[Deep neck space infections: a retrospective cohort study of surgical risk factors].

Objective: To explore risk factors affecting treatment for deep neck space infections (DNSIs) so as to provide guidance for appropriate early managements. Methods: A retrospective cohort study was conducted on inpatients with DNSIs admitted to the Department of Otolaryngology, Head and Neck Surgery, Affiliated Hospital of Qingdao University from March 2013 to February 2021. Patients were divided into surgical and non-surgical groups based on whether they had surgery or not. Information collected included demographic data, disease-related signs and symptoms, treatment history, systemic comorbidities, imaging data and laboratory indicators. Hypothesis testing, univariate Logistic regression and multivariate Logistic regression were used for data processing. Resuts A total of 61 patients were included, including 37 males and 24 females, aged 6-96 years. There were 35 cases (57.4%) in the surgical group and 26 cases (42.6%) in the non-surgical group. Multivariate analysis showed that risk factors for surgery as followings: neck dyskinesia (OR=0.03, 95%CI: 0.00-0.24), dysphagia (OR=0.10, 95%CI: 0.02-0.72), serum white blood cell count≥16.74×109/L (OR=1.18, 95%CI: 1.01-1.39) and interspace gas (OR=0.03, 95%CI: 0.00-0.30). Conclusion: Clinicians should be alert to these risk factors for surgery in the course of treatment and timely surgical treatment for patients who meet the conditions.

[Current status and progress in the standardized surgical management of specimens after radical gastric cancer surgery].

Standardized surgical management of postoperative specimens of gastric cancer is an important part of the standardized diagnosis and treatment of gastric cancer. It can reflect the accurate number and detailed distribution of lymph nodes in the specimen and lay the foundation for accurate and standardized pathological reports after surgery. Meanwhile, it can evaluate the scope of intraoperative lymph node dissection, the safety of cutting edge, and the standardization of surgery (principle of en-bloc dissection), which is an important means of surgical quality control. It also provides accurate research samples for further research and is an important way for young surgeons to train their clinical skills. The surgical management of postoperative specimens for gastric cancer needs to be standardized, including specimen processing personnel, processing flow, resection margin examination, lymph node sorting, measurement after specimen dissection, storage of biological specimens, documentation of recorded data, etc. The promotion of standardized surgical management of specimens after radical gastrectomy can promote the homogenization of gastric cancer surgical diagnosis and treatment in medical institutions and further promote the high-quality development of gastric cancer surgery in China.

[Expression of PTTG1 in gastric adenocarcinoma tissues and its relationship with prognosis].

Objective: To identify new prognostic markers and therapeutic targets for gastric adenocarcinoma. Methods: The public datasets of gastric adenocarcinoma collected from GEO database (GSE33335 and GSE63089) were downloaded for analysis. There were 70 GC tissues and paired normal tissues in the two profile datasets. Differentially expressed genes (DEGs) between GC tissues and normal stomach tissues were selected by the R software. Protein-protein interaction (PPI) of these DEGs were visualized by the Search Tool for the Retrieval of Interacting Genes (STRING). The key gene sets were analyzed by Cytoscape and Molecular Complex Detection (MCODE). The mRNA and protein expression levels of prognosis related genes identified by public database were confirmed by using GC tissues and paired normal tissues collected from July 2019 to September 2019 in Affiliated Hospital of Nantong University. Results: DEGs were identified in the two datasets by using R software. A total of 128 DEGs were detected, including 85 up-regulated genes (log(2)FC>1.2 and FDR<0.01) and 43 down-regulated genes (log(2)FC<-1.2 and FDR<0.01) in the GC tissues. PPI network model and MCODE model were established by using the Online String tool and Cytoscape software, and 27 key genes were obtained, including 25 genes related with prognosis of GC patients (P<0.05). We identified 14 significant DEGs in GC tissues, including cyclin B1 (CCNB1), polo-like kinase 1 (PLK1) and pituitary-tumor transforming gene (PTTG1), which were significantly enriched in the cell cycle pathway through KEGG pathway enrichment analysis. The positive expression rate of PTTG1 in GC tissues was 68.8% (22/32), significantly higher than 18.8% (6/32) in normal gastric tissues (P<0.05). Conclusions: The expression of PTTG1 is different in GC and gastric tissues, implicates it is the key gene in gastric carcinogenesis. The prognoses of GC patients with higher PTTG1 expression are worse. PTTG1 might participate in the development of gastric adenocarcinoma by regulating cell cycle.

[Abnormal expression of insulin-like growth factor-I receptor and inhibitory effect of its transcription intervention on nude mice xenograft tumor].

Objective: To investigate the expression of insulin-like growth factor-I receptor (IGF-IR) in liver cancer and the inhibitory effect of its transcription intervention on nude mice xenograft tumor. Methods: A total of 40 patients with primary liver cancer were enrolled, and 40 samples of cancer lesions, peri-cancerous tissues (with a distance of 2 cm to the margin of cancer lesion), or distal liver tissues (with a distance of 5 cm to the margin of cancer lesion), with a weight of 200 mg, were collected after surgery. Some of these samples were used for pathological examination, and the rest were stored at -85°C. A total of 18 BALB/c nude mice aged 4-6 weeks with a body weight of 18-20 g (9 male and 9 female mice) were randomly divided into control group, negative control group, and co-intervention group, with 6 mice in each group, and fed under specific pathogen-free conditions. The cell line was cultured in the dimethyl sulfoxide complete medium containing 10% fetal bovine serum in a CO2incubator at 37°C. When the cell confluence reached 90% after cell inoculation, shRNA was divided into co-intervention group, negative control group, and untreated control group and were transfected to hepatoma cells using PolyJetTM transfection reagent. Stable cell clones obtained by G418 screening and used for the in vivo study. Immunohistochemistry, Western blotting, and quantitative real-time PCR were used to analyze the expression of IGF-IR in the human hepatoma tissue and cell line. The IGF-IR shRNA eukaryotic expression plasmids were established and screened for the most effective sequence; they were transfected to PLC/PRF/5 hepatoma cells, and the CCK-8 assay was used to analyze the changes in cell proliferation. The stable cell line screened out by G418 was inoculated to establish the subcutaneous xenograft tumor in nude mice. The tumor growth curve was plotted and histological examination was performed. Graphpad Prism 5.0 and SPSS 18.0 were used for plotting and data analysis; the variance test and Q test were used for comparison of means between multiple samples, the t-test was used for comparison of means between any two samples, the chi-square test or Fisher's exact test was used for comparison of rates between samples, and a rank correlation analysis was performed for expression intensity. Results: The liver cancer group had a significantly higher positive rate of IGF-IR than the peri-cancerous group and distal tissue group (82.5% vs 42.5%/10%,χ2= 13.653 and 42.29, bothP< 0.01), as well as significantly higher expression intensity than these two groups (Z= 4.771 and 6.579, bothP< 0.01). IGF-IR was not significantly expressed in the L02 cell line and was strongly expressed in the PLC/PRF/5 hepatoma cells, and the expression intensity of IGF-IR in the PLC/PRF/5 hepatoma cells was 4 and 5 times that in Bel-7404 cells and HepG2 cells, respectively. After the PLC/PRF/5 hepatoma cells were transfected with shRNA4 with the best co-intervention effect, the mean inhibition rate of tumor cell growth reached 63.9% at 72 hours, and the mean inhibition rate of IGF-IR transcription reached 59.6%. Tumor cells were arrested in G1 phase, and there was a significant increase in apoptosis rate. As for the subcutaneous hepatoma xenograft in nude mice, the intervention group had significantly slower tumor growth than the blank control group and negative control group (143±24 mm3 vs 372±46 mm3/350±50 mm3,t= 10.776 and 9.142, bothP< 0.01); the intervention group had significantly downregulated IGF-IR expression, which was significantly lower than that in the blank control group and negative control group (t= 11.184 and 9.450, bothP< 0.01). Conclusion: Intervention of IGF-IR transcription can effectively inhibit the growth of xenograft tumor in nude mice, suggesting that IGF-IR gene might become a new potential target for the treatment of liver cancer.

Serum concentration and mRNA expression in milk somatic cells of toll-like receptor 2, toll-like receptor 4, and cytokines in dairy cows following intramammary inoculation with Escherichia coli.

The objective of the current study was to investigate the toll-like receptors (TLR), including the soluble forms sTLR2 and sTLR4, involved in innate immune responses of dairy cows to experimentally induced Escherichia coli mastitis. Six clinically healthy Holstein dairy cows received an intramammary inoculation of E. coli O111:K58 between 63 and 83 d postpartum. Concentrations of sTLR2 and sTLR4, the proinflammatory cytokines IL-6 and tumor necrosis factor-α (TNF-α), and acute phase proteins serum amyloid A (SAA) and haptoglobin (Hp) in blood were measured by ELISA. Furthermore, 10mL of milk was collected from challenged quarters immediately before inoculation and at 6, 12, 24, 48, and 72 h after inoculation, and mRNA expression of selected genes, including TLR2, TLR4, IL-1ß, IL-6, TNF-α, and IL-8, was quantified by real-time PCR. Escherichia coli intramammary infection elicited a decrease in the circulating levels of leukocytes. Rectal temperature was elevated at 6h postinoculation (PI). Similarly, the serum concentrations of TNF-α, IL-6, and SAA increased at 6h PI. However, serum concentrations of sTLR2, sTLR4, and Hp did not differ after challenge. The mRNA expression of TLR2, IL-1ß, and IL-8 in milk somatic cells increased at 12h PI, whereas a decreased IL-6 mRNA expression was detected from 6 to 48 h PI. In conclusion, we found that TLR2 mRNA expression increased in milk somatic cells collected from infected quarters of cows challenged with E. coli, whereas the concentrations of sTLR2 and sTLR4 remained unchanged after challenge. Thus, sTLR2 and sTLR4 may protect the host by sequestrating pathogen-associated molecular patterns during E. coli mastitis.

Enhanced visible photoluminescence of V(2)O(5) via coupling ZnO/V(2)O(5) composite nanostructures.

V(2)O(5) films capped by a thin ZnO layer had been prepared by sputtering method at room temperature. The initial smooth films transferred to porous composite nanocrystals and nanorods after annealed at 500 degrees C, and enhanced visible light emission was observed. This tremendous enhancement was attributed to the coupling between V(2)O(5) nanorods and ZnO nanoparticles as well as the improved V(2)O(5) crystallinity. Prominent vibrational fine structures of the photoluminescence spectra were also observed.

Defects in cAMP-pathway may initiate carcinogenesis in dividing nerve cells: a review.

The mechanisms of carcinogenesis in nervous tissues are not well understood. It is now established that adenosine 3,',5'-cyclic monophosphate (cAMP)-pathway plays a crucial role in initiating differentiation in transformed and embryonic cells of neuronal and glial origin. Therefore, we propose that defects in the cAMP-pathway may initiate the first phase of carcinogenesis (immortalization). Subsequent genetic abnormalities in oncogenes, anti-oncogenes or other cellular genes individually or in combination may lead to transformation (cancer phenotype). This hypothesis is derived from the fact that an elevation of the cAMP level in murine NB cells induces terminal differentiation in many of these cells in spite of the fact that they are highly aneuploid. Additional changes in cAMP-regulated genes responsible for initiating differentiation may make these cells resistant to cAMP or may make the cAMP-effect on differentiation reversible. Indeed, cAMP-resistant cells exist in NB cell populations, and the cAMP-effect on differentiation is reversible in glioma cells. Identification of genes that initiate, promote and maintain terminal differentiation and those which prevent differentiation following elevation of cAMP in NB cells may increase our understanding of the mechanisms of carcinogenesis. This review illustrates the following: (a) historical background leading to the discovery of cAMP as an inducer of differentiation in nerve cells; (b) identification of potential sites in cAMP-pathway that may play a crucial role in initiating the first phase of carcinogenesis (immortalization) and potential gene targets in immortalized cells whose alterations may cause neoplastic transformation of nerve cells. It is interesting to note that the cAMP pathway remains responsive to an elevated cAMP level in inducing differentiation in NB cells in spite of chromosomal anomalies and genetic changes associated with the maintenance of a cancer phenotype.

Estrogen receptors alpha and beta: prevalence of estrogen receptor beta mRNA in human vascular smooth muscle and transcriptional effects.

BACKGROUND: Estrogens have vascular effects through the activation of estrogen receptors (ERs). In addition to ERalpha, the first ER to be cloned, a second subtype called ERbeta has recently been discovered. METHODS AND RESULTS: Using a reverse-transcriptase polymerase chain reaction assay that employs the same primer pair to simultaneously amplify ERalpha and ERbeta transcripts, we found that ERbeta is the ER form that is predominantly expressed in human vascular smooth muscle, particularly in women. The transcriptional effects of the 2 ERs in transfected HeLa cells differed. In response to 17beta-estradiol, ERalpha is a stronger transactivator than ERbeta at low receptor concentrations. However, at higher receptor concentrations, ERalpha activity self-squelches, and ERbeta is a stronger transactivator. Tamoxifen has partial agonist effects with ERalpha but not with ERbeta. CONCLUSIONS: The protective effects of estrogens in the cardiovascular system of women may be due to the genomic effects of ERbeta in vascular tissue.

An exochelin of Mycobacterium tuberculosis reversibly arrests growth of human vascular smooth muscle cells in vitro.

Proliferation of vascular smooth muscle cells (VSMC) is characteristic of restenosis following balloon angioplasty. We show here that a low concentration of a novel iron chelator, desferri-exochelin 772SM, reversibly arrests the growth of human VSMC in vitro, specifically in G(0)/G(1) and S phases. The lipophilic desferri-exochelin is effective more rapidly and at a 10-fold lower concentration than the nonlipophilic iron chelator deferoxamine. Treatment of growth-synchronized VSMC with the desferri-exochelin results in down-regulation of cyclin E/ Cdk2 and cyclin A/Cdk2 activity but does not affect the cyclin D/Cdk4/retinoblastoma phosphorylation pathway. Both DNA replication and RNA transcription are inhibited in exochelin-treated cells, but protein synthesis is not. The ability of desferri-exochelin 772SM to reversibly block the growth of VSMC in vitro with no apparent cytotoxicity suggests that the exochelin may be useful as a therapeutic agent to limit restenosis in injured vessels.

Ochre suppressor transfer RNA restored dystrophin expression in mdx mice.

The mdx mouse is an animal model for human Duchenne muscular dystrophy. The lack of dystrophin in mdx mice is caused by an ochre mutation in exon 23 of the dystrophin gene. This study tested the feasibility of inhibiting translational termination as an approach for genetic therapy for diseases caused by nonsense mutations. We evaluated both the in vitro and in vivo efficiencies of readthrough of ochre codons in 2 genes with the tRNA suppressor gene. The first target was a CAT reporter gene bearing an ochre mutation at the 5' end (CATochre). The second target was the dystrophin gene in mdx mice. The readthrough efficiencies were about 20% in COS cells and 5.5% in rat hearts. At four weeks after a direct injection of plasmid DNA encoding the tRNA suppressor into mdx mice, dystrophin positive fibers were detected by sarcolemmal immunostaining. This is the first convincing data that a tRNA suppressor gene might be a useful in vivo treatment for the genetic disorders caused by nonsense mutations.

Oxidized low-density lipoproteins stimulate adhesion of monocytes to endothelial cells.

AIM: To study the effects of oxidized low-density lipoproteins (ox-LDL) on the adhesiveness of monocytes to endothelial cells. METHODS: LDL was obtained from healthy human plasma by ultracentrifugation, and oxidized by CuSO4 10 mumol.L-1. The assay of adhesion was performed using cultured bovine aortic endothelial cells (BAEC) and human peripheral blood monocytes. RESULTS: Pretreatment BAEC with ox-LDL enhanced monocyte adhesion to BAEC in time- and dose-dependent manner. ox-LDL as little as 10 mg.L-1 and 30 min of preincubation stimulated monocyte adhesion. Cycloheximide (Cyc, a protein synthesis inhibitor) 1 mg.L-1 and staurosporine (Sta, a PKC inhibitor) 20 nmol.L-1 abolished the effect of ox-LDL (60 mg.L-1), but dextran sulfate 20 mg.L-1 had no effect on monocyte adhesion. Phorbol 12-myristate 13-acetate (PMA) 1 nmol.L-1 and lysophosphatidylcholine (Lys) 6 mumol.L-1 mimicked the effects of ox-LDL and potentiated monocyte adhesion. Sta also suppressed the augmentative effects of Lys and PMA. CONCLUSION: ox-LDL enhances the adhesion of monocytes to BAEC through the activation of PKC.

Meta-analysis of phenotype and genotype of NAT2 deficiency in Chinese populations.

Data on both the incidence of slow acetylator phenotype of probe drugs isoniazid, sulfadimidine or sulfamethazine, caffeine and dapsone in mainland or overseas Chinese, and the distribution of NAT2 genotypes and the frequency of NAT2 alleles in the Chinese populations were summarized and reanalysed using a meta-analysis method. Frequency of the slow acetylator phenotype in 3516 healthy Han Chinese gave an overall mean of approximately 19.9 +/- 4.0%, with the range of the combined data being between 15.8% and 25.5%. In addition, frequencies of the slow acetylator phenotype differ between the different minorities in Chinese populations and the range was between 3.2% and 50.6%, with a mean value of 20.6 +/- 12.9% in a total of 1842 individuals from 17 Chinese minorities. In addition, there was no significant heterogeneity in overseas Chinese between the probe drugs isoniazid and sulfadimidine or sulfamethazine (chi 2 = 5.97, df = 4; p > 0.05), and the mean value of slow acetylator phenotype incidence was 24.5% (119/485; 95% CI: 20.7-28.3%), consistent with that of the native Chinese. As expected, frequency of the slow acetylator genotypes in Chinese populations was 25.4% (112/441; 95% CI: 21.3-29.5%), which was in accordance with that of the slow acetylator phenotype in native or overseas Chinese. For all genotypes, *4/*4 (29.9%, 132/441), *4/*6A (27.4%, 121/441), *4/*7A (12%, 53/441) and *6A/*6A (11.3%, 50/441) occupied 80.6%, but *5A/*7A (0.2%, 1/441), *5A/*5A (1.1%, 5/441) and *7A/*7A (1.8%, 8/441) were not frequently found. From this report, the genotype frequencies of homozygous rapid acetylator, heterozygous rapid acetylator, and homozygous slow acetylator were found to be 0.299 (132/441), 0.447 (197/441) and 0.254 (112/441), respectively. Furthermore, both *4 (52.3%; 95% CI: 49-56%) and *6A (30.5%; 95% CI: 28-34%) were major NAT2 alleles, while *7A (11.2%; 95% CI: 9-13%) and *5A (6%; 95% CI: 4-8%) were uncommonly present. Frequency of the mutant alleles was observed at 0.477 (421/882 alleles). The *7A constituted 23.5% t(99/421) of slow acetylator alleles in Chinese populations, showing that this point mutation exists not only in Oriental or Asiatic, but also in Chinese populations. According to the Hardy-Weinberg equilibrium, in the phenotyped Chinese populations, the mean estimate of predicted allelic frequencies of the genotypes RR, Rr, and rr was 0.294, 0.496, and 0.210 for the Chinese, and the expected frequency of the deficient gene r was 0.458. By comparison, the predicted values are in complete agreement with the observed ones. In conclusion, this meta-analysis determined the accurate population frequencies of phenotype and genotype of the NAT2 genetic deficiency in healthy Chinese subjects.

Prostacyclin analog (cicaprost) protects against damage by hydrogen peroxide to rabbit cardiac sarcoplasmic reticulum.

The membrane fluidity of the cardiac sarcoplasmic reticulum in rabbit was monitored by measuring changes in steady-state fluorescence anisotropy (rs) using diphenylhexatriene as a probe. The Ca(2+)-ATPase activity of the sarcoplasmic reticulum was measured by assaying the amount of inorganic phosphate released from ATP. Hydrogen peroxide caused damage to the sarcoplasmic reticulum, as reflected by decreases in membrane fluidity and Ca(2+)-ATPase activity. The damage caused by hydrogen peroxide was completely prevented by 20 micrograms/ml catalase. Cicaprost (240 nM) provided an effective protection against injury to the sarcoplasmic reticulum caused by exposure to hydrogen peroxide. The rs value was significantly decreased from 0.154 +/- 0.014 to 0.122 +/- 0.005 (p < 0.01). Ca(2+)-ATPase activity was increased from 3.1 +/- 1.31 to 18.87 +/- 2.11 microM phosphate/mg protein/hour (p < 0.01). The protection given by cicaprost was dose dependent. We conclude that cicaprost protects against damage produced by hydrogen peroxide in cardiac sarcoplasmic reticulum in the rabbit. The mechanism of the effect of cicaprost remains to be elucidated.

Effects of hydrogen peroxide on membrane fluidity and Ca(2+)-transporting ATPase activity of rabbit myocardial sarcoplasmic reticulum.

This study was to investigate the effects of hydrogen peroxide on membrane fluidity and Ca(2+)-ATPase activity of rabbit myocardial sarcoplasmic reticulum (SR). The membrane fluidity of SR was monitored by measuring the changes in the steady state fluorescence anisotropies (rs) using diphenylhexatriene as a probe. The Ca(2+)-ATPase activity was determined by assaying the amount of inorganic phosphate (Pi) released from ATP. It was found that the membrane fluidity (rs: 0.154 +/- 0.014 vs 0.113 +/- 0.010, P < 0.01) and Ca(2+)-ATPase activity (3.1 +/- 1.3 vs 25.3 +/- 2.4 mumol Pi.h-1/mg protein, P < 0.01) were reduced in SR exposed to H2O2 (2 mmol.L-1) for 40 min. Catalase 20 micrograms.ml-1 completely prevented the SR damages caused by H2O2. H2O2 jeopardized the SR in a concentration- and time-dependent manner as measured by changes in rs values and Ca(2+)-ATPase activities, which were negatively correlated (r = 0.981, P < 0.01). These results suggest that H2O2 produces dysfunctions of the rabbit myocardial SR, and that the alteration of membrane fluidity may be one of the mechanisms responsible for the decrease of Ca(2+)-ATPase activity.

[Protective effects of gypenosides on experimental myocardial infarction].

In the model of myocardial infarction produced by occlusion of left anterior descending coronary artery (LAD) in rabbit, gypenosides (GP 100 but not 50 mg/kg, ip) reduced myocardial infarct size and decreased serum free fatty acid (FFA). In rat model of myocardial infarction, GP and the fractions of GP of non ginsenosides (FGNG) both in dose of 100 mg/kg, ip, protected significantly myocardial superoxide dismutase (SOD) activity and decreased the myocardial malondialdehyde (MDA). The results indicate that the protective effect of GP on myocardial infarction may be correlated with its prevention of myocardial lipid peroxidation, and attributed to the amelioration of FFA metabolic deterioration.