Objective To investigate the species of sandflies and the prevalence of Leishmania infections in sandflies from selected areas of northern and northwestern China, so as to provide insights into identification of leishmaniasis vectors and assessment of epidemiological trends of leishmaniasis in China. Methods Sandfly samples were collected from Mentougou District of Beijing Municipality, Xiangning County in Linfen City of Shanxi Province, Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, and Payzawat County of Karamay District of Karamay City, Gaochang District of Turpan City in Xinjiang Uygur Autonomous Region from July 2023 to July 2024. Approximately 100 intact female sandfly samples were randomly selected from each site and the species of sandflies was identified according to morphological characteristics and molecular assays. Female sandflies originating from the same habitat were grouped into pools of 10 individuals. Leishmania infection was detected using polymerase chain reaction (PCR) assay targeting the internal transcribed spacer 1 (ITS-1) gene, and the prevalence of Leishmania infection was calculated in sandflies from different sampling sites using the minimum infection rate (MIR) method. In addition, positive amplicons were sequenced and subjected to phylogenetic analysis. Results A total of 6 155 sandflies were collected from different environments at sampling sites across the six aforementioned regions from July 2023 to July 2024. Phlebotomus chinensis (96.00%) was the dominant sandfly species in Mentougou District, Beijing Municipality, with a small proportion of Ph. sergenti (4.00%), and only Ph. chinensis was found in Xiangning County, Linfen City, Shanxi Province. Ph. wui was the only sandfly species detected in Ejin Banner, Alxa League, Inner Mongolia Autonomous Region, and Payzawat County, Kashgar City, Xinjiang Uygur Autonomous Region, and Ph. caucasicus (97.70%) was the dominant sandfly species in Karamay District, Karamay City, Xinjiang Uygur Autonomous Region, with a small proportion of Ph. wui (2.30%), while Ph. alexandri was the only species in Gaochang District, Turpan City, Xinjiang Uygur Autonomous Region. A total of 40, 60, 34, 18, 18, and 22 pools of sandfly samples were tested from Mentougou District in Beijing Municipality, Xiangning County in Linfen City of Shanxi Province, Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, Payzawat County in Kashgar City, Karamay District in Karamay City, and Gaochang District in Turpan City of Xinjiang Uygur Autonomous Region, respectively. L. infantum was detected in Ph. chinensis samples from Mentougou District in Beijing Municipality, and Xiangning County of Linfen City in Shanxi Province, with MIR of 0.25% to 1.00%, and L. donovani was detected in Ph. wui from Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, and Payzawat County in Kashgar City of Xinjiang Uygur Autonomous Region, with MIR of 0.56% to 0.88%; however, no Leishmania infection was detected in Ph. caucasicus from Karamay District in Karamay City or Ph. alexandri from Gaochang District in Turpan City of Xinjiang Uygur Autonomous Region. Phylogenetic analysis showed that the Leishmania ITS-1 gene sequences obtained from Mentougou District in Beijing Municipality and Xiangning County in Linfen City of Shanxi Province were clustered into the same clade with the reference sequences of L. infantum ITS-1 gene, while the Leishmania ITS-1 gene sequences obtained from Ejin Banner in Alxa League of Inner Mongolia Autonomous Region and Payzawat County in Kashgar City of Xinjiang Uygur Autonomous Region were clustered into the same clade with the reference sequences of L. donovani ITS-1 gene. Conclusions There are variations in sandfly species in selected areas of northern and northwestern China, and variations in the species of Leishmania infecting sandflies. Improved surveillance of sandfly vectors and targeted control strategies with adaptations to geographical features and leishmaniasis vectors are recommended.

BACKGROUND:Prolyl hydroxylase domain 2(PHD2)inhibitors can regulate bone metabolism and relieve osteoporosis in ovariectomized rats.cpd17 is a small molecule oral PHD2 inhibitor newly developed by China Pharmaceutical University.It is effective in the treatment of renal anemia with few side effects,but its effect on bone formation and bone resorption is still unclear. OBJECTIVE:To investigate the effects of cpd17 on mouse osteogenic precursor cells. METHODS:Osteogenic precursor cells were treated with cpd17.Alkaline phosphatase activity and extracellular matrix mineralization were measured,and the expression levels of osteogenesis-and osteoclastogenesis-related markers,as well as PHD2 and hypoxia-inducible factor 1α,were detected.After inhibition of the hypoxia-inducible factor 1α pathway using LW6(a hypoxia-inducible factor 1α pathway inhibitor),alkaline phosphatase activity and extracellular matrix mineralization were detected again,as well as the expression levels of osteogenesis-and osteoclastogenesis-related markers,PHD2 and hypoxia-inducible factor 1α. RESULTS AND CONCLUSION:cpd17 significantly enhanced alkaline phosphatase activity and extracellular matrix mineralization,up-regulated the expression of osteogenesis-related markers,down-regulated the expression of osteoclastogenesis-related markers,up-regulated the expression of hypoxia-inducible factor 1α,down-regulate the expression of PHD2.However,cpd17's effects were significantly attenuated by LW6.To conclude,the PHD2 inhibitor cpd17 promotes osteogenic differentiation and inhibits osteoclastic differentiation through activation of the hypoxia-inducible factor 1α signaling pathway.

Objective To investigate the effects of laparoscopic radiofrequency ablation(LRFA)and percutaneous radiofrequency ablation(PRFA)on anti-tumor immunity,complication rate and recurrence rate in patients with primary liver cancer.Methods A total of 81 patients with primary liver cancer treated in Dazhou Central Hospital from January 2020 to August 2022 were selected and divided into observation group(LRFA,n=42)and control group(PRFA,n=39)according to the treatment plan.Compare the total ablation rate,postoperative complication rate,recurrence rate of the two groups,as well as tumor necrosis factor-α(TNF-α),carbohydrate antigen 199(CA199),interleukin-6(IL-6),Golgi protein 73(GP73),C-reactive protein(CRP),alpha-fetoprotein(AFP)and peripheral blood T lymphocyte subpopulation levels before and after surgery.Results There was no significant difference between the observation group(95.24％)and the control group(92.31％)(P＞0.05).At 1 d postoperatively,IL-6 was(124.63±45.41)pg/ml and(168.28±51.26)pg/ml,CRP was(19.14±5.03)ng/L and(28.26±7.47)ng/L,and TNF-α was(94.32±18.49)pg/ml and(108.41±20.11)pg/ml;at 3 d postoperatively,IL-6 was(92.37±24.11)pg/ml and(105.83±27.45)pg/ml in the observation group and the control group,respectively,CRP was(14.87±4.37)ng/L and(17.25±5.06)ng/L,and TNF-α was(75.41±12.10)pg/ml and(82.64±16.83)pg/ml,which were all higher than that of preoperative period(P＜0.05).At 7 d postoperatively,CD3+in the observation group and control group were(66.27±7.82)％and(65.14±7.63)％,AFP was(156.23±30.27)μg/mland(160.84±32.33)μg/ml,GP73 was(65.21±10.26)μg/L and(67.44±11.03)μg/L,CA199 was(44.89±11.41)U/L and(45.12±13.07)U/L,CD4 was(32.02±6.03)％and(31.53±6.11)％,and CD4+/CD8+was(1.31±0.39)and(1.29±0.37)respectively;at 14 d postoperatively,CD3+was(71.25±6.83)％and(70.89±6.76)％,AFP was(48.52±18.31)μg/ml and(50.11±19.12)μg/ml,GP73 was(48.25±8.46)μg/L and(49.12±10.12)μg/L,CA199 was(19.27±5.16)U/L and(20.07±5.39)U/L,and CD4 was(38.25±7.7)U/L and(20.07±5.39)U/L,respectively,in the observation and control groups.g/L,CA199 was(19.27±5.16)U/L and(20.07±5.39)U/L,CD4 was(38.25±7.45)％and(37.61±7.92)％,and CD4+/CD8+was(1.49±0.42)and(1.47±0.45),respectively,which were higher than that of preoperative period(P＜0.05),but the difference between the two groups was not statistically significant(P＞0.05).The postoperative complication rate of 42.86％and recurrence rate of 2.38％in the observation group were lower than 66.67％and 17.95％in the control group(P＜0.05).The 12-month postoperative survival rate of 97.62％in the observation group was not statistically significant compared with 94.87％in the control group(P＞0.05).Conclusion The efficacy of LRFA and PRFA in the treatment of primary hepatocellular carcinoma is comparable,which can effectively improve the body's anti-tumor immunity and reduce the release of serum tumor markers;however,LRFA has less stressful reaction,reduces the occurrence of postoperative complications,and has a lower recurrence rate,which is especially advantageous in the treatment of hepatocellular carcinoma at special sites.

Objective To observe the value of SimGrid(SG)and S-Enhance(SE)for improving image quality of low-dose X-ray films in children.Methods Data of 344 children in intensive care unit who underwent 410 times bedside X-ray examinations,including 290 times of chest X-ray,51 of abdominal X-ray and 69 of chest and abdominal combined X-ray were enrolled.SG and SE were respectively used for post-processing,and the quality of post-processed images were analyzed.Results Among 410 SG post-processing images,250 images were classified as 2-point,147 as 1-point and 13 as 0-point.SG could significantly improve image quality of children≥1 year and body mass≥10 kg(all P＜0.05),with better ability for displaying bones,trachea,peripheral blood vessels,foreign objects,psoas major muscle and intestinal gas(all P＜0.05).Among 410 SE post-processing images,250 images were classified as 2-point,58 as 1-point and 102 of 0-point.SE could significantly improve image quality of children≥0.5 years and with body mass＞4 kg(all P＜0.05),with better ability for displaying bones,trachea,large blood vessels,peripheral vessels,heart posterior blood vessels and foreign objects(all P＜0.05).Conclusion SG could significantly improve displaying of bones,trachea,peripheral blood vessels,foreign objects,psoas major muscle and intestinal gas in children≥1 year and body mass≥10 kg,while SE could improve displaying of bones,trachea,large blood vessels,peripheral blood vessels,heart posterior blood vessels and foreign objects in children aged≥0.5 years and body mass＞4 kg on low-dose X-ray films.

Objective:To explore the mechanism of osteoclast stimulatory transmembrane protein (OC-STAMP) overexpression on epithelial-mesenchymal transition (EMT) .Methods:In April 2021, mice alveolar type Ⅱ epithelial cells MLE-12 were divided into five groups: overexpression control group (NC group), Ocstamp overexpression group (over- Ocstamp group), Fasudil intervention group (over- Ocstamp+Fasudil group), silence control group (si-NC group), Ocstamp silence group (si- Ocstamp group). The protein expressions of OC-STAMP, epithelial marker protein-E-cadherin (E-cad), interstitial marker protein-α-smooth muscle actin (α-SMA), Ras homolog gene family member A (RhoA), Rho GDP dissociation inhibitor α (Rho GDIα), Rho-associated protein kinase (ROCK), phosphate myosin phosphatase (p-MYPT) were examined by Western blotting and Immunocytochemical staining. The filamentous actin (F-actin) was detected by Phalloidin method. t test was used to compare the relative expression of each protein between the two groups. Results:Western blotting and Immunocytochemical staining showed that compared with the NC group, the expression level of E-cad was down-regulated, while the expression levels of α-SMA, Rho GDIα, RhoA, ROCK, p-MYPT were increased, and F-actin expression was enhanced in the over- Ocstamp group. The differences were statistically significant ( P<0.05). There were no significant differences in E-cad and α-SMA protein expression in si- Ocstamp group compared with si-NC group ( P>0.05). Compared with over- Ocstamp group, the expression level of E-cad protein in over- Ocstamp+Fasudil group was up-regulated, the expression levels of α-SMA, Rho GDIα, RhoA, ROCK and p-MYPT protein were decreased, and F-actin expression was weakened, with statistical significance ( P<0.05) . Conclusion:OC-STAMP overexpression in alveolar type Ⅱ epithelial cells may induce actin cytoskeleton remodeling through activation of Rho GDIα/RhoA/ROCK signaling pathway, thus promoting EMT.

Objective:To explore the mechanism of osteoclast stimulatory transmembrane protein (OC-STAMP) overexpression on epithelial-mesenchymal transition (EMT) .Methods:In April 2021, mice alveolar type Ⅱ epithelial cells MLE-12 were divided into five groups: overexpression control group (NC group), Ocstamp overexpression group (over- Ocstamp group), Fasudil intervention group (over- Ocstamp+Fasudil group), silence control group (si-NC group), Ocstamp silence group (si- Ocstamp group). The protein expressions of OC-STAMP, epithelial marker protein-E-cadherin (E-cad), interstitial marker protein-α-smooth muscle actin (α-SMA), Ras homolog gene family member A (RhoA), Rho GDP dissociation inhibitor α (Rho GDIα), Rho-associated protein kinase (ROCK), phosphate myosin phosphatase (p-MYPT) were examined by Western blotting and Immunocytochemical staining. The filamentous actin (F-actin) was detected by Phalloidin method. t test was used to compare the relative expression of each protein between the two groups. Results:Western blotting and Immunocytochemical staining showed that compared with the NC group, the expression level of E-cad was down-regulated, while the expression levels of α-SMA, Rho GDIα, RhoA, ROCK, p-MYPT were increased, and F-actin expression was enhanced in the over- Ocstamp group. The differences were statistically significant ( P<0.05). There were no significant differences in E-cad and α-SMA protein expression in si- Ocstamp group compared with si-NC group ( P>0.05). Compared with over- Ocstamp group, the expression level of E-cad protein in over- Ocstamp+Fasudil group was up-regulated, the expression levels of α-SMA, Rho GDIα, RhoA, ROCK and p-MYPT protein were decreased, and F-actin expression was weakened, with statistical significance ( P<0.05) . Conclusion:OC-STAMP overexpression in alveolar type Ⅱ epithelial cells may induce actin cytoskeleton remodeling through activation of Rho GDIα/RhoA/ROCK signaling pathway, thus promoting EMT.

The obstruction of post-insulin receptor signaling is the main mechanism of insulin-resistant diabetes. Progestin and adipoQ receptor 3 (PAQR3), a key regulator of inflammation and metabolism, can negatively regulate the PI3K/AKT signaling pathway. Here, we report that gentiopicroside (GPS), the main bioactive secoiridoid glycoside of Gentiana manshurica Kitagawa, decreased lipid synthesis and increased glucose utilization in palmitic acid (PA) treated HepG2 cells. Additionally, GPS improved glycolipid metabolism in streptozotocin (STZ) treated high-fat diet (HFD)-induced diabetic mice. Our findings revealed that GPS promoted the activation of the PI3K/AKT axis by facilitating DNA-binding protein 2 (DDB2)-mediated PAQR3 ubiquitinated degradation. Moreover, results of surface plasmon resonance (SPR), microscale thermophoresis (MST) and thermal shift assay (TSA) indicated that GPS directly binds to PAQR3. Results of molecular docking and cellular thermal shift assay (CETSA) revealed that GPS directly bound to the amino acids of the PAQR3 NH₂-terminus including Leu40, Asp42, Glu69, Tyr125 and Ser129, and spatially inhibited the interaction between PAQR3 and the PI3K catalytic subunit (P110α) to restore the PI3K/AKT signaling pathway. In summary, our study identified GPS, which inhibits PAQR3 expression and directly targets PAQR3 to restore insulin signaling pathway, as a potential drug candidate for the treatment of diabetes.

Objective:To screen the serum exosomal microRNAs differentially expressed in early pancreatic cancer patients and evaluate the diagnostic value of exosomal hsa-let-7f-5p.Methods:From January 2019 to January 2020, 19 patients with early pancreatic cancer (early pancreatic cancer group) and 16 patients with chronic mass-forming pancreatitis (pancreatitis group) were selected from Affiliated Hospital of Nanjing University of Chinese Medicine who underwent surgery and were confirmed by pathology. Serum samples of the two groups of patients were collected. At the same time, serum samples of 19 healthy volunteers were selected as the normal control group. The exoEasy Maxi Kit was used to isolate serum exosomes. The structural characteristics of exosomes were observed by transmission electron microscopy (TEM). The particle size of exosomes was observd by nanoparticle tracking analysis. CD 63 and CD 81, the specific protein marker on the surface of exosomes, were identified by western blotting. The total RNA of exosomes was extracted by the miRNeasy Serum/Plasma Kit, and a small RNA library was constructed after quality inspection. With reference to the small RNA database, the differentially expressed exosomal microRNAs in early pancreatic cancer group, pancreatitis group and normal control group were filtered out. The miRNA candidates were validated by quantitative polymerase chain reaction (qPCR) and different expressions of them were analyzed. The role of target genes and metabolic pathways of candidate miRNAs in the occurrence and development of early pancreatic cancer were analyzed by gene ontology (GO) and Kyoto Encyclopeda of Genes and Genomes(KEGG) enrichment pathway. Results:The isolated serum exosomes can be seen to have cup-like vesicle with the double lipid layer by TEM. The main peak of the particle size of target exosomes was about 150 nm. The expression of exosome specific protein markers CD 63 and CD 81 was positive. Comparing the expression of miRNAs among early pancreatic cancer group, pancreatitis group and normal control group, the specific tumor marker exosomal hsa-let-7f-5p was screened out in this study, and its expression in early pancreatic cancer group was significantly higher than that in pancreatitis group and normal control group (both P values <0.05). Receiver operating characteristic curve analysis (ROC) showed that the area under curve (AUC) of exosomal hsa-let-7f-5p to distinguish pancreatic cancer from pancreatitis was 0.843 (95% CI 0.640-1.000). The sensitivity and specificity were 100% and 81.82% respectively. The AUC for distinguishing pancreatic cancer from normal controls was 1.000 (95% CI 1.000-1.000), and both sensitivity and specificity were 100%. The diagnostic efficiency of exosomal hsa-let-7f-5p was equivalent to that of CA19-9 ( P>0.05). The GO analysis results showed that target genes of exosomal hsa-let-7f-5p were mainly involved in complement activation lectin pathway in biological processes, and the proteins expressed by target genes were mainly distributed in cilium, and molecules mostly functioned by combining with nitric-oxide synthase. The KEGG pathway enrichment analysis showed that the target genes were closely related to MAPK signaling pathway. Conclusions:Serum exosomal hsa-let-7f-5p has the potential to be a diagnostic biomarker for early pancreatic cancer.

Objective:To explore the prognostic factors of patients with Vibrio vulnificus sepsis. Methods:The clinical data of 67 patients with Vibrio vulnificus sepsis from January 2008 to December 2019 in the First Affiliated Hospital of Wenzhou Medical University were retrospectively analyzed. Univariate analysis was used to compare the differences in general information, clinical manifestations, admission laboratory indicators, antibiotics and surgery between the death group and the cured group. Then the factors with significant difference in univariate analysis were included in multivariate analysis, and the factors of prognosis were obtained. Results:Univariate analysis showed that there were significant difference in liver disease, admission with hypotension shock, multiple limb injuries; admission leukocytes, platelets, pH value, albumin, lactic acid, aspartate aminotransferase, creatinine, procalcitonin, creatine kinase, activated partial thromboplastin time, prothrombin time between the death group and the cured group (all P <0.05). Multivariate analysis showed that admission lactate ( OR=0.628, 95% CI: 0.461-0.855, P=0.003), albumin ( OR=1.330, 95% CI:1.062-1.667, P=0.013), creatine kinase ( OR=0.999, 95% CI: 0.998-1.000, P=0.016) and admission surgery time ( OR=0.118, 95% CI: 0.015-0.938, P=0.043) were risk factors of the prognosis. Patients with high lactate, creatine kinase and low albumin at admission indicate poor prognosis; patients with admission surgery time≤ 12 h have better prognosis. Conclusion:For the treatment of patients with Vibrio vulnificus sepsis, medical staff should dynamically evaluate these prognostic factors in the early stage, and early surgical treatment should be adopted to improve the prognosis of patients.

Objective:To investigate the status and influencing factors of health-promoting lifestyle in young and middle-aged people with high normal blood pressure, so as to provide the basis for primary prevention about hypertension.Methods:From July to December 2020, a convenience sampling method was used to select 280 cases of young and middle-aged people with high normal blood pressure in the First Affiliated Hospital of Wenzhou Medical University were investigated by self-administered general information questionnaire,and Health Promotion Lifestyle Profile Ⅱ(HPLP-Ⅱ).Results:The HPLP-Ⅱ total score in the young and middle-aged with high normal blood pressure was 146.79 ± 29.57. Multiple linear regression analysis showed that different occupation,income, family history of hypertension and learning-willingness were the influencing factors of their health-promoting lifestyle( P<0.05). Conclusions:The health-promoting lifestyle among the young and middle-aged with high normal blood pressure is at a medium level, health care providers should strengthen guidance to improve their health-promoting lifestyle through health education and other ways for individuals without stable employment, poor income level, no family history of hypertension and lack in willingness to learn.