Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective:To investigate the correlation between the expression level of glypican-6 (GPC6) in gliomas and the prognosis of patients, as well as the effect of GPC6 on the proliferation of glioma cells in vitro.Methods:The transcriptome sequencing (RNA-seq) data of GPC6 gene and clinical data of glioma patients were downloaded from The Cancer Genome Atlas (TCGA) database. A total of 667 patients were enrolled, and the median FPKM value of GPC6 RNA-seq was used to distinguish the low and high expression of GPC6 gene. Kaplan-Meier method was used to analyze the overall survival (OS) of GPC6 high and low expression groups in 664 patients with survival data, and log-rank test was used for inter group comparison. The postoperative tumor tissue chips were retrospectively collected from glioma patients who underwent surgery from February 2008 to October 2011, immunohistochemistry (IHC) was used to detect the expression of GPC6 protein in 147 samples, and semi-quantitative scoring ≥ 4 points or not was used to distinguish the high and low expression of GPC6 protein based on the proportion of positive cells and staining intensity. The distributions of patients with high and low expression of GPC6 protein in tumor tissues were compared between different clinical pathological features in 147 patients, and Kaplan-Meier method was used to analyze the overall survival and disease-free survival (DFS) of patients in the GPC6 protein high and low expression groups; the risk factors that affect poor OS and DFS in patients were analyzed using univariate and multivariate Cox proportional hazards models. The human glioma U251 cells were infected with lentivirus packaged with GPC6 small interfering RNA and lentivirus packaged with unrelated control sequences, namely ShGPC6 group and ShCtrl group, respectively; green fluorescent cells were counted daily and the cell proliferation fold was calculated for 5 days, and the cell proliferation curve was plotted.Results:In TCGA database, the median FPKM value of 667 glioma patients was 225.66. The median FPKM value of GPC6 gene in 515 World Health Organization (WHO) classification low-grade glioma patients was lower than that in 152 high-grade glioma patients (201.10 vs. 347.92), and the difference was statistically significant ( Z = 5.36, P < 0.001); the OS of patients in GPC6 low expression group (332 cases) was better than that in GPC6 high expression group (332 cases), and the difference was statistically significant ( P < 0.001). IHC staining of the tissue chip showed that GPC6 protein was mainly expressed in the cytoplasm and nucleus of glioma cells; high expression of GPC6 protein accounted for 31.3% (46/147) of patients. Among patients who died, relapsed, had high grading, had survival time ≤ 84 months, and had positive epidermal growth factor receptor, the proportion of patients with GPC6 protein high expression was relatively higher, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the proportions of GPC6 protein high expression patients among patients with different genders, age ≤ 43 years old, Ki-67 positivity, and programmed death receptor ligand 1 positivity (all P > 0.05); compared to the GPC6 low expression group (101 cases), the GPC6 protein high expression group (46 cases) had worse overall survival (median OS time: 57.0 months vs. not reached) and DFS (median DFS time: 33.0 months vs. not reached), and the differences were statistically significant (both P < 0.001). Multivariate Cox regression analysis showed that high expression of GPC6 protein (high expression vs. low expression, HR = 1.86, 95% CI: 1.04-3.31, P = 0.036) was the independent risk factor for poor OS in glioma patients, but the expression level of GPC6 protein was not the independent influencing factor for DFS (high expression vs. low expression, HR = 1.55, 95% CI: 0.95-2.53, P = 0.077). After infecting U251 cells with recombinant lentivirus for 3 days, reverse transcription polymerase chain reaction detection showed that the relative expression level of GPC6 mRNA in the ShGPC6 group was lower than that in the ShCtrl group, and the difference was statistically significant ( P < 0.001), the knockdown efficiency of GPC6 reached 79.32%; under the fluorescence microscope, the number and intensity of fluorescence positive U251 cells in the ShCtrl group increased day by day after infection with recombinant lentivirus, while there was no significant change in the ShGPC6 group; starting from the second day, the proliferation rate of U251 cells in the ShCtrl group was higher than that in the ShGPC6 group, and the differences were statistically significant (all P < 0.01). Conclusions:High GPC6 expression level may associate with the high risk of death in glioma patients, and high GPC6 expression may promote the proliferation of glioma cells.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

AIM To investigate the protective effects of the aqueous extract of Bulbophyllum kwangtungense Schltr.on D-galactose(D-gal),an oxidative stress-induced liver damage in rats.METHODS 60 Male SD rats were randomly divided into the normal control group,the model group,the resveratrol group(20 mg/kg),and low-dose,medium-dose,and high-dose B.kwangtungense aqueous extract groups(750,1 500 and 3 000 mg/kg).Except for those of the normal control group,the rats of the other groups were subcutaneously injected with D-gal(200 mg/kg)in the back of neck to establish a liver damage model.Simultaneously,each group underwent the corresponding drug administration by gavage(10 mL/kg)once daily for 42 days.After the treatment period,the rats had their peripheral blood and liver tissues collected for the calculation of the liver index;their serum AST and ALT activities,hepatic SOD and GSH-Px activities,and MDA levels measured;their hepatic pathological changes observed using HE staining;their hepatic mRNA expressions of Nrf2,NQO-1 and HO-1 detected by RT-qPCR;and their hepatic protein expressions of Nrf2,NQO-1,HO-1,Bax,Bcl-2 and cleaved Caspase-3 measured by Western blot.RESULTS Compared with the model group,the groups intervened with medium-dose or high-dose B.kwangtungense aqueous extract showed increased liver index level(P＜0.01);decreased serum AST and ALT activities(P＜0.01);increased hepatic SOD and GSH-Px activities(P＜0.05,P＜0.01);reduced MDA levels(P＜0.01);alleviated pathological liver damage and increased hepatic mRNA and protein expressions of Nrf2,NQO-1 and HO-1(P＜0.05,P＜0.01);and reduced protein expressions of cleaved Caspase-3 and the Bax/Bcl-2 ratio as well(P＜0.05,P＜0.01).CONCLUSION The aqueous extract of B.kwangtungense is protective of rats with D-gal-induced liver injury,and the underlying mechanism may associate with the inhibition of oxidative stress and the suppression of hepatocellular apoptosis mediated by the activation of the Nrf2/HO-1/NQO-1 signaling pathway.

Objective:To investigate the correlation between the expression level of glypican-6 (GPC6) in gliomas and the prognosis of patients, as well as the effect of GPC6 on the proliferation of glioma cells in vitro.Methods:The transcriptome sequencing (RNA-seq) data of GPC6 gene and clinical data of glioma patients were downloaded from The Cancer Genome Atlas (TCGA) database. A total of 667 patients were enrolled, and the median FPKM value of GPC6 RNA-seq was used to distinguish the low and high expression of GPC6 gene. Kaplan-Meier method was used to analyze the overall survival (OS) of GPC6 high and low expression groups in 664 patients with survival data, and log-rank test was used for inter group comparison. The postoperative tumor tissue chips were retrospectively collected from glioma patients who underwent surgery from February 2008 to October 2011, immunohistochemistry (IHC) was used to detect the expression of GPC6 protein in 147 samples, and semi-quantitative scoring ≥ 4 points or not was used to distinguish the high and low expression of GPC6 protein based on the proportion of positive cells and staining intensity. The distributions of patients with high and low expression of GPC6 protein in tumor tissues were compared between different clinical pathological features in 147 patients, and Kaplan-Meier method was used to analyze the overall survival and disease-free survival (DFS) of patients in the GPC6 protein high and low expression groups; the risk factors that affect poor OS and DFS in patients were analyzed using univariate and multivariate Cox proportional hazards models. The human glioma U251 cells were infected with lentivirus packaged with GPC6 small interfering RNA and lentivirus packaged with unrelated control sequences, namely ShGPC6 group and ShCtrl group, respectively; green fluorescent cells were counted daily and the cell proliferation fold was calculated for 5 days, and the cell proliferation curve was plotted.Results:In TCGA database, the median FPKM value of 667 glioma patients was 225.66. The median FPKM value of GPC6 gene in 515 World Health Organization (WHO) classification low-grade glioma patients was lower than that in 152 high-grade glioma patients (201.10 vs. 347.92), and the difference was statistically significant ( Z = 5.36, P < 0.001); the OS of patients in GPC6 low expression group (332 cases) was better than that in GPC6 high expression group (332 cases), and the difference was statistically significant ( P < 0.001). IHC staining of the tissue chip showed that GPC6 protein was mainly expressed in the cytoplasm and nucleus of glioma cells; high expression of GPC6 protein accounted for 31.3% (46/147) of patients. Among patients who died, relapsed, had high grading, had survival time ≤ 84 months, and had positive epidermal growth factor receptor, the proportion of patients with GPC6 protein high expression was relatively higher, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the proportions of GPC6 protein high expression patients among patients with different genders, age ≤ 43 years old, Ki-67 positivity, and programmed death receptor ligand 1 positivity (all P > 0.05); compared to the GPC6 low expression group (101 cases), the GPC6 protein high expression group (46 cases) had worse overall survival (median OS time: 57.0 months vs. not reached) and DFS (median DFS time: 33.0 months vs. not reached), and the differences were statistically significant (both P < 0.001). Multivariate Cox regression analysis showed that high expression of GPC6 protein (high expression vs. low expression, HR = 1.86, 95% CI: 1.04-3.31, P = 0.036) was the independent risk factor for poor OS in glioma patients, but the expression level of GPC6 protein was not the independent influencing factor for DFS (high expression vs. low expression, HR = 1.55, 95% CI: 0.95-2.53, P = 0.077). After infecting U251 cells with recombinant lentivirus for 3 days, reverse transcription polymerase chain reaction detection showed that the relative expression level of GPC6 mRNA in the ShGPC6 group was lower than that in the ShCtrl group, and the difference was statistically significant ( P < 0.001), the knockdown efficiency of GPC6 reached 79.32%; under the fluorescence microscope, the number and intensity of fluorescence positive U251 cells in the ShCtrl group increased day by day after infection with recombinant lentivirus, while there was no significant change in the ShGPC6 group; starting from the second day, the proliferation rate of U251 cells in the ShCtrl group was higher than that in the ShGPC6 group, and the differences were statistically significant (all P < 0.01). Conclusions:High GPC6 expression level may associate with the high risk of death in glioma patients, and high GPC6 expression may promote the proliferation of glioma cells.

This study aimed to compare the ameliorative effects of Lycii Fructus and Chrysanthemi Flos at different ratios on retinal damage in mice and to elucidate the underlying mechanisms. A retinal injury model was established by intraperitoneal injection of sodium iodate(NaIO_3) solution. The mice were divided into the following groups: blank group, model group, positive drug(AREDS 2) group, low-and high-dose groups of Lycii Fructus and Chrysanthemi Flos at 1∶1, low-and high-dose groups at 3∶1, and low-and high-dose groups at 1∶3. Administration was carried out 15 days after modeling. The visual acuity of the mice was assessed using the black-and-white box test. The fundus was observed using an optical coherence tomography device, and retinal thickness was measured. HE staining was used to observe the morphology and pathological changes of the retina. The levels of oxidative factors in serum and ocular tissues were measured using assay kits. The levels of inflammatory factors in serum and ocular tissues were detected by enzyme-linked immunosorbent assay(ELISA), and the expression of Nrf2, HO-1, and NF-κB proteins in ocular tissues was analyzed by Western blot. The results showed that after administration of Lycii Fructus and Chrysanthemi Flos at different ratios, the model group showed improved retinal thinning and disordered arrangement of retinal layers, elevated content of SOD and GSH in the serum and ocular tissues, and reduced levels of MDA, TNF-α, IL-1β, and IL-6. Lycii Fructus and Chrysanthemi Flos at 1∶1 and 1∶3 showed better improvement effects. The combination significantly upregulated the expression levels of Nrf2 and HO-1 and downregulated the expression of NF-κB p65. These results indicate that Lycii Fructus and Chrysanthemi Flos at different ratios can improve retinal damage, reduce oxidative stress, and alleviate inflammation in both the body and ocular tissues of mice. The mechanism may be related to the regulation of the Nrf2/HO-1 and NF-κB signaling pathways in ocular tissues. These findings provide a theoretical basis for the clinical application of Lycii Fructus and Chrysanthemi Flos in the treatment of dry age-related macular degeneration.
Animals ; Mice ; Retina/injuries* ; Male ; Lycium/chemistry* ; Drugs, Chinese Herbal/administration & dosage* ; Chrysanthemum/chemistry* ; NF-kappa B/genetics* ; Humans ; Retinal Diseases/metabolism* ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress/drug effects* ; Flowers/chemistry* ; Heme Oxygenase-1/genetics*

The holistic view is the key in the study of traditional Chinese medicine(TCM). The component structure theory is based on the holistic view to investigate the correlation between material basis and efficiency, which enriches the holistic "component-effect" research of TCM. Gintonin is a newly isolated non-saponin component of ginseng. Compared to ginsenosides, gintonin has many different pharmacological activities, and it provides new knowledge for the holistic research of ginseng. Thus, taking the discovery of gintonin from ginseng as an example, this paper explored the linkage between ginsenosides and gintonin from the perspective of "component-effect" correlations and systematically sorted out the similarities and differences between them in terms of structural characteristics, modes of action, and pharmacological activities. Starting from the collaborative interaction of TCM compounds, the study discussed the application and value of the holistic view in TCM "component-effect" research in the light of the component structure theory to provide new thoughts for the development of modern TCM research.
Panax/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Medicine, Chinese Traditional ; Humans ; Ginsenosides/pharmacology* ; Animals

Jiuwei Xifeng Granules have become a Chinese patent medicine in the market. Because the formula contains Os Draconis, a top-level protected fossil of ancient organisms, the formula was to be improved by replacing Os Draconis with Ostreae Concha. To evaluate whether the improved formula has the same effectiveness and safety as the original formula, a randomized, double-blind, parallel-controlled, equivalence clinical trial was conducted. This study enrolled 288 tic disorder(TD) of children and assigned them into two groups in 1∶1. The treatment group and control group took the modified formula and original formula, respectively. The treatment lasted for 6 weeks, and follow-up visits were conducted at weeks 2, 4, and 6. The primary efficacy endpoint was the difference in Yale global tic severity scale(YGTSS)-total tic severity(TTS) score from baseline after 6 weeks of treatment. The results showed that after 6 weeks of treatment, the declines in YGTSS-TSS score showed no statistically significant difference between the two groups. The difference in YGTSS-TSS score(treatment group-control group) and the 95%CI of the full analysis set(FAS) were-0.17[-1.42, 1.08] and those of per-protocol set(PPS) were 0.29[-0.97, 1.56], which were within the equivalence boundary [-3, 3]. The equivalence test was therefore concluded. The two groups showed no significant differences in the secondary efficacy endpoints of effective rate for TD, total score and factor scores of YGTSS, clinical global impressions-severity(CGI-S) score, traditional Chinese medicine(TCM) response rate, or symptom disappearance rate, and thus a complete evidence chain with the primary outcome was formed. A total of 6 adverse reactions were reported, including 4(2.82%) cases in the treatment group and 2(1.41%) cases in the control group, which showed no statistically significant difference between the two groups. No serious suspected unexpected adverse reactions were reported, and no laboratory test results indicated serious clinically significant abnormalities. The results support the replacement of Os Draconis by Ostreae Concha in the original formula, and the efficacy and safety of the modified formula are consistent with those of the original formula.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Double-Blind Method ; Drugs, Chinese Herbal/therapeutic use* ; Tic Disorders/drug therapy* ; Treatment Outcome

This study identified 8 members including NjBIN2 of the GSK3 family in Nardostachys jatamansi by bioinformatics analysis. Moreover, the phylogenetic tree revealed that the GKS3 family members of N. jatamansi had a close relationship with those of Arabidopsis. RT-qPCR results showed that NjBIN2 presented a tissue-specific expression pattern with the highest expression in roots, suggesting that NjBIN2 played a role in root growth and development. In addition, the application of epibrassinolide or the brassinosteroid(BR) synthesis inhibitor(brassinazole) altered the expression pattern of NjBIN2 and influenced the photomorphogenesis(cotyledon opening) and root development of N. jatamansi, which provided direct evidence about the functions of NjBIN2. In conclusion, this study highlights the roles of BIN2 in regulating the growth and development of N. jatamansi by analyzing the expression pattern and biological function of NjBIN2. It not only enriches the understanding about the regulatory mechanism of the growth and development of N. jatamansi but also provides a theoretical basis and potential gene targets for molecular breeding of N. jatamansi with improved quality in the future.
Brassinosteroids/metabolism* ; Steroids, Heterocyclic/metabolism* ; Gene Expression Regulation, Plant/drug effects* ; Plant Proteins/metabolism* ; Phylogeny ; Nardostachys/metabolism* ; Plant Growth Regulators/pharmacology* ; Plant Roots/drug effects*