Sonodynamic therapy (SDT) can potentially induce immunogenic cell death in tumor cells, leading to the release of ATP, and facilitating the initiation of an immune response. Nevertheless, the enzymes CD39 and CD73 can swiftly convert ATP into immunosuppressive adenosine (ADO), resulting in an immunosuppressive tumor microenvironment (TME). This study introduced a nanomedicine (QD/POM1@NP@M) engineered to reprogram TME by modulating the CD39/CD73/ADO pathway. The nanomedicine encapsulated sonosensitizers silver sulfide quantum dots, and the CD39 inhibitor POM1, while also incorporating homologous tumor cell membranes to enhance targeting capabilities. This integrated approach, on the one hand, stimulates the release of ATP via SDT, thereby initiating the immune response. In addition, it reduced the accumulation of ADO by inhibiting CD39 activity, which ameliorated the immunosuppressive TME. Upon administration, the nanomedicine demonstrated substantial anti-tumor efficacy by facilitating the infiltration of anti-tumor immune cells, while reducing the immunosuppressive cells. This modulation effectively transformed the TME from an immunologically "cold" state to a "hot" state. Furthermore, combined with the checkpoint inhibitor α-PDL1, the nanomedicine augmented systemic anti-tumor immunity and promoted the establishment of long-term immune memory. This study provides an innovative strategy for combining non-invasive SDT and ATP-driven immunotherapy, offering new ideas for future cancer treatment.

ObjectiveTo assess the level of internal exposure to PCP in a community population in Shanghai， to investigate the factors affecting the level of PCP， and to analyze the correlation between the exposure and thyroid hormone levels. MethodsA total of 464 residents of a community in Shanghai were selected as the study subjects. A questionnaire survey was conducted to obtain the demographic information， dietary situation， lifestyle and behavioral habits， and disease history of the individuals， and blood samples were collected. Gas chromatography-electron trap was applied to determine the PCP levels in serum. Multicategorical logistic regression analysis was used to analyze the possible influencing factors of PCP exposure in humans. Thyroid hormone levels were used as the dependent variable and serum PCP as the independent variable. Multiple linear regression analysis was used to assess the association between PCP and thyroid hormones in the community population after controlling the confounding factors such as age， gender， literacy， annual personal income， and chronic diseases. ResultsThe detection rate of serum PCP in 464 subjects was 90.3%， and the median serum PCP level was 0.43 μg·L^-1. The differences in PCP levels among different age groups were statistically significant. There were no significant differences in PCP levels among different gender and BMI groups. The study of PCP exposure factors showed that age， frequency of using plastic products， consumption of freshwater fish， type of occupation， annual income， and consumption of tea or coffee were the potential influencing factors for PCP exposure. Among them， age， frequency of using plastic products， consumption of tea or coffee， and consumption of freshwater fish were positively associated with PCP levels， and annual personal income was negatively associated with it. The results of multiple linear regression analysis showed that among men， PCP levels were positively correlated with TSH （b=0.105， 95%CI：0.017‒0.313） and negatively correlated with FT4 （b=-0.026， 95%CI：-0.057‒0.004）， and among women， PCP levels were positively correlated with TSH （b=0.092， 95%CI：-0.211‒0.904） and FT3 （b=0.017， 95%CI：-0.058‒0.230） and negatively correlated with FT4 （b=-0.013， 95%CI：-0.011‒0.037）. ConclusionSerum PCP detection is common among community residents in Shanghai. Different demographic characteristics or behavioral habits may increase or decrease PCP exposure. PCP exposure then affects human thyroid hormone levels.

AIM:To investigate the effects of sodium selenite(SS)on viability,migration and angiogenesis of human non-small-cell lung cancer(NSCLC)H520 and A549 cells.METHODS:The H520 cells,A549 cells,and hu-man umbilical vein endothelial cells(HUVEC)were divided into control group(0 μmol/L SS),low dose group(5 μmol/L SS),middle dose group(10 μmol/L SS),and high dose group(20 μmol/L SS).Cell viability was assessed using the CCK-8 assay,and the half-maximal inhibitory concentration(IC50)was calculated.Cell migration and invasion abilities were determined through wound healing and Transwell assays.The regulatory effects of SS on angiogenesis,vasculogenic mimicry and"mosaic"vascular formation between HUVEC and NSCLC cells were detected using vessel forming assays.The expression of vascular endothelial growth factor(VEGF)in the supernatant of lung cancer cells in each group was de-tected using chemiluminescence.RT-qPCR was used to assess the mRNA expression of VEGF,vascular endothelial growth factor receptor 2(VEGFR2)and angiotensin II(Ang II).Western blot was used to examine the protein levels of VEGF,p-PI3K,and p-Akt in H520 and A549 cells.RESULTS:The IC50 values of SS to HUVEC,A549 cells and H520 cells for 48 h were 6.762,9.003 and 7.356 μmol/L,respectively.Compared with control group,the wound healing rate was significantly decreased in each group treated with SS for 48 h(P<0.01).In HUVEC,the number of migrating cells in middle dose and high dose groups decreased(P<0.01),whereas in lung cancer cells,the number of migrating cells in each group decreased after SS treatment(P<0.01).The mRNA expression levels of VEGF,VEGFR2 and Ang II were lower in high-dose SS group than those in control group(P<0.05 or P<0.01).In H520 cells,compared with control group,the protein levels of VEGF,p-PI3K and p-Akt in SS treatment groups were significantly decreased(P<0.05 or P<0.01).CONCLUSION:Sodium selenite inhibits the viability and migration of HUVEC,H520 cells and A549 cells,and inhibits the formation of vasculogenic mimicry and mosaic vessels in NSCLC cells.This mechanism may be related to the inhibition of PI3K-Akt signaling pathway activation and regulation of VEGF.

Objective:To explore the distribution pattern of TCM syndromes in patients with pulmonary tuberculosis combined with coronary heart disease in Kunming area.Methods:A survey was conducted by research of questionnaires to the general information and TCM four diagnostic information of pulmonary tuberculosis patients (116 cases) with coronary heart disease admitted to our hospital from April 2019 to December 2020, and principal component analysis and clustering analysis were conducted. Frequency analysis and severity analysis methods were used for data processing.Results:The TCM symptoms of pulmonary tuberculosis patients with coronary heart disease were mainly cough [89.66%(104/116)], expectoration [73.28% (85/116)], dry mouth [70.69% (82/116)], chest tightness [64.66% (75/116)], fatigue [56.03%(65/116)], hot flashes [52.59% (61/116)], shortness of breath [50.00% (58/116)]; tongue color was more common with light red [43.10% (50/116)] and red [47.41% (55/116)]; the tongue shape was more common with cracked tongue [37.07% (43/116)], punctured tongue [27.59% (32/116)] and old tongue [23.28%(27/116)]; tongue coating with little or no coating [48.28%(56/116)], yellow and greasy coating [21.55% (25/116)] was more common; the pulse was usually thin (number) [54.31% (63/116)] and slippery (number) [25.86% (30/116)]. The principal component analysis method used a load coefficient >0.40 as the threshold to screen out the four diagnostic information of 10 principal components; the clustering analysis results were divided into three categories of TCM types: deficiency syndrome, excess syndrome, and mixed deficiency and excess syndrome. There were significant differences in TCM syndrome types among patients of different genders, ages, disease courses, and educational levels ( P<0.05). Deficiency syndrome was more common in male patients [41.18% (21/51)], and mixed syndrome of deficiency and excess was more common in female patients [63.08% (41/65)]; 43-59 years old patients were more likely to have excess syndrome [36.36%(24/66)], and ≥60 years old patients were more likely to have mixed syndrome [70.00% (35/50)]; patients with a course of disease <3 months had more excess syndrome [41.30% (19/46)], patients with a course of 3-12 months had more deficiency syndrome [57.14% (20/35)], and patients with a course of more than 1 year had more mixed syndrome of deficiency and excess [74.29% (26/35)]. male patients [54.17%(26/48)] were more serious in excess syndrome, and female patients [53.33% (24/45)]; were more serious in deficiency syndrome; deficiency syndrome [43-59 years old was 54.17% (42/84), ≥60 years old was 54.17% (12/24)] more serious in patients of different age groups; patients with a course of disease <3 months [56.86% (29/51)] were more serious in excess syndrome, and patients with a course of disease ≥3 months were more serious in mixed syndromes [3~12 months 52.38%(22/42), >1 year 53.33% (24/45)]. Conclusion:The pathogenesis of TCM in patients with pulmonary tuberculosis complicated with coronary heart disease in Kunming is deficiency in root and excess in superficiality; the syndrome is based on yin deficiency and qi deficiency, with blood stasis, phlegm heat, phlegm stasis and phlegm turbidity as the symptoms.

Inducing the degradation of KRAS represents a novel strategy to combat cancers with KRAS mutation. In this study, we identify ubiquitin-specific protease 2 (USP2) as a novel deubiquitinating enzyme of KRAS in multiple myeloma (MM). Specifically, we demonstrate that gambogic acid (GA) forms a covalent bond with the cysteine 284 residue of USP2 through an allosteric pocket, inhibiting its deubiquitinating activity. Inactivation or knockdown of USP2 leads to the degradation of KRAS, resulting in the suppression of MM cell proliferation in vitro and in vivo. Conversely, overexpressing USP2 stabilizes KRAS and partially abrogates GA-induced apoptosis in MM cells. Furthermore, elevated USP2 levels may be associated with poorer prognoses in MM patients. These findings highlight the potential of the USP2/KRAS axis as a therapeutic target in MM, suggesting that strategically inducing KRAS degradation via USP2 inhibition could be a promising approach for treating cancers with KRAS mutations.

ObjectiveTo investigate the characteristics of serum perfluoroalkyl substances （PFASs） exposure and potential influencing factors among community residents in Songjiang District， Shanghai. MethodsIn August 2021， residents who underwent routine health checkups in a community in Songjiang District， Shanghai were recruited as study subjects. The inclusion criteria were adult residents who had lived in the area for more than 3 years， had no occupational exposure history， no underlying diseases， were not pregnant， and were able to complete the questionnaire independently and sign the informed consent form. A questionnaire survey was conducted and venous blood samples were collected. The concentrations of 15 PFASs in serum were determined using ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry （UPLC-MS/MS）. Ordered multi-class logistic regression， interquartile range （IQR）， and odds ratio （OR） were used to explore the influencing factors and risk magnitude of serum PFASs concentrations. ResultsOf the 15 PFASs， 14 were detected， and the detection rates of 7 PFASs were higher than 50%. The highest detected concentrations among the PFASs were PFOS （perfluorooctane sulfonate）， PFOA （perfluorooctanoic acid）， and PFHxS （perfluorohexane sulfonate）， with median concentrations of 48.61 μg∙L^-1， 37.29 μg∙L^-1， and 36.51 μg∙L^-1， respectively. The strongest correlation was between PFDA and PFUnDA （r=0.93， P<0.05）， followed by PFOS and PFDA （r=0.86， P<0.05）. Age， frequency of plastic product use， time spent indoors per day， personal annual income， tea consumption， and daily water intake were potential factors for exposure to PFASs. Among them， age was positively correlated with PFASs； tea consumption was positively correlated with PFNA and PFOA； PFHpA was negatively correlated with the frequency of plastic product use and personal annual income； and PFOS was negatively correlated with the time spent indoors per day. ConclusionThe exposure to serum PFASs among community residents in Songjiang District was relatively serious， and the main components were traditional PFOA， PFOS， and PFHx. Different sociodemographic characteristics had varying degrees of influence on the concentrations of PFASs in serum. The impact of PFASs exposure on the health of community residents deserves further investigation.

CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.
Artemisinins ; Fermentation ; Saccharomyces cerevisiae/metabolism* ; Saccharomyces cerevisiae Proteins/metabolism*

Objective:To identify the pathogens isolated in the subperiosteal abscesses from the pediatric patients with acute osteomyelitis and to investigate the characteristics of bacterial drug resistance.Methods:A retrospective study was conducted on children with acute septic osteomyelitis who were hospitalized in the Children's Hospital, Zhejiang University School of Medicine from January 1, 2011 to March 1, 2018. The results of bacterial cultures isolated from the subperiosteal abscesses or bone marrow fluid were collected. The Merier automatic bacterial identification system (i.e., Vitek) was used to identify the bacteria and to assess the drug sensitivity.Results:(1) A total of 104 pediatric patients were included and 60 (57.7%) were male. Sixty-six strains of pathogens were isolated from 65 patients (62.5%). Among them, 53 strains (51.0%) were Staphylococcus aureus; 3 strains were Escherichia coli; 2 strains (1.9%) were Pseudomonas aeruginosa; 2 strains (1.9%) were Streptococcus pneumoniae, 2 strains (1.9%) were Ochrobactrum anthropi, and 4 strains (3.8%) were other bacteria. Pathogens were not found in 39 patients (37.5%). (2) Staphylococcus aureus accounted for 81.5% (53/65) of the pathogen-positive cases. Among them, 23 strains (43.4%) were methicillin-resistant Staphylococcus aureus (MRSA). Aureus-positive children were statistically significantly older ( P=0.028), heavier ( P=0.040) and had higher C-reactive protein (CRP) level ( P=0.038) than the aureus-negative children. (3) All the 53 Staphylococcus aureus strains were resistant to penicillin and 56.6% of them were only sensitive to benzocillin. The resistance rates to compound sulfamethoxazole, tetracycline, clindamycin and erythromycin were 11.3%, 30.2%, 67.9% and 69.8%, respectively. The sensitivity rate of the strains to furantoin was 90.2%. All strains were sensitive to quinupristin/dalfopristin, linezolid, rifampicin, tigecycline, levofloxacin, moxifloxacin, ciprofloxacin and vancomycin. There was 69.8% of the strains resistant to three or more different types of antibiotics. Conclusions:Staphylococcus aureus is the most common pathogen that causes the acute septic osteomyelitis in children, and the resistance rate to Benzocillin is relatively high. Therefore, Benzocillin and Clindamycin, as the traditionally-used drugs, should not be considered as the first choice when empirically using intravenous antibiotics. In the present study, pathogens in 39 patients (37.5%) were not detected in their subperiosteal abscesses or bone marrow fluid, so further effort should be made to investigate the etiology of these patients.

Objective: To investigate the immunomodulatory activity of polysaccharides from the roots of Brassica rapa. Methods: The crude polysaccharide from roots of B. rapa (BRP) was extracted and purified to further investigate the active fraction of BRT for inducing macrophage phagocytosis. Results: Effects on RAW264.7 cells demonstrated that BRP behaved better phagocytic capacity and had potent immunomodulatory activity, including increasing production of nitric oxide (NO), tumor necrosis factor α (TNFα) and upregulating mRNA levels of inducible NO synthase (iNOS) and TNFα. Furthermore, modulation of macrophage by BRP was indicated to be mediated via the activation of Akt and nuclear factor-kappa B (NF-κB). Conclusion: The beneficial effects of BRP could be used as an immunotherapeutic adjuvant in treatment of inflammatory diseases.

Objective:To investigate the correlations of melanin concentration hormone (MCH) in cerebrospinal fluid (CSF) and serum with sleep disorder, memory dysfunction and prognoses in patients with cerebral ischemic stroke (CIS).Methods:One hundred elderly CIS patients, admitted to Department of Neurology, He'nan Provincial People's Hospital from June 2021 to January 2022 were enrolled as CIS group, and 50 subjects collected from Physical Examination of the same hospital during the same period were enrolled as control group. MCH levels in the CSF and serum were detected by ELISA. Sleep quality was assessed by polysomnography and Pittsburgh Sleep Quality Index (PSQI). Memory function was assessed by Rivermead Behavioral Memory Test 2 nd Edition (BMT-II). Prognoses were assessed by modified Rankin Scale (mRS) 3 months after discharge. The clinical data and MCH levels of the two groups were compared; the differences in MCH levels among CIS patients with different degrees of sleep disorder, and different memory functions and prognoses were compared. Correlations of MCH level and sleep parameters with RBMT-II scores in these CIS patients were analyzed. Results:Compared with that in the control group, the proportion of patients with hypertension in CIS group was significantly higher ( P<0.05). Compared with the control group ([42.39±16.11] pg/mL), the serum MCH level in CIS group ([36.89±15.19] pg/mL) was statistically lower ( P<0.05). In CIS patients, patients with mild or severe sleep disorder had significantly decreased CSF MCH level compared with patients without sleep disorder ( P<0.05), patients with severe sleep disorder had significantly decreased CSF MCH level compared with patients with mild sleep disorder ( P<0.05); patients with severe sleep disorder had significantly decreased serum MCH level compared with patients without sleep disorder ( P<0.05); CSF MCH level was negatively correlated with PSQI scores, sleep latency and wake frequency ( P<0.05), and positively correlated with percentage of rapid eye movement ( P<0.05); serum MCH level in CIS patients was negatively correlated with PSQI scores and wake frequency ( P<0.05). In CIS patients, the CSF and serum MCH levels in patients with memory dysfunction was significantly lower compared with those with normal memory function ( P<0.05); a positive correlation was noted between RBMT-II scores and CSF MCH level ( P<0.05). In CIS patients, patients with poor prognosis had statistically lower CSF and serum MCH levels compared with those with good prognosis ( P<0.05). Conclusion:The serum MCH level in CIS patients is significantly decreased, which is closely related to the occurrence of sleep disorder and memory dysfunction after stroke; and they further affects the prognoses.