation time of IL-17. The mouse astrocytes were respectively stimulated by 10, 20, 50, 100 and 200 μg/L IL-7 for 6 hours, and similarly, the relative mRNA expression levels of IL-6 and tumor necrosis factor-α were detected to determine the optimal concentration of IL-17. At 6 hours after IL-17 (50 μg/L) stimulation, the mRNA expression levels of miR-136-5p and A20 in mouse astrocytes were detected by RT- PCR, and the protein expression level of A20 was detected by western blot assay. In addition, the lentiviral expression vector (miR-136-5p-inhibition) was constructed and transfected into the mouse astrocytes that were also stimulated by IL-7 to detect the expression levels of miR-136-5p, A20 mRNA and A20 protein. RESULTS AND CONCLUSION: Compared with the blank control group, the expression level of miR-136-5p in the miR-136-5p-inhibition group was significantly decreased after 6-hour IL-17 stimulation (P < 0.05). The expression levels of A20 mRNA and protein in each group were significantly decreased after 6-hour IL-17 (50 μg/L) stimulation (P < 0.05). The expression levels of A20 mRNA and protein in the miR-136-5p-inhibition group were significantly higher than those in the blank control group (P < 0.05), while there were no significant differences in the expression level of A20 protein between blank control and negative groups (P > 0.05). To conclude, miR-136-5p makes certain effect on the expression of A20 protein in astrocytes after IL-17 stimulation.