BACKGROUND:Osteoarthritis is now considered a metabolic disease.Previous studies have shown that glycolysis plays an important role in the occurrence and development of osteoarthritis.Compound 3k,as a novel small molecule inhibitor of glycolysis,has anti-inflammatory and anti-tumor effects.Therefore,it can target glycolysis and is expected to provide new ideas for the treatment of osteoarthritis. OBJECTIVE:To explore the role of Compound 3k in osteoarthritis caused by glycolytic overactivity based on the hypoxia-inducible factor 1 alpha(HIF-1α)/reactive oxygen species(ROS)pathway. METHODS:ATDC5 chondroblasts at logarithmic growth phase were taken to induce osteoarthritis in an in vitro cellular model by the action of 10 ng/mL interleukin-1β for 24 hours.The cytotoxicity of Compound 3k at different concentrations(0.25,0.5,1,2.5,5,10,15 μmol/L)was detected by cell counting kit-8 assay,and the appropriate concentrations were selected for the subsequent experiments.The chondrocytes were randomly divided into control,model and treatment groups.The model group was induced with 10 ng/mL interleukin 1β,and the treatment group was pre-stimulated with Compound 3k for 2 hours and then co-cultured with interleukin 1β.The proliferation of the cells in each group was detected by the cell counting kit-8 assay;the inflammatory level of the cells in each group was detected by the ELISA kit;the ROS,extracellular lactate and glucose contents were detected using the kit;qRT-PCR and western blot were used to detect the levels of related inflammatory factors,interleukin-6 and tumor necrosis factor-α,glycolysis-related genes glucose transporter protein-1,glyceraldehyde 3-phosphate dehydrogenase,monocarboxylate transporter protein-1 and HIF-1α. RESULTS AND CONCLUSION:Compared with the control group,the model group showed a decrease in cell proliferative activity,active glycolysis level,manifested by an increase in extracellular lactate content(P<0.001)and a decrease in glucose content(P<0.001),interleukin-6(P<0.000 1)and tumor necrosis factor-α(P<0.001).The expression levels of glycolysis-related genes glucose transporter protein-1(P<0.001),glyceraldehyde 3-phosphate dehydrogenase(P<0.001),monocarboxylic acid transporter protein-1(P<0.001)and HIF-1α(P<0.001)in the model group were all up-regulated,accompanied by oxidative stress and overproduction of ROS.Compared with the model group,Compound 3k treatment effectively increased cell proliferation activity and inhibited the level of overactive glycolysis(P<0.001),while suppressing the expression of genes related to inflammation(P<0.001)and glycolysis in osteoarthritic chondrocytes,inhibiting oxidative stress,downregulating the expression level of HIF-1α(P<0.000 1)and decreasing the content of ROS.To conclude,Compound 3k inhibits interleukin-1β induced chondrocyte inflammation,and its mechanism may be related to glycolysis and HIF-1α/ROS mediated oxidative stress.

Objective:To explore the feasibility and efficacy for the dissection and ligation of the superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer.Methods:Eight cadaveric heads were selected, and the laryngopharynxes were harvested. The positions of the superior laryngeal arteries entering the larynxes were dissected and observed under endoscopic vision, and their anatomical characteristics were summarized. Twenty-nine patients (all were male, aged 39-74 years old) with hypopharyngeal cancer who underwent transoral endoscopic surgery at the Department of Otorhinolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were selected, and the patients were randomly divided into two groups by drawing lots, namely, the superior laryngeal artery was actively dissected and occluded during surgery in observation group ( n=15) or not in control group ( n=14). The differences in surgical time, bleeding volume, postoperative complications, and postoperative disease-free survival rate were compared between the two groups. Statistical analysis was conducted using SPSS 25.0 software. Results:The entry point of the superior laryngeal artery into the larynx was approximately at the level of the superior edge of the thyroid cartilage, and entered the larynx at the posterior one-third of the lateral wall of the pyriform fossa. The superior laryngeal artery might be determined through endoscopic exploration in all patients of observation group. The endoscopic surgery time [(40.00±7.56) minutes] and intraoperative bleeding volume [(24.00±8.28) ml] in the observation group were respectively less than those [(48.57±14.06) minutes and (42.86±15.41) ml] in the control group, and the differences were statistically significant ( t=-2.064, P=0.049; t=-4.064, P=0.001). There was no case with postoperative bleeding in the observation group, but with one case of postoperative bleeding in the control group. Total disease free survival rate was 86.2% and there was no significant difference in disease free survival rates between the two groups during a follow-up period of at least 36 months ( P=0.986). Conclusion:Dissection of the superior laryngeal artery during endoscopic surgery for hypopharyngeal cancer is feasible, and pre-management and occlusion of the superior laryngeal artery can effectively reduce intraoperative bleeding.

Ultrasound has no radiation, portable, low price advantages, has been widely applied in the areas of general surgery, gynecology and obstetrics,however, there are some restrictions on ultrasound in bone imaging, its application in spine surgery are rare. In recent years, the development of technology and equipment has broadened the application scenarios of ultrasound in spine surgery. In terms of diagnosis, musculoskeletal ultrasound can clearly show the anatomical structure of muscles, tendons, joints and other tissues, which is of great significance for the early diagnosis of spinal diseases. In terms of treatment, ultrasound guided interventional therapy for spinal pain has been relatively mature, including selective nerve root block and facet joint block, which has shown high accuracy, safety and efficacy. In recent years, the application of ultrasound-assisted endoscopic spinal surgery has increased, which can reduce the radiation exposure faced by the operators and patients. Based on the evaluation of the integrity of the pedicle screw track and the distance between the inner wall of the screw track and the cortical bone by ultrasound, researchers have further explored its application in spinal pedicle screw placement, and developed new pedicle pathfinder. However, there are still controversies about the parameter settings of ultrasound equipment. In addition, intraoperative ultrasound can assist in locating tumors, assessing the extent of resection, and evaluating the effect of decompression and spinal cord perfusion during spinal tumor surgery. Although ultrasound has a broad application prospect in spine surgery, there are still some problems, such as low image clarity, low resolution accuracy, and limited bone imaging. In conclusion, the application of ultrasonic technology in spine surgery has shown great potential, with the progress of technology and the improvement of the doctor skills, the future will play a more important role in the field of spine surgery.

Objective:This study was performed to investigate the feasibility of preservation of internal branch of superior laryngeal nerve(ibSLN) during transoral endoscopic surgery for hypopharyngeal squamous cancer(HSCC) and the influence on patient′s swallowing function after operation.Methods:From May 2020 to June 2021, the data of 29 HSCC patients who required for transoral endoscopic surgery in the Department of Otorhinolaryngology Head and Neck Surgery, the Second Xiangya Hospital of Central South University were prospectively included, and the included patients were divided into two groups randomly by lottery. According to whether ibSLN was actively dissected during operation, they were divided into ibSLN preservation group ( n=15) and control group ( n=14, without ibSLN preservation). Operation time, intraoperative hemorrhage, intraoperative neck dissection, postoperative radiotherapy, postoperative recurrence within 1 year, retention and swallowing function, the recovery of oral soft diet and the quality of life were compared between two groups. SPSS 25.0 software was used for statistical analysis. Results:The study included 29 eligible patients, including 25 males and 4 females.The age ranged from 42 to 67 (56.07±5.93) years. There were no significant differences( P>0.05) between 2 groups in the following data,including age( t=-0.56), gender( χ2=0.01), TNM stage(T stage χ2=0.29, N stage χ2=0.02), pathological diagnosis( χ2=0.03), preoperative swallowing function( χ2=0.00) and M. D. Anderson Dysphagia Inventory(MDADI) score(global t=0.55, emotional t=0.16, functional t=0.60, physical t=0.64), operation time( t=1.62) and intraoperative hemorrhage( t=-1.46), intraoperative neck dissection( χ2=0.01), postoperative radiotherapy( χ2=0.32), postoperative recurrence within 1 year( P>0.050). The swallowing function was evaluated by water swallowing test after operation. The swallowing function of ibSLN preservation group was better than control group, and the difference between two groups was statistically significant on the 1st ( χ2=4.44, P=0.035), 5th ( χ2=4.24, P=0.039) and 7th ( χ2=4.55, P=0.033) day after operation. On the 14th day after operation, the MDADI scores of patients in the ibSLN preservation group were higher than those in the control group in global ( t=2.45, P=0.021), functional ( t=2.54, P=0.017) and physical ( t=2.24, P=0.034) dimensions, except for emotional dimension ( t=1.89, P=0.070). The median time of oral soft diet( U=23.00, P<0.001), normal oral diet( U=21.00, P<0.001) and the nasogastric tube removal time ( U=18.50, P<0.001) in ibSLN preservation group was 2 days, 5 days and 6 days respectively, earlier than that in control group, which had statistically significant difference. Conclusion:Our results show that it is feasible to preserve the ibSLN during HSCC transoral endoscopic surgery, which can achieve rapid recovery of postoperative swallowing function.

The emergence of regular short repetitive palindromic sequence clusters (CRISPR) and CRISPR- associated proteins 9 (Cas9) gene editing technology has greatly promoted the wide application of genetically modified pigs. Efficient single guide RNA (sgRNA) is the key to the success of gene editing using CRISPR/Cas9 technology. For large animals with a long reproductive cycle, such as pigs, it is necessary to screen out efficient sgRNA
Animals ; CRISPR-Cas Systems/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; Gene Editing ; RNA, Guide/genetics* ; Swine

Objective:To analyze the mutations in desmosomal protein genes of arrhythmogenic right ventricular cardiomyopathy (ARVC) in relatives of Yunnan unexplained sudden death (YUSD) cases in Nanjian County, Dali Prefecture, Yunnan Province, and provide a basis for etiological hypothesis and control measures.Methods:The blood samples of YUSD case relatives ( n = 7) and control villagers ( n = 7) were collected, and basic situation investigation and electrocardiography (ECG) examination were performed at the same time. Blood DNA was extracted as a template for PCR amplification, and Sanger method was used to perform plakophilin 2 (PKP2), desmoglein 2 (DSG2), desmocollin 2 (DSC2), desmoplakin (DSP), and junction plakoglobin (JUP) five ARVC desmosomal protein genes sequencing of a total of 97 exons, and comprehensive analysis of gene mutations was carried out. Results:Five of YUSD case relatives carried genetic mutation sites, including DSP gene heterozygous synonymous mutations about exon 20 c.2862 C>T (p.Cys954Cys) and exon 24F c.7122 C>T (p.Thr2374Thr), DSC2 gene heterozygous missense mutation about exon 15 c.2326 A>G (p.Ile776Val), and all the five people were single heterozygous mutation carriers. Among them, two case relatives of the father-son carried the same site mutation of the DSC2 gene; the abnormal ECGs of three YUSD case relatives were ST-T change or clockwise rotation. However, the mutation sites of PKP2, DSG2, DSC2, DSP and JUP genes in control villagers were not detected.Conclusions:YUSD case relatives in Nanjian County carry ARVC desmosomal protein genes DSP and DSC2 mutations. Pathogenic mutation of DSC2 gene c.2326 A>G (p.Ile776Val) is may related to the incidence of some YUSD cases.

Objective: To investigate the effects of antimicrobial peptides merecidin on the biological functions of human lung adenocarcinomaA549 cells and the potential signaling pathways and targets that involved in promoting apoptosis, by studying the changes of phosphorylation levels of proteins in A549 cells after merecidin treatment. Methods: The antibacterial peptide mericidin (9 μmol/L) was applied to treat A549 cells for 6 h, and the total protein was collected and extracted. The peptide was digested by trypsin and labeled with TMT, and then fractionated by HPLC. The phosphorylated peptides were enriched with IMAC-Fe, and finally subjected to mass spectrometry analysis. Library identification and quantification of phosphorylated peptides obtained by mass spectrometry were processed using MaxQuant software, to further analyze the functions and pathways of differentially expressed phosphorylated proteins by combining with bioinformatic analysis. Results: Through IPA analysis of phosphorylated proteins in the normal control group and the antibacterial peptide merecidin treatment group, 753 differentially phosphorylated proteins in mericidin treatment group were screened out under the conditions of |Fold Change|≥2 and P<0.05, including 229 significantly up-regulated genes and 417 down-regulated genes. Among them, the differentially expressed proteins related to apoptosis included RB1, MAPK1, ARAF, PTK2, FOXO, MARCKSandsoon.Theresultsofbiologicalprocessanalysisshowedthatdifferentiallyexpressed phosphorylated proteins were mainly concentrated in cell signal transduction, degradation and transport of nucleic acid, and cellular energy metabolism, protein translation and synthesis, and cytoskeleton formation etc. The enrichment results showed that the differentially expressed phosphorylated proteins were mainly involved in apoptosis-related MAPK, ErbB, PI3K-Akt, and Ras signaling pathways. Protein-protein interaction analysis indicated the associations among apoptosis-related proteins PTK2, PRKCA, MA2PK2, MAPK1, and LMNA. Conclusion: The antibacterial peptide merecidin may induce apoptosis and alteration of other cell functions by affecting a variety of genes and signaling pathways such as RB1, MAPK1,ARAF, PTK2, FOXO and MARCKS etc.

Objective:To evaluate the feasibility and efficacy of partial superficial parotidectomy with V-shaped incision by comparing with the Blair incision and hairline N-shaped incision.Methods:From January 2015 to January 2016, 60 patients (47 males and 13 females, with an age range of 25- 63 years) required for superficial partial parotid gland resection were randomly divided into three groups: V-shaped incision (VI) group, Blair incision (BI) group and hairline N-shaped incision (NI) group, with 20 cases in each group.Intraoperative, postoperative and follow-up indexes were compared between three groups. Operative time and drainage volume in the surgery of tumors at different sites in VI group were compared. SPSS18.0 software was used for statistic analysis.Results:There were no statistically significant differences among the three groups in operative time, drainage volume, postoperative hospital stay, periauricular numbness, fistulas, pain score, facial palsy, and scar score at the 3rd month after surgery ( P>0.05). For appearance satisfaction score at the 6th month after surgery, VI group was better than BI group or NI group, with significant differences(VI group vs. BI group: 9.00[8.00, 9.00] vs. 5.00[4.00, 5.25], χ 2=6.629, P<0.001; VI group vs. NI group: 9.00[8.00, 9.00] vs. 7.00[6.00, 8.00], χ 2=2.942, P=0.010; BI group vs. NI group: 5.00[4.00, 5.25] vs. 7.00[6.00, 8.00], χ 2=-3.687, P=0.001). For tumors located in the front, upper and middle of parotid gland, there were no statistically significant differences in operative time and drainage volume between the three groups ( P>0.05). For tumors located at the lower part of parotid gland, the difference in operative time between the three groups was statistically significant ( F=7.278, P=0.01). With pairwise comparison, operative time in VI group was longer than that in BI group or NI group, but there was no significant difference between BI group and NI group (VI group vs. BI group: (181.00±22.89) min vs. (132.50±9.01) min, t=3.694, P=0.004; VI group vs. NI group:(181.00±22.89) min vs. (149.00±15.94) min, t=2.585, P=0.025; BIgroup vs. NI group, (132.50±9.01) min vs. (149.00±15.94) min, t=1.257, P=0.235). For tumors located at the lower part of parotid gland, the differences in intraoperative drainage volume were not statistically significant between three groups ( P>0.05). There were no statistically significant differences in operative time and drainage volume in the surgery of tumors at different sites in VI group ( P>0.05). Conclusions:By use of V-shaped incision for the surgery of benign parotid gland tumors, the operation time of tumors located only in the lower part of the parotid gland will be prolonged. For tumors in different sites without increasing surgical complications, this modality can get good cosmetic effect.

Objective: To investigate blood pressure and vascular remodeling of OSAS by establishing the chronic-intermittent hypoxia model in rat. Methods: Experiments were performed on 35 adult male Sprague-Dawley rats. Animals were randomly divided into four groups: unhandled control group (with 5 rats in it), CIH group at 9/6/3 weeks (with 10 ratsin each group). Rats in CIH group went through 8-hour intermittent hypoxia everyday, and those in control group were raising normally. After 9-week experiment, blood pressure was measured. The changes of the following indexes were observed: pathological changes of aorta and the middle aorta thickness (HE staining), the collagen of aorta wall (Masson staining). The experimental data were analyzed by SPSS 24.0 statistical software. The variance was analyzed by one-way analysis of variance, and the irregularity was selected using the calibration t test. Results: The systolic and diastolic blood pressures of the CIH9, 6, and 3 weeks groups and the control group were: (127±13) and (79±9), (124±11) and (81±7), (101±11) and (75±9), (91±10) and (65±9) mmHg (1 mmHg=0.133 kPa). The systolic blood pressure and diastolic blood pressure of the rats in the week of CIH 9 and 6 weeks were significantly higher than the control group (F=14.64, P=0.000; F=6.81, P=0.000). There was no significant difference in the mean blood pressure between the three groups of CIH and the control group. Membrane thickness in CIH9, 6 and 3 weeks and control group were: (20±2), (19±2), (14±2), (13±3) μm. Compared with the control group, the aortic pathology and thickness of the middle layer of the CIH9 and 6 weeks group were significantly thicker (F=20.24, P=0.000), but there was no significant difference between the CIH3 week group and the control group; the collagen deposition was unchanged compared with the control group. Conclusion Intermittent hypoxia for 6 weeks or more in rats resulted in the increasement of blood pressure, morphological changes of aorta and vascular remodeling in thickened media.