Effects and mechanisms of substance P on the proliferation and angiogenic differentiation of bone marrow mesenchymal stem cells: Bioinformatics and in vitro experiments.

The slight release of substance P (SP) from the end of peripheral nerve fibers causes a neurogenic inflammatory reaction, promotes vascular dilation and increases vascular permeability. However, whether SP can promote the angiogenesis of bone marrow mesenchymal stem cells (BMSCs) under high glucose conditions has not been reported. This study analyzed the targets, biological processes and molecular mechanisms underlying the effects of SP on BMSCs. BMSCs cultured in vitro were divided into a normal control group, high glucose control group, high glucose SP group and high glucose Akt inhibitor group to verify the effects of SP on BMSCs proliferation, migration and angiogenic differentiation. SP was found to act on 28 targets of BMSCs and participate in angiogenesis. Thirty-six core proteins, including AKT1, APP, BRCA1, CREBBP and EGFR, were identified. In a high glucose environment, SP increased the BMSCs proliferation optical density value and cell migration number and reduced the BMSCs apoptosis rate. In addition, SP induced BMSCs to highly express the CD31 protein, maintain the wall structure integrity of the matrix glue mesh and promote increases in the number of matrix glue meshes. These experiments showed that in a high glucose environment, SP acts on 28 targets of BMSCs that encode core proteins, such as AKT1, APP and BRCA1, and improves BMSCs proliferation, migration and angiogenic differentiation through the Akt signaling pathway.

Establishment of a modified Kyoto classification scoring model and its significance in the diagnosis of Helicobacter pylori current infection.

BACKGROUND AND AIMS: We aimed to establish a modified model of the Kyoto classification score and verify its accuracy for predicting Helicobacter pylori (HP) infection during endoscopy. METHODS: Patients who underwent gastroscopy from June 2020 to March 2021 were included in this study. Atrophy, intestinal metaplasia, hypertrophy of the gastric fold, nodularity, diffuse redness, sticky mucus, spotty redness, xanthoma, map-like redness, fundic gland polyp, and regular arrangement of collecting venules (RAC) were recorded according to the Kyoto classification of gastritis. The HP infection status of participants was determined by a 13C breath test, anti-HP antibody, and histopathologic hematoxylin and eosin staining. The modified Kyoto classification scoring model was established based on univariate analysis and logistic regression analysis. The modified scoring model was used to judge the status of HP infection in patients undergoing gastroscopy from July to September 2021 and to evaluate the accuracy of the prediction. RESULTS: Of 667 participants in the derivation dataset, 326 cases had HP infection and 341 cases did not. Atrophy, hypertrophy of the gastric fold, nodularity, diffuse redness, sticky mucus, and spotty redness were associated with HP current infection. Thus, a new scoring model, termed the modified Kyoto classification scoring model, was constructed that included atrophy, hypertrophy of the gastric fold, nodularity, diffuse redness, sticky mucus, spotty redness, fundic gland polyp, and RAC as indicators. To test the model, 808 subjects, including 251 HP-positive patients, comprised the validation dataset. CONCLUSIONS: The modified Kyoto classification scoring model improved the accuracy of endoscopic determination of HP current infection and has clinical application potential in the Chinese population.

Double sodium channel mutation, I265T/L1014F, is possibly related to pyrethroid-resistant in Thrips palmi.

Thrips palmi Karny (Thysanoptera: Thripidae) can harm a variety of agricultural crops and transmit plant viruses, causing heavy economic losses. In the Hainan province of China, pyrethroids were sprayed widely to control T. palmi, which leaded to resistance to pyrethroids in T. palmi. The bioassay has shown that the resistance ratio of T. palmi to pyrethroids increases annually. Resistance ratio to λ-cyhalothrin has increased from 10.711 to 23.321 and to cypermethrin has increased from 5.507 to 23.051 for 3 years, 2020-2022. The double mutation (I265T/L1014F) was identified from the field strain for the first time, which were located in the domains I and II of the voltage-gated sodium channel of T. palmi, respectively. The double mutation is probably the reason for the higher resistance of T. palmi in Hainan. The frequencies of the double mutation were 53.33% in HN2020, 70.00% in HN2021, and 96.67% in HN2022. Results indicated that T. palmi had developed different degrees of resistance to pyrethroids in Hainan. This study provides theoretical guidance for the use of insecticides in the field control of thrips.

Indoxacarb resistance-associated mutation of Liriomyza trifolii in Hainan, China.

Liriomyza trifolii, which has been recently prevalent in China, harms more than 300 plant species, especially cowpea in Hainan. This pest also affects the quality and production of vegetables in winter. Indoxacarb is the first commercial oxadiazine pesticide, which is a new efficient insecticide used to control pests of Diptera, including L. trifolii. The unique mechanism of indoxacarb is that indenyl is transformed into N-demethoxycarbonyl metabolite (DCJW) in insects and acts on inactivated sodium channel; DCJW could then destroy the conduction of nerve impulses, which leads to movement disorders, feeding stoppage, paralysis, and eventually the death of pests. The field population of L. trifolii developed resistance by 769 times higher than the sensitive population in Sanya, Hainan. Results revealed the existence of a mutation (i.e., V1848I) in the sixth transmembrane segment of Domain IV of the sodium channel in the field population. The homozygous resistant genotype frequency for the V1848I mutation was 10-15% among the three field-collected populations. This paper reports for the first time the presence of the kdr mutation V1848I in resistant populations of L. trifolii to indoxacarb. The present study will contribute to the understanding of the evolution of indoxacarb resistance and contribute to the development of resistance management practices for winter vegetables in Hainan.

Performance evaluation of four prediction models for risk stratification in gastric cancer screening among a high-risk population in China.

OBJECTIVE: Early detection of gastric cancer (GC) is a critical step for decreasing mortality. The aim of this study was to evaluate the performance of four prediction models for risk stratification in the screening of GC and precancerous lesions among a large, high-risk population in China. DESIGN: This study was a retrospective analysis of data from the Provincial Gastric Cancer Screening Program (Zhejiang, China) spanning the period between October 2016 and April 2019, in which 97,541 individuals from the urban areas of 10 cities in Zhejiang province, China participated in this program. Demographic and clinical characteristics data were collected, and serum pepsinogens I and II, gastrin-17, and anti-H. pylori IgG antibody were detected. Participants were asked to voluntarily undergo gastroscopy. The performance of the ABC method, new ABC method, Tu's prediction model, and Li's prediction model, which stratified participants into low-, medium- and high-risk subgroups, were evaluated using the area under the receiver-operating characteristic (ROC) curve (AUC) and Youden index. RESULTS: Among the participants, 6005 (3447 males and 2558 females, mean age of 58.35 years), voluntarily underwent gastroscopy. Overall, 72 (1.20%) GC cases (30 early and 42 advanced) and 2006 cases with precancerous lesions (270 atrophic gastritis, 1634 intestinal metaplasia, and 102 dysplasia/intraepithelial neoplasia) were identified. Notably, Li's prediction model achieved the greatest AUC and Youden index values (0.708 and 0.319, respectively) for predicting GC, and exhibited the greatest ability to detect precancerous lesions, especially intestinal metaplasia. CONCLUSION: Li's prediction model performs the best for risk stratification in the screening, detection, and diagnosis of GC and precancerous lesions, whereas the overall performance of the other three models is similar ( www.chictr.org.cn , ChiCTR2100043363).

Circular RNA circ_001350 regulates glioma cell proliferation, apoptosis, and metastatic properties by acting as a miRNA sponge.

Glioma is an aggressive malignancy with increasing incidence and threatens people's health worldwide. Accumulating evidence revealed that circular RNAs (circRNAs) play important functions in cancers. A previous study demonstrated that circ_001350 was elevated in glioma tissue samples than nontumorous tissue specimens screened by high-throughput microarray. The level of circ_001350 in glioma tissue specimens and cell lines was detected by quantitative real-time polymerase chain reaction. The Fisher exact test was carried out to estimate the correlation of circ_001350 level with clinical characteristics. Cell proliferation, apoptosis, and motility abilities were detected using cell counting kit-8, clonogenic, flow cytometry, and transwell experiments, respectively. The potential target of circ_001350 was identified by the luciferase assay. circ_001350 level was significantly enhanced in glioma tissue specimens and cells. Further, elevated expression of circ_001350 was closely linked to patients' clinical severity. Knockdown of circ_001350 could inhibit cell proliferation and metastatic properties and increase apoptotic cells. circ_001350 could directly bind to miR-1236 and regulate its expression to exert oncogenic functions. Collectively, circ_001350 directly sponges miR-1236, thus contributing to malignant progression of glioma.

Roles of High-resolution Manometry in Predicting Incomplete Bolus Transit in Patients With Dysphagia.

BACKGROUND: High-resolution manometry (HRM) is used to assess esophageal motility diseases. Abnormalities in a number of HRM parameters have been reported in patients with dysphagia. However, it is unclear whether some of abnormal HRM parameters are predictive of dysphagia. The aim of this retrospective study was to investigate the roles of HRM parameters in predicting incomplete bolus clearance (IBC) in patients with dysphagia using high-resolution impedance manometry. METHODS: A total of 644 wet swallows were reviewed and analyzed in 63 patients with symptoms of dysphagia or reflux who underwent a clinical high-resolution impedance manometry test. IBC was defined based on impedance measurement. The relationship of each of abnormal HRM parameters with IBC was analyzed and their roles in predicting IBC were determined. RESULTS: Patients with symptoms of both dysphagia and reflux showed the highest IBC rate, and patients with symptoms of reflux had the lowest IBC rate. The IBC was more prevalent in the distal esophagus. Premature contractions and peristalsis with large breaks were associated with a higher IBC rate in the proximal esophagus (P<0.05); large breaks, ineffective peristalsis, and abnormalities of the esophageal gastric junction functions were associate with higher IBC rates in the distal esophagus (P<0.05). Abnormalities in a number of motility parameters were able to predict IBC with high specificities and/or high sensitivity, such as pan esophageal pressurization, ineffective peristalsis, and large breaks. Abnormal integrative relaxation pressure of the lower esophageal sphincter with concurrent pan esophageal pressurization, ineffective peristalsis, or large breaks is predictive of IBC with nearly 100% of specificity. CONCLUSIONS: Abnormalities in a number of HRM parameters are not only useful in diagnosing esophageal motility diseases, but also valuable in predicting IBC during swallowing.

Anti-inflammatory effects and mechanisms of vagal nerve stimulation combined with electroacupuncture in a rodent model of TNBS-induced colitis.

The purpose of this study was to determine the effects and mechanisms of vagal nerve stimulation (VNS) and additive effects of electroacupuncture (EA) on colonic inflammation in a rodent model of IBD. Chronic inflammation in rats was induced by intrarectal TNBS (2,4,6-trinitrobenzenesulfonic acid). The rats were then treated with sham ES (electrical stimulation), VNS, or VNS + EA for 3 wk. Inflammatory responses were assessed by disease activity index (DAI), macroscopic scores and histological scores of colonic tissues, plasma levels of TNFα, IL-1ß, and IL-6, and myeloperoxidase (MPO) activity of colonic tissues. The autonomic function was assessed by the spectral analysis of heart rate variability (HRV) derived from the electrocardiogram. It was found that 1) the area under curve (AUC) of DAI was substantially decreased with VNS + EA and VNS, with VNS + EA being more effective than VNS (P < 0.001); 2) the macroscopic score was 6.43 ± 0.61 in the sham ES group and reduced to 1.86 ± 0.26 with VNS (P < 0.001) and 1.29 ± 0.18 with VNS + EA (P < 0.001); 3) the histological score was 4.05 ± 0.58 in the sham ES group and reduced to 1.93 ± 0.37 with VNS (P < 0.001) and 1.36 ± 0.20 with VNS + EA (P < 0.001); 4) the plasma levels of TNFα, IL-1ß, IL-6, and MPO were all significantly decreased with VNS and VNS + EA compared with the sham ES group; and 5) autonomically, both VNS + EA and VNS substantially increased vagal activity and decreased sympathetic activity compared with sham EA (P < 0.001, P < 0.001, respectively). In conclusion, chronic VNS improves inflammation in TNBS-treated rats by inhibiting proinflammatory cytokines via the autonomic mechanism. Addition of noninvasive EA to VNS may enhance the anti-inflammatory effect of VNS.NEW & NOTEWORTHY This is the first study to address and compare the effects of vagal nerve stimulation (VNS), electrical acupuncture (EA) and VNS + EA on TNBS (2,4,6-trinitrobenzenesulfonic acid)-induced colitis in rats. The proposed chronic VNS + EA, VNS, and EA were shown to decrease DAI and ameliorate macroscopic and microscopic damages in rats with TNBS-induced colitis via the autonomic pathway. The addition of EA to VNS provided a significant effect on the behavioral assessment of inflammation (DAI, CMDI, and histological score) but not on cytokines or mechanistic measurements, suggesting an overall systemic effect of EA.View this article's corresponding video summary at https://youtu.be/-rEz6HMkErM.

The effects of bufadienolides on HER2 overexpressing breast cancer cells.

HER2 is a proto-oncogene frequently amplified in human breast cancer, its overexpression is correlated with tamoxifen resistance and decreased recurrence-free survival. Arenobufagin and bufalin are homogeneous bufadienolides of cardiac glycosides agents. In this research, we studied the effects of arenobufagin and bufalin on cellular survival and proliferation of HER2 overexpressing breast cancer cells and the mechanism under the results including the direct effect on HER2 downstream pathways. Our results showed that arenobufagin and bufalin could significantly inhibit the proliferation and survival of HER2 overexpressing breast cancer cells, along with the declination of SRC-1, SRC-3, nuclear transcription factor E2F1, phosphorylated AKT, and ERK. And the combination of each bufadienolide in low dose with tamoxifen could significantly enhance the inhibitory effect of tamoxifen on HER2 overexpressing breast cancer cells. All above suggest that arenobufagin and bufalin may be potential therapy adjuvants for HER2 overexpressing breast cancer therapy.

Electrical neuromodulation at acupoint ST36 normalizes impaired colonic motility induced by rectal distension in dogs.

Electroacupuncture (EA) has been shown to improve impaired gastric motility and slow waves in both humans and animals. However, its effects on colonic motility have rarely been investigated. The aim of this study was to investigate the effects and underlying mechanisms of EA on impaired colonic motility induced by rectal distension (RD)in dogs. Colon contractions and transit were measured in various sessions with and without EA in hound dogs chronically placed with a colonic cannula. Colonic contractile activity was assessed by motility index (MI). Autonomic functions were determined by the spectral analysis of the heart rate variability derived from the electrocardiogram. It was found 1) RD suppressed colonic motility by 40.5% (10.8 ± 0.9 with RD vs. 6.4 ± 0.8 at baseline, P < 0.002). EA at ST36 normalized colonic contractions suppressed by RD (12.9 ± 2.8, P < 0.002 vs. RD and P = 0.1 vs. control). 2) Administration of atropine blocked the ameliorating effect of EA on colon motility. 3) RD also delayed colonic transit (65.0 ± 2.0% with RD vs. 86.0 ± 1.9% without RD, P < 0.001) that was restored with EA (84.0 ± 1.9%, P = 0.178 vs. control). 4) EA increased vagal activity suppressed by RD (0.37 ± 0.07 with RD + EA vs. 0.09 ± 0.03 with RD without EA, P < 0.001). In conclusion, RD inhibits colonic contractions and delays colonic transit in dogs; EA at ST36 restores the RD-induced impairment in both colonic contraction and transit by enhancing vagal activity and mediated via the cholinergic pathway.

Evaluation of the Expanded Criteria of Endoscopic Submucosal Dissection for Early Gastric Cancers: A Meta-Analysis.

BACKGROUND: In 2010, undifferentiated early-stage gastric cancers (EGCs; ≤1 cm and confined to the mucosa) were included in the expanded criteria for endoscopic submucosal dissection (ESD), as established by the Japanese Gastric Cancer Association. OBJECTIVE: To evaluate the safety and efficacy of the newly expanded criteria of ESD for EGCs. METHODS: PubMed, Embase, and the Cochrane Library were searched for relevant studies published between January 1, 2010 and July 10, 2014. These studies were collected by 2 authors and then analyzed with RevMan 5.0. RESULTS: Six studies including 6,687 patients were identified. Compared to the standard group (SG), the expanded group (EG) had higher bleeding (OR 1.66; 95% CI 1.02-2.68), perforation (OR 2.03; 95% CI 1.25-3.31), positive lateral margin rates (OR 16.88; 95% CI 3.83-74.5) and lower en bloc resection rate (OR 0.48; 95% CI 0.34-0.70). The local recurrence (OR 2.46; 95% CI 0.31-19.23) and total survival rates (OR 0.92; 95% CI 0.76-1.12) between the 2 groups were not statistically different. CONCLUSIONS: Considering the similar rates of local recurrence and total survival between the SG and EG, the use of ESD for the management of EGCs that meet the expanded criteria could be considered a safe and effective treatment method.

[Curcuma wenyujin Diterpenoid compound C fought against gastric cancer: an experimental study].

OBJECTIVE: To study the role of nuclear factor (NF)-kappaB pathway with p38MAPK in Curcuma wenyujin diterpenoid compound C (CDCC) fighting against inflammation and inducing gastric cancer cell apoptosis by stimulating gastric cell SGC7901 with tumor necrosis factor-alpha (TNF-alpha). METHODS: Human umbilical vein endothelial cells (HUVECs) and human gastric cancer SGC-7901 cells were in vitro acted by CDCC in different concentrations at different time points. Their growth inhibition ratios were measured by MTT assay. The apoptosis rate of gastric cancer cells was detected by Annexin V-FITC/PI double staining. Nuclear translocation of p65 was detected by cell immunofluorescence. Expression levels of p38MAPK/P-p38MAPK, p65/P-p65, and Caspase 3/P-Caspase 3 were measured by Western blot. RESULTS: CDCC had significant inhibitory effect on the proliferation of SGC-7901. It also could effectively induce the apoptosis of gastric cancer cell SGC-7901. It also could reduce nuclear translocation of p65 in gastric cancer cell SGC-7901. Results of Western blot indicated that expression levels of p38MAPK and p65 were reduced and the expression level of Caspase-3 was elevated along with increased concentrations of CDCC (P<0.05). CONCLUSION: Apoptosis executive protein Caspase 3 activated by regulating p65 via p38MAPK might be one of possible mechanisms for CDCC fighting against inflammation and gastric cancer.

Melatonin attenuates hypoxic pulmonary hypertension by inhibiting the inflammation and the proliferation of pulmonary arterial smooth muscle cells.

Hypoxia-induced inflammation and excessive proliferation of pulmonary artery smooth muscle cells (PASMCs) play important roles in the pathological process of hypoxic pulmonary hypertension (HPH). Melatonin possesses anti-inflammatory and antiproliferative properties. However, the effect of melatonin on HPH remains unclear. In this study, adult Sprague-Dawley rats were exposed to intermittent chronic hypoxia for 4 wk to mimic a severe HPH condition. Hemodynamic and pulmonary pathomorphology data showed that chronic hypoxia significantly increased right ventricular systolic pressures (RVSP), weight of the right ventricle/left ventricle plus septum (RV/LV+S) ratio, and median width of pulmonary arterioles. Melatonin attenuated the elevation of RVSP, RV/LV+S, and mitigated the pulmonary vascular structure remodeling. Melatonin also suppressed the hypoxia-induced high expression of proliferating cell nuclear antigen (PCNA), hypoxia-inducible factor-1α (HIF-1α), and nuclear factor-κB (NF-κB). In vitro, melatonin concentration-dependently inhibited the proliferation of PASMCs and the levels of phosphorylation of Akt and extracellular signal-regulated kinases1/2 (ERK1/2) caused by hypoxia. These results suggested that melatonin might potentially prevent HPH via anti-inflammatory and antiproliferative mechanisms.

Intrahepatic Gas Caused by Acute Gastroenteritis: Hepatic Portal Venous Gas or Biliary Tract Gas?

Purpose: Hepatic portal venous gas is not a specific disease and is often only an imaging manifestation in patients with acute abdomen. However, its appearance often indicates serious disease and poor prognosis. It is not difficult to distinguish typical portal venous gas from biliary tract gas on computed tomography because of their relatively different distribution within the liver. But the difference is not absolute. Case Description: An 82-year-old female was admitted to the emergency department due to epigastric pain, nausea and vomiting for 1 day. Intrahepatic gas was found on computed tomography (CT), which was initially diagnosed as portal venous gas, and contrast-enhanced abdominal CT was performed 3 hours after the first plain CT scan and revealed a significant reduction of intrahepatic gas, then diagnosed as biliary tract gas. Two days later, enhanced abdominal CT showed that biliary tract gas had disappeared. Continuous gastrointestinal decompression, anti-infection, rehydration and other treatments were given. After treatment, abdominal pain, nausea, vomiting and other symptoms of the patient were gradually relieved. The patient refused gastroenteroscopy and was discharged after 13 days of hospitalization. Conclusion: Portal venous gas and biliary tract gas may have similar CT findings and be misdiagnosed, and enhanced CT examination is necessary to confirm the diagnosis.

Ischemic stroke due to stylocarotid artery syndrome: a case report and review.

Stylocarotid artery syndrome (SAS) is a rare variant of Eagle's syndrome that may lead to transient ischemic attack or stroke. The underlying pathophysiological mechanism involves compression of the internal carotid artery by an elongated styloid process (ESP), potentially resulting in vascular occlusion or dissection. An ESP exceeding 2.5 cm is deemed elongated, with a length of 3.0 cm considered clinically significant. Although the prevalence of ESP ranges from 4.0% to 7.3%, symptomatic cases are rare; symptoms are present in only approximately 4.0% of individuals with an ESP. Unlike the typical symptoms of Eagle's syndrome, SAS may not cause pharyngeal discomfort, the sensation of a foreign body in the throat, dysphagia, or facial pain. This absence of characteristic symptoms as well as the development of central nervous system symptoms often leads patients to seek care from neurologists instead of otolaryngologists, increasing the likelihood of misdiagnosis or underdiagnosis. We herein report a unique case of ischemic stroke caused by SAS and present a literature review on cases of SAS-associated ischemic stroke published in the past decade. The reporting of this study conforms to the CARE guidelines.

Increased expression of FOXQ1 is a prognostic marker for patients with gastric cancer.

Altered expression of forkhead box Q1 (FOXQ1) is observed in various types of human cancers. However, the clinical significance of FOXQ1 expression in gastric cancer (GC) remains largely unknown. The present study aims to explore the clinicopathological significance and prognostic value of FOXQ1 in GC. FOXQ1 messenger RNA (mRNA) and protein expression were determined by quantitative real-time reverse transcriptase-polymerase chain reaction and Western blot in 20 pairs of fresh frozen GC tissues and corresponding noncancerous tissues. Additionally, FOXQ1 expression was analyzed by immunohistochemistry in 158 clinicopathologically characterized GC cases. The correlation of FOXQ1 expression with patients' survival rate was assessed by Kaplan-Meier and Cox regression. Our results showed that the expression levels of FOXQ1 mRNA and protein in GC tissues were both significantly higher than those in non-cancerous tissues. Our results showed that the high expression of FOXQ1 in GC was related to tumor size (P = 0.026), histological grade (P = 0.021), lymph node involvement (P = 0.002), and tumor-node-metastasis stage (P = 0.028). Kaplan-Meier survival analysis showed that a high expression level of FOXQ1 resulted in a significantly poor prognosis of GC patients. Furthermore, Cox multivariates analysis indicated that FOXQ1 expression level was an independent prognostic factor for the overall survival rate of GC patients. In conclusion, overexpression of FOXQ1 is closely related to progression of GC and might be regarded as an independent predictor of poor prognosis for GC.

Production and quality of Hami melon (Cucumis melo var. reticulatus) and pest population of Thrips palmi in UV-blocking film greenhouses.

BACKGROUND: Ultraviolet (UV) interferes with the vision, flight initiation, dispersal, host, and population dispersion of herbivorous insects. Hence, UV-blocking film has recently been developed as one of the most potential tools to control pests under tropical greenhouse conditions. In this study, we investigated the effects of UV-blocking film on the population dynamics of Thrips palmi Karny and the growth status of Hami melon (Cucumis melo var. reticulatus) in greenhouses. RESULTS: By comparing thrips populations in greenhouses covered with UV-blocking films with those covered with ordinary polyethylene films, we found that the UV-blocking film effectively reduced the thrips population within 1 week and continued to control the population, meanwhile the quality and yield of melon in UV-blocking greenhouses also had a substantial increase. CONCLUSION: The UV-blocking film remarkably inhibited the population growth of thrips and greatly improved the yield of Hami melon cultivated in UV-blocking greenhouse compared with the control greenhouse. Overall, UV-blocking film is a very powerful potential tool for green pest control in the field, enhancing the quality of tropical fruits, and providing a new wind vane for sustainable green agriculture in the future. © 2023 Society of Chemical Industry.

Identification and validation of reference genes for normalization of gene expression analysis using qRT-PCR in Megalurothrips usitatus (thysanoptera: thripidae).

Introduction: Gene expression analysis by reverse transcription quantitative polymerase chain reaction (qRT-PCR) has been widely used in research including insects. The selection of appropriate reference genes is the key to obtaining accurate and reliable results from qRT-PCR. However, studies on the expression stability of reference genes in Megalurothrips usitatus remain lacking. Methods: In this study, qRT-PCR was used to analyze the expression stability of candidate reference genes in M. usitatus. The expression levels of six candidate reference gene transcription of M. usitatus were analyzed. GeNorm, NormFinder, BestKeeper, and ΔCt were used to analyze the expression stability of M. usitatus treated with biological factors (developmental period treatment) and abiotic factors (light, temperature, insecticide treatment, respectively). Comprehensive stability ranking of candidate reference genes was recommended by RefFinder. Results and Discussion: Results showed that ribosomal protein S (RPS) was the most suitable expression in insecticide treatment. Ribosomal protein L (RPL) was the most suitable expression at developmental stage and light treatment, whereas elongation factor was the most suitable expression in temperature treatment. RefFinder was used to comprehensively analyze the above four treatments, and the results showed that RPL and actin (ACT) showed high stability in each treatment. Therefore, this study identified these two genes as reference genes in the qRT-PCR analysis of different treatment conditions of M. usitatus. Ourfindings will be beneficial for improving the accuracy of qRT-PCR analysis for future functional analysis of the target gene expression in M. usitatus.

Functional Characterization of Knockdown Resistance Mutation L1014S in the German Cockroach, Blattella germanica (Linnaeus).

The effectiveness of pyrethroid insecticides is seriously threatened by knockdown resistance (kdr), which is induced in insects by inherited single-nucleotide polymorphisms in the voltage-gated sodium channel (VGSC) gene. VGSC's L1014F substitution results in the classic kdr mutation, which is found in many pest species. Other substitutions of the L1014 locus, such as L1014S, L1014C, L1014W, and L1014H, were also reported. In 2022, a new amino acid substitute L1014S of Blattella germanica was first discovered in China. We modified the BgNav1-1 sodium channel from cockroaches with the L1014S mutation to study how pyrethroid sensitivity and channel gating were affected in Xenopus oocytes. The L1014S mutation reduced the half-maximal activation voltage (V1/2,act) from -19.0 (wild type) to -15.5 mV while maintaining the voltage dependency of activation. Moreover, the voltage dependence of inactivation in the hyperpolarizing shifts from -48.3 (wild type) to -50.9 mV. However, compared with wild type, the mutation L1014S did not cause a significant shift in the half activation voltage (V1/2,act). Notably, the voltage dependency of activation was unaffected greatly by the L1014S mutation. Tail currents are induced by two types of pyrethroids (1 µM): type I (permethrin, bifenthrin) and type II (deltamethrin, λ-cyhalothrin). All four pyrethroids produced tail currents, and significant differences were found in the percentages of channel modifications between variants and wild types. Further computer modeling showed that the L1014S mutation allosterically modifies pyrethroid binding and action on B. germanica VGSC, with some residues playing a critical role in pyrethroid binding. This study elucidated the pyrethroid resistance mechanism of B. germanica and predicted the residues that may confer the risk of pyrethroid resistance, providing a molecular basis for understanding the resistance mechanisms conferred by mutations at the 1014 site in VGSC.

A new protective gel to facilitate ulcer healing in artificial ulcers following oesophageal endoscopic submucosal dissection: a multicentre, randomized trial.

There are significant risks of adverse events following oesophageal endoscopic submucosal dissection (ESD), such as stricture, delayed bleeding and perforation. Therefore, it is necessary to protect artificial ulcers and promote the healing process. The current study was performed to investigate the protective role of a novel gel against oesophageal ESD-associated wounds. This was a multicentre, randomized, single-blind, controlled trial that recruited participants who underwent oesophageal ESD in four hospitals in China. Participants were randomly assigned to the control or experimental group in a 1:1 ratio and the gel was used after ESD in the latter. Masking of the study group allocations was only attempted for participants. The participants were instructed to report any adverse events on post-ESD days 1, 14, and 30. Moreover, repeat endoscopy was performed at the 2-week follow-up to confirm wound healing. Among the 92 recruited patients, 81 completed the study. In the experimental group, the healing rates were significantly higher than those in the control group (83.89 ± 9.51% vs. 73.28 ± 17.81%, P = 0.0013). Participants reported no severe adverse events during the follow-up period. In conclusion, this novel gel could safely, effectively, and conveniently accelerate wound healing following oesophageal ESD. Therefore, we recommend applying this gel in daily clinical practice.