[Clinical report of hoding cricoarytenoid joint reduction with visual laryngoscope under intravenous anesthesia].

Objective: To investigate the reduction effect of hoding cricoarytenoid joint reduction with visual laryngoscope under intravenous anesthesia. Methods: The therapeutic effects of 40 patients with arytenoid dislocation(AD)treated by closed reduction in the single center from January 2020 to September 2021 were retrospectively analyzed, including 21 males and 19 females, median age 48 years. The etiology, symptoms, preoperative evaluation methods, reduction mode, reduction times, and the recovery of arytenoid cartilage movement and sound after reduction were evaluated and analyzed. Results: All patients had obvious hoarseness and breath sound before treatment. Under stroboscopic laryngoscope or electronic nasopharyngoscope, different degrees of vocal cord movement disorder and poor glottic closure can be seen. There were 28 cases of left dislocation, 9 cases of right dislocation and 3 cases of bilateral dislocation. The etiology of dislocation of cricoarytenoid joint: 25 cases (62.5%) of tracheal intubation under general anesthesia were the most common causes, was as follows by laryngeal trauma, gastroscopy, cough, vomiting and so on. Among them, 28 cases of reduction were initially diagnosed in our department, and 12 cases were diagnosed later after failure of reduction treatment. Of the 40 patients, 6 underwent reduction 24 hours after dislocation; 18 cases from 3 days to 1 month; 7 cases from 1 to 3 months; 6 cases were reset in 3~6 months; Over 6 months in 3 cases. After one reduction, 10 cases (10/40, 25%) recovered normal pronunciation, 14 cases (14/40, 35%) recovered normal pronunciation after two reduction, 10 cases (10/40, 25%) recovered normal pronunciation after three times, 2 cases (2/40, 5%) recovered normal pronunciation after four times, and 1 case (2.5%) recovered normal pronunciation after five times. Thin slice CT scan of larynx and cricoarytenoid joint reconstruction showed the types of AD: subluxation in 37 cases (92.5%) and total dislocation in 3 cases; 28 cases of left dislocation, 9 cases of right dislocation and 3 cases of bilateral dislocation; 29 cases (72.5%) had posterior dislocation and 11 cases (27.5%) had anterior dislocation. All patients were treated by intravenous anesthesia with arytenoid cartilage clamped by cricoarytenoid joint reduction forceps under visual laryngoscope. The curative effect was evaluated by stroboscopic laryngoscope and/or voice analysis at 1-2 weeks after operation. The vocal cord movement returned to normal and the pronunciation was good in 37 cases (92.5%). Conclusions: Hoding cricoarytenoid joint reduction with the vision laryngoscope under intravenous anesthesia is easy to operate and the reduction effect is more stable. It is a effective method for AD.

KLF15 reduces the level of apoptosis in mouse liver induced by sepsis by inhibiting p38MAPK/ERK1/2 signaling pathway.

OBJECTIVE: Sepsis-induced acute liver injury (ALI) involves multiple systems in the body. The disease is acute and critical, with various symptoms, including extensive necrosis of liver cells. There is currently no effective treatment to deal with ALI in a timely manner. This study verified the therapeutic effect of Krüppel-like factor 15 (KLF15) on ALI by studying its anti-apoptotic effect on the liver. MATERIALS AND METHODS: We induced ALI in mice with lipopolysaccharide (LPS)/D-galactosamine (D-GaIN). Recombinant mouse KLF15 was used to treat mice to examine the protective effects of KLF15 on mouse liver and the effects of apoptosis-related molecules. In addition, we cultured Kupffer cells and determined the anti-inflammatory and anti-apoptotic effects of KLF15 and its mechanism by overexpressing KLF15. RESULTS: Exogenous KLF15 effectively reduced the levels of TIBL, ALT, AST, and inflammatory factors (COX-2, MCP-1, IL-1ß, and TNF-α) in mouse serum. The results of HE staining also demonstrate that KLF15 improves the morphology of liver tissue. In addition, the expression of KLF15 in LPS-induced Kupffer cells was significantly reduced and KLF15 increased the viability of Kupffer cells and decreased the level of inflammation in Kupffer cells. In both in vivo and in vitro experiments, KLF15 reduced the level of apoptosis in hepatocytes or Kupffer cells and inhibited the activity of the p38MAPK/ERK1/2 signaling pathway. CONCLUSIONS: KLF15 reduces the apoptosis and inflammation levels of liver and Kupffer cells by inhibiting the p38MAPK/ERK1/2 signaling pathway and alleviates LPS/D-GaIN-induced ALI.

Effect of lncRNA CRNDE on sepsis-related kidney injury through the TLR3/NF-κB pathway.

OBJECTIVE: The aim of this study was to detect the expression of long non-coding ribonucleic acid (lncRNA) colorectal neoplasia differentially expressed gene (CRNDE) in the kidney tissues of mice with sepsis-induced acute kidney injury (AKI) and its effect on KI, and to further explore its mechanism. MATERIALS AND METHODS: A total of 60 male C57 mice were randomly divided into 3 groups based on a random number table, including the control group (Sham group, n=20), sepsis-related KI group [lipopolysaccharide (LPS) group, n=20] and CRNDE inhibition group [LPS + CRNDE small interfering ribonucleic acid (siRNA) group, n=20]. Mice in LPS and LPS + CRNDE siRNA groups were intraperitoneally injected with 5 mg/kg LPS, while the tail vein was injected with 5 µL CRNDE siRNAs. After 12 h, the expression level of lncRNA CRNDE in kidney tissues of mice in each group was detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR). At the same time, 2 mL of orbital blood was collected from each mouse, and the levels of creatinine and blood urea nitrogen were detected. Subsequently, kidney tissue samples were collected from mice in each group. Periodic acid Schiff (PAS) staining was used to assess the injury of renal tubulointerstitium, followed by scoring. Hematoxylin and eosin (H&E) staining was applied to detect cell injury in kidney tissues. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was adopted to examine the apoptosis in kidney tissues in mice of each group. Meanwhile, the distribution and expression of p65 in kidney tissues of mice in each group were determined via immunohistochemical staining. Finally, the expression of Toll-like receptor 3 (TLR3)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway in kidney tissues of mice in each group was detected using Western blotting. RESULTS: Compared with the Sham group, lncRNA CRNDE level in the kidney of the LPS group was remarkably up-regulated (p<0.05). The levels of creatinine and blood urea nitrogen in LPS + CRNDE siRNA group were notably lower than those of the LPS group (p<0.05). PAS staining results manifested that renal tubulointerstitial injury in the LPS group was significantly more serious than that of the LPS + CRNDE siRNA group (p<0.05). According to H&E staining results, serious edema, rupture and necrosis observed in kidney tissue cells of the LPS group. However, after the intervention of CRNDE siRNA, cell edema and necrosis were markedly relieved. In addition, TUNEL staining results indicated that the apoptotic level of kidney tissue cells in the LPS + CRNDE siRNA group was significantly lower than that of the LPS group (p<0.05). Subsequent immunofluorescence staining demonstrated that p65 expression in the LPS group increased significantly, which was markedly inhibited by CRNDE siRNA intervention (p<0.05). Furthermore, Western blotting displayed that CRNDE siRNA could effectively inhibit the activation of TLR3 and p65 in mouse kidney tissue induced by LPS (p<0.05). CONCLUSIONS: Inhibition of CRNDE can reduce sepsis-induced KI by blocking the activation of the TLR3/NF-κB pathway. Moreover, CRNDE is expected to become a target for clinical treatment of sepsis-related KI.

[The significance of circulating tumor cells in head and neck squamous cell carcinoma: a preliminary study].

Objective: To investigate the significance of circulating tumor cells (CTC) in squamous cell carcinoma of the head and neck (HNSCC). Methods: Twenty-four patients with HNSCC treated between October 2016 and July 2017 in our department were selected (experimental group), including 23 males and 1 females, aged 47-81 years. There were 14 cases of squamous cell carcinoma of larynx and 10 cases of hypopharynx, including I-Ⅱ stage (5 cases) and Ⅲ- Ⅳ stage (19 cases). All patients were primary and/or relapsed after treatment. Nine healthy volunteers were selected as control group. A novel in vivo capture technique (CellCellector system) was used to detect CTC. SPSS23.0 was used for statistical analysis. Results: The total capture rate of CTC in patients with HNSCC before treatment was 70.8% (17/24), with 40% (2/5) for patients at I-Ⅱ stage, and 78.9% (15/19) for patients at Ⅲ- Ⅳ stage, and was 0 in patients of control group. The total capture rate of CTC in patients with HNSCC after treatment was 50% (8/16). There was no significant correlation between CTC and age, sex, location of tumor or lymph node metastasis (P>0.05). CTC was related to tumor staging and tumor differentiation (P<0.05). The positive rate of EGFRVⅢ in CTC was 26.3% (5/19). Conclusions: The CellCollector system is a very efficient way of detecting CTC, and CTC plays an important role in the occurrence, progression and metastasis of HNSCC.

[Clinical effect of alanyl glutamine in the treatment of patients with gastrointestinal function obstacle caused by severe phorate poisoning].

Objective: To observe the therapeutic efficacy of alanyl glutamine injection on patients with gastrointestinal function obstacle caused by severe phorate poisoning. Methods: A total of 80 eligible patients with gastrointestinal function obstacle caused by severe phorate poisoning were randomly divided into the control group (n=40) and treatment group (n=40) . The control group was treated with the conventional therapy, which included forbidden diet, atropine, pralidoxime iodide, anti-inflammatory, albumin infusion, ω-3 fish oil fat emulsion, protection of organs function, blood perfusion, and Fat Emulsion, Amino Acids (17) and Glucose Injection. The treatment group was treated with alanyl glutamine injection plus the conventional therapy. To observe the time of recovering to normal of gastrointestinal function between the two groups, compared the AChE activity and changes of prealbumin, albumin and total protein of the two groups respectively. Furthermore, the total atropine dosage, the total pralidoxime iodide dosage and ICU stay time between the two groups were also compared. Results: The gastrointestinal function recovery time of patients in the treatment group was less than the control group, the difference was statistically significant (P<0.05) . From the third day of treatment, the serum cholinesterase activity of the treatment group was higher than the control group, the difference was statistically significant (P<0.05) . On the 5th day and 10th day of the treatment, the prealbumin, albumin and total protein of the treatment group were significantly higher than these indexes of the control group in the same period, the difference were statistically significant (P<0.05) . The total atropine dosage, the total pralidoxime iodide dosage and ICU stay time in the treatment group were lower than the control group, the difference were statistically significant (P<0.05) . Conclusion: Alanyl glutamine injection has a great therapeutic effect for gastrointestinal function obstacle patients caused by severe phorate poisoning.

[Enrichment and detection of circulating tumor cells and its application in head and neck squamous cell carcinoma].

Circulating tumor cells are tumor cells from the primary tumor or metastatic tumor to fall off into the circulatory system. They are closely related to the recurrence and metastasis of tumor. Detection of circulating tumor cells as a "liquid biopsy" , to get the tumor information through non-invasive sampling method. It has the advantages of short time of tumor diagnosis, repeatable operation, and has gradually replaced the traditional biopsy. In this paper, the enrichment and detection of circulating tumor cells and its application in head and neck squamous cell carcinoma were reviewed.

[Preliminary study on the expression of EGFRvâ ¢ and CXCR4 in head and neck squamous cell carcinoma].

Objective:The aim of this study is to detect the expression of EGFRvⅢ and CXCR4 in HNSCC and to explore its possible mechanism. Method:We selected 60 cases of HNSCC from May 2013 to November 2016 in our hospital for surgical treatment, including 44 cases of laryngeal squamous cell carcinoma, 14 cases of hypopharyngeal squamous cell carcinoma, and 2 cases of nasal squamous cell carcinoma. pTNM staging according to AJCC 2010, Ⅲ stage 17 cases, Ⅳa stage 40 cases, and Ⅳb stage 3 cases; according to tumor differentiation degree: 13 cases of high differentiation, moderately differentiation in 31 cases, and 16 cases of low differentiated; primary tumor, positive lymph node metastasis as the experimental group, and 10 cases of adjacent tissues are taken as control (control group). The expression of EGFRvⅢ and CXCR4 in primary tumor, metastasis and control group was observed by immunohistochemistry. The immunohistochemical results are semi quantitatively analyzed by Image-Pro Plus. All analyses were conducted using the SPSS 23.0 and Graphpad-Prism 5.0. Result:The expression of EGFRvⅢ and CXCR4 in HNSCC is higher than control, which was not correlated with age, gender, location, degree of differentiation, T grading, and TNM stage (P>0.05). But there was a significant correlation with N grading (P<0.05). The pathology result shows lymph node metastasis in cancer tissues were high expression, but the difference in expression between primary and metastatic tumors are not statistically significant. There is no significant positive correlation between EGFRvⅢ and CXCR4 (r=0.144, P>0.05). Conclusion: Both EGFRvⅢ and CXCR4 promote the invasion and metastasis of advanced HNSCC, however, there may be some synergistic effect between them.

[The role of CD4 ⁺ CD25 ⁺ T regs and CCL17, CCL22 in the pathogenesis of head and neck squamous cell carcinoma].

Objective:To investigate the role of CD4 ⁺ CD25 ⁺ T regs and CCL17 and CCL22 in the pathogenesis of HNSCC.Method:Twenty cases of HNSCC were enrolled. All patients were primary or recurrent after treatment (chemotherapy, surgery). The primary tumor was taken as the experimental group, and the adjacent normal tissues from the primary tumor 1-3 cm were taken as control group. CD4 ⁺ /Foxp3 and CD25⁺/Foxp3 were detected by immunofluorescence, while CCL17 and CCL22 were detected by ELISA. The difference and correlation between the amount of CD4⁺,CD25⁺ and the expression of CCL17, CCL22 were observed and analyzed.Result:The difference of mean optical density between CD4⁺/Foxp3 and CD25⁺/Foxp3 was statistically significant between the experimental group and the control group (P<0.05). The concentration of CCL17 and CCL22 was statistically different between the two groups (P<0.01). There was a positive correlation between CD25⁺and CCL17,CCL22(r=0.595, 0.720,P<0.01).Conclusion:CD4⁺CD25⁺T regs and CCL17,CCL22 played an important role in the pathogenesis of head and neck squamous cell carcinoma,both of which interacted with each other,and promoted the recurrence and metastasis of HNSCC.

[Preliminary study on the expression of CD4⁺CD25⁺Tregs and Foxp3 in peripheral blood of patients with head and neck squamous cell carcinoma].

Objective:This paper discusses the expression and significance of CD4⁺CD25⁺ Tregs and Foxp3 in peripheral blood of patients with head and neck squamous cell carcinoma. Method:We have collected 40 cases of head and neck squamous cell carcinoma patients with newly diagnosed or relapse after treatment, all of them underwent surgery, 39 males and 1 females, aged 41-79 years, in our department from January 2014 to December 2015. At the same time, 10 healthy volunteers are enrolled as control group. 2 ml peripheral blood has been detected by flow cytometry, and the ratio of CD4⁺CD25⁺/CD4⁺ and CD4⁺CD25⁺Fxop3⁺/CD4⁺ are calculated, respectively. SPSS 23.0 is used for statistical analysis. Result:CD4⁺CD25⁺ Tregs is highly expressed in head and neck tumors, compared with that in the healthy control, and the difference is statistically significant (P<0.01). There is significant difference between the early and late stage (P<0.05). The positive rate of Foxp3+ is higher in CD4⁺CD25⁺ Tregs positive cells than in control group (P<0.01). The difference of positive rate between late stage and early stage head and neck tumors is statistically significant (P<0.05). There is a significant positive correlation between CD4⁺CD25⁺ Tregs and Foxp3 (r=0.95). Conclusion:CD4⁺CD25⁺ Tregs and Foxp3 are highly expressed in the peripheral blood of patients with head and neck squamous cell carcinoma. Through the inhibition of the immune system in patients with head and neck squamous cell carcinoma, the development of carcinoma were promoted.

[Association between genetic polymorphisms of HIF-2α gene and high altitude pulmonary hypertension in Han population].

Objective: To test the hypothesis that the single nucleotide polymorphisms (SNPs) in HIF-2α gene were associated with the susceptibility of high altitude pulmonary hypertension (HAPH) in Han population in China. Methods: Those Han population who emigrated to plateau (average altitude 3 300 m) and have been lived here for more than 20 years were included as the research subjects, the method of cluster random sampling was used to enroll 49 HAPH patients as the case group (HAPH group) and 39 free of HAPH people as the control group. The Sanger chain termination method was used to detect the SNPs of rs1562453, rs1867785, rs4953361, rs7598371 and rs11125068 in HIF-2α gene. Results: The genotype and frequencies of rs1562453 were CC (49.0%), CT (46.9%) and TT (4.1%) in the HAPH group, and the locus's genotype and frequencies were CC (76.9%), CT (17.9%) and TT (5.1%) in the control group. The alleles and frequencies of rs1562453 were C (72.4%) and T (27.6%) in the HAPH group, and the locus's alleles and frequencies were C (85.9%) and T (14.1%) in the control group. Their genotypes and alleles frequencies were of significant difference between the two groups (genotypes P=0.017; allele P=0.031). Meanwhile, The genotype and frequencies of rs1867785 were AA (46.9%), AG (46.9%) and GG (6.1%) in the HAPH group, and the locus's genotype and frequencies were AA (74.4%), AG (25.6%) and GG (0.0%) in the control group. The alleles and frequencies of rs1867785 were A (70.4%) and G (29.6%) in the HAPH group, and the locus's alleles and frequencies were A (87.2%) and G (12.8%) in the control group. Their genotypes and alleles frequencies were of significant difference between the two groups (genotypes P=0.020; allele P=0.008). Genotypes frequencies of the rs1562453 and rs1867785 were analyzed by Logistic Regression which showed that the genotypes frequencies of rs1562453 were also of significant difference between two groups (Wald=9.561, P=0.008), CT vs CC (ß=1.720, OR=5.580, P=0.011). Conclusion: The SNPs of rs1562453 in HIF-2α gene may be associated with the development of HAPH among Chinese Han population, and individuals with the genotype CT may be more vulnerable to HAPH than those who carry genotype CC, and allele T may be a risk factor for HAPH.

miR-133 inhibits pituitary tumor cell migration and invasion via down-regulating FOXC1 expression.

Many studies have shown that microRNA (miR)-133 functions as a tumor suppressor in a variety of metastatic cancers, including breast cancer, gastric cancer, and liver fibrosis. However, the influence of miR-133 on pituitary tumor malignancy has not yet been reported. The purpose of this study was to explore the role of miR-133 in pituitary tumor cell migration and invasive ability and the molecular mechanisms involved. Our findings suggest that in pituitary adenoma cell lines, through direct targeting and negative control of forkhead box C1 (FOXC1), miR-133 can inhibit pituitary adenoma cell migration and invasion. In addition, epithelial-to-mesenchymal transition can be induced by miR-133. Additionally, a negative correlation was found between FOXC1 and miR-133 expression when comparing their expression levels between cancerous tissue and adjacent normal tissue. This suggests that miR-133 can inhibit cell migration and invasion by directly targeting FOXC1, implying that miR-133 could be a potential therapeutic target for treatment of invasive pituitary adenoma.

MDM2 can promote the ubiquitination, nuclear export, and degradation of p53 in the absence of direct binding.

MDM2 can bind the N terminus of p53 and promote its ubiquitination and export from the nucleus to the cytoplasm, where p53 can then be degraded by cytoplasmic proteasomes. Several studies have reported that an intact MDM2 binding domain is necessary for p53 to be targeted for ubiquitination, nuclear export, and degradation by MDM2. In the current study, we examined whether the MDM2 binding domain of p53 could be provided in trans through oligomerization between two p53 molecules. p53 proteins mutated in their MDM2 binding domains were unable to bind MDM2 directly and were resistant to MDM2-mediated ubiquitination, nuclear export, and degradation when expressed with MDM2 alone. However, these same p53 mutants formed a complex with MDM2 and were efficiently ubiquitinated, exported from the nucleus, and degraded when co-expressed with MDM2 and wild-type p53. Moreover, this effect required MDM2 binding by wild-type p53 as well as oligomerization between wild-type p53 and the MDM2 binding-deficient p53 mutants. Taken together, these results support a model whereby MDM2 binding-deficient forms of p53 can bind MDM2 indirectly through oligomerization with wild-type p53 and are subsequently targeted for ubiquitination, nuclear export, and degradation. These findings may have important implications regarding the DNA damage response of p53.

Downregulation of MDM2 stabilizes p53 by inhibiting p53 ubiquitination in response to specific alkylating agents.

p53 is stabilized in response to DNA damaging stress. This stabilization is thought to result from phosphorylation in the N-terminus of p53, which inhibits p53:MDM2 binding, and prevents MDM2 from promoting p53 ubiquitination. In this report, the DNA alkylating agents mitomycin C (MMC) and methylmethane sulfonate (MMS), as well as UV radiation, stabilized p53 in a manner independent of phosphorylation in p53 N-terminus. This stabilization coincided with decreased levels of MDM2 mRNA and protein, and a corresponding decrease in p53 ubiquitination. Importantly, MDM2 overexpression inhibited the stabilization of p53 and decrease in ubiquitination following MMC, MMS, and UV treatment. This indicates that downregulation of MDM2 contributes to the stabilization of p53 in response to these agents.

MDM2-dependent ubiquitination of nuclear and cytoplasmic P53.

Wild-type p53 is stabilized and accumulates in the nucleus of DNA damaged cells. The effect of stabilizing p53 is to inhibit cell growth, either through a G1 cell cycle arrest or apoptotic cell death. MDM2 can inhibit p53 activity, in part, by promoting its rapid degradation through the ubiquitin proteolysis pathway. In the current study, MDM2-mediated degradation of p53 was partially inhibited in cells treated with leptomycin B (LMB), a specific inhibitor of nuclear export. In contrast, levels of ubiquitinated p53 increased in LMB-treated cells, indicating that nuclear export is not required for p53 ubiquitination. To investigate this further, p53 mutants were generated which localize to either the nucleus or cytoplasm, and their susceptibility to MDM2-mediated ubiquitination was assessed. p53 mutants that localized to either the nucleus or the cytoplasm were efficiently ubiquitinated, and their steady-state levels decreased, when coexpressed with MDM2. In addition, an MDM2-mutant that localized to the cytoplasm was able to ubiquitinate and degrade a p53 mutant which was similarly localized in the cytoplasm. Our results indicate that nuclear export is not required for p53 ubiquitination, and that p53 proteins that localize to either the nucleus or cytoplasm can be ubiquitinated and degraded by MDM2.

The MDM2 RING-finger domain is required to promote p53 nuclear export.

MDM2 can bind to p53 and promote its ubiquitination and subsequent degradation by the proteasome. Current models propose that nuclear export of p53 is required for MDM2-mediated degradation, although the function of MDM2 in p53 nuclear export has not been clarified. Here we show that MDM2 can promote the nuclear export of p53 in transiently transfected cells. This activity requires the nuclear-export signal (NES) of p53, but not the NES of MDM2. A mutation within the MDM2 RING-finger domain that inhibits p53 ubiquitination also inhibits the ability of MDM2 to promote p53 nuclear export. Finally, inhibition of nuclear export stabilizes wild-type p53 and leads to accumulation of ubiquitinated p53 in the nucleus. Our results indicate that MDM2-mediated ubiquitination, or other activities associated with the RING-finger domain, can stimulate the export of p53 to the cytoplasm through the activity of the p53 NES.

Immunological analysis of beta-thalassemic mouse intestinal proteins reveals up-regulation of sucrase-isomaltase in response to iron overload.

Maintenance of iron homeostasis must balance the demand for iron due to heme synthesis, which is driven by hematopoiesis, and the restricted intestinal uptake of iron, which otherwise limits absorption of this toxic element. The consequences of perturbed iron homeostasis are witnessed in inherited forms of beta-thalassemia in which erythroid hyperplasia results in enhanced intestinal iron absorption despite tissue iron overload. To gain a better understanding of intestinal factors that are induced when iron homeostasis is disrupted, a panel of monoclonal antibodies that recognize intestinal microvillous membrane proteins of the beta-thalassemic Hbbd(th3)/Hbbd(th3) mouse was established. The monoclonal antibodies were screened by differential Western blotting against normal and beta-thalassemic mouse intestine to identify antigens modulated in the disease state. Here we report the initial characterization of one immunoreactive species that is up-regulated in beta-thalassemic mouse intestine and the tentative identification of this antigen as sucrase-isomaltase. Studies in Caco-2 cells revealed the rather unexpected finding that expression of this intestinal hydrolase is increased in response to iron toxicity.