[Estimation of Early Postmortem Interval of Asphyxial Death Rats at Different Ambient Temperatures by GC-MS-Based Metabolomics].

ABSTRACT: Objective To establish the orthogonal partial least square （OPLS） model for the estimation of early postmortem interval （PMI） of asphyxial death rats in four ambient temperatures based on gas chromatography-mass spectrometry （GC-MS） metabolomics. Methods The 96 rats were divided into four temperature groups （5 ℃, 15 ℃, 25 ℃ and 35 ℃）. Each temperature group was further divided into 3 h, 6 h, 12 h and 24 h after death, and 6 other rats were taken as the control group. The cardiac blood was collected at the set time points for the four temperature groups and 0 h after death for the control group for the metabolomics analysis by GC-MS. By OPLS analysis, the variable importance in projection （VIP）>1 and the result of Kruskal-Wallis test P<0.001 were used to screen out the differential metabolite related to PMIs in the cardiac blood of rats of different temperature groups. Then OPLS regression models of different temperature groups were established with these metabolites. At the same time, a prediction group for investigating the prediction ability of these models was set up. Results Through the analysis of OPLS, 18, 15, 24 and 30 differential metabolites （including organic acids, amino acids, sugars and lipids） were screened out from the rats in groups of 5 ℃, 15 ℃, 25 ℃ and 35 ℃, respectively. The prediction results of the four temperature group models showed that the prediction deviation of 5 ℃ model was larger than that of other groups. The prediction results of other temperature groups were satisfactory. Conclusion There are some differences in the changes of metabolites in cardiac blood of rats at different ambient temperatures. The influence of ambient temperature should be investigated in the study of PMI estimation by metabolomics, which may improve the accuracy of PMI estimation.

Ineffectiveness of the Tobacco Hazards Prevention Act in patients with severe and persistent mental illness.

OBJECTIVES: The effect of the Tobacco Hazards Prevention Act (THPA) on the reduction of tobacco use in the general population in Taiwan is understood. However, there has been little research on how these policies affect people with severe and persistent mental illness (SPMI). Hence, the influence of the THPA on daily cigarette use by patients with SPMI was investigated in a 3-year follow-up. The risk and protective factors in smoking behavior and cessation were investigated. STUDY DESIGN: This is an observational study. METHODS: A total of 144 participants (78 of whom had smoked >100 cigarettes throughout their lifetime) were randomly selected from a healthcare network. Then, 100 of 144 patients with SPMI, included 50 smokers and 50 non-smokers, have agreed with 1st year follow-up. Eighty-two patients with SPMI, consisted of 44 smokers and 38 non-smokers, have agreed with 2nd year follow-up. RESULTS: Although women were less likely to smoke than men, those who did smoke reduced the number of cigarettes less than men over the 3-year period. Less-educated patients were more likely to smoke than those who were more educated but reduced the number of daily cigarettes more after implementation of the THPA. Maternal overprotection was a risk factor and paternal care a protective factor for smoking. Addiction, a perception of pleasure, interpersonal relationships, and self-destructive behavior increased cigarette smoking. Patients perceived that smoking alleviated their depressive symptoms. CONCLUSIONS: The THPA was ineffective for patients with SPMI. Some factors had different effects on the development of smoking behavior and the cessation process. Understanding the etiology of smoking can help policymakers establish more effective programs for smoking prevention and cessation.

Genetic polymorphisms in the prostaglandin pathway genes and risk of head and neck cancer.

OBJECTIVE: Previous studies examining the association between genetic variations in prostaglandin pathway and risk of head and neck cancer (HNC) have only included polymorphisms in the PTGS2 (COX2) gene. This study investigated the association between genetic polymorphisms of six prostaglandin pathway genes (PGDS, PTGDS, PTGES, PTGIS, PTGS1 and PTGS2), and risk of HNC. METHODS: Interviews regarding the consumption of alcohol, betel quid, and cigarette were conducted with 222 HNC cases and 214 controls. Genotyping was performed for 48 tag and functional single-nucleotide polymorphisms (SNPs). RESULTS: Two tag SNPs of PTGIS showed a significant association with HNC risk [rs522962: log-additive odds ratio (OR) = 1.42, 95% confidence interval (CI): 1.01-1.99 and dominant OR = 1.58, 95% CI: 1.02-2.47; rs6125671: log-additive OR = 1.49, 95% CI: 1.08-2.05 and dominant OR = 1.96, 95% CI: 1.16-3.32]. In addition, a region in PTGIS tagged by rs927068 and rs6019902 was significantly associated with risk of HNC (global P = 0.007). Finally, several SNPs interacted with betel quid and cigarette to influence the risk of HNC. CONCLUSIONS: Genetic variations in prostaglandin pathway genes are associated with risk of HNC and may modify the relationship between use of betel quid or cigarette and development of HNC.

[Research advances on the role of cell senescence in chronic wound healing].

Chronic wounds bring huge pressure and difficulties to patients, their families, and society due to their long-term refractory characteristics and serious poor prognosis. Recently, more and more evidences have proven that cell senescence exists in chronic wounds and affects wound healing. This article reviews the characteristics of cell senescence in chronic wounds, discusses the relationship between cell senescence and chronic wound healing, and summarizes current series of anti-senescence strategies to bring out the possibility of treating chronic wounds with cell senescence as a potential target.

Does the use of tranexamic acid increase the risk of VTE in patients with hemoptysis?

OBJECTIVE: The aim of this study was to create a predictive nomogram that can accurately identify the risk factors of venous thromboembolism (VTE) in hospitalized patients exhibiting hemoptysis. PATIENTS AND METHODS: The present study gathered clinical and demographic data of 1,052 hospitalized patients with hemoptysis at Dongyang Hospital between January 2016 and January 2021 through the Lejiu database. The patients were categorized into two groups: the thrombotic event group (n=123) and the non-thrombotic event group (n=929), based on the presence of VTE before discharge. The study utilized univariable and multivariable logistic regression analyses to identify the independent risk factors for VTE, with the occurrence of thrombotic events serving as the dependent variable. Furthermore, a nomogram prediction model was formulated to verify the findings. RESULTS: In hospitalized patients with hemoptysis, the risk of VTE was found to be independently associated with the administration of tranexamic acid (TXA), the presence of D-dimer, and the Charlson Comorbidity Index (CCI) score (p<0.05). CONCLUSIONS: A nomogram model was constructed to evaluate the probability of VTE in patients hospitalized with hemoptysis. This model allows for the timely detection of early VTE warning signs, which may ultimately reduce its occurrence.

A survey on the management of acute rhinosinusitis among Asian physicians.

BACKGROUND: Based on the `European Position Paper on Rhinosinusitis and Nasal polyps (EP3OS 2007)`, this study aimed to investigate general practitioners (GPs) and other specialists` understanding when managing patients with acute rhinosinusitis (ARS) in Asia. METHODOLOGY: Among a total of 2662 questionnaires completed, 2524 (94.8%) were valid for analysis. There were 1308 GPs (51.8%), 989 otolaryngologists (39.2%) and 227 paediatricians (9%) from Mainland China, Hong Kong, Indonesia, India, Malaysia, Pakistan, Philippines, Singapore, Thailand and Taiwan. RESULTS: ARS is affecting an estimated 6 - 10% of patients seen in a daily out-patient practice. The EP3OS criteria are well supported by Asian physicians (94.1%). Most physicians (62.7%) agreed that radiological investigation is not needed to diagnose ARS. However, even for mild ARS (common cold), medical treatments were still recommended by 87% of GPs, 83.9% of otolaryngologists, and 70% of paediatricians. The top three first-line treatments prescribed were antihistamines (39.2%), nasal decongestants (33.6%), and antibiotics (29.5%). Antibiotics usage increased as the first line treatment of moderate (45.9%) and severe (60.3%) ARS. CONCLUSION: ARS is commonly managed by GPs, otolaryngologists, and paediatricians in Asia. However, understanding of the management of ARS needs further improvement to minimize unnecessary use of radiological investigations, overuse of antibiotics, and under use of nasal corticosteroids.

ETV4 promotes the progression of gastric cancer through regulating KDM5D.

OBJECTIVE: This study was aimed to investigate the expression characteristics of ETS variant 4 (ETV4) in gastric cancer (GCa), and to further explore whether it promotes the development of GCa by regulating KDM5D. PATIENTS AND METHODS: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression of ETV4 in 35 pairs of tumor tissue and paracancerous tissue specimens collected from GCa patients, and the interplay between ETV4 expression and clinical indexes, as well as the prognosis of GCa patients, were analyzed. Meanwhile, the expression of ETV4 in GCa cell lines was verified using qRT-PCR assay. Furthermore, ETV4 knockdown model was constructed using lentivirus in GCa cell lines including AGS and BGC-823, and then, the transwell invasion and cell wound healing assays were applied to analyze the effect of ETV4 on the biological function of GCa cells. In addition, an in-depth study of the relationship between ETV4 and KDM5D was conducted. RESULTS: The results of qRT-PCR showed that the expression level of ETV4 in GCa tissue samples was remarkably higher than that in adjacent tissues, and the difference was statistically significant. Compared with patients with low expression of ETV4, the patients with high ETV4 expression had a higher occurrence rate of lymph node or distant metastasis and a lower overall survival rate. Similarly, the metastasis ability of GCa cells in the ETV4 expression knockdown group (sh-ETV4) was remarkably decreased when compared with the sh-NC group. In addition, qRT-PCR results indicated that the protein expression of KDM5D was significantly increased after the knockdown of ETV4. Therefore, it was demonstrated that ETV4 might be able to regulate the malignant progression of GCa via modulating KDM5D expression. Finally, the results of the cell reverse experiment confirmed that the silence of ETV4 could reverse the malignant progression of GCa induced by the downregulation of KDM5D. CONCLUSIONS: ETV4 expression was found remarkably elevated in GCa tissues and was significantly associated with the occurrence of lymph node or distant metastasis and poor prognosis. In addition, ETV4 might promote GCa cell metastasis by modulating KDM5D.

The significance of HBD-3 and fluorescent composite carriers in the processof bone formation in rats infected with Staphylococcus aureus.

OBJECTIVE: The objective of the present study was to explore the significance of human ß-defensin 3 (HBD-3) through establishment and evaluation of the model of implant-related biofilm infection of the femoral condyle of the outer knee using Sprague-Dawley (SD) rats. MATERIALS AND METHODS: Age-matched SD rats were divided into three groups, the HBD-3 group, HBD-3 fluorescent liposome group, and the HBD-3 liposome-microbubble fluorescent composite carrier group. After biofilm infection for 24 h, the fluorescent composite vector was injected intraperitoneally 2 times/day. After the first injection, rats in each group were sacrificed on the 7th, 14th, and 28th day. The lower end of the femur bone was harvested after removing the surrounding soft tissue. H&E and immunohistochemical staining were applied and light microscopy was used for observation. Fluorescent markers including tetracycline and calcein were used to follow the formation of new bone in vivo. Undecalcified specimens were embedded in epoxy resin (thickness of roughly 150 m), and confocal microscopy was used for observation. RESULTS: By assessing cell proliferation with cell counting kit-8, the proliferation ability of cells in the HBD-3 liposome-microbubble fluorescent composite carrier group was significantly increased compared with the other groups (p<0.05). qPCR was used to measure the levels of alkaline phosphatase (ALP), type I collagen, osteocalcin (OCN), osteopontin (OPN), and bone sialoprotein (BSP) in each group. The levels of these genes in the HBD-3 liposome-microbubble fluorescent composite carrier group were significantly higher than those in other groups (p<0.05). CONCLUSIONS: The application of the HBD-3 liposome-microbubble fluorescent composite carrier can significantly promote osteogenesis in rats infected with Staphylococcus aureus, and increase the expression levels of ALP, type I collagen, OCN, OPN, and BSP.

Triterpenoid glycosides from Adina rubella.

Two new triterpenoid glycosides, quinovic acid-3 beta-O-beta-D- glucopyranosyl-(1-->4)-alpha-L-rhamnopyranoside and quinovic acid-3 beta-O-beta-D-glucopyranosyl-(1-->3)-beta-D-fucopyranoside (named rubelloside A and B, respectively), were isolated from roots of Adina rubella. Their structures were elucidated by spectral and chemical means. Rubelloside B exhibited immunological enhancement.

Analysis of the expression of Fas-L in nasopharyngeal carcinoma tissues.

Nasopharyngeal carcinoma (NPC) is an epithelial cancer with a high incidence in Southeast Asia. How it escapes attack from the host immune system is not fully understood. Recently, pieces of evidence show that Fas-ligand (Fas-L)-mediated apoptosis may be involved in immune privilege of tumours. To determine whether a similar mechanism may exist in NPC, the expression of Fas-L was analysed. Biopsy specimens of the nasopharynx were taken from 27 NPC patients. Histologically, they were either non-keratinizing or undifferentiated carcinomas. Nasopharyngeal biopsies of 11 other patients that proved to have no tumour served as control. The transcripts of Fas-L were detected by reverse transcription-polymerase chain reaction. Localization of Fas-L protein was performed with immunohistostaining using an antibody recognizing human Fas-L. All nasopharyngeal tissues have a similar amount of transcripts of Fas-L. However, the Fas-L protein was detected exclusively on the cell surface of malignant epithelial cells of NPC. The present findings suggest that Fas-L protein may be involved in evading immune attack of NPC.

Lectin immunohistochemistry study of nasal inverted papilloma and associated neoplasms.

BACKGROUND: We aimed to investigate the tumor biology of nasal IP and identify the biological characteristics associated with early diagnosis and malignant transformation. MATERIALS AND METHODS: Lectins immunohistochemistry were performed on inverted papilloma (IP), and associated neoplasms. IP synchronized with polyp or squamous cell carcinoma (SCC) and IP metachronized with SCC were analyzed and correlated to clinical information. RESULTS: Canavalia ensiformis (ConA) and Arachis hypogaea (PNA) with neuraminidase pretreatment (NA-PNA) showed similar staining in both the IP and SCC portions of the IP synchronized with SCC. The IP and polyp portions of an IP synchronized polyp had positive NA-PNA staining, while papilloma and polyps alone had negative staining. Strong NA-PNA staining in the IP (transformed to SCC) showed significant differences from IP. CONCLUSION: These biological characteristics define IP as a premalignant neoplasm. NA-PNA staining may be helpful for an early detection of IP. Strong NA-PNA staining in IP may predict malignant transformation.

Transformation of mucosal secretory elements in chronic maxillary sinusitis after endoscopic sinus surgery.

To evaluate the secretory element transformation of maxillary sinus mucosa after endoscopic sinus surgery (ESS), I enrolled 20 patients. Five normal antral mucosae were used as controls, and 15 antral mucosae were investigated according to pathologic and secretory patterns. The post-ESS mucosa was evaluated at the 16th week, when gross recovery appeared. Specimens from the rear wall of the antrum were observed under a scanning electron microscope (x2,000). A significant decrease of goblet cells and increase of glands was illustrated in the sinusitis cases, especially the polypoid and purulent groups. In post-ESS cases, the number of goblet cells is about the same as in controls, but the number of gland openings is higher. My conclusions were that 1) the repair of the antral mucosa requires more time than gross recovery and 2) post-ESS follow-up should be longer than 16 weeks to prevent recurrence.

Overexpression of Fas-ligand in human nasal polyps.

Apoptosis mediated through the Fas/Fas-L system is essential in regulating immune function, developing organs, and conferring immune privilege. To illustrate the role of the Fas/Fas-L system in the pathogenesis of human nasal polyps, we investigated the transcripts and protein level of the Fas-L gene in 8 human nasal polyp tissues and 7 nasal turbinate mucosa specimens using reverse transcription-polymerase chain reaction and Western blotting. Localization of Fas-L was performed with immunohistochemistry. The transcripts of the Fas-L gene were detected at similar levels in both polyps and nasal mucosa. There was a significant overexpression of Fas-L protein on nasal polyps compared to nasal mucosa. Fas-L-positive cells were localized on the epithelial layers of cystically dilated glands and the down-growing epithelium of nasal polyps. Fas-L may play an important role in the pathogenesis of human nasal polyps, including cystic degeneration of submucosal glands and conferring of immune privilege to nasal polyp formation.

Branchiogenic squamous cell carcinoma: a case report.

Branchiogenic carcinoma is extremely rare. A 41-year-old man presented with a well-defined, fluctuant, painless mass on his right neck and received complete tumour excision. The pathological diagnosis was branchial cleft squamous cell carcinoma, which was characterized microscopically by a branchial cleft cyst with a tumour and a non-tumour transitional zone. In the past, several hundred cases were reported erroneously because of the rather complicated diagnostic criteria. The modified criteria of Khafif introduced in 1989 are more practical, and all were met in this case.

Presence of neuropeptides in human nasal polyps.

The pathophysiological effects of non-cholinergic, non-adrenergic neuropeptides are well known in the nasal mucosa, but unclear in the polyps. Since the pathophysiological roles of neuropeptides depend on their presence in the target tissue, specimens of nasal polyps were removed from 20 patients and examined for the presence of vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), dopamine-beta-hydroxylase (DBH), substance P (SP) and calcitonin gene-related peptide (CGRP). To visualize these neuropeptide fibers, immunohistochemical staining by the peroxidase-anti-peroxidase method and color reaction by Nickel enhancement of diaminobenzidine (DAB) were used. Fine varicose neuropeptides immunostained fibers were predominantly distributed in the pedicle of the polyps. No neuropeptides were found in the mucosal epithelium and subepithelium. NPY fibers were predominantly seen around the thick wall vessels, whereas the VIP fibers were mainly to be found in close proximity to the submucosal glands and fairly close to the vessels. SP or CGRP fibers were not found in the polyps. VIP and NPY in the pedicle of the polyps may be present in connection with mucosal inflammation, tissue edema and cystic degeneration of the glands in the early stage of polyp formation. Thus these neuropeptides may contribute to the development and growth of nasal polyps.

Distribution and quantity of neuroendocrine markers in allergic rhinitis.

Neuroendocrine components exist in the human nasal mucosa. However, the pathophysiological and neuroimmunological roles of the regulatory peptides in allergic rhinitis (AR) require further investigation. To analyse the functional morphology and quantify the tissue concentration of regulatory peptides in the nasal mucosa of AR subjects, human inferior turbinate mucosa specimens from 25 patients with AR, 20 patients with non-allergic rhinitis and 10 patients without any nasal diseases were investigated. Using immunohistochemistry and radioimmunoassays, we detected the presence, distribution and concentrations of various neuropeptides [vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), substance P (SP) and calcitonin gene-related peptide (CGRP)] and general neuroendocrine markers (neuron-specific enolase and chromogranin A). Quantitative analysis of the stained fibres and cells was performed using a graphic AutoCAD program. The presence and distribution of NPY, CGRP and SP nerve fibres and neuroendocrine cells were similar among the three subject groups. AR subjects had significantly higher tissue concentrations of VIP and SP. AR subjects had increased numbers of VIP fibres which predominantly innervated vessels. Thus, VIP and SP play important neuroimmunological roles in the pathogenesis of AR.

Macrophages in degradation of collagen/hydroxylapatite(CHA), beta-tricalcium phosphate ceramics (TCP) artificial bone graft. An in vivo study.

The macrophages mediated biodegradation of two biomaterials, collagen/hydroxylapatite (CHA) and beta-tricalcium phosphate ceramics (TCP), was studied in 24 male Kunming mice and 20 male C57BL/6 mice with histopathologic, histochemical and ultrastructural observation. It was demonstrated that macrophages infiltrated after CHA, TCP were implanted. The macrophages could be differentiated from fibroblasts and the other infiltrated cells for special cellular profile and strong acid phosphatase activity. Morphologically, monocyte-macrophages and infused multinuclear giant cell degraded CHA and TCP by phagocytosis and extracellular resorption. The carbonic anhydrase activity of macrophages was demonstrated by histochemical technique. It suggested that macrophages secreted H+ and accomplished the decalcification of calcium phosphate compound of CHA and TCP. We conclude that macrophages are the main mediating cells which degraded CHA and TCP intracellularly and extracellularly.

Normalization of maxillary sinus mucosa after FESS. A prospective study of chronic sinusitis with nasal polyps.

To observe the normalization of antral mucosa after FESS, 71 patients suffering from chronic sinusitis with nasal polyps were enrolled in this study. Pre-operative profiles including history, allergy skin test, and saccharin time test were done. Endoscopic findings of the antral mucosa were divided into 3 groups: polyposis, oedema, and thickening. The time sequence required for normalization of the mucosa was observed through the antromeatal opening. A patient's history of more than seven years, a saccharin time longer than 36 min, and polyposis of antral mucosa are ominous signs for recovery. The most frequent endoscopic finding of maxillary sinusitis is oedema and many of the sinus units (42%) gained complete recovery during the third month. Pre-operative profiles and endoscopic findings during surgery can serve as a reference of recovery time and treatment modality.

Recovery of non-invasive Aspergillus sinusitis by endoscopic sinus surgery.

No previous data regarding non-invasive Aspergillus sinusitis (NIAS) treated by endoscopic sinus surgery (ESS) has stated any relevance to pre-operative evaluations, disease entities, and the reversibility of symptoms and the sinus mucosa. This prospective study of 31 patients with NIAS and treated by ESS (Strammberger's method) was designed to tackle the above problems. All patients were followed post-operatively by endoscopy once a week. The most refractory symptom was post-nasal dripping. The other symptoms such as foul odour showed progressive improvement by the second post-operative week. Prolonged saccharin time and disease history longer than two years showed the warning signs of a poor recovery. Most patients belonged to the chronic indolent sinusitis group with 55.5% having a complete recovery within the 9th to 12th post-operative weeks. Most patients of allergic Aspergillus sinusitis manifested a recurrent course within months following a post-ESS silent period. All patients of aspergilloma completely recovered prior to the 8th post-operative week.

Assessment of cell surface glycoconjugates in normal, benign and malignant human nasal mucosa.

Aberrant glycosylation of proteins is a common characteristic of neoplastic changes. No reports exist relating cell surface glycoconjugates to normal, benign and malignant human nasal mucosa. Using lectin affinity histochemistry, glycoconjugate reactivities for peanut agglutinin (PNA), concanavalin A (Con A), Griffonia simplicifolia agglutinin II (GSA-II), soy bean agglutinin (SBA) and Ulex europaeus agglutinin l (UEA-I) were analysed in the following groups: normal, benign (polyp, papilloma, and inverted papilloma) and malignant (squamous cell carcinoma (SCC) alone, SCC arising in inverted papilloma, and adenocarcinoma). The positive rate of lectin staining was evaluated using a quantitative AutoCAD programme. We correlated glycoconjugate expression to clinical features, diagnosis, and malignant transformation. The positive rate of PNA after neuraminidase pre-treatment (NA-PNA) staining was higher in inverted papilloma, while all-negative in polyp and papilloma. NA-PNA staining may be used as a differential diagnostic tool. Both inverted papilloma portions and SCC portions of the SCC synchronized with inverted papilloma subjects showed similar Con A and NA-PNA staining patterns. The biological characteristics define inverted papilloma as a pre-malignant neoplasm. The positive rate of PNA staining was significantly higher in inverted papilloma (inverted papilloma transformed to SCC) compared to inverted papilloma alone. Hence, PNA staining may predict malignant transformation of inverted papilloma. However, further investigations are required to prove this possibly worthwhile prognostic marker.