Photoredox-Catalyzed Synthesis of C-Benzoselenazolyl/Benzothiazolyl Glycosides from 2-Isocyanoaryl Selenoethers/Thioethers and Glycosyl Bromides.

Molecules containing heteroatoms, such as Se and S, play an indispensable role in the discovery and design of pharmaceuticals, whereas Se has been less studied. Here, we described a photoredox strategy to synthesize C-benzoselenazolyl (Bs) glycosides from 2-isocyanoaryl selenoethers and glycosyl bromides. This reaction was carried out under mild conditions with high efficiency. C-Benzothiazolyl (Bt) glycosides could also be synthesized from 2-isocyanoaryl thioethers using this strategy. This method can access novel seleno/thiosugars, which will benefit Se/S-containing drug discovery.

Improving the mapping accuracy of soil heavy metals through an adaptive multi-fidelity interpolation method.

Soil heavy metal pollution poses a serious threat to environmental safety and human health. Accurately mapping the soil heavy metal distribution is a prerequisite for soil remediation and restoration at contaminated sites. To improve the accuracy of soil heavy metal mapping, this study proposed an error correction-based multi-fidelity technique to adaptively correct the biases of traditional interpolation methods. The inverse distance weighting (IDW) interpolation method was chosen and combined with the proposed technique to form the adaptive multi-fidelity interpolation framework (AMF-IDW). In AMF-IDW, sampled data were first divided into multiple data groups. Then one data group was used to build the low-fidelity interpolation model through IDW, while the other data groups were treated as high-fidelity data and used for adaptively correcting the low-fidelity model. The capability of AMF-IDW to map the soil heavy metal distribution was evaluated in both hypothetical and real-world scenarios. Results showed that AMF-IDW provided more accurate mapping results compared with IDW and the superiority of AMF-IDW became more evident as the number of adaptive corrections increased. Eventually, after using up all data groups, AMF-IDW improved the R2 values for mapping results of different heavy metals by 12.35-24.32%, and decreased the RMSE values by 30.35%-42.86%, indicating a much higher level of mapping accuracy relative to IDW. The proposed adaptive multi-fidelity technique can be equally combined with other interpolation methods and provide promising potential in improving the soil pollution mapping accuracy.

Research on Medical Security System Based on Zero Trust.

With the rapid development of Internet of Things technology, cloud computing, and big data, the combination of medical systems and information technology has become increasingly close. However, the emergence of intelligent medical systems has brought a series of network security threats and hidden dangers, including data leakage and remote attacks, which can directly threaten patients' lives. To ensure the security of medical information systems and expand the application of zero trust in the medical field, we combined the medical system with the zero-trust security system to propose a zero-trust medical security system. In addition, in its dynamic access control module, based on the RBAC model and the calculation of user behavior risk value and trust, an access control model based on subject behavior evaluation under zero-trust conditions (ABEAC) was designed to improve the security of medical equipment and data. Finally, the feasibility of the system is verified through a simulation experiment.

An Infusion Containers Detection Method Based on YOLOv4 with Enhanced Image Feature Fusion.

The detection of infusion containers is highly conducive to reducing the workload of medical staff. However, when applied in complex environments, the current detection solutions cannot satisfy the high demands for clinical requirements. In this paper, we address this problem by proposing a novel method for the detection of infusion containers that is based on the conventional method, You Only Look Once version 4 (YOLOv4). First, the coordinate attention module is added after the backbone to improve the perception of direction and location information by the network. Then, we build the cross stage partial-spatial pyramid pooling (CSP-SPP) module to replace the spatial pyramid pooling (SPP) module, which allows the input information features to be reused. In addition, the adaptively spatial feature fusion (ASFF) module is added after the original feature fusion module, path aggregation network (PANet), to facilitate the fusion of feature maps at different scales for more complete feature information. Finally, EIoU is used as a loss function to solve the anchor frame aspect ratio problem, and this improvement allows for more stable and accurate information of the anchor aspect when calculating losses. The experimental results demonstrate the advantages of our method in terms of recall, timeliness, and mean average precision (mAP).

Molecular epidemiology analysis of fowl adenovirus detected from apparently healthy birds in eastern China.

BACKGROUND: Fowl adenovirus is of major concern to the poultry industry worldwidely. In order to monitor the prevalent status of Fowl adenovirus in China, a total of 1920 clinical samples from apparently healthy birds in the 25 sites of poultry flocks, Slaughterhouse and living bird markets from 8 provinces in eastern China were collected and detected by PCR, sequencing, and phylogenetic analysis. RESULTS: The epidemiological survey showed that Fowl adenoviruses were detected in living bird markets, and circulating in a variety of fowl species, including chickens, ducks, goose and pigeons. Among the 1920 clinical samples, 166 samples (8.65%) were positive in the fowl adenovirus PCR detection. In this study, totally all the 12 serotypes (serotypes of 1, 2, 3, 4, 5, 6, 7, 8A, 8B, 9, 10 and 11) fowl adenoviruses were detected, the most prevalent serotype was serotype 1. Phylogenetic analysis indicated that 166 FAdVs of 12 serotypes were divided into 5 fowl adenovirus species (Fowl aviadenovirus A, B, C, D, E). CONCLUSIONS: In the epidemiological survey, 8.65% of the clinical samples from apparently healthy birds were positive in the fowl adenovirus PCR detection. Totally all the 12 serotypes fowl adenoviruses were detected in a variety of fowl species, which provided abundant resources for the research of fowl adenoviruses in China. The newly prevalent FAdV serotypes provides valuable information for the development of an effective control strategy for FAdV infections in fowls.

SERS detection of thiram using polyacrylamide hydrogel-enclosed gold nanoparticle aggregates.

The development of sensitive and long-term signal-stable plasmonic substrates is vital to the in-field application of the surface-enhanced Raman spectroscopy (SERS) technique. The colloidal gold nanoparticles (AuNPs) system is commonly used in SERS detection, but it shows less signal stability and reproducibility due to the uncontrollable aggregation of nanoparticles by adding aggregating agents in SERS detection. In this study, we developed a new SERS detection platform based on polyacrylamide hydrogel-enclosed plasmonic gold nanoparticle aggregates (PAH-AuANs). In the system, the formation of PAH can rapidly stabilize the gold nanoparticle aggregates, avoiding the over-aggregation or precipitation of AuNPs. With the PAH concentration in the range of 6-10 % and AuNPs at the concentration of 0.2 nM, the resulting PAH-AuNAs platform exhibited both sensitive SERS activity and excellent SERS signal stability. The relative standard deviation of the 4-MBA probe SERS signal collected from the PAH-AuNAs platform was lower than 3 %. The limit of detection for the pesticide thiram was down to 0.38 µg/L with a handheld Raman spectrometer. Moreover, the procedure for preparing the PAH-AuNAs platform was easy to handle, offering a new strategy for in-field detection of environmental contaminants with a handheld Raman spectrometer in the future.

Duraplasty with autologous nuchal ligament fascia to reduce postoperative complications in pediatric patients undergoing neoplasia resection with a suboccipital midline approach.

OBJECTIVE: The authors sought to explore the safety and efficacy of an autologous nuchal ligament for dural repair in pediatric patients undergoing tumor resection through a suboccipital midline approach. METHODS: Pediatric patients diagnosed with posterior fossa neoplasia who underwent surgery through a suboccipital midline approach were retrospectively reviewed. The patients were divided into artificial graft and autograft groups according to whether artificial duraplasty material or autologous nuchal ligament was used to repair the dura. Postoperative complications were reviewed and analyzed, including CSF leak, pseudomeningocele, and meningitis, during hospitalization and follow-up. Univariate and multivariate logistic regression analyses were used to investigate the relationship between duraplasty material and postoperative complications, as well as other risk factors for postoperative complications. Furthermore, multinomial logistic regression analysis was used to clarify which postoperative complications the autologous nuchal ligament tended to reduce. RESULTS: This retrospective study included 66 pediatric patients who underwent tumor resection through a suboccipital midline approach. The clinical baseline characteristics were comparable between the two groups. The results showed that the autograft group had significantly fewer postoperative complications, especially pseudomeningocele, compared with the artificial graft group. Moreover, the time required to repair the dura in the autograft group was significantly less than that in the artificial graft group. Further results revealed that the duraplasty material, ependymoma, preoperative severe hydrocephalus requiring an external ventricular drain (EVD), and postoperative hydrocephalus exacerbation were independent risk factors for postoperative complications. In particular, the autologous fascia of the nuchal ligament tended to reduce pseudomeningocele more than CSF leak and meningitis. However, compared with pseudomeningocele and CSF leak, both ependymoma and postoperative hydrocephalus exacerbation were more likely to increase the occurrence of meningitis. In contrast, preoperative severe hydrocephalus requiring EVD led to increased rates of postoperative complications. CONCLUSIONS: For pediatric patients with intracranial tumors who need to undergo resection through a suboccipital midline approach, dural repair using the nuchal ligament is safe, cost-effective, and time saving and significantly reduces postoperative complications.

Antibiotic resistance genes and bacterial community distribution patterns in pig farms.

Antibiotic-resistant pathogens pose high risks to human and animal health worldwide. In recent years, many studies have been carried out to investigate the role of gut microbiota as a pool of antibiotic resistance genes (ARGs) in human and animals. Both the structure and function of the gut bacterial community and related ARGs in pig remain unknown. In this study, we characterized the gut microbiomes and resistomes of fecal samples collected from sixteen pig farms located in sixteen cities of Shandong Province by metagenomic sequencing. Alpha diversity indicated that fecal samples from Dezhou (DZ) and Jinan (J) showed higher alpha diversity, and the lowest was from pig farms of Rizhao (RZ). Other pig farms showed similar alpha diversity. Besides, we found that the composition of gut bacterial among these pig farms varied greatly. Helcococcus massiliensis was the dominant bacterial species in pig farms of RZ and Zibo (ZB), while Prevotella sp. P5-92 occupied a superior proportion in Binzhou (BZ) and Yantai (YT). The proportion of Lactobacillus johnsonii was similar among farms of Qingdao (QD), Linyi (LY), Taian (TA), Weifang (WF), Weihai (WH), and YT. In total, 1112 ARGs were obtained and classified into 69 groups from 48 fecal samples. ARG abundance was higher in farms of Dongying (DY) and WH than others, while the lowest farms in BZ and ZB. Interestingly, it is found that BZ pig farm was exclusive, so the tetQ gene showed a higher abundance. In contrast, the load of APH(3') - IIIa in fecal samples from DY, J-1, LC, WF, and WH was high. Meanwhile, the most relevant ARGs and the corresponding microbes were screened out. Our metagenomic sequencing data provides new insights into the abundance, diversity, and structure of bacterial community in pig farms. Meanwhile, we screened ARG-carrying bacteria and explored the correlation between ARGs and bacterial community, which provide a comprehensive view of the pig fecial ARGs and microbes in different farms of Shandong.

First Report of Target Spot Caused by Rhizoctonia solani AG-6 in Tobacco in China.

Tobacco is an annual and solanaceous crop, which is widely produced in China. In July 2020, tobacco target spot was observed on 50% of tobacco plants in a 5-ha commercial field of Bijie (27.32° N, 105.29° E), Guizhou province, China. Typical symptoms firstly appeared on the old leaves as round watery spots. Then the spots became a diameter of 2 to 20 cm, with concentric ring lines and dead spots. Fifteen small pieces (5 × 5 mm) of leaf tissue were cut from the edge of the lesions, surface sterilized and placed on potato dextrose agar (PDA) medium amended with kanamycin (0.1 mg/ml). Isolate J136, one of five isolates with similar morphology, was selected for pathogen identification. The culture of the isolate on PDA was brown and exhibited radial mycelial growth after incubation at 28 oC in darkness for 5 days. Hyphae of the fungus were white at the beginning, turned light brown to brown at the later stages, and finally became thick and separated. Sclerotia were brown and produced on PDA after 25 days of incubation in the dark. These characteristics were similar to the colony characteristics of R. solani. The genomic DNA of Isolate J136 was extracted using the CTAB method. PCR analyses were conducted using the following primers specifically designed for the detection of individual AGs or subgroups of R. solani: AG-1 IA, IB and IC (Kuninaga 2003), AG-2-1, AG-2-2, IIIB, IV and LP (Carling et al. 2002), AG-3 PT (Misawa 2015), AG-4 HG-I and HG-II (Kuninaga 2003), and AGs-5-6 (Arakawa and Inagaki 2014). Among the 12 specific primer pairs, only AG-6-specific primers amplified a fragment of ca. 230 bp product, indicating that the tested strain belonged to R. solani AG-6. The sequence was deposited in GenBank with accession no. MZ379468. Using BLASTN search, the sequence of the gene was aligned with the voucher specimen, R. solani AG-6. A phylogenetic tree was constructed based on these sequences. After wards, Isolate J136 was tested for hyphal anastomosis reaction using the R. solani AG-6 standard strain according to the method described by Ogoshi (1987). The hyphal diameter at the point of anastomosis was reduced, with obvious anastomosis point, and the death of adjacent cells, indicating their anastomosis reactions (Anderson 1982). Thus, based on the morphological and genetic analyses, the fungus was identified as R. solani AG-6. To verify its pathogenicity, six plants (cv. Yunyan87) at the 5-to-6 leaf stage were inoculated with mycelial PDA plugs (5 mm in diameter). Leaves inoculated with PDA-only plugs served as the controls. Treated tobacco plants were maintained at a temperature range of 15 to 25 oC in a greenhouse with 85% relative humidity. After 5 days inoculation, typical symptoms were observed on the inoculated leaves, whereas no symptoms were observed on the control leaves. Koch's postulates were fulfilled by re-isolation of the pathogen from the diseased leaves. R. solani AG-2-2 is the only previously reported group of R. solani, which causes tobacco target spot in the field (Gonzalez et al. 2011). Therefore, to our knowledge, this is the first report of R. solani AG-6 causing target spot of tobacco in the field in China. Since considerable losses caused by the disease have frequently happened in this region, addition of this new group pathogen in the disease pool can be more problematic. Proper disease control strategies are in need to be developed to prevent further losses.

A novel sampling-free algorithm for subsurface data assimilation using Gaussian process-derived sensitivities.

Accurate characterization of hydraulic parameters is vital for modeling subsurface flow and transport. In the past decade, ensemble-based methods have been widely applied in estimating unknown parameters from state measurements. However, these methods require sufficiently large ensemble sizes to guarantee the accuracy of the ensemble averaged parameter sensitivities, leading to heavy computational burdens especially in large-scale problems. Although different surrogates have been introduced to alleviate the computational burden, the sensitivity information therein is still calculated by sampling the surrogate. Therefore, the sampling error is still inevitable. In this study, we propose an adaptive Gaussian process (GP) based iterative smoother (GPIS) algorithm in which the parameter sensitivity indices are analytically derived from the GP surrogate. During the iterations, the GP surrogate is adaptively refined by taking the updated parameter realizations as new base points. Both numerical and experimental cases are conducted to test the effectiveness of GPIS. We also compare its performance in estimating the heterogeneous hydraulic conductivity field with that of its prototype iterative ensemble smoother (IES) and our previously developed GP based iterative ensemble smoother (GPIES). Results show that, using the GP-derived sensitivity indices, GPIS shows advantages over GPIES in terms of both estimation accuracy and computational efficiency. Although subsurface flow and transport problems are considered in this work, the proposed method can be equally applied in other hydrological problems.

Characterizations of the endolysin Lys84 and its domains from phage qdsa002 with high activities against Staphylococcus aureus and its biofilms.

Staphylococcus aureus (S. aureus), particularly methicillin-resistant S. aureus (MRSA), and its biofilms are great threats in the food industry. Bacteriophage-encoded endolysins are promising tools to inhibit pathogens and to eliminate their biofilms. In this work, a virulent phage qdsa002 against S. aureus ATCC43300 (MRSA) was isolated, and the phage's endolysin (Lys84) and its domains were expressed and purified. Morphological and genome analyses demonstrated that qdsa002 is a Twort-like phage from Myoviridae. Lys84 contains two catalytic domains (CHAP and Amidase_2) and one cell binding domain (SH3b). This endolysin exhibits a strong lytic activity against S. aureus and has a wider bactericidal spectrum than qdsa002. Moreover, Lys84 exceed 10 µM effectively removed around 90 % of the biofilms of S. aureus. Besides, CHAP and Amidase_2 domains remained 61.20 % and 59.46 % of lytic activity as well as 84.31 % and 70.11 % of anti-biofilm activity of Lys84, respectively. The lytic and anti-biofilm activities of the combination of CHAP and Amidase_2 were close to 90 % of those of Lys84. These results indicated that Lys84 and its domains might be alternative antimicrobials for controlling S. aureus and its biofilms.

Systematical quantitative evaluation of left ventricular mechanical dysfunction in patients with cirrhosis using ultrasonic layer-specific strain imaging / 中华超声影像学杂志

Objective:To assess the left ventricular (LV) myocardial mechanical dysfunction in patients with cirrhosis using ultrasonic layer-specific strain imaging and to explore its value in clinical application.Methods:A total of 80 consecutive cirrhosis patients without cardiovascular diseases were prospectively enrolled from October 2020 to March 2021 in Sichuan Provincial People′s Hospital, 39 of whom were assigned to the compensated group and 41 were assigned to the decompensated group according to the occurrence of portal hypertension. Forty-three healthy volunteers during the same period were randomly recruited as the control group. Transthoracic echocardiography was performed to assess the LV configuration and functional parameters. LV global longitudinal strain in endocardial, middle and epicardial myocardium (GLSendo, GLSmid, GLSepi), and longitudinal strain (LS) in basal, middle and apical segments, and peak strain dispersion (PSD) were obtained using ultrasonic layer-specific strain imaging. ΔLS was calculated by the formula of GLSendo-GLSepi. Then, the differences of related parameters among three groups were compared.Results:①Conventional echocardiography: compared with the control group, the interventricular septum end-diastolic thickness (IVSTd), left ventricular posterior wall end-diastolic thickness (LVPWd), left ventricular mass (LVM) and LVM index (LVMI) were increased in compensated and decompensated groups (all P<0.05), while no significant differences in conventional echocardiographic parameters were identified between the two cirrhosis groups (all P>0.05). ②Global layer-specific strain: compared with the control group, GLSendo, GLSmid, GLSepi and ΔLS were decreased and PSD was increased in compensated and decompensated groups (all P<0.05); Moreover, the decompensated group showed a more impaired GLSendo, GLSmid and GLSepi than compensated group (all P<0.05), whereas there were no significant differences of ΔLS and PSD between the two groups(all P>0.05). ③Segmental layer-specific strain: compared with the control group, LS values of three layers in compensated and decompensated groups were reduced at basal, middle and apical levels (all P<0.05); Compared with the compensated group, LS values of three layers in decompensated group tended to be reduced at above there levels, but only apical segments had significant differences (all P<0.05). Conclusions:There are different degrees of LV mechanical dysfunction in patients with variable severity of cirrhosis. Ultrasonic layer-specific strain imaging has the potential to quantitatively assess the state of cardiac involvement in patients with cirrhosis and to provide visual evidence for the early and accurate diagnosis of myocardial injuries.

SLC19A1 May Serve as a Potential Biomarker for Diagnosis and Prognosis in Osteosarcoma.

BACKGROUND: Osteosarcoma is the most frequent primary malignant tumor of bone. SLC19A1 has been explored as a novel biomarker in some cancers. In this research, the diagnostic and prognostic value of SLC19A1 expression in osteosarcoma was evaluated by bioinformatics analysis. Data were sourced from the Gene Expression Omnibus (GEO) database. METHODS: Gene expression data and clinical materials of patients with osteosarcoma were collected from GSE42352 and GSE21257 datasets. The mRNA expression of SLC19A1 was compared between osteosarcoma cells and mesenchyme stem cells with the Wilcoxon rank-sum test. Moreover, receiver operating characteristic (ROC) curve analysis was performed to determine the diagnostic merit of SLC19A1 for osteosarcoma. The relationship between SLC19A1 and clinicopathological characteristics was analyzed using logistic regression. Besides, the correlation between SLC19A1 and survival rate was assessed using Kaplan-Meier and Cox regression. The biological functions of SLC19A1 were annotated and evaluated through gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA). RESULTS: SLC19A1 was significantly highly expressed in osteosarcoma cells (p < 0.001). The ROC curve showed an area under the curve of 0.899, which indicated a high diagnostic value. High SLC19A1 expression showed a negative correlation with Huvos grade [odds ratio (OR) = 0.09 for III vs. I, p = 0.014]. Kaplan-Meier survival analysis showed that the overall survival (OS) of the patients with high SLC19A1 expression was significantly poorer than the low SLC19A1 expression group (p = 0.016). The univariate analysis revealed that high SLC19A1 expression was associated with poor OS [p = 0.013, hazard ratio (HR) = 6.74, 95% CI = 1.49 - 30.46]. The multivariate analysis revealed that SLC19A1 expression (p = 0.014, HR = 8.03, 95% CI = 1.52 - 42.51) was independently correlated with OS. GSEA showed that genes in high expression group of SLC19A1 were enriched in KEGG pathways, including "Glyoxylate and dicarboxylate metabolism", "Oxidative phosphorylation", "Aminoacyl tRNA biosynthesis", "Base excision repair", "Pyrimidine metabolism" and "Proteasome". GSVA further suggested their importance in the progression of osteosarcoma. CONCLUSIONS: SLC19A1 may be a potential biomarker for diagnosis and prognosis in osteosarcoma.

[Mid-term follow-up efficacy of interspinous dynamic stabilization system for lumbar degenerative diseases].

OBJECTIVE: To explore mid-term clinical efficacy and effect on adjacent segment degeneration of Wallis and Coflex interspinous implants for lumbar degenerative diseases. METHODS: From January 2011 to January 2013, 55 patients with L4, 5 degenerative lumbar spine diseases treated with interspinous devices were retrospectively analyzed, including 31 males and 24 females aged from 25 to 67 years old with an average of 43.3 years old; 21 patients were lumbar spinal stenosis and 34 patients were lumbar disc herniation. All patients were divided into Wallis group (33 cases) and Coflex group (22 cases) according to the interspinous fixation system. Visual analogue scale(VAS) was used to evaluate low back pain and lower limb pain, Japanese Orthopedic Association(JOA) score and Oswestry Disability Index(ODI) score were used to evaluate lumbar function. Surgical segment and adjacent segments, range of motion(ROM), disc height and the Pfirrmann grade of upper the adjacent segments were compared before and after operation. RESULTS: Fifty-five patients were followed up from 48 to 72 months with an average of 60.4 months. VAS score of low back pain and lower limb pain, JOA and ODI score of lumbar at 48 months after operation were improved than before operation between two groups(P<0.01), but there was no statistical difference for group comparisons(P>0.05). ROM, disc height of surgical segments were significantly lower than those before operation between two groups (P<0.05), while ROM of the upper and lower adjacent segments and disc height did not change significantly (P>0.05). There was no significant difference in ROM and disc height for group comparisons(P>0.05). There was no change in Pfirrmann grade of the upper adjacent segment degeneration between two groups(P>0.05). Four patients with primary lumbar disc herniation had a recurrence of 1 to 3 years after operation, including 3 in Wallis group and 1 in Coflex group, with an average age of 35.2 years old. CONCLUSIONS: Wallis and Coflex interspinous implants have the similar mid-term efficacy for the treatment of lumbar degenerative diseases, and could delay adjacent segment degeneration, but could not prevent recurrence of disc herniation.

[Ribosomal Protein S9 Expression in Multiple Myeloma and Its Effect on Cell Biological Function].

Objective To identify the expression of ribosomal protein S9(RPS9)in multiple myeloma(MM)and explore its effect on the biological characteristics of myeloma cells and the corresponding mechanisms. Methods Bone marrow mononuclear cells were harvested in 10 healthy volunteers(CON group)and bone marrow CD138 +cells from 30 MM patients(CD138+group).Quantitative polymerase chain reaction(qPCR)was performed to detect RPS9 expression at mRNA level.In three cases from CON group and 11 cases from CD138+group,Western blot was performed to detect RPS9 at protein level.GSE19784 dataset was employed to detect the relationships of RPS9 expression with the overall survival rate,nuclear factor-κB(NF-κB),small ubiquitin-like modifier(SUMO),and ubiquitin pathway.After the RPS9 knock-down vector was constructed,flow cytometry was performed to detect the infection efficiency and qPCR and Western blot to detect the knock-down efficiency.RPMI8226 was divided into CON group and RPS9-short hairpin RNA(shRNA)group,in which annexin V allophycocyanin/propidium iodide(PI)double staining was performed to detect the change of apoptosis,CCK8 to detect the proliferation change,and PI staining to detect cell cycle change.After sentrin-specific protease 1(SENP1)overexpression vector was constructed,Western blot was performed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)from NF-κB pathway in CON,RPS9-shRNA,and RPS9-shRNA-SENP1 cells;in addition,annexin V/PI double staining was also performed to detect the apoptosis in these three cells. Results The relative expression of RPS9 in CON group and CD138+group was(1.00±0.12)and(5.45±0.71),respectively(t=4.291,P=0.0036).Western blot showed RPS9 expression was high in most myeloma CD138+cells.The high expression of RPS9 was associated with both extramedullary invasion and overall survival in GSE19784 dataset.After RPMI8226 was infected with CON or RPS9-shRNA lentivirus for 48 hours,flow cytometry confirmed that the infection efficiencies were above 90% in both groups.qPCR and Western blot confirmed that RPS9 expression was inhibited at both mRNA and protein levels.After RPMI8226 CON and RPS9-shRNA infected with virus for 48 hours,the proportion of annexin V-positive cells in CON and RPS9-shRNA cells was(3.47±0.37)% and(18.60±64.00)%(t=9.015,P=0.0008).The proliferation index significantly differed between CON group and RPS9-shRNA group at 72 hours(t=6.846,P=0.0024).When CON and RPS9-shRNA were infected with virus for 48 hours,the proportion of G2 phase cells was(29.28±3.42)% and(10.43±1.43)%,respectively(t=9.329,P=0.0007).The RPS9 expression was positively correlated with SENP1 in GSE19784 dataset and negatively correlated with IκBα coding gene NFKBIA.Western blot further confirmed that RPS9 knockdown inhibited the expression of SENP1,inhibited the phosphorylation of NF-κB subunit P65 and inhibitor IκBα,and promoted the expression of IκBα.Overexpression of SENP1 not only impeded this effect but also reduced RPS9-induced apoptosis. Conclusions RPS9 is highly expressed in MM CD138+cells and is associated with overall survival and extramedullary infiltration.Inhibition of RPS9 can promote apoptosis,cell cycle arrest,and proliferation of myeloma cells.RPS9 can affect the activation of NF-κB pathway and cell apoptosis through SENP1,suggesting that SENP1 may be a key factor in the biological effect of RPS9.

[Effect of Denticleless E3 Ubiquitin Protein Ligase on Proliferation and Colone Formation of Multiple Myeloma Cells].

Objective To determine the effect of denticleless E3 ubiquitin protein ligase(DTL)on the proliferation and clone formation of multiple myeloma(MM)cells and investigate the related mechanism. Methods Mononuclear cells were extracted from 34 MM patients.Mononuclear cells harvested from 14 healthy volunteers were used as controls.Quantitative polymerase chain reaction was used to detect the change of DTL at mRNA level.Furthermore,12 MM patients and 2 controls were selected,in whom the change of DTL at protein level was detected by Western blot.Human MM cell line RPMI8226 was divided into control(CON)group and DTL-short hairpin RNA(DTL-shRNA)group,which was infected with the CON and DTL-shRNA virus,respectively,for 48 hours.The infection efficiency was detected by using flow cytometry,the knock-down efficiencies at mRNA and protein levels were detected by quantitative polymerase chain reaction and Western blot,the change of cell counts in the next 0,24,48,72,96 hours were measured with CCK8 assay.The CON and DTL-shRNA cells were cultured in semisolid medium.Ten days later,inverted phase microscopy was used to measure the number of colones that contain more than 50 cells,annexin V/propidium iodide double staining to detect apoptosis,and propidium iodide staning to detect cell cycle.Finally,Western blot was empoyed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)in nuclear factor-κB(NF-κB)pathway and electrophoretic mobility shift assay(EMSA)to detect the NF-κB transcriptional ability. Results The DTL expression was(1.00±0.12)and(9.36±3.71),respectively in the bone marrow mononuclear cells of healthy volunteers and in the CD138+cells of MM patients(t=3.65,P=0.0024).DTL was also highly expressed in MM CD138+positive cells at protein level.After RPMI8226 was infected by CON and DTL-shRNA virus for 48 hours,green fluorescent protein-positive cells accounted for more than 90%.The relative expression of DTL was(1.00±0.01)and(0.21±0.04)(t=33.19,P<0.0001)at mRNA level and(0.52±0.13)and(0.11±0.02)at protein level(t=5.399,P=0.0057).CCK8 revealed that CON and DTL-shRNA cells proliferated by(1.00±0.03)vs.(1.00±0.02),(2.19±0.28)vs.(1.47±0.13),(3.50±0.14)vs.(2.24±0.19),(5.43±0.41)vs.(3.08±0.14),(7.42±0.17)vs.(4.29±013)after 0,24,48,72,and 96 hours(F=24.58,P=0.001).The number of colone containing more than 50 cells was in 76±4 in CON group and 0 in DTL-shRNA group(P<0.01).The proportion of G1 stage cells was(28.61±8.64)% in CON group and(57.25±10.37)% in DTL-shRNA group(t=3.675,P=0.0213).The proportion of annexin V+in CON and DTL-shRNA groups was(3.21±0.89)% vs.(34.71±18.68)%(t=2.895,P=0.0443).After RPMI8226 was infected with CON or DTL-shRNA virus for 48 hours,the relative expression of phosphorylation P65 was(1.52±0.14)vs.(0.82±0.11)(t=6.81,P=0.0024),the P65 relative expression was(0.25±0.04)vs.(0.24±0.08)(t=0.19,P=0.85),the CON and DTL-shRNA phosphorylation-IκBα relative expression was(0.19±0.03)vs.(0.13±0.02)(t=2.882,P=0.0449),and the IκBα was(0.22±0.05)vs.(1.01±0.06)(t=17.52,P<0.0001).Detection of the transcriptional ability of DTL-shRNA NF-κB by EMSA further confirmed the down-regulation of DTL suppressed the NF-κB transcriptional ability. Conclusions DTL is highly expressed in MM cells,and down-regulation of DTL suppresses the cell proliferation,inhibit the colony formation,and induce cell apoptosis and cell cycle arrest.The effect of DTL on the biological functions of MM cells is related to the change of NF-κB pathway.

Human Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles Inhibit Endometrial Cancer Cell Proliferation and Migration through Delivery of Exogenous miR-302a.

MicroRNAs (miRNAs) are potential therapeutic targets in endometrial cancer, but the difficulties associated with their delivery to tumor target cells have hampered their applications. Human umbilical cord mesenchymal stem cells (hUCMSCs) have a well-recognized tumor-homing ability, emphasizing the capacity of tumor-targeted delivery of extracellular vesicles. hUCMSCs release extracellular vesicles rich in miRNAs, which play a vital role in intercellular communication. The purpose of this study was to verify a potential tumor suppressor microRNA, miR-302a, and engineered hUCMSC extracellular vesicles enriched with miR-302a for therapy of endometrial cancer. Here, we observed that miR-302a was significantly downregulated in endometrial cancer tissues when compared with adjacent tissues. Overexpression of miR-302a in endometrial cancer cells robustly suppressed cell proliferation and migration. Meanwhile, the proliferation and migration were significantly inhibited in endometrial cancer cells when cultured with miR-302a-loaded extracellular vesicles derived from hUCMSCs. Importantly, our data showed that engineered extracellular vesicles rich in miR-302 significantly inhibited the expression of cyclin D1 and suppressed AKT signaling pathway in endometrial cancer cells. These results suggested that exogenous miR-302a delivered by hUCMSC-derived extracellular vesicles has exciting potential as an effective anticancer therapy.

Bayesian Monitoring Design for Streambed Heat Tracing: Numerical Simulation and Sandbox Experiments.

Heat tracing methods have been widely employed for subsurface characterization. Nevertheless, there were very few studies regarding the optimal monitoring design for heat tracing in heterogeneous streambeds. In this study, we addressed this issue by proposing an efficient optimal design framework to collect the most informative diurnal temperature signal for Bayesian estimation of streambed hydraulic conductivities. The data worth (DW) was measured by the expected relative entropy between the prior and posterior distributions of the conductivity field. An adaptively refined Gaussian process surrogate was employed to alleviate the computational burden, resulting in at least three orders of magnitude of speed-up. The applicability of the optimal experimental design framework was evaluated by both numerical and sandbox experimental cases. Results showed that the most informative locations centered in the transition zones among the main patterns of the hydraulic conductivity field, while the most informative times centered in a short period after the minimum/maximum temperature appeared. With the fixed number of measurements, extending the calibration period was more beneficial than increasing the monitoring frequency in improving the estimation results. To our best knowledge, this work is the first study on Bayesian monitoring design for streambed characterization with the heat tracing method. The method and results can provide guidance on selecting monitoring strategies under budget-limited conditions.

The epidemiological characteristics and the source of infection of reemerge human rabies from 2012 to 2017 in Qinghai / 中华疾病控制杂志

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.

Ribosomal Protein S9 Expression in Multiple Myeloma and Its Effect on Cell Biological Function / 中国医学科学院学报

Objective To identify the expression of ribosomal protein S9(RPS9)in multiple myeloma(MM)and explore its effect on the biological characteristics of myeloma cells and the corresponding mechanisms. Methods Bone marrow mononuclear cells were harvested in 10 healthy volunteers(CON group)and bone marrow CD138 +cells from 30 MM patients(CD138+group).Quantitative polymerase chain reaction(qPCR)was performed to detect RPS9 expression at mRNA level.In three cases from CON group and 11 cases from CD138+group,Western blot was performed to detect RPS9 at protein level.GSE19784 dataset was employed to detect the relationships of RPS9 expression with the overall survival rate,nuclear factor-κB(NF-κB),small ubiquitin-like modifier(SUMO),and ubiquitin pathway.After the RPS9 knock-down vector was constructed,flow cytometry was performed to detect the infection efficiency and qPCR and Western blot to detect the knock-down efficiency.RPMI8226 was divided into CON group and RPS9-short hairpin RNA(shRNA)group,in which annexin V allophycocyanin/propidium iodide(PI)double staining was performed to detect the change of apoptosis,CCK8 to detect the proliferation change,and PI staining to detect cell cycle change.After sentrin-specific protease 1(SENP1)overexpression vector was constructed,Western blot was performed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)from NF-κB pathway in CON,RPS9-shRNA,and RPS9-shRNA-SENP1 cells;in addition,annexin V/PI double staining was also performed to detect the apoptosis in these three cells. Results The relative expression of RPS9 in CON group and CD138+group was(1.00±0.12)and(5.45±0.71),respectively(t=4.291,P=0.0036).Western blot showed RPS9 expression was high in most myeloma CD138+cells.The high expression of RPS9 was associated with both extramedullary invasion and overall survival in GSE19784 dataset.After RPMI8226 was infected with CON or RPS9-shRNA lentivirus for 48 hours,flow cytometry confirmed that the infection efficiencies were above 90% in both groups.qPCR and Western blot confirmed that RPS9 expression was inhibited at both mRNA and protein levels.After RPMI8226 CON and RPS9-shRNA infected with virus for 48 hours,the proportion of annexin V-positive cells in CON and RPS9-shRNA cells was(3.47±0.37)% and(18.60±64.00)%(t=9.015,P=0.0008).The proliferation index significantly differed between CON group and RPS9-shRNA group at 72 hours(t=6.846,P=0.0024).When CON and RPS9-shRNA were infected with virus for 48 hours,the proportion of G2 phase cells was(29.28±3.42)% and(10.43±1.43)%,respectively(t=9.329,P=0.0007).The RPS9 expression was positively correlated with SENP1 in GSE19784 dataset and negatively correlated with IκBα coding gene NFKBIA.Western blot further confirmed that RPS9 knockdown inhibited the expression of SENP1,inhibited the phosphorylation of NF-κB subunit P65 and inhibitor IκBα,and promoted the expression of IκBα.Overexpression of SENP1 not only impeded this effect but also reduced RPS9-induced apoptosis. Conclusions RPS9 is highly expressed in MM CD138+cells and is associated with overall survival and extramedullary infiltration.Inhibition of RPS9 can promote apoptosis,cell cycle arrest,and proliferation of myeloma cells.RPS9 can affect the activation of NF-κB pathway and cell apoptosis through SENP1,suggesting that SENP1 may be a key factor in the biological effect of RPS9.