Objective This study reports on the obligatory parasitism of water mites Arrenurus sp. on Anopheles messeae at the Manzhouli Port, Inner Mongolia, China. Methods Duing July 2024, a survey on the mosquito diversity was conducted at the Manzhouli Port. Captured mosquitoes and their ectoparasites were identified to species level. Results A total of 1840 adult mosquitoes were collected, representing species from three genera: Culex(Cx. modestus, Cx. pipiens pallens), Aedes(Ae. dorsalis, Ae. flavidorsalis, Ae. flavescens), and Anopheles (An. messeae). Among all the mosqutioes specimens,3 out of 150 captured An. messeae were found to carry ectoparasitic mites, with number of 2,4,27 mites separately. Morphological and molecular identification reached the same result as water mites(Hydrachnidiae, Hydracrina). COI gene sequence showed 94% similarity with the closest species Arrenurus truncatellus. Conlusions Literature review suggests water mites are host-specific parasitism of mosquito species and herein with the first record of Arrenurus sp. parasiting on An. Messeae in the most high-latitude region globally.

Objective To evaluate the diagnostic efficacy of different pathogen detection methods in active tuberculosis (ATB) of kidney transplant recipients (KTRs) and analyze their diagnostic value in different types of clinical specimens. Methods A retrospective observational study was conducted, including adult KTRs with suspected ATB from the Eighth Medical Center of Chinese People's Liberation Army General Hospital from January 2020 to January 2025. Baseline data of the recipients were collected and analyzed, and the diagnostic criteria were based on a composite reference standard (including clinical diagnosis and bacteriological confirmation). The single-detection efficacy of acid-fast staining smear, mycobacterium tuberculosis culture, Xpert test for the detection of mycobacterium tuberculosis nucleic acid and resistance to rifampicin (Xpert MTB/RIF), as well as tuberculosis or non-tuberculosis mycobacterial DNA detection was compared. The differences in sensitivity among different methods were analyzed. Results A total of 95 KTRs with suspected ATB and 136 specimens were included. And 52 cases were diagnosed as ATB, among which 48 cases were confirmed by bacteriological means (92%). The sensitivity of Xpert MTB/RIF and DNA detection (0.55 and 0.52, respectively) was significantly higher than that of acid-fast staining smear (0.30) and mycobacterium tuberculosis culture (0.35) (P < 0.01). There was a statistically significant difference in the bacteriological diagnosis rate among different specimen types (P = 0.035). The detection sensitivity of each method was the highest in pus, puncture tissue and bronchoalveolar lavage fluid, followed by sputum and pleural and abdominal effusions, and the lowest in cerebrospinal fluid. Conclusions The sensitivity of nucleic acid amplification technology for immunosuppressed KTRs is slightly reduced, but it should still be recommended as the preferred initial screening tool. The comprehensive strategy of "multi-site sampling (prior to obtaining high bacterial load specimens) combined with multiple method detection" may improve the diagnostic efficacy.

Ten ursolic acid derivatives were designed from the lead compound ursolic acid by introducing 1,2,3-triazole at C-3 and C-28. The target compounds were synthesized and characterized by ¹H NMR and ¹³C NMR. MTT assay was used to study the antitumor activity of these compounds in human cancer cells with high expression (MCF-7 and SGC-7901). The results showed that the antitumor activity of all compounds on MCF-7 and SGC-7901 tumor cells was significantly higher than that of ursolic acid. The compound II₄ exhibited significant antitumor activity which was equivalent to the positive control drug nilotinib, molecular docking showed that the compound II₄ have high binding ability with c-Kit, which deserves further research.

Objective:To investigate the completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients in the national multicenter real-world database.Methods:The prospective real-world study was conducted. The clinicopathological data of 1 074 patients who underwent surgical treatment for mid and low rectal cancer in 47 national medical institutions, including Peking Union Medical College Hospital et al, from May 12,2023 to May 11,2024 were collected. Observation indicators: (1) clinical characteristics of patients with mid and low rectal cancer; (2) initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer; (3) initial imaging evaluation of patients with mid and low rectal cancer; (4) imaging evaluation after neoadjuvant therapy for patients with mid and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3). Count data were described as absoluter numbers and/or percentages. Results:(1) Clinical characteristics of patients with mid and low rectal cancer. Of the 1 074 patients, there were 713 males and 361 females, aged 63(56,70)years. The body mass index of 1 074 patients was 24(21,26)kg/m 2.For American Society of Anesthesiologists classification, there were 147 cases of stage Ⅰ, 641 cases of stage Ⅱ, 157 cases of stage Ⅲ, 2 cases of stage Ⅳ, and there were 127 cases missing data. (2) Initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer. Of the 1 074 patients, there were 787 cases (73.28%) undergoing complete colonoscopy, and there were only 197 cases (18.34%) undergoing immunohistochemical evaluation of all four mismatch repair proteins. (3) Initial imaging evaluation of patients with mid and low rectal cancer. Of the 1 074 patients, there were 842(78.40%) patients completing magnetic resonance imaging (MRI) or ultrasound evaluation, and there were 914(85.10%) patients completing chest, abdomen, and pelvis enhanced computed tomography (CT) evaluation. In the 149 patients completing rectal ultrasound evaluation, there were 122 cases (81.88%) comple-ting T staging evaluation, and there were 81 cases (54.36%) completing N staging evaluation. In the 808 patients completing rectal MRI evaluation, there were 708 cases (87.62%) completing T staging evaluation, and there were 590 cases (73.02%) completing N staging evaluation. (4) Imaging evalua-tion after neoadjuvant therapy for patients with mid and low rectal cancer. Of the 388 patients with neoadjuvant therapy, there were 332 patients (85.57%) completing MRI or ultrasound evaluation, and there were 327 patients (84.28%) completing chest, abdomen, and pelvis enhanced CT evalua-tion. In the 70 patients completing rectal ultrasound evaluation, there were 65 cases (92.86%) com-pleting T staging evaluation, and there were 49 cases (70.00%) completing N staging evaluation. In the 327 patients completing rectal MRI evaluation, there were 246 cases (75.23%) completing T staging, and there were 228 cases (69.72%) completing N staging evaluation. Conclusion:The com-pletion rate of tumor imaging evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients on a national scale is relatively good.

Aim To explore the role and mechanism of epimedokoreanin B(EKB)in HepaRG cell pyroptosis through endoplasmic reticulum stress and NLRP1-me-diated pyroptosis pathway.Methods The effect of EKB on the viability of HepaRG cells at different con-centrations was determined by MTT assay,and the cell growth status was recorded by Incucyte.Four groups of HepaRG cells were set up.The control group was cul-tured with complete medium for 24 h;the drug admin-istration group was cultured with three concentration gradients of 6.25,12.5 and 25 μmol·L-1 of EKB for 24 h.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins and pyroptosis-related proteins in the cells of each group.Results HepaRG cells showed cytotoxicity at a concentration of 6.25 μmol·L-1 for 24 h,and the half maximal inhibitory concentration(IC50)was 12.41 μmol·L-1.Incucyte recordings of the cell growth status showed that the cells in the control group were in good growth status,and the vesicular pyropto-sis cells appeared in the different concentrations of EKB and the cells swelled and ruptured after 24 h.Western blot showed that the protein expression levels of endoplasmic reticulum stress-related proteins pERK,eIF-2α,ATF-4,GRP78,and CHOP significantly in-creased in HepaRG cells at 25 μmol·L-1 of EKB compared with the control group.The proteins of the classical pathway of cellular pyroptosis mediated by NLRP1,caspase-1,cleaved caspase-1,GSDMD,GS-DMD-N significantly increased in HepaRG cells.Con-clusion EKB administration induces HepaRG cell py-roptosis,and EKB activates HepaRG cells to undergo endoplasmic reticulum stress and activates the NLRP1/caspase-1/GSDMD-mediated pyroptosis pathway.

Objective:To investigate the effects of exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) during osteogenic differentiation on the polarization of Raw264.7 macrophages and to elucidate the underlying mechanisms.Methods:PBS, uninduced BMSCs conditioned medium (CM), and osteogenic induction BMSCs CM for 7 d, 14 d, and 21 d were respectively added to Raw264.7 macrophages. After 48 h of treatment, Western blotting was used to detect and compare the expression of M1 markers of macrophages [inducible nitric oxide synthase (iNOS) and CD86] and M2 markers of macrophages [arginase-1 (ARG-1) and CD163] in each group. In addition, Raw264.7 macrophages were divided into three groups: the PBS group (only PBS added), the BMSCs-exo group (exosomes derived from uninduced BMSCs were added), and 7D-BMSCs-exo (exosomes derived from BMSCs were added after 7 days of osteogenic induction). The absorbance values of Raw264.7 cells in each group at 24 h, 48 h and 72 h were detected by an enzyme-linked immunosorbent assay (ELISA) reader. Western blotting was performed to assess changes of M1 or M2 marker proteins in Raw264.7 macrophages treated by exosome. The supernatants of the three groups of Raw264.7 macrophages were then co-cultured with BMSCs. Alizarin Red staining was used to quantify the formation of mineralized nodules, and alkaline phosphatase (ALP) staining was used to evaluate the osteogenic activity, and the expression levels of osteogenesis-related proteins Runx2, ALP, osteopontin (OPN), and osteocalcin (OCN) were detected. The migration ability of endothelial progenitor cells (EPCs) was detected by scratch assay for migration distance, and the angiogenesis ability was detected by in vitro tube formation assay for the number of vascular rings formed. The expressions of vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF) were detected by Western blot. The activation of the MAPK signaling pathway was evaluated by measuring phosphorylated and total P38 and ERK1/2 levels.Results:Western blot analysis showed that CM from BMSCs after 7 days of osteogenic induction (7D-BMSCs-CM) induced the strongest M2 polarization in macrophages. Compared with the PBS group, 7D-BMSCs-CM induced the most significant polarization of Raw264.7 macrophages to the M2 type, and increased ARG-1 and CD163 expression to 1.36±0.09 and 1.69±0.09, respectively ( P<0.05), while decreasing iNOS and CD86 to 0.21±0.03 and 0.29±0.03 ( P<0.05). The absorbance values of macrophages in the 7D-BMSCs-exo group were significantly higher than those in the PBS group at 24 h, 48 h, and 72 h ( P<0.05). Compared with BMSCs-exo group, 7D-BMSCs-exo upregulated the expressions of CD163 and ARG-1 while inhibited the expressions of iNOS and CD86 ( P<0.05). Alizarin red staining showed enhanced mineral deposition and a higher degree of mineralization in the 7D-BMSCs-exo group, the staining intensity of ALP also increased simultaneously, the Western-blot results showed that the protein expressions of Runx2, ALP, OPN and OCN were respectively higher than those in the PBS group ( P<0.05). The results of the scratch assay showed that the relative migration distance of EPCs cells in the 7D-BMSCS-exo group at 24 h reached 2.30±0.05 μm, which was higher than that in the PBS group 1.10±0.02 μm ( P<0.05). The Tube formation experiment showed that the number of vascular rings in the EPCs group was higher than that in the PBS group (30.3±2.5 and 15.0±1.0, P<0.05), and the protein expressions of VEGF-A and PDGF were upregulated. Furthermore, the levels of phosphorylated P38 and ERK1/2 were significantly reduced in the 7D-BMSCs-exo group 0.40±0.06 and 0.25±0.06 compared with the PBS group ( P<0.05). Conclusions:Exosomes secreted by BMSCs during osteogenic differentiation promote the M2 polarization of Raw264.7 macrophages, with the most pronounced effect observed at the 7th day. M2-polarized macrophages, in turn, enhance the osteogenic differentiation of BMSCs and the angiogenic capacity of EPCs. These processes are closely associated with the suppression of the MAPK signaling pathway.

Objective To explore the feasibility and safety of ultrasound-guided percutaneous microwave ablation for unresectable pancreatic cancer.Methods Totally 84 patients who underwent ultrasound-guided percutaneous microwave ablation for unresectable pancreatic cancer were enrolled,and the technical success rate,complete ablation rate,complication rate,pain relief rate and survival time,etc.were observed.Results The median age of 84 cases was 61.5 years.Totally 86 tumors,including 44.19％(38/86)at the head/neck and 55.81％(48/86)at the body/tail of pancreas were detected,and a total of 85 ablation sessions were performed with the median ablation energy applied per tumor of 9.90(1.08,21.60)kJ and the complete ablation rate of 42.86％(36/84).The technical success rate was 100％(85/85).Thirty-nine complication events occurred in 25 cases,no ablation-related death.Among 34 patients underwent ablation mainly for pain symptoms,the pain score decreased from(6.22±1.12)points before treatment to(1.94±1.64)points after treatment(P＜0.001).During 6.8(3.3,12.9)months' follow-up,the mean survival time was(8.5±6.7)months,and all 47 patients died due to tumor progression.Conclusion Ultrasound-guided percutaneous microwave ablation was safe and feasible for unresectable pancreatic cancer.

This study evaluated the potential of OPC-167832 as a new method for the treatment of Mycobacterium fortuitum infec-tion.Drug sensitivity tests were conducted with the broth microdilution method to determine the minimum inhibitory concentration(MIC)of OPC-167832 against standard strains of M.fortuitum and 44 clinical isolates of M.fortuitum.A DprE1 overexpression strain was constructed,and the effect in the MIC of OPC-167832 against M.fortuitum were explored.Intracellular germicidal tests and checkerboard tests were conducted to verify the ability of OPC-167832 to kill intracellular M.fortuitum,and its interaction with five drugs:amikacin,clarithromycin,imipenem,moxifloxacin,and clofazimine.The MIC50 and MIC90 against 44 clinical isolates of M.fortuitum were 0.031 25 μg/mL and 0.062 5μg/mL,respectively.The epidemiological cut-off value(ECOFF)was 0.062 5 μg/mL.Overexpression of DprE1 led to resistance to OPC-167832 in M.fortuitum.After 24 hours of incubation,the intracellular bacterial in-hibition rate of OPC-167832 at a 1 μg/mL concentration was 81.37%,exceeding the 74.05%inhibition rate of amikacin at a 1 μg/mL concentration.OPC-167832 showed strong inhibitory activity against M.fortuitum in vitro and in macrophages,and might provide a promising treatment for M.fortuitum infection.

Objective:To investigate the effects of exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) during osteogenic differentiation on the polarization of Raw264.7 macrophages and to elucidate the underlying mechanisms.Methods:PBS, uninduced BMSCs conditioned medium (CM), and osteogenic induction BMSCs CM for 7 d, 14 d, and 21 d were respectively added to Raw264.7 macrophages. After 48 h of treatment, Western blotting was used to detect and compare the expression of M1 markers of macrophages [inducible nitric oxide synthase (iNOS) and CD86] and M2 markers of macrophages [arginase-1 (ARG-1) and CD163] in each group. In addition, Raw264.7 macrophages were divided into three groups: the PBS group (only PBS added), the BMSCs-exo group (exosomes derived from uninduced BMSCs were added), and 7D-BMSCs-exo (exosomes derived from BMSCs were added after 7 days of osteogenic induction). The absorbance values of Raw264.7 cells in each group at 24 h, 48 h and 72 h were detected by an enzyme-linked immunosorbent assay (ELISA) reader. Western blotting was performed to assess changes of M1 or M2 marker proteins in Raw264.7 macrophages treated by exosome. The supernatants of the three groups of Raw264.7 macrophages were then co-cultured with BMSCs. Alizarin Red staining was used to quantify the formation of mineralized nodules, and alkaline phosphatase (ALP) staining was used to evaluate the osteogenic activity, and the expression levels of osteogenesis-related proteins Runx2, ALP, osteopontin (OPN), and osteocalcin (OCN) were detected. The migration ability of endothelial progenitor cells (EPCs) was detected by scratch assay for migration distance, and the angiogenesis ability was detected by in vitro tube formation assay for the number of vascular rings formed. The expressions of vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF) were detected by Western blot. The activation of the MAPK signaling pathway was evaluated by measuring phosphorylated and total P38 and ERK1/2 levels.Results:Western blot analysis showed that CM from BMSCs after 7 days of osteogenic induction (7D-BMSCs-CM) induced the strongest M2 polarization in macrophages. Compared with the PBS group, 7D-BMSCs-CM induced the most significant polarization of Raw264.7 macrophages to the M2 type, and increased ARG-1 and CD163 expression to 1.36±0.09 and 1.69±0.09, respectively ( P<0.05), while decreasing iNOS and CD86 to 0.21±0.03 and 0.29±0.03 ( P<0.05). The absorbance values of macrophages in the 7D-BMSCs-exo group were significantly higher than those in the PBS group at 24 h, 48 h, and 72 h ( P<0.05). Compared with BMSCs-exo group, 7D-BMSCs-exo upregulated the expressions of CD163 and ARG-1 while inhibited the expressions of iNOS and CD86 ( P<0.05). Alizarin red staining showed enhanced mineral deposition and a higher degree of mineralization in the 7D-BMSCs-exo group, the staining intensity of ALP also increased simultaneously, the Western-blot results showed that the protein expressions of Runx2, ALP, OPN and OCN were respectively higher than those in the PBS group ( P<0.05). The results of the scratch assay showed that the relative migration distance of EPCs cells in the 7D-BMSCS-exo group at 24 h reached 2.30±0.05 μm, which was higher than that in the PBS group 1.10±0.02 μm ( P<0.05). The Tube formation experiment showed that the number of vascular rings in the EPCs group was higher than that in the PBS group (30.3±2.5 and 15.0±1.0, P<0.05), and the protein expressions of VEGF-A and PDGF were upregulated. Furthermore, the levels of phosphorylated P38 and ERK1/2 were significantly reduced in the 7D-BMSCs-exo group 0.40±0.06 and 0.25±0.06 compared with the PBS group ( P<0.05). Conclusions:Exosomes secreted by BMSCs during osteogenic differentiation promote the M2 polarization of Raw264.7 macrophages, with the most pronounced effect observed at the 7th day. M2-polarized macrophages, in turn, enhance the osteogenic differentiation of BMSCs and the angiogenic capacity of EPCs. These processes are closely associated with the suppression of the MAPK signaling pathway.

Objective To investigate the effect of intravenous injection of remifentanil on the comfort level of birth-giving women with scarred uterus undergoing cesarean section.Methods A total of 82 birth-giving women with scarred uterus who underwent cesarean section in the Suqian Hospital of Jiangsu Provincial People's Hospital from October 2021 to September 2023 were selected and randomly divided into the observation group and the control group(with 41 cases in each group).Before skin resection of cesarean section,the birth-giving women of the observation group were injected with 0.05 μg·kg-1·min-1 remifentanil intravenously until the end of the operation,and these in the control group was injected with the same amount of normal saline.The vital signs and pain at different time points,traction reaction and occurrence of maternal and infant complications were compared between the two groups.Results The mean arterial pressure(MAP),heart rate(HR),pain visual analogue scale(VAS)score and incidence of traction reactions at abdominal exploration in the observation group were significantly lower than those in the control group(P<0.05).There was no significant difference in the neonatal umbilical vein pH value,umbilical vein blood pulse oximetry saturation(SPO2),Apgar scores 1 minute and 5 minutes after birth of newborn,or nausea,vomiting and respiratory depression of birth-giving women between the two groups(P>0.05).Conclusion For birth-giving women with scar uterus who underwent cesarean section,intravenous injection of 0.05 μg·kg-1·min-1 of reifentanil can reduce the fluctuation of vital signs,significantly relieve the traction reaction at abdominal exploration,with a few maternal and infant complications,which is conducive to improving the comfort level of the birth-giving women.