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 evaluate and compare the long-term efficacy of endoscopic radiofrequency ablation(RFA)and photodynamic therapy(PDT)combined with biliary stenting for the treatment of unresectable extrahepatic cholangiocarcinoma.Methods Clinical data of patients with cholangiocarcinoma who received endoscopic RFA or PDT treatment from February 2018 to February 2023 were retrospectively collected.The patients were divided into RFA group(n=30,received endoscopic RFA combined with biliary stent placement)and PDT group(n=20,received PDT combined with biliary stent placement).The frequency of treatment,stent patency time,overall survival time and adverse events incidence were counted.The factors affecting the survival time of patients were analyzed.Results The overall survival time was 14.0(95%CI:11.8～16.2)months in RFA group and 18.0(95%CI:15.4～20.6)months in PDT group,the median patency time of stent was 4.0(95%CI:2.7～5.3)months in RFA group and 3.5(95%CI:2.3～4.7)months in PDT group,the differences were not statistically significant(P>0.05).Multivariate Cox regression analysis showed that the H＾R of patients with≥2 endoscopic RFA or PDT treatments was 2.417,which was a protective factor affecting overall survival(P=0.018),while the H＾R of TNM stage Ⅲ to Ⅳ was 0.300,which was a risk factor affecting the overall survival period(P=0.002).No significant difference was found in clinical success rate(both 100.00%)and adverse events incidence between the two groups[28.13%(9/32)vs.23.81%(5/21)],the difference was not statistically significant(P>0.05).Conclusion The long-term efficacy of endoscopic RFA or PDT combined with biliary stenting in the treatment of unresectable extrahepatic cholangiocarcinoma is comparable,while the sequential treatment of endoscopic RFA or PDT≥2 times can effectively prolong the overall survival of patients.

Objective:To observe the effects of tetrandrine tablets combined with montelukast sodium on respiratory function,inflammatory mediators,serum matrix metalloproteinase-9(MMP-9)and basic fibroblast growth factor(bFGF)levels in pneumoconiosis patients.Methods:120 pneumoconiosis patients who were admitted to our hospital from January 2023 to June 2024 were selected,they were divided into control group(montelukast sodium treatment,n=60)and study group(tetrandrine tablets combined with montelukast sodium treatment,n=60)by random number table method.The pulmonary function indexes[first second forced expiratory volume(FEV1),forced vital capacity(FVC),maximum ventilation volume(MVV)],6-minute walking test(6MWT)disitance,St.George Respiratory Questionnaire(SGRQ)score,Improved Medical Research Council Dyspnea(mMRC)index,and inflammatory mediators[Cystatin C(Cys C),interleukin-2 receptor(IL-2R),C-reactive protein(CRP)]and serum MMP-9,bFGF levels were compared between the two groups.The occurrence of adverse reactions were observed.Results:FEV1,FVC,MVV,6MWT distance in the study group at 6 months after treatment were higher than those in the control group,and SGRQ score,mMRC index,CysC,IL-2R,CRP,MMP-9,bFGF were lower than those in the control group(P＜0.05).There was no significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion:Tetrandrine tablets combined with montelukast sodium in the treatment of pneumoconiosis patients can effectively improve respiratory function,increase exercise endurance,regulate inflammatory mediators and serum MMP-9,bFGF levels,which is safe and reliable.

Objective To evaluate and compare the long-term efficacy of endoscopic radiofrequency ablation(RFA)and photodynamic therapy(PDT)combined with biliary stenting for the treatment of unresectable extrahepatic cholangiocarcinoma.Methods Clinical data of patients with cholangiocarcinoma who received endoscopic RFA or PDT treatment from February 2018 to February 2023 were retrospectively collected.The patients were divided into RFA group(n=30,received endoscopic RFA combined with biliary stent placement)and PDT group(n=20,received PDT combined with biliary stent placement).The frequency of treatment,stent patency time,overall survival time and adverse events incidence were counted.The factors affecting the survival time of patients were analyzed.Results The overall survival time was 14.0(95%CI:11.8～16.2)months in RFA group and 18.0(95%CI:15.4～20.6)months in PDT group,the median patency time of stent was 4.0(95%CI:2.7～5.3)months in RFA group and 3.5(95%CI:2.3～4.7)months in PDT group,the differences were not statistically significant(P>0.05).Multivariate Cox regression analysis showed that the H＾R of patients with≥2 endoscopic RFA or PDT treatments was 2.417,which was a protective factor affecting overall survival(P=0.018),while the H＾R of TNM stage Ⅲ to Ⅳ was 0.300,which was a risk factor affecting the overall survival period(P=0.002).No significant difference was found in clinical success rate(both 100.00%)and adverse events incidence between the two groups[28.13%(9/32)vs.23.81%(5/21)],the difference was not statistically significant(P>0.05).Conclusion The long-term efficacy of endoscopic RFA or PDT combined with biliary stenting in the treatment of unresectable extrahepatic cholangiocarcinoma is comparable,while the sequential treatment of endoscopic RFA or PDT≥2 times can effectively prolong the overall survival of patients.

Objective : To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis. Methods : An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression. Results : Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01). Conclusion Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.

Objective:To observe the effects of tetrandrine tablets combined with montelukast sodium on respiratory function,inflammatory mediators,serum matrix metalloproteinase-9(MMP-9)and basic fibroblast growth factor(bFGF)levels in pneumoconiosis patients.Methods:120 pneumoconiosis patients who were admitted to our hospital from January 2023 to June 2024 were selected,they were divided into control group(montelukast sodium treatment,n=60)and study group(tetrandrine tablets combined with montelukast sodium treatment,n=60)by random number table method.The pulmonary function indexes[first second forced expiratory volume(FEV1),forced vital capacity(FVC),maximum ventilation volume(MVV)],6-minute walking test(6MWT)disitance,St.George Respiratory Questionnaire(SGRQ)score,Improved Medical Research Council Dyspnea(mMRC)index,and inflammatory mediators[Cystatin C(Cys C),interleukin-2 receptor(IL-2R),C-reactive protein(CRP)]and serum MMP-9,bFGF levels were compared between the two groups.The occurrence of adverse reactions were observed.Results:FEV1,FVC,MVV,6MWT distance in the study group at 6 months after treatment were higher than those in the control group,and SGRQ score,mMRC index,CysC,IL-2R,CRP,MMP-9,bFGF were lower than those in the control group(P＜0.05).There was no significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion:Tetrandrine tablets combined with montelukast sodium in the treatment of pneumoconiosis patients can effectively improve respiratory function,increase exercise endurance,regulate inflammatory mediators and serum MMP-9,bFGF levels,which is safe and reliable.

Aim To analyze the differences in plasma biomarkers and metabolic pathways between Atractylodes chinensis and Atractylodes coreana after intervention in spleen deficiency rats, and discuss the spleen strengthening mechanism of the two from a non targeted metabolomics perspective. Methods A spleen deficiency model was established in SD rats using a composite factor method of improper diet, excessive fatigue, and bitter cold diarrhea. To determine the content of gastrointestinal and immunological indicators, UHPLC-QE-MS technology was used, combined with principal component analysis (PC A) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) methods to search for biomarkers in plasma of spleen deficiency rats, and metabolic pathways were induced using the Pathway database. Results After administration of Atractylodes chinensis and Atractylodes coreana, various indicators in plasma of spleen deficiency rats showed varying degrees of regression. Metabolomics analysis showed that Atractylodes chinensis and Atractylodes coreana respectively recalled 70 and 82 plasma differential metabolites. Atractylodes chinensis mainly regulated two metabolic pathways : "Glycine, serine, and threonine metabolism, and "Thiamine metabolism". Atractylodes coreana mainly regulated five metabolic pathways, "Glycine, serine, and threonine metabolism", "Thiamine metabolism, "Pyrimidine metabolism", "Butanoate metabolism", and "Riboflavin metabolism". Conclusions Both Atractylodes chinensis and Atractylodes coreana have certain regulatory effects on spleen deficiency rats, and their mechanism of action may be related to regulating metabolic pathways such as "Glycine, serine, and threonine metabolism, and "Thiamine metabolism"in spleen deficiency.

AIM To establish the HPLC fingerprints for freeze-dried powder of Yinhuo Decoction,and to determine the contents of rehmannitin D,methylophiopogonanone A,verbascoside,protocatechuic acid,catalpol,schisandrol A,schisandrin B,schisandrin A.METHODS The fingerprint esablishment was performed on a 30℃thermostatic Supersil AQ18 column(4.6 mm×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 230 nm.The UPLC-MS/MS content determination was performed on a 30℃thermostatic Alphasil VC-C18 column(2.1 mm×100 mm,2.5 μm),with the mobile phase comprising of acetonitrile-0.1%formic acid flowing at 0.3 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.RESULTS There were 17 common peaks in the fingerprints for 10 batches of freeze-dried powder with the similarities of more than 0.90.and piloside,crystal blue glycoside,deacetylated caryoside schisandrin A were identified.Eight constituents showed good linear relationships within their own ranges(R2＞0.990 6),whose average recoveries were 98.7%-101.1%with the RSDs of 2.0%-5.2%.CONCLUSION The combination of fingerprints and content determination can completely characterize the quality of Yinhuo Decoction reference sample,thus provide a reference for the quality evaluation of its key chemical properties.

AIM To establish the quality standard of the zhujieshen Formula Granules based on standard decoction.METHODS The contents and transfer rates of ginsenoside Ro and chikusetsu saponin Ⅳa in standard decoction and formula granules were determined by HPLC,after which the transfer rates were calculated.The HPLC characteristic chromatograms of standard decoctions were established,after which cluster analysis and principal component analysis were adopted.Then the HPLC characteristic chromatograms of formula granules were established.RESULTS Nine common peaks were found in the HPLC characteristic chromatograms of seventeen batches of standard decoctions with the similarities of more than 0.9(except for S6,S12),which were clustered into two categories,and the accumulative variance contribution rate of three principal components reached 86.7％.The contents of ginsenoside Ro in three batches of formula granules were 83.1-88.6 mg/g,and the transfer rates were 53.1％-55.5％.The contents of chikusetsusaponin Ⅳa were 14.8-15.0 mg/g,and the transfer rates were 47.4％-48.1％.Nine common peaks were found in the HPLC characteristic chromatograms of three batches of formula granules with the similarities of 0.998,0.998 and 0.999,respectively.CONCLUSION This reasonable and reliable method can comprehensively evaluate the quality of Zhujieshen Formula Granules,and provide a reference for the quality control.