The midgut epithelium of mosquitoes adjusts cell proliferation and endoreplication to respond to physiological challenges.

BACKGROUND: Hematophagous mosquitoes transmit many pathogens that cause human diseases. Pathogen acquisition and transmission occur when female mosquitoes blood feed to acquire nutrients for reproduction. The midgut epithelium of mosquitoes serves as the point of entry for transmissible viruses and parasites. RESULTS: We studied midgut epithelial dynamics in five major mosquito vector species by quantifying PH3-positive cells (indicative of mitotic proliferation), the incorporation of nucleotide analogs (indicative of DNA synthesis accompanying proliferation and/or endoreplication), and the ploidy (by flow cytometry) of cell populations in the posterior midgut epithelium of adult females. Our results show that the epithelial dynamics of post-emergence maturation and of mature sugar-fed guts were similar in members of the Aedes, Culex, and Anopheles genera. In the first three days post-emergence, ~ 20% of cells in the posterior midgut region of interest incorporated nucleotide analogs, concurrent with both proliferative activity and a broad shift toward higher ploidy. In mature mosquitoes maintained on sugar, an average of 3.5% of cells in the posterior midgut region of interest incorporated nucleotide analogs from five to eight days post-emergence, with a consistent presence of mitotic cells indicating constant cell turnover. Oral bacterial infection triggered a sharp increase in mitosis and nucleotide analog incorporation, suggesting that the mosquito midgut undergoes accelerated cellular turnover in response to damage. Finally, blood feeding resulted in an increase in cell proliferation, but the nature and intensity of the response varied by mosquito species and by blood source (human, bovine, avian or artificial). In An. gambiae, enterocytes appeared to reenter the cell cycle to increase ploidy after consuming blood from all sources except avian. CONCLUSIONS: We saw that epithelial proliferation, differentiation, and endoreplication reshape the blood-fed gut to increase ploidy, possibly to facilitate increased metabolic activity. Our results highlight the plasticity of the midgut epithelium in mosquitoes' physiological responses to distinct challenges.

Retraction Note: The carcinogenic complex lncRNA FOXP4-AS1/EZH2/LSD1 accelerates proliferation, migration and invasion of gastric cancer.

The article "The carcinogenic complex lncRNA FOXP4-AS1/EZH2/LSD1 accelerates proliferation, migration and invasion of gastric cancer, by R.-Y. Chen, Q. Ju, L.-M. Feng, Q. Yuan, L. Zhang, published in Eur Rev Med Pharmacol Sci 2019; 23 (19): 8371-8376-DOI: 10.26355/eurrev_201910_19148-PMID: 31646567" has been retracted by the authors. After publication, the authors raised concerns about the reliability of the data used to conduct the study by stating that some data cannot be repeated by further research. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19148.

[Survey on tobacco use and associated factors in population in Shandong province, 2016-2017].

Objective: To understand the status of tobacco use and related influencing factors in population in Shandong province. Methods: Stratified multi-stage cluster sampling was used to select respondents from 216 villages (communities) of 36 districts (counties) in Shandong province. Influencing factors on smoking were analyzed by logistic regression model. Results: The adequate sample size was 6 271 participants. After complex weighted, the current smoking prevalence was 23.93%,45.58% in males and 1.18% in females. The Public's smoking rates varied widely among different groups in the population. 45-64 age group had the highest smoking rate (25.07%). Smoking rates were substantially different in education levels, with the highest in junior high school (28.94%). The rate was higher in the rural area (24.98%) than that in the urban areas (23.08%). The average daily smoking rate was 20.23%. The average age of initiating smoking was 21.21 years. The average daily cigarette intake was 16.31 cigarettes. Among all the former and current smokers, the quitting rate was 20.79%. Multiple logistic regression model analysis showed that gender, age, occupation, region, and health knowledge score were correlated with smoking behavior. The current smoking rate of men was much higher than that of women (OR=49.625, 95%CI: 37.832-65.093). The current smoking rate in 45-64 age group was higher than that in the 15-24 age group (OR=1.830, 95%CI: 1.048-3.194). The current smoking rate of medical (OR=0.403, 95%CI: 0.187-0.866) and retired personnel (OR=0.648, 95%CI: 0.481-0.873) were lower than those engaging in agriculture, forestry, animal husbandry and the fishery ,respectively. The prevalence of residents living in the central part showed lower rate on current smoking than that in the eastern region (OR=0.724, 95%CI: 0.606-0.865). The current smoking prevalence of smoke hazard in 1-3 score group was higher than that in the group with 4-6 score (OR=1.432, 95%CI: 1.240-1.654). Conclusions: Smoking rate in adults in Shandong remained stable and at a high level. Comprehensive intervention measures such as tobacco control and health education should be carried out to reduce the smoking rate.

A multi-center study on recurrent inguinal hernias: assessment of surgeons' compliance to guideline-based repair and evaluation of short-term outcomes.

INTRODUCTION: As patients with recurrent inguinal hernia (RIH) are at a higher risk of perioperative complications, international guidelines have been developed to mitigate these risks by recommending the reverse approach for repair. We aim to study the characteristics of RIH, compliance to guidelines-based repair (GR) and determine factors influencing compliance to guidelines. METHODS: A retrospective study of patients with RIH was carried out at two tertiary institutions in Singapore, over 10 years from January 2010 to 2020. RESULTS: There were 16 patients with bilateral recurrences and 214 patients with unilateral recurrences. The characteristics of patients with non-guidelines-based repair (NGR) versus GR were similar, p > 0.05. GR was performed for 128 (52.1%) hernias as compared to NGR for 118 (47.9%) hernias. The open approach was more common in NGR than GR, 115/118 (89.8%) versus 58/128 (45.3%), p < 0.001. Forty (n = 40, 16.3%) RIH presented emergently, of which 37 underwent NGR while 3 underwent GR, p < 0.0001. More consultants were present during GR 103/128 (80.5%) as compared to NGR 78/118 (66.1%), p = 0.018. Emergency presentation of hernia recurrence, OR 7.74 (CI 6.11-9.20), p = 0.005, and open repair during the index repair were significantly associated with NGR, OR 6.63 (CI 4.42-8.84), p = 0.01. Median length of stay was shorter in the GR 1 day (IQR 1-2 days) versus 2 days (IQR 2-5 days) in the NGR group, p = 0.02. CONCLUSION: The compliance rate of GR for RIH is 52%. NGR for RIH had acceptable short-term outcomes. For elective presentation of RIH, GR should be encouraged given a shorter length of hospital stay.

[Prevalence and associated factors regarding electronic cigarette smoking among adults in Shandong province].

Objective: To understand the awareness, current or past behavior on electronic cigarette smoking (ECS) and associated factors, among adults aged ≥15 years old in Shandong province. Relationship between ECS and cessation of smoking was also studied. Methods: Using the global standard questionnaire, a cross-sectional survey was conducted among 6 401 adults selected by stratified multi-stage cluster sample method. Electronic data was collected by handheld computer. Data were weighted and analyzed by SPSS 19.0 complex program. Results: The prevalence rates of awareness, ever or current smoking of e-cigarettes appeared as 37.6%, 3.8% and 0.6% respectively. Data from logistic regression analysis showed that risk factors for ECS were living in urban area, at younger age, being current or former cigarettes smoker and with knowledge on the risks of smoking or being encouraged to stop smoking. Among the electronic cigarettes smokers, those who tried to quit smoking (30.1%) had a significantly higher rate than that of the non-smokers (19.5%) (P<0.05), however, the successful quitting rate was much lower than that in the non-ECS users (P<0.05). Conclusions: Rates on the awareness and uptake of electronic cigarettes are on the rise. It is necessary to strengthen and standardize the monitoring programs to provide evidence for setting up targeting intervention strategies.

The carcinogenic complex lncRNA FOXP4-AS1/EZH2/LSD1 accelerates proliferation, migration and invasion of gastric cancer.

OBJECTIVE: To clarify the role of lncRNA FOX4-AS1 in the progression of gastric cancer (GC) via interacting with EZH2/LSD1. PATIENTS AND METHODS: Relative level of FOXP4-AS1 in GC tissues and adjacent normal tissues was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The potential influences of FOXP4-AS1 on cellular behaviors of GC cells were evaluated via a series of functional experiments. Bioinformatics prediction, RNA immunoprecipitation (RIP) assay, and Western blot were conducted to verify the potential of EZH2/LSD1 as a target of FOXP4-AS1. RESULTS: FOXP4-AS1 was upregulated in GC tissues relative to controls. Its level was higher in GC patients with stage III-IV than those with stage I-II. The survival rate was lower in GC patients presenting the high expression of FOXP4-AS1 compared with those presenting low expression. Transfection of sh-FOXP4-AS1 1# or sh-FOXP4-AS1 2# attenuated proliferative, migratory, and invasive abilities of AGS and BGC7901 cells. FOXP4-AS1 could bind to LSD1 and EZH2, and positively regulated their expression levels. Transfection of sh-LSD1 or sh-EZH2 reduced the proliferative ability of GC cells. CONCLUSIONS: FOXP4-AS1 binds to EZH2/LSD1 to form a carcinogenic complex, thus accelerating GC cells to proliferate, migrate and invade.

[Study on the relationship between smoking behavior and health literacy among grade 4-6 primary students in Shandong province].

Objective: To investigate the associations between smoking and literacy on health among 4-6 grade primary school students. Methods: A questionnaire survey was conducted to research on health literacy and smoking among 4-6 grade pupils in Shandong province, through a multi-stage stratified cluster random sampling method. Sociodemographic characteristics, health literacy level and smoking rate were collected from respondents. Binary logistic regression analysis was used to evaluate the association of smoking and health literacy. Results: A total of 9 240 questionnaires were distributed, with the rate of valid response as 99.7%. The current smoking rate of the students was 2.6%, on higher in boys (3.1%) than in girls (2.0%). 60.8% of 4-6 grade students were found to have adequate health literacy level. Levels of literacy health in both boy and girl school students appeared 56.7% and 64.9%, respectively. Results indicated that health literacy in smokers (14.4%) was lower than that in non-smokers (62.0%). Results from the binary logistic regression analysis showed that the independent influencing factors would include grade, father's education level, economic situation of the family, self-assessment on the school record and literacy on health (P<0.01). After controlling the other independent variables, the smoking rate was 8.62 (1/0.116) times in students with low literacy level on health, than those with high literacy level. Conclusions: Literacy on health was significantly associated with smoking in the 4-6 grade pupils of Shandong province.

Immunohistochemical studies and fluorodeoxyglucose uptake on positron emission tomography in pharyngeal cancer for predicting radiotherapy-based treatment outcomes.

OBJECTIVES: This study correlated immunohistochemical studies with fluorodeoxyglucose (FDG) uptake on positron emission tomography-computed tomography (PET-CT) and identified prognostic factors for radiotherapy (RT)-based treatment outcomes in patients with squamous cell carcinoma of the oropharynx and hypopharynx. METHODS: Genomic data from pre-treatment biopsy specimens (Glut1, CAIX, VEGF, HIF-1α, EGFR, Ki-67, Bcl-2, CLAUDIN-4, YAP-1, c-Met and p16) of 76 patients were analysed using tissue microarrays. FDG uptake was evaluated using the maximum standardised uptake value (SUVmax), metabolic tumour volume (MTV) and total lesion glycolysis (TLG). RESULTS: The overexpression of Glut1 positively associated with increased values of the SUVmax, MTV and TLG, whereas VEGF and HIF-1α expression with the MTV and TLG, respectively. A VEGF immunoreactive score (IRS) >2 (P = 0.001, hazard ratio [HR] = 3.94) and an MTV defined by an SUV of 2.5 (MTV2.5) >14.5 mL (P = 0.004, HR = 3.31) were prognostic factors for low cause-specific survival, whereas a VEGF IRS >2 (P = 0.02, HR = 2.83) for low primary relapse-free survival. CONCLUSION: The overexpression of Glut1, VEGF and HIF-1α associated with increased FDG uptake. For patients with pharyngeal cancer requiring RT, the treatment outcome can be stratified by VEGF and MTV2.5.

Novel association of thymic carcinoid with a germline mutation in a kindred with multiple endocrine neoplasia 1 (MEN1).

Multiple endocrine neoplasia 1 (MEN1) is an autosomal dominant syndrome characterized by a triad of endocrine (parathyroid, enteropancreatic and pituitary) tumors. Familial MEN1 is defined by one first-degree relative having at least one of these 3 main tumors, and is associated with germline mutations in the MEN1 gene on 11q13 in a large proportion of cases. MEN1 patients may also develop non-endocrine tumors, notably thymic carcinoid. These are rare tumors found predominantly in men, and are a major cause of death in MEN1 due to their insidious nature, lack of effective treatment and unpredictable recurrence. Prophylactic thymectomy has been advocated for prevention but continued surveillance for recurrence is necessary. Although genotype-phenotype correlation in MEN1-related thymic carcinoid is inconsistent, there is a high prevalence of truncating mutations in this condition. We describe a father and son with MEN1, associated with thymic carcinoid (father) and the truncating mutation R29X (son), which was not previously reported in MEN1-related thymic carcinoid, and review the literature about thymic carcinoids in MEN1. Our cases illustrate the importance of a high index of suspicion for early diagnosis and lifelong surveillance in MEN1, and the utility of genetic analysis in defining surveillance for MEN1-related thymic carcinoid.

Biopsy forceps is inadequate for the resection of diminutive polyps.

BACKGROUND AND STUDY AIMS: Cold biopsy forceps polypectomy (CBP) is often used for the removal of diminutive polyps. The efficacy of the technique has not been thoroughly assessed. The aim of this study was to prospectively assess the efficacy of CBP for removing diminutive polyps. PATIENTS AND METHODS: This was a prospective study from St Vincent's Hospital, a tertiary referral hospital in Melbourne, Australia. A total of 143 patients were screened and 52 patients with ≥ 1 diminutive polyps were enrolled. CBP was used to resect diminutive polyps until no polyp tissue was visible. The polyp base was then resected using endoscopic mucosal resection (EMR) with a 1 - 2-mm margin. The CBP and EMR samples were compared to assess completeness of the resection. RESULTS: Overall 39 % (21 / 54) of diminutive polyps were completely resected using CBP. After binary logistic regression analysis, polyp histology was found to be predictive of resection, with complete resection of 62 % (13 / 21) for adenomas and 24 % (8 / 33) for hyperplastic polyps (odds ratio 5.1; P = 0.008). The size and number of bites taken with the forceps were not predictive of complete response. CONCLUSIONS: Within the limitations of a modest sample size, CBP appears to be inadequate treatment for the removal of diminutive polyps.

Two new mutations in the keratin 4 gene causing oral white sponge nevus in Chinese family.

OBJECTIVE: We investigated white sponge nevus (WSN) in a Chinese family, and tried to find new mutation and demonstrated that this mutation is the causative mutation for WSN in this family and this condition affects a functionally important segment of the keratin 4 protein. MATERIALS AND METHODS: We studied the affected family with the 32-year-old female patient, her mother, her younger sister and her daughter. Pathologic examinations were performed. DNA was extracted from peripheral blood lymphocytes, K4 and K13 genes were amplified by polymerase chain reaction (PCR) and sequenced. RESULTS: Direct sequencing of PCR products revealed two new mutations in the keratin 4 gene, the heterozygous missense mutation 1829G-->A in exon 2B, and 2324A-->G in non-coding region. No any mutation was found in the keratin 13 gene. CONCLUSIONS: We found two new mutations in the keratin 4, which may be related with the development of WSN.

Diagnosis of solid pancreatic masses by endoscopic ultrasound-guided fine-needle aspiration.

BACKGROUND: Endoscopic ultrasound (EUS) with fine-needle aspiration (FNA) is increasingly being used in the staging algorithm for pancreatic carcinoma. This allows for a tissue diagnosis, which was previously difficult to obtain. The aim of this study is to assess the utility of EUS-FNA in establishing the diagnosis of solid pancreatic mass lesions in an Australian population. METHODS: A retrospective review of the EUS databases of St Vincent's Hospital Melbourne and Western Hospital, Melbourne from November 2002 to May 2006 was undertaken. The focus was on patients with a solid pancreatic mass who underwent EUS-FNA. Surgical pathology or long-term follow up was used to identify false-positive or false-negative results. RESULTS: EUS was undertaken to investigate a solid pancreatic or distal common bile duct mass lesion in 155 patients. Seventy-two of these underwent EUS-guided FNA. Mean age was 68 years. A positive tissue diagnosis of malignancy could be made in 55 (76%). Nine (13%) had benign histology, with 8 (11%) having inadequate tissue obtained from FNA. A later tissue diagnosis of carcinoma was made in eight of those with either benign or inadequate histology, although in all cases there were EUS features diagnostic of malignancy, with FNA limited by technical difficulties. The overall utility of EUS-FNA showed a sensitivity of 87%, specificity 100%, positive predictive value 100%, negative predictive value 52% and overall accuracy 89%. CONCLUSION: EUS-FNA gives a high return for histological diagnosis of solid pancreatic mass lesions and should be part of the standard management algorithm for pancreatic carcinoma.

Synthesis and biological evaluation of novel naphthoquinone fused cyclic aminoalkylphosphonates and aminoalkylphosphonic monoester.

A series of novel naphthoquinone fused cyclic alpha-aminophosphonates, 2-alkoxy-3,4-dihydro-2H-naphtho[2,3-e][1,4,2]oxazaphosphinane-5,10-dione 2-oxide 3-17 and naphthoquinone fused cyclic alpha-aminophosphonic monoester 18 were synthesized for the first time. These cyclic alpha-aminophosphonates were evaluated for antitumor activity on four human tumor cell lines, and three of them showed significant cytotoxicity (IC(50): 0.019-5.15 microM) comparable to that of the reference drug doxorubicin. Furthermore, inhibition assays for topoisomerase II-mediated relaxation of supercoiled DNA indicated that the naphthoquinone fused cyclic aminophosphonates were catalytic inhibitors of topoisomerase II.

The accuracy of EUS-FNA in assessing mediastinal lymphadenopathy and staging patients with NSCLC.

Optimal management of nonsmall cell lung cancer (NSCLC) depends on tissue diagnosis and accurate staging. Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is minimally invasive and provides cytological confirmation of malignant mediastinal disease. The aim was to assess the accuracy of EUS-FNA in cases of enlarged mediastinal lymphadenopathy (LN) of unknown aetiology and in the staging of NSCLC. A total of 52 consecutive patients with stage I-IIIb NSCLC or enlarged mediastinal LN of unknown aetiology underwent EUS-FNA. Negative results were confirmed with a surgical procedure: mediastinoscopy, video-assisted thoracic surgery (VATS) or lobectomy with systematic mediastinal lymph node dissection. In total, 34 patients had EUS-FNA performed for diagnosis, whilst the remaining 18 had EUS-FNA for staging. The overall sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy (95% confidence interval) were 93% (77-99), 100% (78-100), 100% (87-100), 88% (63-99) and 95% (84-99), respectively. When EUS-FNA was used in patients with NSCLC, the sensitivity, specificity, PPV, NPV and accuracy were 92% (73-99), 100% (69-100), 100% (85-100), 83% (51-98) and 94% (80-99), respectively. For mediastinal LN of unknown aetiology, no malignant disease was missed. Endoscopic ultrasound-guided fine-needle aspiration is an accurate tool for assessing mediastinal lymph node involvement in nonsmall cell lung cancer and in the diagnosis of unexplained mediastinal lymphadenopathy. Endoscopic ultrasound-guided fine-needle aspiration is a minimally invasive procedure that can be used as an adjunct or alternative to mediastinoscopy.

Chronic hepatitis B: recommendations for therapy based on the natural history of disease in Australian patients.

BACKGROUND: Chronic hepatitis B infection (CHB) is a major health problem in Australia and worldwide. CHB is associated with significant long-term morbidity and mortality. Well tolerated treatment is now available, however the development of resistance is common and the optimal timing of treatment is yet to be determined. Identifying the factors that influence the natural history of CHB may help determine which patients need treatment and when to start it. OBJECTIVE: To determine the demographics, clinical features and virological profile of Australian patients infected with CHB and the influence of these factors on disease activity and severity. STUDY DESIGN: Review of prospectively collected demographic, clinical and virological features of all patients positive for hepatitis B surface antigen (HBsAg) for more than 6 months who were referred to St. Vincent's Hospital liver clinics. Age, sex and ethnicity were correlated with hepatitis B e antigen status (HBeAg), HBV replication status (ALT and HBV DNA), genotype and liver histology. RESULTS: 703 chronic hepatitis B surface antigen positive patients were identified. The patients were predominantly male with an average age of 44. Eighty two percent of patients were born overseas, primarily from Asian (65%) and Mediterranean countries (14%). Two thirds (426) had an elevated ALT (median 79) at presentation. HBeAg was positive in 37%. Active viral replication, defined as abnormal ALT or positive HBVDNA, was present in 74%, 48% of whom were HBeAg negative. In a subset of 103 patients genotyped, 8% had genotype A, 29% B, 41% C and 22% D. Genotype correlated with ethnicity; patients infected with genotypes A were predominantly Caucasian, B and C were Asian, and D were Mediterranean. Of 296 (42%) patients who underwent liver biopsy, 76 (27%) had advanced fibrosis. Advanced fibrosis was associated with increasing age and Mediterranean ethnicity. CONCLUSION AND RECOMMENDATIONS: Perinatal or early childhood transmission is predominant mode of infection in Australia. Two thirds of this cohort had active replication and were at increased risk of developing cirrhosis and/or hepatoma. Advanced disease was associated with age and ethnicity. HBeAg negative CHB accounts for almost half of all those with active viral replication. This parallels the rise in this form of CHB in Asia and the Mediterranean basin. Screening should be offered to people born in, or with parents born in areas of high endemnicity. To detect the development of active disease, patients with positive HBsAg but normal ALT should have liver function tests done 6 monthly and those with elevated ALT should be referred for consideration of therapy, irrespective of HBeAg status.

Five new diprenylated flavonols from the leaves of Broussonetia kazinoki.

Five new diprenylated flavonols, broussonol A (1), broussonol B (2), broussonol C (3), broussonol D (4), and broussonol E (5), along with two known compounds, were isolated from an ethanolic extract of the leaves of Broussonetia kazinoki. Their structures were elucidated by chemical and spectral methods. Cytotoxic activities were evaluated against several different cell lines.

Use of an overtube for enteroscopy--does it increase depth of insertion? A prospective study of enteroscopy with and without an overtube.

BACKGROUND AND STUDY AIMS: Although a stiffening overtube is commonly used with push enteroscopy, in the belief that this will allow increased insertion into the small intestine, there is no prospective data to support this view. The aim of this study is to prospectively study the depth of insertion into the small intestine at enteroscopy with and without an overtube. PATIENTS AND METHODS: A total of 38 patients referred for enteroscopy were prospectively studied. Alternate enteroscopies were performed with or without an overtube; therefore 19 patients had enteroscopy with and 19 without an overtube. The groups were well matched for age, sex, indication, use of fluoroscopy, and dedicated anesthetic assistance. Depth of insertion was assessed by advancing the enteroscope as far as possible, then straightening the enteroscope until the tip began withdrawing. The difference between the straightened insertion depth and the distance from the incisors to the pylorus was recorded as the insertion depth beyond the pylorus. This was considered the major end point. Statistical analysis was performed using the Mann-Whitney test for nonparametric data. RESULTS: The median straightened total insertion depth from the incisors was greater when enteroscopy was performed with an overtube compared with enteroscopy without an overtube (125 cm vs. 110 cm, P=0.05). The median straightened insertion depth beyond the pylorus was significantly greater with overtube use (70 cm vs. 50 cm, P = 0.01). No significant difference between the groups was observed in terms of the likelihood of significant findings at enteroscopy. CONCLUSIONS: Use of an overtube for push enteroscopy results in significantly deeper insertion into the small intestine. Although a larger study would be needed to demonstrate an increase in diagnostic yield and to confirm the safety of overtube use, this study does provide the first objective evidence of an advantage in terms of insertion depth.

New annonaceous acetogenins from the roots of Uvaria calamistrata.

Five new annonaceous acetogenins, calamistrins C-G (1-5), were isolated from an ethanolic extract of the roots of Uvaria calamistrata. Compounds 1-3 were mono-THF ring acetogenins; compounds 4 and 5 were bis-THF acetogenins, with the THF rings from C-18 to C-25. The absolute configurations of 3, 4, and 5 as well as the partial absolute configurations of 1 and 2 were determined by (13)C NMR spectroscopy and advanced Mosher methodology.